Methods of Soil Analysis

Published 1996
METHODS OF SOIL ANALYSIS
PART3
Chemical Methods
Soil Science Society of America Book Series
Books in the series are available from the Soil Science Society of America, 677 South
Segoe Road, Madison, WI 53711 USA.
1. MINERALS IN SOIL ENVIRONMENTS. Second Edition. 1989.

J.B. Dixon and S. B. Weed, editors R. C. Dinauer, managing editor
2. PESTICIDES IN THE SOIL ENVIRONMENT: PROCESSES, IMPACTS,

AND MODELING. 1990.
H. H. Cheng, editor S. H. Mickelson, managing editor
3. SOIL TESTING AND PLANT ANALYSIS. Third Edition. 1990.

R. L. Westerman, editor S. H. Mickelson, managing editor
4. MICRONUTRIENTS IN AGRICULTURE. Second Edition. 1991.

J. J. Mortvedt et al., editors S. H. Mickelson, managing editor
5. METHODS OF SOIL ANALYSIS: PHYSICAL AND MINERALOGICAL

METHODS. Part 1. Second Edition. 1986.
Arnold Klute, editor R. C. Dinauer, managing editor
METHODS OF SOIL ANALYSIS: CHEMICAL AND MICROBIOLOGICAL

PROPERTIES. Part 2. Second Edition. 1982.
A. L. Page et al., editor R. C. Dinauer, managing editor
METHODS OF SOIL ANALYSIS: CHEMICAL METHODS. Part 3. 1996.

D. L. Sparks, editor J.M. Bartels, managing editor
Methods of
Soil Analysis
Part3
Chemical Methods
Editorial Committee: D. L. Sparks

A. L. Page
P.A. Helmke
R. H. Loeppert
P. N. Soltanpour
M. A. Tabatabai
C. T. Johnston
M. E. Sumner
Managing Editor: J.M. Bartels
Editor-in-Chief SSSA: J. M. Bigham
Number 5 in the Soil Science Society of America Book Series
Published by: Soil Science Society of America, Inc.

American Society of Agronomy, Inc.
Madison, Wisconsin, USA
1996
Copyright © 1996 by the Soil Science Society of America, Inc.
ALL RIGHTS RESERVED UNDER THE U.S. COPYRIGHT LAW OF

1978 (P. L. 94-553)
Any and all uses beyond the "fair use" provision of the law require written
permission from the publishers and/or author(s); not applicable to contri-
butions prepared by officers or employees of the U.S. Government as part
of their official duties.
Second printing 1999.

Third printing 2001.
Soil Science Society of America, Inc.

677 South Segoe Road, Madison, Wisconsin 53711 USA
Library of Congress Cataloging-in-Publication Data
Library of Congress Catalog Card Number: 96-70096
Printed in the United States of America

CONTENTS
Page
Foreword ix
Preface xi
Contributors .. .. ... . ... ... .... .. .... ... .... .... .... .... ... .. ... .. .. ... ... ... ..... ... .. .... ... .. .. ... ... .. .... .. xiii
Conversion Factors for SI and non-SI Units .......... ...................... ........ ........ xvii
1 Sampling
Roger G. Petersen and Lyle D. Calvin ...................................... 1
2 Quality Assurance and Quality Control

E. J. Klesta, Jr. and J. K. Bartz .................................................. 19
3 Dissolution for Total Elemental Analysis

L. R. Hossner .................................. ...... .................... .... ........ ..... 49
4 Atomic Absorption and Flame Emission Spectrometry

Robert J. Wright and Tomasz I. Stuczynski .............................. 65
5 Inductively Coupled Plasma Emission Spectrometry

and Inductively Coupled Plasma-Mass Spectroscopy
Parviz N. Soltanpour, Greg W. Johnson,
Stephen M. Workman, J. Benton Jones, Jr.,
and Robert 0. Miller ................................................................. 91
6 Neutron Activation Analysis

Philip A. Helrnke .................. .................. .... ..... ......... .... ....... ... ... 141
7 Elemental Analysis by X-Ray Fluorescence Spectroscopy

A. D. Karathanasis and Ben F. Hajek ....................................... 161
8 Liquid Chromatography
M. A. Tabatabai and W. T. Frankenberger, Jr. .......................... 225
9 Differential Pulse Voltammetry

Larry M. Shuman ..................................................... ................. 247
10 Fourier Transform Infrared and Raman Spectroscopy

C. T. Johnston and Y. 0. Aochi ................................................. 269
V
vi CONTENTS
11 Electron Spin (or Paramagnetic) Resonance Spectroscopy

Nicola Senesi ............................................................................. 323
12 X-Ray Photoelectron Spectroscopy

R. K. Vempati, T. R. Hess, and D. L. Cocke ............................ 357
13 X-Ray Absorption Fine Structure Spectroscopy

Scott Fendorf and Donald L. Sparks ......................................... 377
14 Salinity: Electrical Conductivity and Total Dissolved Solids

J. D. Rhoades............................................................................. 417
15 Carbonate and Gypsum

Richard H. Loeppert and Donald L. Suarez .............................. 437
16 Soil pH and Soil Acidity

Grant W. Thomas....................................................................... 475
17 Lime Requirement
J. Thomas Sims.......................................................................... 491
18 Aluminum
Paul M. Bertsch and Paul R. Bloom ......................................... 517
19 Lithium, Sodium, Potassium, Rubidium, and Cesium

Philip A. Helmke and Donald L. Sparks ................................... 551
20 Beryllium, Magnesium, Calcium, Strontium, and Barium

Donald L. Suarez ....................................................................... 575
21 Boron
R. Keren ..................................................................................... 603
22 Silicon
R. Lewis Jones and Gary B. Dreher .......................................... 627
23 Iron
Richard L. Loeppert and W. P. Inskeep ..................................... 639
24 Manganese
R. P. Gambrell ........................................................................... 665
25 Chromium
Richmond J. Bartlett and Bruce R. James................................. 683
26 Copper and Zinc

Stewart T. Reed and D.C. Martens............................................ 703
CONTENTS vii
27 Molybdenum and Cobalt

John L. Sims ................ .............................................................. 723
28 Nickel, Cadmium, and Lead

Michael C. Amacher ........... ....................................................... 739
29 Mercury
James G. Crock .......................................................................... 769
30 Selenium and Arsenic

P. M. Huang and Roger Fujii ................... ... ...... .................... ..... 793
31 Bromine, Chlorine, and Fluorine

W. T. Frankenberger, Jr., M.A. Tabatabai,
D. C. Adriano, and H. E. Doner ................................................ 833
32 Phosphorus
Shiou Kuo ........ ............................................................. ..... ........ 869
33 Sulfur
M. A. Tabatabai ........... .......................................................... .... 921
34 Total Carbon, Organic Carbon, and Organic Matter

Darrell W. Nelson and Lee E. Sommers ................................... 961
35 Organic Matter Characterization

Roger S. Swift ........................................................................... 1011
36 Extraction of Organic Chemicals

Brij L. Sawhney ......................................................................... 1071
37 Nitrogen-Total
John M. Bremner ....................................................................... 1085
38 Nitrogen-Inorganic Forms
R. L. Mulvaney .......................................................................... 1123
39 Nitrogen-Organic Forms
F. J. Stevenson ........................................................................... 1185
40 Cation Exchange Capacity and Exchange Coefficients

Malcolm E. Sumner and William P. Miller ........ ....................... 1201
41 Charge Analyses of Soils and Anion Exchange

Lucian W. Zelazny, Liming He,
and An M. Vanwormhoudt ........................................................ 1231
viii CONTENTS
42 Redox Measurements of Soils

W. H. Patrick, Jr., R. P. Gambrell, and S. P. Faulkner .............. 1255
43 Kinetic Methods and Measurements

Donald L. Sparks, Theodore H. Carski, Scott E. Fendorf,
and Charles V. Toner, IV .................................. .............. ........... 1275
44 Equilibrium Modeling in Soil Chemistry

S. V. Mattigod and J.M. Zachara .............................................. 1309
FOREWORD
Analytical methods are the foundation of a scientific discipline. This was
recognized by the Soil Science Society of America when an effort was initiated
in 1957 to give recognition to the body of analytical methods developed specifi-
cally to characterize soil composition and properties. Publication of the first edi-
tion of the "Methods of Soil Analysis" in 1965, under the editorship of Dr. C.A.
Black, marked a milestone in the development of the field of soil science.
Although there existed several books on soil analysis prior to 1965, this publica-
tion was the first authoritative treatise collectively authored by soil scientists
under the joint sponsorship of the American Society of Agronomy and American
Society of Testing and Materials, and published as volumes in the Agronomy
Monograph series. The publication quickly became the primary reference book
on methods for analyzing many soil physical, chemical, and microbiological
properties.
After the Soil Science Society of America created the Book Series, the
Boards of Directors of the American Society of Agronomy and Soil Science
Society of America reached an agreement in 1993 "to publish all future reprints,
revised editions, and new versions of Methods of Soil Analysis and all subse-
quent parts as part of the SSSA Book Series." The third edition of Methods of
Soil Analysis will now have three volumes. The volume covering the microbio-
logical and chemical methods was published in 1995. The current volume will
cover the chemical methods, and the volume on physical and mineralogical meth-
ods is under preparation. This volume includes coverage of newer methods for
characterizing soil chemical properties as well as several methods for character-
izing soil chemical processes. It should continue to serve as the primary reference
on analytical methods and provide soil and environmental scientists additional
tools to advance our knowledge of soil properties and soil processes.
H. H. Cheng, president
Soil Science Society of America
ix
PREFACE
The second edition of Methods of Soil Analysis, Part 2, Chemical and
Microbiological Properties was published in 1982. It was edited by AL. Page,
R.H. Miller, and D.R. Keeney. The 2nd edition is recognized as the benchmark
reference on chemical and microbiological analyses of soils. It has been used
widely by soil scientists and professionals in other fields. More than 11,000
copies have been sold. Due to major advances in analytical equipment and
methodology, the desire to include new chapters on analyses of soil chemical
processes, and the need to include additional material on microbiological analy-
ses, Part 2 has been divided into two parts and revised. The first book, Methods
of Soil Analysis, Microbiological and Biochemical Properties (Part 2), was pub-
lished in 1995 as SSSA Book Series No. 5. This book, Methods of Soi/Analysis:
Chemical Methods, is Part 3 of SSSA Book Series No. 5.
This book contains 44 chapters, written by 70 authors from throughout the
world. A new chapter on quality assurance and quality control is included.
Updated chapters are included on the principles of various instrumental methods
and their applications to soil analysis. Additionally, new chapters are included on
Fourier transform infrared, Raman, electron spin resonance, x-ray photoelectron,
and x-ray absorption fine structure spectroscopies. The application of these meth-
ods to analyzing soil chemical reactions is currently one of the major research
areas in the soil and environmental sciences.
Chapters are included on analyses of soil chemical properties including soil
salinity, carbonate and gypsum, soil pH and acidity, lime requirement, cation and
anion exchange capacities, and organic matter. Methods for the analyses of solu-
ble, sorbed, and total concentrations of 34 elements are also included.
Additionally, these chapters include useful background information on the chem-
istry of the elements. A new chapter on methods for organic chemical extraction
is included.
A new aspect of this book is the addition of procedures for analyzing
important soil chemical processes. These include redox and surface charge
(points of zero charge) analyses, and kinetic methods and measurements. Chapter
44, the last chapter, discusses equilibrium modeling in soil chemistry.
The editorial committee, that was instrumental in the planning of the book
and in the reviewing process, was composed of:
D.L. Sparks, Chairman, University of Delaware, Newark, DE.
AL. Page, University of California, Riverside, CA.
P.A. Helmke, University of Wisconsin, Madison, WI.
R.H. Loeppert, Texas A&M University, College Station, TX.
P.N. Soltanpour, Colorado State University, Fort Collins, CO.
M.A. Tabatabai, Iowa State University, Ames, IA.
C.T. Johnston, Purdue University, West Lafayette, IN.
M.E. Sumner, University of Georgia, Athens, GA.
xi
xii PREFACE
The editorial committee expresses its sincerest gratitude to the anonymous

reviewers for their careful and thoughtful evaluation of the manuscripts. Many
thanks are extended to Jon Bartels of the SSSA Headquarters Staff for his out-
standing editorial assistance.
Methods of Soil Analysis: Chemical Properties should be useful not only
to students and professionals in soil science, but to those in allied fields such as
engineering, chemistry, geosciences, and marine studies, who are increasingly
interested in soils.
D.L. Sparks, Editor-in-Chief

University of Delaware
Newark, Delaware 19717-1303
AL. Page, Associate Editor

University of California, Riverside
Riverside, California 92521
P.A. Helmke, Associate Editor

University of Wisconsin
Madison, Wisconsin 53706-1299
R.H. Loeppert, Associate Editor

Texas A&M University
College Station, Texas 77843
CONTRIBUTORS
Domy C. Adriano Professor and Head, Biogeochemical Ecology Division, Savannah
River Ecology Laboratory, Drawer E, Aiken SC 29802
Michael C. Amacher Research Soil Chemist, USDA-FS, Forestry Sciences Laboratory, 850
N. 1200 E., Logan UT 84321
Y.O.Aochi Staff Research Associate, Department of Soil and Environmental

Sciences, University of California, Riverside CA 92521
Richmond J. Bartlett Professor of Soil and Environmental Chemistry, Department of Plant

and Science, Hills Building, University of Vermont, Burlington VT
05405
J. K. Bartz Chemist, Washington State Department of Ecology, 1315 W. 4th Ave-

nue, Kennewick, WA 99336
Paul M. Bertsch Professor, Advanced Analytical Center for Environmental Science,

University of Georgia, Savannah River Ecology Laboratory, Drawer E,
Aiken SC 29802
Paul R. Bloom Professor of Soil Science, Department of Soil, Water, and Climate,
University of Minnesota, 1991 Upper Buford Circle, St. Paul MN
55108
John M. Bremner Distinguished Professor Emeritus, Department of Agronomy, Iowa

State University, Ames IA 50011-1010
L.D.Calvin Professor Emeritus of Statistics, Department of Statistics, Oregon

State University, Corvallis OR 97331
Theodore H. Carksi Research Associate/Supervisor, Agricultural Products, Stine-Haskell

Research Center, E.I. duPont de Nemours and Company, Building
210N, Room 204, Elkton Road, P.O. Box 30, Newark DE 19714
D.LCocke J.M. Gill Professor of Chemistry, Department of Chemistry, Lamar

University, P.O. Box 10022, Beaumont TX 77710
James G. Crock Analytical Geochemist, Branch of Geochemistry, Analytical Chemis-

try Services Group, U.S. Geological Survey, Denver Federal Center,
Mail Stop 973, Building 20, Box 25046, Denver CO 80225-6046
Harvey E. Doner Professor of Soil Chemistry, Division of Ecosystem Sciences, ESPM,

University of California, 101 Giannini Hall, Berkeley CA 94720-3110
Gary B. Dreher Head, Analytical Geochemistry Section, Illinois State Geological

Survey, 615 E. Peabody Drive, Champaign IL 61820
S.P. Faulkner Assistant Professor, Wetland Biogeochemistry Institute, Louisiana

State University, Baton Rouge LA 70803-7511
Scott Fendorf Assistant Professor of Soil Science, Soil Science Division, University
of Idaho, Moscow ID 83844-2339
xiii
xiv CONTRIBUTORS
W.T. Frankenberger, Jr. Professor of Soil Microbiology and Biochemistry, Department of Soil
and Environmental Sciences, University of California, Riverside CA
92521-0424
Roger Fujii Research Chemist, Water Resources Division-California District, U.S.

Geological Survey, 2800 Cottage Way, Sacramento CA 95825
R.P. Gambrell Professor, Wetland Biogeochemistry Institute, Louisiana State Univer-

sity, Baton Rouge LA 70803-7511
Ben F. Hajek Professor, Department of Agronomy and Soils, 201 Funchess Hall,
Auburn University, Auburn AL 36849
Liming He Research Assistant, Scripps Institute of Oceanography, University of

California, La Jolla CA 92093-0202
Philip A. Helmke Professor of Soil Science, Department of Soil Science, University of

Wisconsin, 1525 Observatory Drive, Madison WI 53706-1299
T. R. Hess Gill Chair, Department of Chemistry, Lamar University, P.O. Box

10022, Beaumont TX 77710
L. R. Hossner Professor of Soil Chemistry, Department of Soil and Crop Sciences,

Texas A&M University, College Station TX 77843-2474
P.M.Huang Professor of Soil Science, Department of Soil Science, University of

Saskatchewan, 51 Campus Drive, Saskatoon, SK S7N 5A8 Canada
William P. Inskeep Professor, Department of Plant, Soil and Environmental Sciences,

Montana State University, Bozeman MT 59717
Bruce R. James Associate Professor of Soil Chemistry, Department of Agronomy,

University of Maryland, College Park MD 20742
Greg W. Johnson Spectroscopist, Matheson Gas Products, 1861 Lefthand Circle Drive,
Longmont CO 80501
C.T. Johnston Associate Professor of Soil Chemistry, Department of Agronomy,

Purdue University, 1150 Lilly Hall, West Lafayette IN 47907-1150
J. Benton Jones, Jr. Vice President, Micro-Macro International, Inc., 183 Paradise Boule-
vard, Athens GA 30607
R. Lewis Jones Professor of Soil Mineralogy and Ecology, Department of Natural

Resources and Environmental Sciences, University of Illinois, 1102 S.
Goodwin Avenue, Urbana IL 61801
A.O. Karathanasis Professor of Soil Mineralogy and Pedology, Department of Agronomy,

University of Kentucky, N-122 Agricultural Sciences Building North,
Lexington KY 40546-0091
Rami Keren Director, Institute of Soils and Water, The Volcani Center, ARO, P.O.
Box 6, Bet Dagan 50250, Israel
E. J. Klesta, Jr. Director of Technology, Cozz Iron & Metal, Inc., 605 Alexandria
Drive, Naperville IL 60565
ShiouKuo Soil Scientist, Department of Crop and Soil Sciences, Washington State
University, 7612 Pioneer Way E., Puyallup WA 98371-4998
Richard H. Loeppert Professor, Department of Soil and Crop Sciences, Texas A&M Uni-
versity, College Station TX 77843-2474
CONTRIBUTORS xv
D. C. Martens W.G. Wysor Professor of Agriculture and Life Sciences, Department of

Soil and Environmental Sciences, Virginia Polytechnic Institute and
State University, Blacksburg VA 24061-0404
S. V. Mattigod Battelle Pacific Northwest Laboratory, P.O. Box 999, Richland WA

99352
Robert O. Miller Soil Scientist/Extension Soil Specialist, Department of Land, Air and
Water Resources, Hoagland Hall, University of California, Davis CA
95616-8627
William P. Miller Professor of Soil Science, Department of Crop and Soil Sciences, 3111
Miller Plant Sciences Building, University of Georgia, Athens GA
30602-7503
Richard L. Mulvaney Professor of Soil Science, Department of Natural Resources and

Environmental Sciences, University of Illinois, 1102 S. Goodwin
Avenue, Urbana IL 61801
Darrell W. Nelson Dean and Director, Agricultural Research Division, 207 Agricultural
Hall, University of Nebraska, Lincoln NE 68583-0704
W. H. Patrick, Jr. Boyd Professor and Director, Wetland Biogeochemistry Institute

Louisiana State University, Baton Rouge LA 70803-7511
Roger G. Petersen Professor Emeritus of Statistics, Department of Statistics, Oregon State

University, Corvallis OR 97331
Stewart T. Reed Assistant Professor of Soil Science, CESTA, 306E South Perry Paige
Building, Florida A&M University, Tallahassee FL 32307
J. D. Rhoades Laboratory Director, USDA-ARS, U.S. Salinity Laboratory, 450 W.

Big Springs Road, Riverside CA 92507
Brij L Sawhney Soil Chemist, The Connecticut Agricultural Experiment Station, 123
Huntington Street, P.O. Box 1106, New Haven CT 06504
Nicola Senesi Professor of Soil Chemistry and Director, Istituto di Chimica Agraria,
Universita di Bari, Via Amendola 165/A, Bari-70126, Italy
Larry M. Shuman Professor of Soil Chemistry, Department of Crop and Soil Sciences,
Georgia Agricultural Experiment Station, University of Georgia,
Griffin GA 30223-1797
John L. Sims Professor Emeritus of Agronomy, Department of Agronomy, N-122

Agricultural Science Building North, University of Kentucky,
Lexington KY 40546-0091
J. Thomas Sims Professor of Soil Science, Department of Plant and Soil Science, 149
Townsend Hall, University of Delaware, Newark DE 19717-1303
Parviz N. Soltanpour Professor of Soil Science, Department of Soil and Crop Sciences, Cll 7
Plant Sciences Building, Colorado State University, Fort Collins CO
80523
Lee E. Sommers Professor of Soil Science and Department Head, Department of Soil
and Crop Sciences, Colorado State University, Fort Collins CO 80523
Donald L. Sparks Distinguished Professor of Soil Science, Department of Plant and Soil
Sciences, University of Delaware, Newark DE 19717-1303
xvi CONTRIBUTORS
Frank J. Stevenson Professor Emeritus, Department of Agronomy, University of Illinois,

1102 S. Goodwin Avenue, Urbana IL 61801-4798
Tomasz I. Stuczynski Soil Scientist, Institute of Soil Science and Plant Cultivation, Osada
Palacowa, 24-100 Pulawy, Poland
Donald L Suarez Research Leader, USDA-ARS, U.S. Salinity Laboratory, 450 W. Big
Springs Road, Riverside CA 92507
Malcolm E. Sumner Regents' Professor of Soil Science, Department of Crop and Soil
Sciences, 3111 Miller Plant Sciences Building, University of Georgia,
Athens GA 30602
Roger S. Swift Chief, CSIRO, Division of Soils, PMB 2, Glen Osmond 5064,
Australia
M. A. Tabatabai Professor of Soil Chemistry, Department of Agronomy, Iowa State

University, Ames IA 50011-1010
Grant W. Thomas Professor of Agronomy, Department of Agronomy, University of

Kentucky, Lexington KY 40546-0091
Charles V. Toner, IV Graduate Research Fellow, Department of Plant and Soil Sciences,
University of Delaware, Newark DE 19717-1303
An M. Vanwormhoudt 49 Lillibrooke Crescent, Maidenhead, Berkshire SL6 3XJ U.K.
R. K. Vempati Department of Chemistry, Lamar University, P.O. Box 10022,

Beaumont TX 77710
Stephen M. Workman Technical Manager, Analytical Technologies, Inc., 225 Commerce

Drive, Fort Collins CO 80524
Robert J. Wright Research Soil Scientist, USDA-ARS, Environmental Chemistry

Laboratory, Building 007, Room 224, BARC-West, Beltsville MD
20705
J.M. Zachara Chief Scientist, Pacific Northwest Laboratory, P.O. Box 999, Mail Stop
K3-61, Richland WA 99352
Lucian W. Zelazny Thomas B. Hutchison, Jr., Professor of Soil Chemistry/Mineralogy,

Department of Crop and Soil Environmental Science, Virginia
Polytechnic Institute and State University, Blacksburg VA 24061-0404
Conversion Factors for SI and non-SI Units
xvii
Conversion Factors for SI and non-SI Units ;_
==
To convert Column 1 To convert Column 2
into Column 2, into Column 1,
multiply by Column 1 SI Unit Column 2 non-SI Units multiply by
Length
0.621 kilometer, km (103 m) mile, mi 1.609
1.094 meter, m yard, yd 0.914
3.28 meter, m foot, ft 0.304
1.0 micrometer, µm (10-6 m) micron,µ 1.0
3.94 X 10-2 millimeter, mm (10-3 m) inch, in 25.4
10 nanometer, nm (10-9 m) Angstrom,A 0.1 ("'.)
0
Area
z
<
t"1
2.47 hectare, ha acre 0.405
247 square kilometer, km2 (103 m)2 acre 4.05 X 10-3 0
-
0.386 square kilometer, km2 (103 m)2 square mile, mi2 2.590 z
2.47 X 10-4 square meter, m2 acre 4.05 X loJ ::
("'.)
10.76 square meter, m2 square foot, ft 2 9.29 X 10-2
1.55 X 10-3 square millimeter, mm2 (l0-3 m)2 square inch, in 2 645
""l
Volume 0
:,::,
9.73 X 10-3 cubic meter, m 3 acre-inch 102.8 rr,
35.3 cubic meter, m3 cubic foot, ft3 2.83 X 10-2 -
6.10 X 104 cubic meter, m3 cubic inch, in3 1.64 X 10-5 i::,
2.84 X 10-2 liter, L (10-3 m3) bushel, bu 35.24 z
1.057 liter, L (10-3 m3) quart (liquid), qt 0.946 0
3.53 X 10-2 liter, L (10-3 m3)
z
cubic foot, ft 3 28.3 00
0.265 liter, L (10-3 m3) gallon 3.78 -
33.78 liter, L (10-3 m3) ounce (fluid), oz 2.96 X 10-2 z
2.11 liter, L (10- 3 m 3) pint (fluid), pt 0.473 --3
rr,
-
Mass ("'J
0
2.20 X 10-3 gram, g (10-3 kg) pound, lb 454 z
<
t"l
3.52 X 10-2 gram, g (10-3 kg) ounce (avdp), oz 28.4
2.205 kilogram, kg pound, lb 0.454 ....
0.01 kilogram, kg quintal (metric), q 100 0
907
z
1.10 X 10-3 kilogram, kg ton (2000 lb), ton
1.102 megagram, Mg (tonne) ton (U.S.), ton 0.907 ("'J
1.102 tonne, t ton (U.S.), ton 0.907
d
,,
Cl.l
Yield and Rate ""l
0
,,
0.893 kilogram per hectare, kg ha- 1 pound per acre, lb acre- 1 1.12
7.77 X 10-2 kilogram per cubic meter, kg m- 3 pound per bushel, lb bu- 1 12.87 ....Cl.l
1.49 X 10-2 kilogram per hectare, kg ha- 1 bushel per acre, 60 lb 67.19 ;i.-
1.59 X 10-2 kilogram per hectare, kg ha- 1 bushel per acre, 56 lb 62.71
z
c::,
1.86 X 10-2 kilogram per hectare, kg ha- 1 bushel per acre, 48 lb 53.75 z
9.35 0
0.107 liter per hectare, L ha- 1 gallon per acre z
893 tonnes per hectare, t ha- 1 pound per acre, lb acre- 1 1.12 X 10-3 ....en
893 megagram per hectare, Mg ha- 1 pound per acre, lb acre- 1 1.12 X 10-3 c::
0.446 megagram per hectare, Mg ha- 1 ton (2000 lb) per acre, ton acre- 1 2.24 ....z
0.447 '"'3
2.24 meter per second, m s- 1 mile per hour Cl.l
Specific Surface
10 square meter per kilogram, m2 kg- 1 square centimeter per gram, cm2 g- 1 0.1
1000 square meter per kilogram, m2 kg- 1 square millimeter per gram, mm 2 g- 1 0.001
Pressure
9.90 megapascal, MPa (106 Pa) atmosphere 0.101

10 megapascal, MPa (106 Pa) bar 0.1
1.00 megagram, per cubic meter, Mg m-3 gram per cubic centimeter, g cm- 3 1.00
2.09 X 10-2 pascal, Pa pound per square foot, lb ft- 2 47.9
1.45 X 10-4 pascal, Pa pound per square inch, lb in-2 6.90 X 103
~-
><
(continued on next page)
Conversion Factors for SI and non-SI Units
To convert Column 1 To convert Column 2
into Column 2, into Column 1,
multiply by Column 1 SI Unit Column 2 non-SI Units multiply by
Temperature
1.00 (K- 273) Kelvin, K Celsius, °C 1.00 (°C + 273)
(9/5 °C) + 32 Celsius, °C Fahrenheit, °F 5/9 (°F- 32)
Energy, Work, Quantity of Heat

9.52 X 10-4 joule, J British thermal unit, Btu 1.05 X 103 (")
0.239 joule, J calorie, cal 4.19 0
107 joule, J erg 10-7 z
0.735 joule, J foot-pound
<
trl
1.36
2.387 X 10-5 joule per square meter, J m-2 calorie per square centimeter (langley) 4.19 X 104
105 newton, N dyne 10-s
...0
1.43 X 10-3 watt per square meter, W m-2 calorie per square centimeter 698
z
minute (irradiance), cal cm-2 min- 1 (")
Transpiration and Photosynthesis ,::,

rJl
3.60 X 10-2 milligram per square meter second, gram per square decimeter hour, 27.8
mg m-2 s-1 g dm-2 h-1 6,::,
5.56 X 10-3 milligram (H20) per square meter micromole (H20) per square centi- 180 rJl
second, mg m-2 s- 1 meter second, µmo! cm-2 s-1
...
10-4 milligram per square meter second, milligram per square centimeter 104
mg m-2 s- 1 second, mg cm-2 s-1 =
z
35.97 milligram per square meter second, milligram per square decimeter hour, 2.78 X 10-2 0
mg m-2 s-1 mg dm-2 h-1 rJl
...?,
Plane Angle z
57.3 radian, rad 0 rJl
degrees (angle), 1.75 X 10-2
("')
Electrical Conductivity, Electricity, and Magnetism 0
z
siemen per meter, S m- 1 millimho per centimeter, mmho cm-1 0.1 <
t"l
10 10--4
104 tesla, T gauss, G [I.)
0
-
Water Measurement z
cubic meter, m3 acre-inches, acre-in 102.8 ("')
9.73 X 10-3 ..,
9.81 X 10-3 cubic meter per hour, m3 h- 1 cubic feet per second, ft 3 s-1 101.9 0
cubic meter per hour, m3 h-1 U.S. gallons per minute, gal min- 1 0.227 [I.)
4.40
hectare-meters, ha-m acre-feet, acre-ft 0.123 "'l
8.11 0
hectare-meters, ha-m acre-inches, acre-in 1.03 X 10-2
97.28
acre-feet, acre-ft 12.33 [I.)
8.1 X 10-z hectare-centimeters, ha-cm
Concentrations
-
i:::,
centimole per kilogram, cmol kg- 1 milliequivalents per 100 grams, meq 1 z
1 0
100 g-1 z
10 c',i
0.1 gram per kilogram, g kg- 1 percent,%
milligram per kilogram, mg kg- 1 parts per million, ppm 1 c::
-
1 z
:::i
[I.)
Radioactivity
2.7 X 10-II becquerel, Bq curie, Ci 3.7 X 1010

2.7 X 10-2 becquerel per kilogram, Bq kg- 1 picocurie per gram, pCi g- 1 37
gray, Gy (absorbed dose) rad, rd 0.01
100
sievert, Sv (equivalent dose) rem (roentgen equivalent man) O.Gl
100
Plant Nutrient Conversion
Elemental Oxide
p PzOs 0.437
2.29
K KzO 0.830
1.20
Ca Cao 0.715
1.39
Mg MgO 0.602
1.66 ~-
Published 1996
Chapter1
Samplingl
R. G. PETERSENAND L. D. CALVIN, Oregon State University,

Corvallis, Oregon
INTRODUCTION
The purposeof any soil sampleis to obtain information about a particular soil.
The sampleitself is seldom,if ever, the entire soilmassin which we are interest-
ed. This larger aggregateof material, in which we are ultimately interested,is
called the "population" or "universe."Information from the sampleis of interest
only insofar as it yields information about the population, and the information
mayor may not be representative,dependingon how the sampleis selected.
The populationitself may be large or small, or even a part of what would
ordinarily be considereda larger population.For example,a populationmight be
all the soil in a field to a depthof 1 m, the clay portion in the top 15 cm of a field
plot, or the organicmatterin the B horizon over severalfields of a commonsoil
type. It is that portion of the soil for which we want additional information.
For any population,there are certain characteristics whichdescribeit. For
soils, thesemay include the thicknessof eachhorizon in a soil, the percentorgan-
ic matter,the amountof solublesalts,or the pH. The true value of eachsuchchar-
acteristicin the populationis called a parameter.The purposeof samplingis to
estimatetheseparameterswith an accuracythat will meetour needsat the lowest
possiblecost. All practical studiesare limited in funds, and sampling therefore
becomesa necessityin nearly all scientific investigations.If the population is
relatively homogeneous,a very small samplemay tell us all we desire to know
about the population.With soils, however,variation and heterogeneityseemthe
rule ratherthan the exception.
VARIATION OF SOILS
Soils are characterizedby severaltypesof variation, and if we are to do an

adequatejob of representinga particularsoil populationby meansof a sample,it
1 Reprintedfrom Methods of Soil Analysis, Part 1. Physical and Mineralogical Methods-SSSA

Book Seriesno. 5.
Copyright © 1996 Soil ScienceSociety of America and American Society of Agronomy, 677 S.
SegoeRd., Madison,WI 53711,USA. Methods ofSoil Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
1
2 PETERSEN& CALVIN
is necessarythat we considerthe natureof thesetypes of variation. The soil is

not a homogeneousmass but a rather heterogeneousbody of material. And
becauseof this heterogeneity,systemshavebeenset up that attemptto delineate
soil classification units which approachhomogeneitywithin themselves,but
which, at the sametime, are distinctly different from all other units. Thus, one
type of soil variation is the variation amongthe severalunits which have been
classifiedas homogeneous.Differencesamongtheseunits may be large or small
depending,among other things, on the differential effect of the factors which
formed the soils. Poorly drainedsoils formed from recentalluvium, for example,
are usually different in most of their propertiesfrom well-drainedsoils formed
from residual parent material. The variation in propertiesamong soils formed
from the sameparentmaterial undersimilar conditions,on the other hand,may
be rathersmall even though the soils be classifiedas different soils.
Because of the nature of soil-forming processes,distinct boundaries
betweensoil classification units are rare. Although the modal profiles of two
adjacentsoils seriesmay be distinctly different, thereis usually a gradualtransi-
tion, in the field, betweenone seriesand another.Superimposedon this pattern
of slowly changingcharacteristics,however, we may find rather marked local
variations.
Theselocal variationsmay result from naturalcauses,suchas sharp vege-
tative or topographicvariations, or from man-madevariation. Available phos-
phorus,for example,may vary widely within a seriesbecauseof differential fer-
tilization or liming practicesin the past.A similar patternof variation is found in
the subsoil.
Soil propertiesvary not only from one location to anotherbut also among
the horizons of a given profile. The horizon boundariesmay be more distinct
than are the surfaceboundariesof a soil classificationunit. Here, also, however,
zonesof transition are found betweenadjacenthorizons.Furthermore,consider-
able local variation may occur within a particularhorizon.
Thesecharacteristicsshouldbe kept in mind when samplingsoils. The soil
populationto be sampledshouldbe subdivided,both horizontally and vertically,
into sampling strata which are as homogeneousas possible, and the several
sourcesof variation within the populationshouldbe sampledif valid inferences
are to be madeabout the populationfrom the sample.
The intensity with which a soil must be sampledto estimatewith given
accuracysomecharacteristicwill dependon the magnitudeof the variationwith-
in the soil populationunderconsideration.The more heterogeneous the popula-
tion, the more intensemust be the samplingrate to attain a given precision.Few
dataare available,however,from which the magnitudesof the severalsourcesof
variation in soils may be estimated.In general,although differenceshave been
found to exist among soil classification units, considerablevariation may be
expectedwithin the units for such characteristicsas pH, availableP, exchange-
able K, exchangeableNa, conductivity, volume weight, permeability,and poros-
ity (Allmaras and Gardner,1956; Masonet aI., 1957; Olson et aI., 1958; Sayegh
et aI., 1958). In some instances,the variation within contiguousclassification
units isso greatthat it is not feasibleto estimatedifferencesamongthe units with
any satisfactorydegreeof precision. For most characteristics,the variation, both
SAMPLING 3
within and amongunits, decreases, with increasingdepth in the profile. Hence,

fortunately, it is usually necessaryto samplethe subsoil at a lower rate than the
topsoil to attain comparableaccuracy.
SAMPLING PLANS
When a sampleis actually drawn from the population, it is necessaryto

think of the populationas composedof a numberof separateunits. Thesemay
be the numberof cores,7 cm squareand 15 cm long, that can be taken from a
field, or the numberof spadeloads which would comprisethe areaunderstudy.
They may be naturalunits of the population,suchas an individual plant; or they
may be artificial ones,suchas fields within a soil series. The essentialfeatureis
that they be separateand distinct units and that their total numbercomprisethe
entire population.
The sampling plan designateswhich units of the population are to be
includedin the sample. There are alwaysa numberof different plansor designs
which can be used. Someare more precisethan others(provide a smallererror),
and somemay be carriedout at a much lower cost. In general,the bestdesignis
one that providesthe maximumprecisionat a given costor that providesa spec-
ified precision(error) at the lowest cost.
The principlesof soil samplingwere outlined by Cline (1944) and havenot
changedmaterially. Therehasbeena greaterappreciationof the systematicsam-
ple since then, and new methodshave been suggestedfor estimatingsampling
error; however,very few studies havebeenmadeuponwhich to basesoundsam-
pling procedures.Perhapsthe cost of making intensivestudieshas discouraged
suchwork, but if progressis to be forthcoming, further investigationof the reli-
ability and efficiency of different samplingplans is needed.
JudgmentSample
The researchworker ordinarily knows somethingabouthis populationand

would like to use this information in obtaining a representativesample. Effort
hassometimesbeendirectedtowardthe useof his judgmentin selectingthe most
"typical" sitesfrom which to draw the sample. These"typical" sitesare obtained
by selectingthe sitesthat, in the samplersjudgment,presenta representativepic-
ture of the population. Becausethe samplingunits are selectedwith different but
unknownprobabilities,samplesselectedin this mannerare biased. As an exam-
ple, someworkersarevery careful to include the extremesof the population(and
may therefore oversamplethe extremes),while others attempt to exclude all
extremesas being unrepresentative of the bulk of the population.
Unfortunately,unlessadditional data are availableor the true characteris-
tics are known for the population,there is no way of assessingthe accuracyof
the results from such a sample.The samplemay representthe populationvery
well or it may not; any confidencein the resultsmust rest entirely on faith in the
sampler'sjudgment.
If a small sampleis to be taken,a judgmentsamplemay havea lower error
than a randomsample.As samplesize increases,however,the error with a ran-
4 PETERSEN& CALVIN
dom samplebecomesprogressivelysmaller, whereaswith a judgmentsample,

the error drops much more slowly and quickly becomeslarger than with a ran-
dom sample.Unlessthe researchworker is particularly skillful in selecting"typ-
ical" sites,the error with a randomsamplebecomessmallerthan that with a judg-
ment sample at a relatively small sample size. As sample size increases,the
selection of "typical" sites also becomesmore difficult and time-consuming.
However, if low accuracyis satisfactoryand no estimateof precisionneeded,a
judgmentsamplemay be satisfactoryfor the purpose.
Simple Random Sample
If n units are to be selectedfrom the population,a simple randomsample

is definedas a sampleobtainedin sucha mannerthat eachpossiblecombination
of n units has an equal chanceof being selected.In practice,it is usually drawn
by selectingeachunit separately,randomly,and independentlyof any units pre-
viously drawn. This, of course,requiresthat eachunit can be listed or potential-
ly listed. With a field, for example,it is not necessaryto list all possiblecore
sampleswhich can be taken, althoughit could be done if necessary.
In soil investigations,the unit to be includedin the sampleis usually a vol-
ume of soil, althoughit may be an areaof ground,a plant, or a volume of water.
If the uni~s are listed, a randomsamplecan easily be taken by the useof a table
of randomnumbers(Fisherand Yates, 1967). Often, however,it is more conve-
nient to spot the field location by selectingrandomdistanceson a coordinatesys-
tem and using the intersectionof the two randomdistancesas the point at which
the unit is to be taken.This systemworks well for fields of both regularand irreg-
ular shape,sincethe points outsidethe areaof interestare merely discardedand
only the points inside the areaare usedin the sample.
The locationof the point at which eachsamplingunit is to be takenis often
reachedin two steps.The two stepsprovide a fairly simple method to use and
also ensureobjectivity, to preventa selectionbias which can occur when only a
single step is used. The first step is to reach an approximatepoint in the field.
This can be accomplishedby (i) mappingthe areaand establishingtwo baselines
at right anglesto eachother, which intersectat an arbitrarily selectedorigin, say
the southwestcomer; (ii) establishinga scaleinterval to be usedin locating the
selectedpoint, e.g., a pace, yard, chain, or 0.1 mile; (iii) drawing two random
numbersrepresentingthe numberof basicscalingunits to be measuredalong the
two directions; and (iv) locating the intersectionof two distancesin the field as
an approximatepoint for sampling.Using this point as the origin of a new coor-
dinate system,a secondrandomcoordinateis measuredoff to provide an exact
point for the samplingunit. The samplingunit to be taken is then defined as the
soil unit for which the selectedpoint is someprescribedpoint, say the centerof
the southwestcomerof the samplingtool. The scaleinterval for the secondran-
dom coordinateshouldbe of the samedimensionsas the samplingtool, with the
maximum value of the secondcoordinateequal to the scaleinterval of the first
coordinatesystem.For example(seeFig. 1-1), if the first scaleinterval is a pace
and a 10- by lO-cm samplingunit is to be used,then the secondset of random
distancesshould be selectedfrom numbers0 to 9 (9 10-cm units per pace).This
SAMPLING 5
95
paces
16 ________ ,J+
I
O~----~I----------------~
o 37 127
paces
Fig. 1-1. Selectionof randomly selectedpoints in a field by a two-stepprocedure.Randomlengths
were 37 and 16 paces alongthe eastand north axes,respectively,to establishthe approximatepoint
for sampling(x). RandomlO-cm units drawn were 2 and 8 units to the eastand north, respective-
ly, to establishan exactpoint (+) for locationgthe southwestcornerof the 10- by lO-cm sampling
tool. Subsequentsamplingunits are selectedin a like manner.Note: The lO-cm units are not drawn
to scale,but are exaggeratedto illustrate the procedure.
procedureprovidesan unbiasedmethodof selectingthe sample,sinceevery unit

has an equal chanceof being included in the sample.With large fields or areas,
it is sometimesadvisableto use a three-stepprocedureas an easiermethod of
locating the samplingunit. In this casethe first scaleinterval may be in kilome-
ters. For very small areas,a onestepproceduremay be used. When a circular
samplingtool is used,this procedurecould conceivablyintroducesomebias into
the estimates;however,this is not thought to be important in practice.
Sometimesa sampleis taken from a one-dimensionalpopulation,such as
a list of cards,a row of pots, or distancesalong a line. A simple randomsample
of this type can be taken in the samemanneras the sampleof distancesin one
direction only in the coordinateor two-dimensionalsystem.Ordinarly only a sin-
gle-stepprocedureis used,drawing one randomnumberfor eachunit to be sam-
pled. Sampling is usually without replacement,i.e., no unit is used more than
once in the sample.
If more than one samplingunit is includedin the sample,the randomsam-
ple providesan estimateof the samplingerror. Estimatesof the meany and vari-
anceV( y) of the meanare given by
y = (I: l=1 Yi)/n [1]
V(.Y) = I:7=1 (Yi - yf/n(n - 1) = s2/n [2]
whereYi is the value observedfor the ith samplingunit and n is the numberof
samplingunits in the sample.If more than 10% of the population is included in
the sample,an adjustmentshould be madein the estimateof the varianceof the
mean. Discussionsof this correction for sampling from finite populationsand
other details of analysisare ably presentedby severalauthors(Cochran, 1977;
Deming, 1950; Hansenet aI., 1953; Hendricks,1956).
Once the varianceof the mean has been estimated,the usual confidence
limits may be placedaroundthe meanby the relationship
6 PETERSEN& CALVIN
[3]
in which L is the confidencelimit, ta , is the Student'st with (n - 1) degreesof

freedom(which is tabulatedin most textbookson statistics)at the a probability
level, and s2/n is as previouslydefined.
If an estimateof the variance is available from previoussamplesof the
populationor can be arrived at from knowledgeof the population,then an esti-
mate of the numberof samplesnecessaryin future samplingto obtain a given
precisionwith a specifiedprobability may be obtainedin the following manner
from Eq. [2] and [3]:
[4]
whereD is the specifiedlimit.

As an example of the application of these equations,supposewe have
taken 10 coresat randomfrom the 0- to 15-cm layer of soil in a particularfield
and have obtained the following estimatesof exchangeablepotassium(K) in
ppm:
59,47,58,80,57,58,62,52,50,47.
The meanexchangeableK level, estimatedusing Eq. [1], would be
y =(59 + 47 + ... + 47)/10 =570/10=57.0 ppm.

The varianceof the meanwould be, from Eq. [2],
V( Y) =(834.00)/(10)(9)=92.67/10=9.267.
The 95% confidencelimits are then obtainedusing Eq. [3]:
L = 57.0 ± 2.26(9.267)1/2= 57.0 ± (2.26)(3.04)

L= 57.0 ± 6.87.
Thus, we can say that unlessa 1 in 20 chancehasoccurredin sampling,the true

populationmeanlies within the range50.13 to 63.87 ppm K; that is, the proba-
bility is 0.95 that the true meanlies within the range50.13 to 63.87 ppm K.
Supposewe are interestedin determining the number of observations
requiredin future samplingof the populationto estimatethe meanwithin ±5.00
ppm, at the 95% probability level. The necessarynumberof sampleswould be
estimatedusing Eq. [4]:
n = (2.26)2 (92.67)/(5.00f= (5.11)(92.67)125.00

n = 18.94.
Hence,to obtain the desiredprecisionit is estimatedthat the samplingrate

shouldbe increasedfrom 10 to 19 in future sampling.
SAMPLING 7
Stratified Random Sample
With stratified randomsamplingthe populationis broken into a numberof

subpopulations,and a simple randomsampleis taken from eachsubpopulation;
e.g., a field may be mappedby series (subpopulations)and a random sample
taken fromeachseries.The reasonsfor samplingsoils in this mannerinclude the
desire(i) to makestatementsabouteachof the subpopulationsseparatelyand (ii)
to increasethe precisionof estimatesover the entire population.For the first rea-
son, the samplingcan be considereda seriesof separatesamplingstudieson a
numberof different populations.The secondreason,however, raisessomespe-
cial considerations,both from the standpointof designand analysis.
If the stratified randomsampleis to have greaterprecisionthan the simple
randomsample,stratificationmust eliminatesomeof the variation from the sam-
pling error. Since every subpopulation,or stratum, is sampled,the differences
amongthe stratummeansare eliminatedfrom the error, and only the within-stra-
tum variation contributesto the samplingerror. If strataare constructedin such
a manneras to make the units within eachstratummore homogeneousthan the
variation among the stratum means, the stratified random sample will have
greaterprecisionthan the simple randomsample.The basisfor making effective
stratificationmay be anothervariable relatedto or correlatedwith the character-
istic of interest,previousknowledgeabout the distribution of the characteristic,
or merely geographicproximity. Any prior information which will aid in making
homogeneousgroupsof units of the populationcan be used.
In general, the more stratification the greater the increasein precision.
There are several precautionsthat should be considered,though, to prevent
excessivestratification. Precision increasesat a decreasingrate as strata are
divided more and more, until a point is reachedwhere no further gain in preci-
sion is obtained.The additional strata also complicate the analysisso that the
gain in precisionmust be consideredin relation to the effort requiredto obtain it.
Since at least two units must be sampledin eachstratumto have an estimateof
the samplingerror from that stratum,it is usually advisableto keep the number
of stratasmall enoughto allow satisfactoryestimatesof error.
The total numberof samplingunits is often allocatedto the stratapropor-
tionately; e.g., if a stratumcontains20% of the populationthen 20% of the sam-
pling units will be taken fromthat stratum.This allocation is not optimum in the
sense thatthe varianceof the meanwill be a minimum; however,unlessthe vari-
ation within the stratadiffers markedly from stratumto stratumit will be nearly
asgood as optimum allocation.Optimum allocation,both with equaland unequal
costs per unit in different strata, is discussedat length by Cochran(1977) and
Hansenet al. (1953).
y,
The estimateof the meanover all strata, and the varianceof this mean,
V( y), are given by
[5]
8 PETERSEN& CALVIN
L
V(y) = (1/N2) LN~(S~/nh) [6]
h=1
whereNh is the total numberof units in the hth stratum,L is the total numberof
strata,N is the total numberof units in all strata,and
as given in "Simple RandomSample."It may be noted that NhlN is the propor-

tion of all samplingunits in the hth stratum;hencethe proportionof the popula-
tion in the hth stratummay be usedin the equationratherthan the total number.
When the allocationis proportionaland all stratahavea commonvariance,
which is a commonoccurrencein soil sampling,then the estimationof the mean
and varianceis simplified:
y= ( i Yi) In
1=1
[7]
[8]
s;
wheren is the total numberof units in the sampleand is the pooledvariance
within strata(s; measuresthe variation amongsampleswithin strata).
If more than 10% of the units in any stratum are included in the sample,
use should be made of the finite population correction referred to in "Simple
RandomSample."
To illustrate the computationsinvolved in stratified randomsampling,sup-
posewe are interestedin estimatingthe cation-exchangecapacityof the surface
soil in a field which containsthree soil types:A, Band C. Supposefurther that
type A represents1/6 of the total areaof the field, type B 1/3, and C 1/2 of the
total area.We might stratify the field with respectto soil type and draw a simple
randomsampleof coresfrom eachstratum,the numberof coresfrom eachstra-
tum beingproportionalto the fraction of the total areaoccupiedby eachsoil type.
That is, we might take 2 samplesfrom typeA, 4 from type B, and 6 from type C.
The resultsof sucha samplingplan are shown in Table I-I.
The meanexchangecapacitywould be estimatedusing Eq. [7] as
y = (11.6 + 13.4 + ... + 17.1 + 19.50/12= 16.62meq per 100 g.

Assuming a common within-stratum variance, the variance of the mean
could be estimatedusing an analysisof varianceof the sampledata, shown in
Table 1-2.
The varianceof the meanV( y) would then be estimatedusing the "with-
in-strata" meansquareand Eq. [8]:
V(y) =2.94/12=0.24.
SAMPLING 9
Table 1-1. Cation-exchange

capacityof soil samplesfrom a field stratified accordingto soil type.
Cation-exchangecapacity(meq/100g) of samplesform indicatedtype
A B C
11.6 19.0 14.2

13.4 21.4 16.5
1B.1 15.7
17.B 15.2
17.1
19.5
Total 25.0 76.3 9B.2
Mean 12.50 19.0B 16.37
Table 1-2. Analysis of varianceof cation-exchangecapacity (meq/100g) of soil samplesin Table

1-1.
Sourceof variation Degreesof freedom Mean square
Among strata 2 29.22
Among sampleswithin strata 9 2.94
And, by analogyto simple randomsampling,we might placethe 95% confidence

limits about the meanusing Eq. [3]:
L = 16.62 ± 2.26(0.24)1/2 = 16.62 ± (2.26)(.49)= 16.62 ± 1.11.
Thus we can say that the true populationmean lies within the rangeof 15.51 to
17.73 meq per 100 g with probability 0.95.
SystematicSample
As the stratified randomsampleis an attempt to ensurethat better cover-

age of the population is obtainedthan with a simple randomsample,so also is
the use of a systematicsamplea further stepin this samedirection. It is a natur-
al extensionand one that is receiving increasingattention.The fact that it is easy
to use in practicehas undoubtedlybeena factor in its wide usage.
The definition of a systematicsamplethat is usedhere coverssamplesin
which the selectedunits are at regulardistancesfrom eachother, either in one or
two dimensions.If the populationis of one dimension(e.g., units along a line, in
a list, or in any linear order), the first unit is assumedto be selectedat random
from the first k units and subsequentunits at eachkth interval (Cochran,1977).
If the populationis of two dimensions,e.g., plots in a field or sectionsof an area,
the surfacecan be consideredto be composedof a numberof strataof common
size and shape.In one stratuma unit is selectedat random,and all units in com-
parablepositionsin all strataare included in the sample.Alternatively, the stra-
tum pattern can be imposedat random on the surfaceand the centerunit from
each stratum included in the sample(see Fig. 1-2). In practice, it is not neces-
sary that the stratumoutlinesactually be designated,althoughthe proceduremust
permit them to be. This definition includes the type of systematicsamplemost
10 PETERSEN& CALVIN
A B
I
{
\ II
r- ......~ ./
Fig. 1-2. Systematicsamplesin two dimensions.Design A is composedof all units at the intersec-
tions of equidistantparallel lines at right anglesto eachother. DesignB is composedof all units at
the intersectionsof equidistantparallel lines, eachset at 60° from vertical. Trianglesare formed
by drawing horizontal lines throughthe intersections.
commonin practice,althoughmany otherscould be designed.This type of sys-

tematicsamplecan also be consideredas a single complexsamplingunit select-
ed at randomfrom suchk units in the population.
For the one-dimensionalcase, systematicsampleshave been compared
with randomand stratified randomsamples,both theoretically(Cochran,1946;
Madow and Madow, 1944; Madow 1949, 1953; Yates, 1948) and empirically
(Finney, 1948; Hasel, 1938; Osborne,1942; Williams, 1956; Yates, 1948). The
resultshave favored systematic samples in nearly all cases.An exceptionpoint-
ed out by severalworkers (Cochran,1953; Finney, 1948; Madow and Madow,
1944; Yates, 1948) may occur when the populationhasa periodic trend, suchas
might be experiencedby a line acrosshills and valleys in a forested area, or
acrossa field with row effects causedfrom previousfertilizer treatment.When
this occurs,the systematicsampleis much less precisethan randomsamplingif
the interval betweenthe samplingunits in the systematicsampleis equal to an
integral multiple of the period, e.g., samplingunits takenon eachrow and none
betweenrows. If the interval is equal to an odd multiple of the half period, sys-
tematic samplingwill be more precise.Milne (1959) statesthat such periodici-
ties seldomoccur in nature;however,Matern (1960) gives severalexamplesbut
indicatesthat a considerablelossof precisionoccursonly when the interval coin-
cidesclosely with the periodicity.
The two-dimensionalcase,applicablefor most field sampling, has been
studied theoretically (Das, 1950; Matern, 1960; Quenouille, 1949; Zubrzycki,
1958) for severaltypesof populations.Empirical studieshave beenvery limited
and confined to crops and forested areas. The theoretical studies have all
assumedthat a correlationexistsbetweenunits close to eachother and that this
correlation decreasesexponentially with the distancebetweenthe units. This
seemsto be a realistic assumption,and it has beensupportedby empirical stud-
ies (Finney, 1948; Milne, 1959; Rigney and Reed,1946).The resultsare similar
to the one-dimensionalcasein that the systematicsamplehasgreaterprecision
SAMPLING 11
than random or stratified random samples, if the correlation between plots

decreasesexponentially with distancebetweenthe plots in both directions. T.
Daleniuset ai. (unpublished)have also shown that the optimum patternfor sev-
eral classesof populationsis the triangulargrid shownin Fig. 1-2B, althoughthe
squaregrid is almost as precise.
The effect of additionaltrendsin the populationhasnot yet beenfully stud-
ied, althoughsuch trendsmay be a more importantfactor than the correlationin
the precision of systematicsampling. It has been pointed out (Madow and
Madow, 1944; Yates, 1948) that the systematicgrid pattern is inefficient when
there is a fertility gradientalong rows or columnsof a field. Since this may be
the casein fields which havea slopeor drainageproblem(or other not suchobvi-
ous conditionsimposinga gradienteffect), it is apparentthat systematicdesigns
cannotbe usedwithout considerationof the form of the populationdistribution.
The empirical comparisonshave shown somewhatconflicting results. In
sampling for crop yield on 14 fields, Haynes(1948) found that the systematic
squaregrid pattern(see Fig. 1-2A) had about the sameprecision as the simple
random sample and less precision than the stratified random sample. Matern
(1960) found that squareand triangular grids gave greater precision than the
stratified random and considerablygreater precision than the random sample
when sampling for percent forested area and percent lake area in a region.
Haynes hypot~esized that a linear trend might be the causeof his results and
showedon the basisof theory that the systematicsampleis indeedlessprecisein
the presenceof such a trend. It can be shown that the precisionalso dependson
relative gradientsin the two directions.In general,when linear trendsare present,
more samplingunits shouldbe selectedin the direction of the strongestgradient.
One of the main problemswith the systematicsamplehasbeenin the esti-
mate of samplingerror from the sampleitself. A numberof methodshave been
suggested(Hansenet aI., 1953; Madow, 1953; Milne, 1959; Osborne, 1942;
Smith, 1938; Williams, 1956), but thosemost commonly usedin practiceare the
following:
1. Assumethat the popUlation was in randomorder before the systemat-
ic samplewas drawn and estimatethe samplingerror as with a simple
randomsample;
2. Block or stratify the sample,assumingthat the variation amongunits
within a block or stratumis only samplingvariation, and estimatethe
samplingerror as with a stratified randomsample;and
3. Take a numberof separatesystematicsamples,eachdrawn at random
from all possiblesystematicsamplesof the sametype, i.e., eachsys-
tematic sample is drawn with a separaterandomly selectedstarting
point.
The sample mean is calculatedfrom each sample, and these meansare
treatedas the data from a simple randomsampleof size equal to the numberof
separatesystematicsamples.Method 3 is the only unbiasedmethodof assessing
the samplingerror without making someassumptionsaboutthe form of the pop-
ulation; however,if the assumptionsmadeundermethods1 and 2 are reasonably
valid, thesemethodsgive greaterprecision.
12 PETERSEN& CALVIN
SOURCESOF ERRORS
Errors arising from soil sampling(Das, 1950) fall into three generalcate-
gories, viz., sampling error, selectionerror, and measurementerror. Each con-
tributesto the total error, and considerationof eachof them is necessaryto ensure
a satisfactorysamplingprocedure.
Samplingerror is the error arising from the fact that the sampleincludes
only the selectedsamplingunits ratherthan the entire population.It is causedby
the inherentvariationamongthe units of the populationand can be avoidedcom-
pletely only by including all of the populationin the sample.
Selectionerror arisesfrom any tendencyto selectsomeunits of the popu-
lation with a greateror lesserprobability than was intended,e.g., a tendencyto
avoid rocky sitesin a field or to oversamplethe bordersof a field. Thereis some
tendency for selection errors to cancel each other; and yet, as sample size
increases,bias resultingfrom the selectionproceduremay becomean increasing
portion of the total error. The twostep procedureoutlined in "Simple Random
Sample"is a methoddesignedto minimize selectionerror, and analogousproce-
durescan usually be devised.
Measurementerror is that error causedby the failure of the observedmea-
surementto be the true value for the unit. It includesboth the randomerrors of
measurement,which tend t9 cancelout with increasedsamplesize, and biases,
which are usually independentof samplesize. Examplesof random errors are
thosecausedby assumingconstantweightsfor coresof soil which have variable
weights andthosecausedby chancevariationsof techniquein the analyticalpro-
cedure. Biases,independentof sample size, would result if tare weights were
ignoredor if a calibrationcurve were offset to one side of the appropriatecurve.
If a biasedtechniqueis used,such as subsamplingfrom the top of a con-
tainerof improperly mixed soil (selectionerror) or using an analytical test which
gives a readingtoo high (measurementerror), then this error, or bias, would not
be includedin the computedsamplingerror. Only constantattentionto technique
can hold thesebiasesto a minimum, and even then no estimateof their magni-
tude is ordinarily available.
Although considerablestudy is neededon the sourcesof error in practical
surveys,some evidencewould point to sampling error generally being larger
than randommeasurement error (Cline, 1944;Hammondet aI., 1958; Rigney and
Reed,1946).
SUBSAMPLING
In many types of soil investigation,the use of subsampling,or multistage

sampling, is advantageous.With this technique,the sampling unit, selectedby
oneof the previouslydescribedmethods,is divided into a numberof smallerele-
ments. The characteristicunder considerationis then measuredon a sampleof
theseelementsdrawn at randomfrom the unit. For example,a sampleof cores
may be taken from a field plot, and a numberof small samplestaken from each
core for chemicalanalysis.
SAMPLING 13
The primary advantageof subsamplingis that it permits the estimationof

somecharacteristicof the larger samplingunit without the necessityof measur-
ing the entire unit. Hence,by using subsampling,the cost of the investigation
might be considerablyreduced.At the same time, however, subsamplingwill
usually decreasethe precisionwith which the characteristicis estimated.At each
stage of sampling, an additional componentof variation, the variation among
smallerelementswithin the largerunits, is addedto the samplingerror. Thus, the
efficient useof subsamplingdependson striking a balancebetweencost and pre-
cision.
To illustrate the statistical considerationsinvolved in subsampling,sup-
posewe draw n samplingunits at randomfrom a population,and from eachunit
we select, at random, m elements.The mean, on a per elementbasis, may be
estimatedfrom
y = ( i=1i: j=1EYij ) /nm [9]
y
in which is the samplemean,Yij is the observationon the jth elementin the ith
unit, n is the numberof units in the sample,andm is the numberof elementssam-
pled per unit.
The varianceof the mean,V(y), on a per elementbasis,is given by
V( y) = CJs/n + CJ~/mn [10]
where CJS is the variation amongunits, CJ~ is the variation amongelementswith-

in units, and nandm are as previously defined.
The componentsof variance,CJt and CJ~, may be estimatedfrom an analy-
sis of varianceof the sampledata,shown in Table 1-3.
The variation amongelements,CJ~ is estimatedby the meansquareamong
elementswithin units, s~.The variation amongunits, CJS is estimatedfrom
[11 ]
If the sampleincludes>10% of the units in the population,an adjustmentshould

be madein the estimateof the varianceof the mean. The proper adjustmentis
adequatelydiscussedby severalauthors(Cochran,1977; Deming, 1950; Hansen
et aI., 1953).
Table 1-3. Analysis of varianceof the sample(on a per elementbasis).

Mean squareis
Sourceof variation Degreeof freedom Mean squaret an estimateof
Among units (n - 1)
Among elementswithin units n(m - 1)
t =mL,{)li - y)Z/(n - 1)
Sb2
=I/f.iYij - Yi)Z/n(m - 1)
sw2
where)Ii =the meanof the ith unit =(Ij!,! Yij)/m.
14 PETERSEN& CALVIN
If estimatesof a; and a6 are availablefrom previoussamplesor can be

arrived at from a knowledgeof the population,we can useequation[10] to pre-
dict the varianceof the meanin future samplingfrom the sametype of popula-
tion. The proposedsamplingand subsamplingrates,nandm, and the estimates
a;
ofthe variancecomponents, and at, aresubstitutedinto the appropriateequa-
tion.
It is apparentfrom Eq. (10] that the varianceof the meanmay always be
reducedat a more rapid rate by increasingthe numberof units sampledthan by
increasingthe samplingratewithin units. This may not be the most efficient pro-
cedure,however, if the cost of taking the sampleis considered.The optimum
samplingand subsamplingrate will dependon the relative cost of samplingthe
unit and that of samplingthe elementwithin the unit. A cost relationshipwhich
hasbeenuseful in many typesof soil investigationis given by
(12]
in which C is the total cost of obtaining an estimateof the mean,Cb, is the cost
per unit, directly assignableto the unit and independentof the numberof ele-
mentsper units, Cwo is the cost per element,and nandm are the numberof units
and the numberof elementsper unit, respectively,in the sample.
When the varianceof the mean is of the form given by Eq. [10] and the
cost relationshipis that given by Eq. [12], it can be shown(Cochran,1977) that
the optimum subsamplingrate, m', may be obtainedfrom
[13]
That is,m' can be shownto give the smallestvariance fora given cost, or alter-
natively, the leastcostfor a given variance.The samplerate, n, is found by solv-
ing eitherEq. [10] or Eq. [12], dependingon whetherthe varianceor the cost has
beenspecified.
In practice,m' will usually not be an integer,and the nearestintegershould
be selected.In addition, the varianceusually changesratherslowly for valuesof
m in the region of the optimum. Thus somelatitude in the selectionof m' can be
tolerated.
As an exampleof the computationsinvolved in subsampling,supposewe
were interestedin estimatingthe effect of continuousgrazingon the bulk densi-
ty of a pasturesoil. And supposethat we drew a randomsampleof five 3- by 3-
m samplingunits, andwithin eachunit we selected,at random,threesubsamples
in the form of 7.5-em cores upon which the bulk density measurements were
made.The resultantobservationsmight appearas in Table 1-4.
The meanbulk density may be estimated fromEq. [9] as
_ (1.53 + 1.48 + ... + 1.58 + 1.50) 22.99

Y=
5x 3
=-15- =1.53.
The analysisof varianceof the datais shown in Table 1-5.

SAMPLING IS
Table 1--4. Bulk density of three subsamplesof soil in eachof five samplingunits.
Bulk density of soil in indicatedsamplingunit
Subsample 2 3 4 5
g/cm2
1 1.53 1.63 1.47 1.59 1.64
2 1.48 1.58 1.41 1.36 1.58
3 1.64 1.67 1.50 1.41 1.50
The variancecomponentsmay then be estimatedas
0; = 0.0061
06 = (0.0168- 0.0061)/3= 0.0036.
Thus from Eq. [10] the varianceof the mean,on a core basis,is found to be
V(y) =0.0036/5+ 0.0061/5x 3 =0.0007+ 0.0004=0.0011.

Now supposewe are interestedin determining the effect on the variance of
reducingthe numberof samplingunits to 3 and increasingthe numberof cores
per unit to 5. Again we would use eq. [10] to obtain
V( y) =0.0036/3+ 0.0061/3x 5 =0.0012+ 0.0004 =0.0016.

Now supposewe wish to estimatewhat the optimum samplingrate would be in
future samplingfrom the sametype of population,and supposethat it costsfive
times as much to samplean additional unit as it doesto take an additional core
in eachunit. From Eq. [13] we would find the optimum samplingrate to be
m' = [(5 x 0.0061)/(1x 0.0036)p/2 =(0.0305/0.0036)112 =(8.47)112

m' = 2.91.
Thus, in samplingfrom a similar population,we would expectthe maximumpre-

cision for a given cost by taking 3 coresper unit.
It should be pointed out that subsamplingneednot be restrictedto a two-
stageprocedure.In some instances,the sample may be taken in three or more
stages.A core of soil, for example,might be subsampledto provide small quan-
tities of soil for chemical analysiswhich, in tum, might be subsampledto pro-
vide aliquots for the final measurement.The principles involved in multistage
samplingare essentiallyextensionsof the principlesdiscussedhere.The statisti-
Table 1-5. Analysis of varianceof bulk density data in Table 1-4.

Sourceof variation Degreesof freedom Mean square
Among units 4 0.0168

Among coreswithin units 10 0.0061
16 PETERSEN& CALVIN
cal considerationsare adequatelypresentedby severalauthors(Cochran,1977;

Deming, 1950; Hansenet aI., 1953; Yates, 1953).
COMPOSITESAMPLES
In many soil investigations,a substantialsaving in total cost can result if

laboratoryanalysesare performedon acompositeof the field samplesratherthan
on the individual samples.The procedureconsistsof taking a numberof field
samplesadequateto representthe population in question, thoroughly mixing
thesesamplesto form one compositeor bulk sample,and performing the labo-
ratory analyseson the compositesampleor on asubsampleof the composite.The
assumption,of course,is that a valid estimateof the meanof somecharacteristic
of the population may be obtainedfrom this single analysis of the composite
sample.This assumptionis true only undercertainconditions.All sampleswhich
form the compositemust be drawn from the populationunderconsideration,and
eachsamplemust contributethe sameamountto the composite.
For example,supposea study is initiated to determinethe changesin avail-
able soil phosphoruswhich have taken place in a field experiment which
involved the annual applicationof varying incrementsof phosphorusto certain
of the plots in the experiment. A composite sample for each plot could be
obtained by pooling a number of borings taken at random from the plot.
Similarly, if no estimateof precision is needed,the mean availablephosphorus
for a given treatmentwould be obtainedfrom a compositeof equal numbersof
samplesfrom all plots with a commontreatment.Forming a compositeby pool-
ing samplestakenfrom severaltreatments,however,is seldomif everjustified.
It should be pointed out that the compositesamplesprovide only an esti-
mate of the meanof the populationfrom which the samplesforming the com-
posite are drawn. No estimateof the varianceof the meanand hencethe preci-
sion with which the meanis estimatedcan be obtainedfrom a single composite
sample.It is not sufficient to analyze two or more subsamplesfrom the same
compositeto obtain an estimateof the variation within the population. Such a
procedurewould permit the estimationof variationamongsubsampleswithin the
composite,but not the variation amongsamplesin the field. Similarly, if com-
positesare formed from sampleswithin different partsof a population,the vari-
ability amongthe parts,but not the variability within the parts,can be estimated.
If an estimateof the variability among sampling units within the population is
required, two or more samplestaken at random within the population may be
analyzedseparately.
The accuracywith which a populationmeanis estimatedfrom a compos-
ite is dependenton the variability amongsampling units within the population
and the numberof such units includedin the composite.Becauseof the differ-
encein variability amongthe severalcharacteristicsof a soil and amongthe sev-
eral typesof samplingunits, no generalizationscan be maderegardingthe num-
ber of samplingunits requiredfor a compositesample.If an estimateof the vari-
ability amongunits is available,it can be usedto determinethe numberof units
to include in the compositeto attain a given precision.The procedureis the same
as that given in "Simple RandomSample"for estimatingthe sizeof a simple ran-
SAMPLING 17
dom sample.If no estimateof variability is available, compositingshould be

avoidedif possible.
Subjectto the foregoing restrictions,compositingmay prove valuablein
soil investigation.It permitsthe precisionwith which the meanis estimatedto be
increasedby increasingthe numberof units included in the samplewithout, at
the sametime, increasingthe costof analysis.
REFERENCES
Allmaras, R. R., and C. O. Gardner. 1956. Soil sampling for moisture determinationin irrigation
experiments.Agron. J. 48:15-17.
Cline, Marlin D. 1944. Principlesof soil sampling.Soil Sci. 58:275-288.
Cochran,W. G. 1946. Relative accuracyof systematicand stratified samplesfor a certain classof
populations.Ann. Math. Statist. 17:164-177.
Cochran,W. G. 1977. Samplingtechniques.3rd ed. JohnWiley and Sons,Inc., New York.
Das, A. C. 1950. Two-dimensionalsystematicsamplingand associatedstratified and randomsam-
pling. Sankhya10:95-108.
Deming, W. E. 1950. Sometheory of sampling.JohnWiley and Sons,Inc., New York.
Finney, D. J. 1948. Randomand systematicsamplingin timber surveys.J. Forestry22:6499.
Fisher,R. A., and F. Yates. 1967. Statisticaltablesfor biological, agricultural,and medical research.
6th ed. Oliver and Boyd, Edinburgh.
Hammond,L. C., W. L. Pritchett,and V. Chew. 1958. Soil samplingin relation to soil heterogeneity.
Soil Sci. Soc. Am. Proc. 22:548-552.
Hansen,M. H., W. N. Hurwitz, and W. G. Madow. 1953. Samplesurveymethodsand theory, Vols. I
and 11. JohnWiley and Sons,New York.
Hasel,A. A. 1938. Samplingerrorsin timber surveys.J. Agric. Res.57:713-736.
Haynes,J. D. 1948.An empirical investigationof samplingmethodsfor an area.M.S. thesis,North
CarolinaStateCollege,Raleigh,NC.
Hendricks,w. A. 1956.The mathematicaltheory of sampling.ScarecrowPress,New Brunswick,NJ.
Madow, W. G. 1949. On the theory of systematicsampling:II. Ann. Math. Statist.20:333354.
Madow, W. G. 1953. On the theory of systematicsampling:III. Ann. Math. Statist.24: 101106.
Madow, w. G., and L. H. Madow. 1944.On the theory of systematicsampling:1. Ann. Math. Statist.
15:1-24.
Mason,D. D., J. F. Lutz, and R. G. Petersen.1957. Hydraulic conductivity as relatedto certainsoil
propertiesin a numberof greatsoil groups-samplingerrorsinvolved. Soil Sci. Soc.Am. Proc.
21:554-560.
Matern, Bertel. 1960. Spatialvariation. Stochasticmodelsand their applicationto someproblemsin
forest surveys and other sampling investigations. Meddelanden Fran Statens
Skogsforskningsinstitut.Band. 49, NR. 5.
Milne, A. 1959.The centricsystemarea-sampletreatedasa randomsample.Biometrics15:270-297.
Olson, R. A., A. F. Drier, and R. C. Sorensen.1958. The significanceof subsoil and soil seriesin
Nebraskasoil testing.Agron. 1. 50:185-188.
Osborne,J. G. 1942. Samplingerrorsof systematicand randomsurveysof cover-typeareas.J. Am.
Statist.Assoc.37:256-264.
Quenouille,M. H. 1949. Problemsin plane sampling.Ann. Math. Statist.20:355-375.
Rigney, 1. A., and J. F. Reed. 1946. Somefactors affecting the accuracyof soil sampling. Soil Sci.
Soc. Am. Proc. 10:257-259.
Sayegh,A. H., L. A. Alban, and R. G. Petersen.1958. A samplingstudy in a saline and alkali area.
Smith, H. F. 1938. An empirical law governingsoil heterogeneity.J. Agric. Sci. 28:1-23.
Williams, R. M. 1956. The varianceof the meanof systematicsamples.Biometrika43:137-148.
Yates,F. 1948. Systematicsampling.Phil. Trans. Roy. Soc. London, Ser. A. 241:345-377.
Yates, F. 1953. Samplingmethodsfor censusesand surveys,2nd ed. CharlesGriffen and Co. Ltd.,
London.
Zubrzycki, S. 1958. Remarkson random,stratified and systematicsamplingin a plane. Colloquium
Mathematicum6:251-264.
Published 1996
Chapter 2
Quality Assurance and Quality Control
EUGENE J. KLESTA, Chemical Waste Management, Inc.
JOAN K. BARTZ, Dames & Moore, Richland, Washington
SCOPE
This section will describethe concepts, principles,and general proceduresof
quality assurance(QA) as it appliesto the analysisof soil samplesand materials
relatedto soil science.Although the primary focus will be on analytical chem-
istry, samplingwill be discussedto someextentbecauseof its importanceto the
overall accuracyof the generateddata. Data producedfrom the analysisof the
sampleare representativeof the population and are reliable only if the original
samplesthemselvesare representativeof the population.
Sampling
Samplingactivities require quality assuranceprinciples such as the devel-
opmentanduseof standardizedsamplingprocedures,training and documentation
of samplingpersonnel,and the creationof traceableand defensibleinformation
throughthe useof labelsand chainsof custody.A samplingplan shouldbe writ-
ten to describethe location, number,size, and frequencyof samplesto be taken.
Specificpreservationor handlingprocedureswhich are not readily inferred must
be usedto preventthe contaminationof the sample.There are a wide variety of
samplingstrategieswhich can be employedto obtain sampleswhich will result
in the accumulationof desiredinformation. When compositingtechniquesare to
be used,specific information regardingthe numberof incrementsto be compos-
ited and the size of eachincrementmust be includedin the samplingplan. When
any deviationsfrom the samplingplan take place,clear, completedocumentation
is required to produce a permanentrecord that some nonconformingpractices
were performed.The reasonfor the deviation also must be included.
Physical Testing
Quality assuranceprinciples are applicableto physical testing. Reliability
of information, preciseand accuratedata, and defensibility are desirableconse-
SegoeRd., Madison,WI 53711,USA. Methods of Soil Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
19
20 KLESTA & BARTZ
quencesthat occurwhen standardizedproceduresare used,recordkeepingprac-

tices include all pertinentinformation, test performersare properly trained,com-
plete training documentationis maintained,and a quality control (QC) systemis
in place to determineerrors, to identify bias, and to require corrective action
when needed.The importanceof physicaltestingis often overlooked.Parameters
suchascompressibility, permeability, or particlesizedistributioncanbe extreme-
ly important for the successor failure of a particularproject. The quality assur-
ancesystemmust include theseareasof testingto be an acceptablesystem.
ChemicalTesting
Chemical testing may be commonly referred to as analysisor analytical

chemistry.The principlesof QA andQC are more establishedandgenerallyprac-
ticed moreoften in the arenaof analyticalchemistrythan in the samplingor phys-
ical testingareas.Regulatoryagenciesand commercialclients haverequiredthat
laboratoriesimplementQA and QC practicesto improve the users' confidence in
the analytical data. The following discussionwill describethe critical compo-
nentsof a quality systemandwaysto implementQC to improve the accuracyand
precisionof analyticaldata.
POLICY STATEMENT
A policy statementis an essentialpart of any quality system.Basically it is

a written statementwhich is developedand communicatedto all participantswho
are responsiblefor the implementationand managementof the quality system.It
is importantto understandthe objectivesof the quality systemand the domainof
its authority. Likewise, the policy statementwill communicate,by their absence,
which areasare not included in this particular quality system. Several policy
statementsmay be neededto cover all areasof concern.The goal to be achieved
by the quality systemparticipantsmust be establishedand communicated.
Authority
Participantsin a quality programshould be able to recognizeclearly that

the management,governingbody, or academicauthority supports,endorses,and
participatesin the program. The scopeof the program should be defined and
delineatedto avoid confusion.Successfulimplementationof the quality program
resultswith support"from the top" and fails without it.
Goal
The goal shouldbe includedin the policy statement.The goal of the qual-
ity systemmust be documentedin terms that allow for measurableevaluation.
One of the key componentsof a quality system is an assessmentphase.The
assessment of the complianceto requirementsof the programcannotbe readily
madeif the ultimate goal is not understood.
QUALITY ASSURANCE & QUALITY CONTROL 21
Responsibility
Membersof the organizationwho are responsiblefor the implementation

of the quality system,assessment of the complianceto the requirements,and del-
egation of authority to take corrective action must be designated.Each partici-
pant'sresponsibilitiesshould be clearly defined. The responsibilitiesof thosein
chargeof the organizationalso should be defined. The more precisethe quality
statementand the accompanyingresponsibilitiesare, the more successfulwill be
the implementationof the program.
DEFINITIONS
Quality Assurance
Quality assuranceis the systemwhich includesrequirements,procedures,

and assessment to ensurethat the goal of the programis achievedand to measure
the level of achievementto that goal. Any specific goals that are expectedfrom
the quality assuranceactivities should be documented.For example,one state-
ment of a major corporationincludesthesewords " ...to provide valid, defensi-
ble data in a timely manner."
Quality Control
Quality control is a systemof proceduresand practiceswhich result in the

increasein precision and the decreasein bias. The use of duplicate analysis,
spiked samples,standardreference materials, and QC check samplesare all
mechanismsusedto demonstratethe control of quality. Quality control is a sub-
set of QA. The principles of QC reach the level of the field technician,bench
chemist,and laboratoryanalyst.
Defensibility
Defensibility is a quality of analytical data, field information, and general

recordkeepingwhich allows for the admissionof the recordsinto a courtof law
to standon their own merits. The lack of ambiguity, the clarity and accuracyof
information, and the completenessof the explanationsby the authorall improve
the defensibility of the data.
Traceability
Traceability is a characteristicwhich is essentialto substantiatethe validi-

ty of analyticaldata,the appropriateness of the sampletested,and the correlation
to the calibrationstandardsused.The defensibility of information relies heavily
on its traceability. The correlation of information is a necessaryattribute of a
quality system.
22 KLESTA & BARTZ
QUALITY ASSURANCE MANUAL
An essentialpart of the quality assurancesystemis the quality assurance

manual.The manualcan take a variety of forms. The main attributesfor devel-
oping an acceptablemanual are: completeness,portability, applicability, and
clarity.
All necessarymaterial must be included in the manual. The usersof the
manualneedto have a copy of the manualnear at hand. Putting togethera huge
tome that will not find its way off the shelf defeatsthe purposeof the manual.
The manualis focusedon the activities taking place in the laboratoryor in field
operations.Specificity of proceduresis necessaryto makethe manualapplicable.
The ability to edit the contentsin a reasonableamountof time is necessary.As
the quality systemdevelops,grows, or changes,the quality assurancemanual
must keeppacewith this progress.The responsibleauthorsof the quality assur-
ancemanualshouldwork at removing ambiguity.andconfusion.
STANDARD OPERATING PROCEDURES
One of the prerequisitesfor having and maintaininga good quality system

rests on the need to have all membersof the staff operating on "common
ground."Writing and using standardproceduresare the first stepstoward achiev-
ing comparability.Peopleinvolved in activities tend to developtheir own way of
doing things. Creativity under theguiseof improvementcan result in changesin
procedureswhich could result in undesirableconsequences and unusabledata.
Managementof the organization must determine where and when Standard
OperatingProcedures(SOPs)are needed.
Organization
A collection of SOPsis necessaryto have an acceptablequality system.A

format shouldbe developedso that all SOPswill be in the sameformat. The stan-
dardizedformat improvesthe easeof use by the reader.Standardoperatingpro-
ceduresare usedfor referenceby personnelin the field operationsand the labo-
ratory operations.Easily finding "key" information in a SOP resultsin an over-
all improvement in efficiency. The reader finds the information quickly and
returnsto the taskat hand.Throughcontinueduseof the SOPs,the technicalstaff
becomesmore familiar with the procedureandgenerallywhat is expectedin day-
to-day operations.Standardoperatingproceduresare essentialelementsfor the
training of new personnel.
Analytical Structure
Standardoperatingproceduresare useful for clarification and standardiza-
tion of analytical methods.All analystsshould be performingthe analyticalpro-
ceduresexactly the sameway. Becausesomeanalytical methodsdo not contain
the specificity neededto ensurecompleteconsistency,it is necessaryfor the lab-

oratory managementto clarify thesepoints of confusion and to specify details
where the methodsare lacking. The developmentof SOPscovering analytical
methodssatisfiesthis requirement.Specificationof calculations,choiceof labo-
ratory equipment, or timelimits for certainstepsnot addressedin the methodare
examplesof points to be included in an analytical SOP. Generallaboratorypro-
cedureswhich can affect the analytical results shouldbe includedin the scopeof
SOPs.Glasscleaningprocedures,safetyconcernsand rules, and samplehandling
proceduresand disposalpracticesare examplesof issuesto be covered.
Quality AssuranceStructure
Standardoperatingproceduresare neededto describethe quality assurance
structure and responsibilities.When the organizationdeterminesthat specific
quality control or quality assurancepracticesare required, then SOPscovering
theseareasmust be written and distributedto all personnel.If a quality assurance
unit or a quality assuranceofficer is the mechanismemployed to overseethe
quality systemand to determineadherenceto the program, then the respective
responsibilities,duties,and authority must be spelledout in the SOPs.
ManagementResponsibility
The managementof a samplingor analytical operationhas a responsibili-
ty to develop an organizedquality system.Leadershipis an essentialfactor to
ensurethat a quality programis successful.The use of standardizedprocedures
resultsin conformity andclarity. The repeatabilityof analysisis greatly enhanced
by conformanceto standardizedprocedures.Managementmust set the "ground
rules" for the rest of the organization. Approval of the standardprocedures
should be done by managementin a systematicprocess.
The quality managementfunction is responsiblefor instituting standard
proceduresfor the QA and QC practicesthat are to be performeduniversally.
Input from the staff performing the sampling and the analysesis an important
considerationfor managementto solicit. The practicality and usefulnessof the
standardprocedureswill be greatly improved by using the input of the staff.
Adherenceto the proceduresfollows readily when staff membershave an inte-
gral part in the developmentof thoseprocedures.
EmployeeResponsibility
Each memberof the samplingor analytical staff is required to be knowl-
edgeableof all SOPswhich impacton their job responsibilities.The currentSOP
mustbe usedto perform the samplingor analyticaltasks.When it is necessaryto
alter the procedurefor any numberof legitimate reasons,the employeehas the
responsibilityto notify the managementthat the SOPis in needof revision. Stan-
dard operatingprocedureswhich are written with input from the employeeare
generallythe most practical and useful. Cooperationin writing and revising the
documentsis one of the key attributesof a successfulquality program.
24 KLESTA & BARTZ
Analytical Methods
Written analyticalmethodsare a necessityto maintainQA at an acceptable
level. The analystsmust follow the stepsof a procedureexactly the way they are
written. The comparabilityof analyticaldatafrom analystto analystdependson
adherenceto written analytical methods.The laboratoryshould maintain a cur-
rent methodsmanualwhich containsall of the methodsthat are being usedin the
laboratory.Methodswhich are no longer usedshouldbe maintainedin historical
files, but should be removedfrom all manualsfound in the laboratory.Records
must be maintainedto documentwhen a changeis madefrom one version of a
methodto a later version of a method.The defensibility of the data dependson
the ability to correlatethe data to the versionof the methodin use at the time of
generation.
StandardMethods
Analytical methodswhich have undergonea rigorous processfor review,
validation, and promulgationare the most desirablemethodsto be used. Soci-
eties andassociationswhich publish methodshavea variety of approvalprocess-
es which are used to quality the methodbefore it is given the final approvalof
the organization.The defensibility of analytical data is improved considerably
when data are generatedusing a standardmethod.The scopeand applicationof
the methodmust be appropriatefor the material to be analyzed.The misuseof a
standardmethodis just as unacceptable as using no standardmethodat all. Some
of the commonissuersof methodsare: AmericanSocietyfor Testingand Mater-
ials (ASTM), American Public Health Association(APHA), Soil ScienceSoci-
ety of America (SSSA), Association of Official Analytical Chemists-Interna-
tional (AOAC), United StatesEnvironmentalProtectionAgency (USEPA).
Validation of Methods
Proceduresfor the validation of analyticalmethodsare neededin the qual-
ity assurancemanual.Whetherthe methodis a standardmethodor one that was
developedwithin the laboratory,it is critical to demonstratedefensiblythat the
analystsare capableof generatingresultsthat meetthe acceptancecriteria of the
method.The numberof replicateanalysesto be performedand the materialto be
usedfor the validation of the methodshould be specifiedclearly in the applica-
ble SOP.
Modification of Methods
Whensomespecificstepor procedureof a standardmethoddoesnot apply
to particular sample matrices or technical improvementscan be made, then a
methodmodification should be written, reviewed,and approved.The natureof
the modification cannotchangethe basicchemistryof the procedure.Technical
concernsshould be reviewed by staff membersfor correctnessand technical
merit. Validation of the modified method should be performedto identify any
potentialbias. Modifications must be in writing and shouldbe placedwithin the
analytical methodsmanual for ready reference.A standardform for modifica-
tions is helpful to ensure that all necessaryissues for a modification to be
QUALITY ASSURANCE & QUALITY CONTROL 2S
approvedare includedin the modification. The methodmodification must not be

usedbefore it receivesapproval.
Approval
An approvalsystemshouldbe put in placeto preventthe unauthorizeduse
of nonstandardmethodsor method modifications. The person responsiblefor
methodapprovalsshouldhavesignificantexperiencein the use and development
of analyticalmethods.The approvalshouldbe madeby the staff memberwith the
most responsibilityfor ensuringthat analyticaldataof the highestquality are pro-
duced. In multilaboratory situations,an appointedofficial should be given the
authority and responsibilityto review and approveall methodmodificationsand
nonstandardmethods.Distribution of the modification to all holdersof the meth-
ods manualalso is the responsibilityof that appointedofficial.
RecordKeeping
Recordkeepingis a very importantaspectof the quality assurancesystem.

Analytical data are defensiblewhen the processwhich was usedto generatethe
datacan be retracedthrough all stepsof the processsuchas test portion amount,
length of time for extraction step, dilutions, operatingconditions of the instru-
ment, and calculations.Data must be recordedand preservedin such a way that
all critical information can be retrievedat somefuture date.
Logbooks
Logbooks for recording information and data should be bound and
designedfor the identificationof the book andthe uniquenessof the pageand line
numbers.Logbooksshould be sequentiallynumbered, traceable from the issuer
to the user, and subject to a system for inventory and archive. Meeting these
requirementswill result in completelydefensiblerecords.
Benchsheets
Various laboratory organization schemes may necessitatethe use of
benchsheetsfor recording analytical data. The practical benefit of using
benchsheetsis that they can be designedand used for specific analytical tests.
Having customizedforms can be very useful in assuringthe completedocumen-
tation of all critical parameters.The analystis somewhatforced to completeall
of the blankson a benchsheet.Times, temperatures,flow rates,and calculations
can be incorporated in the format resulting in improved record keeping.
Benchsheetsshould be prelabeledwith unique sequentialnumbers.When a sig-
nificant numberare accumulated,the benchsheets shouldbe identified and bound
in such a way that they cannotbe lost or stolen.
Data Recording
Data should be recordedin permanentink. Black or blue ink is preferred
becauseit canbe photocopiedand doesnot bleedor becomeillegible as othercol-
26 KLESTA & BARTZ
ors do. Datashouldneverbe obliteratedby crossingout, using opaquecorrection

fluids, or coveredby tape. Correctionsto data may be made as describedin the
sectionon "Data Management."
Archiving
Defensibility of information includesthe ability to reproducethe informa-
tion at somefuture date which may occur days, months,or many yearsafter the
information was first generated.To achievethis objective,a systemfor archiving
must be in place. The proceduresfor the distribution, use, recovery,and storage
of logbooksmust be described.The accumulationand storageof benchsheets and
instrumentprintoutsmust be detailed.Secure,fireproof storageis requiredfor all
paperrecords.An organizedsystemwill facilitate retrieval of information. The
systemcan be set up by date, project, client, or stateas long as a test of the sys-
tem during the quality assessment phaseof the programprovesto be successful.
Proceduresfor magneticmediashould be determinedand clearly documentedin
an SOP.The useof compactdiscs-readonly memory(CD-ROMs) and laserdisks
are becomingmore acceptableeachyearbecauseof the advantagesof size reduc-
tion, searchcapability, and permanence.Use of magneticor optical media may
not be acceptableas the only meansof archiving. The responsibleauthority for
oversight of the work should be consultedbefore implementing an optical or
magneticsystemwhich replacesthe paperrecordscompletely.
Computers
The use of computersto capture,calculate,and store analytical data great-
ly enhancesthe quality and accessibility of the data. The transcriptionof data
from one location to anothercan result in a significant numberof errors. Putting
the information into an electronic medium reducesthe chanceof transcription
errors. Information from a computerizedsystemis retrievedin a fraction of the
time that a "paper" systemwould take. There are somequality assuranceconsid-
erationsthat must be usedwhen developingan electronicsystem.
Security
Accessto the computersystemmust be controlled. The use of passwords
and a hierarchalorganizationadd to the overall security. Managersand supervi-
sorswill haveaccessto a greateramountof information than the analystor tech-
nician. A magneticor electronic"audit trail" or history fill shouldbe includedin
the computersystem.Wheneveraccessis grantedto storedinformation, a sepa-
rate nonaccessiblerecord is madeof the transaction.Wheneverchanges tostored
dataare made,a justification shouldbe requiredby the systembeforethe change
can be completed.Both the original information and the correctedinformation
should be storedpermanently.Instrumentationis becomingmore computerized,
and the transferof databy electronicandmagneticmeansis becomingmore com-
monplace.Becauseof thesedevelopments,securityis an essentialprinciple of the
quality system.
Backup Procedures
Computersystemsrequire a specializedprocedurewhich is not neededin
typical paperrecord systems.Becauseof the possibilitiesof magneticdistortion
or mechanicalfailure, duplicationof essentialdatais requiredto preventthe per-
manentloss of data. The easeof duplication and the relatively small amountof
spaceneededto store the duplicate recordsmake backupproceduressuitableto
computersystems.The conceptof maintaininga backup to all paperrecordsis
impractical and limited by space.The developmentand use of an SOP is essen-
tial to ensurethat the backupproceduresare done at the requiredfrequencyand
in the correctmanner.
Tape. When a tape system is used for archiving and backup procedures,
thereare somespecific requirementsto ensurethe integrity of the tapesfor future
access.Tapesshould be rewound periodically (approximatelyevery 6 mo) and
datashould be transferredto new tapeson a regularbasis(every 5 yr or less). It
also is necessaryto maintainthe hardwareneededto be able to read the tapes.As
technologyimproves,size, speed,and capacityalso change.Storing tapeswhich
cannotbe readbecausethe hardwarehasnot beenmaintainedshould be prevent-
ed. Tapesmust be storedin secure,hardened,fireproof facilities to preventcata-
strophicloss.
Alternate Media. The developmentof alternatemedia is improving the
longevity of archiving in somecondensedform. The use of CD-ROMs or laser
disks which use optical meansrather than magneticmeansto store information
will increasethe amountof time that information can be storedbefore the trans-
fer to new media is needed.Theseoptical media are estimatedto be stable for
approximately20 yr. Developmentsof technologysuchas thesewill improve the
ability to retrieve datawhen long-term storageis needed.
Training
Having qualified personnelis one of the primary componentsof a quality
system.To assurethat the peopleperforming the sampling,analysis,and quality
assessment functions are both competentand conscientious,propertraining must
be conducted and documented.Although technically educated people have
knowledgewhich can be appliedto the tasksat hand,it is very importantthat spe-
cific training in proceduresis conducted. Consistencybetween personnel is
improved through proper training. Specific proceduralsteps or quality control
practicesmay not be readily discernibleeven to a technically educatedperson.
Serialtraining shouldbe avoidedbecauseit allows for "folklore" to creepinto the
system.Combining standardizedprocedureswith a good training program will
result in the highestprobability for preciseand accurateinformation.
Methods
All personnelwho are to perform a specific samplingor analytical method
shouldreceivetraining in that methodand should have documentedevidenceof
28 KLESTA & BARTZ
proficiency in the method.The training scenarioshouldinclude the following: (i)

The traineereadsand understandsthe written method.(ii) The traineeobserves
the trainer perform the method in its entirety. (iii) The trainee performs the
methodwith observationby the trainer.(iv) The traineeperformsthe method a
secondtime using a check sampleor a referencematerial to demonstrateprofi-
ciency. (In the caseof sampling,proficiency is measuredby comparinganalyti-
cal results from the trainee'ssample to those from the trainer's sample.)(v)
Short-termfollow-up by the trainer should occur to ensurethat the method is
being followed as written and to answerany questionswhich may needanswers
or clarification. One of the major sourcesof error in samplingor in the laborato-
ry is the result of nonconformanceto the method.
Quality Assurance/QualityControl Program

The specific proceduresof the quality assurance/qualitycontrol program
also requiretraining. The frequencyof replicatesand spikes,the interpretationof
suspectdata, the use and interpretation of control charts, and the corrective
actionswhich are to be taken when an out of control situation occurscannotbe
left to chance.Knowledgeof the procedures,useof quality control "tools," and a
soundunderstandingof the quality philosophyof the organizationare achievable
goalsof the training program.Managerswill becomemore comfortablewith del-
egatingdecisionsand relying on subordinatesto carry out and correct the quali-
ty system,when a training programincludesthe quality systemas a part of the
curriculum.
Documentation
Ail training recordsmust be kept up-to-dateand complete.Developing a
training plan is one way to assurethat all requiredtraining occursat the proper
frequency.Each sampling or analytical staff membermust have a training file
which includes a record of what training is required for that personand docu-
mentedevidencethat the training hasoccurred.
Certification
Upon completionof sampling,analytical,or quality assurance/quality con-
trol training, the managementof the organizationshouldcertify that the personis
qualified to perform the procedureswithout additional supervision.
Annual recer-
tification shouldbe done for all procedures.
Facilities
Properlaboratoryfacilities will increasethe ability to producehigh-quality

analytical data.The instrumentationmust be capableof generatingresultswhich
meet the needsof the data user. Adequatespacewill aid in limiting crossconta-
mination and loss or breakageof samples.Imperviousmaterialsshould be used
in the laboratory.Adequateamountsof bench spaceand hood spaceshould be
provided to prevent overcrowdingof activities, some of which may have the
potentialto be dangerous.
QUALITY ASSURANCE& QUALITY CONTROL 29
Exposure and Environment

Personalprotectiveequipment(PPE) should be provided to all sampling
and analyticalpersonnel.The useof fume hoodsmust be requiredfor all activi-
ties which may emit toxic or hazardousfumes. Handlingof all "unknown" sam-
plesmustbe doneusingrubberglovesto preventcontactwith the skin. Exposure
has anothermeaningwith regardto the laboratoryequipment.Expensiveequip-
ment shouldnot be exposedto excessiveheator cold, dust, or sourcesof electri-
city which have not beenconditionedor stabilized.Air conditioning and dehu-
midifiers should be used where appropriateto prevent overheatingor damage
from condensation.The quality of analytical data can be directly relatedto the
quality of the instrumentationand conditionsunderwhich it is operated.Specif-
ic safetyrulesshouldbe developedandenforcedto comply with the Occupational
Safetyand Health Act (OSHA) and to preventany injuries to analyticalstaff.
Laboratoryaccessshouldbe controlled.Unauthorizedpersonnelshouldbe
allowedinto laboratoryareasonly when escorted.The integrity of the datais best
demonstrated by havingan environmentwhich is restrictedto staff that havebeen
properly trainedand certified to perform the work.
Instruments and Equipment
Instrumentsand equipmentare primary componentsof a sampling and

analysisquality assurancesystem.Samplingeventsto be performedaccordingto
a written samplingplan cannotbe successfulunlesspropersamplingequipment
is provided in adequatequantities.Analytical procedurescannot be followed
properly without instrumentationcapableof achievingthe desiredsensitivity and
precision.Providingthe materialsis only the first step.Appropriatemaintenance
and calibration are neededto ensurequality data.
Maintenance
Written proceduresand implementationof decontaminationof sampling
equipmentare an integral part of assuringquality. Properuse of equipmentwas
coveredto someextentin the sectionon training. It is essentialto keepsampling
equipmentin properworking orderby replacingexpendablepartsand by follow-
ing a maintenanceplan.
Analytical instrumentsrequireservicingat properintervals.The quality as-
surancemanualshould contain an instrumentmaintenanceplan. Daily, weekly,
monthly, and yearly preventivemaintenanceproceduresshould be enumerated
for eachtype of instrument.Schedulingof preventativemaintenance service calls
by the instrument manufactureralso should be included. A daily instrument
checkcomparedto statisticallybasedcontrol limits shouldbe requiredas part of
the quality control procedures.This will assistin isolating instrumentmalfunc-
tions. Maintainingsparepartsandkeepinga currentinventoryof partswill assure
reductionsin down time and preventusingold or inadequatepartsbecauseof the
pressuresof dataproduction.
30 KLESTA & BARTZ
Calibration
Calibrationis requiredbeforethe generationof sampledatabegins.A SOP
should be written and implementedwhich includesthe frequencyof calibration,
the numberof calibrationstandardsto be used,and the acceptancecriteria for the
calibration curve. The numberof calibration points is inversely proportional to
the linearity of the curve, as describedin the section"Calibration." The rangeof
concentrationto be usedfor acceptablequantitationshould be documentedand
followed. The natureof analytical chemistry is basedprimarily on the compari-
son of the unknown concentrations,i.e., samples,to known concentrations,i.e.,
standards.The use of current, accuratecalibration curvescannotbe overempha-
sized.
List
The samplingand analytical personnelshould maintain a currentlist of all
samplingand analyticalequipment.The instrumentlist shouldcontainthe model
number,serial number, date of purchase,and generalcondition. The list should
be kept current,reflecting recentacquisitionsor retirementof equipment.The list
helps to show current assetsof the organizationin an easymannerand to assist
managementin budgetingfor future replacementof old equipment.
Data Management
A SOP should be written for all phasesof data management. The collec-
tion, calculation,verification, and reportingof data are to be included.A system
for organizationof the data and the processfor recoveryof archiveddatashould
developedand documented.
Primary Data
Primary data are sometimesreferredto as "raw" data. The first record of
the dataconstitutesprimary data.Instrumentsprovide primary datain the form of
printouts,chromatograms,or spectra.All of theseformats must be datedand ini-
tialed by the analyst.Samplingeventsalso include forms of primary data in field
notebooks,chains of custody, and samplelabels. Analytical proceduresthat do
not produceprinted recordsrequire that all primary data be recordedin bound,
numberedlogbooks or on prenumberedbenchsheets that will be subsequently
bound. The accuracyand completenessof primary data are essentialfor having
truly defensiblerecords.
SecondaryData
Secondarydataensueswhen primary data are usedto calculatean analyti-
cal result or when primary dataare copiedinto anotherformat. The SOPon data
managementmust include mechanismsfor the review of secondarydata and the
proceduresfor correctiveaction when errors are found. Quality control limits of
acceptabilityare anotherform of secondarydata.If the acceptancelimits are inac-
curate,a significant amountof analyticaldatacan be generatedthat appearsto be
acceptablewhen, in fact, it is not.
Calculations
All calculationsshould be clearly understandable.Logbook or benchsheet
formats which include the primary data, the analytical factor, the dilution factor,
and the resultsare helpful to ensurethe accuracyof the calculationand to verify
during data review. Dependenceon calculatorprogramsor computerprograms
without independentverification of the accuracyof the programcan lead to the
generationof a significant numberof errors.
Corrections
The SOP should include the correct procedurefor making correctionsin
primary or secondarydata. It is generallyacceptedto draw a single line through
the incorrect information, add the correct information, and then date and initial
the correction. When it is not obvious why the correction is being made, then
additional notes of explanationshould be included. The true test of defensible
data occurswhen the trail from primary data to analytical result can be followed
without any explanationsfrom the personnelinvolved. The recordsshouldtell the
"whole story."
Documentation
Documentationincludestwo concepts:written documentsand the process
of documentation.The number, types, distribution, and revision processfor all
written manuals must be included in the SOP on documentation.These may
include the samplingprocedures,analytical methods,and quality assuranceman-
ual along with any other documentsthat the organizationdeemsnecessary.The
personnelresponsiblefor the generation,distribution, and revision of the manu-
als should be designatedin the SOP.
ASSESSMENT
A functional quality assurancesystemincludesa mechanismfor the assess-
ment of systemimplementationand performance.The quality assurancesystem
usesthe assessment phaseto impart corrective action and subsequentimprove-
mentsare made to the system.Typically, quality assuranceaudits are conducted
to determinethe statusof conformanceand to developaction plans to eliminate
occurrencesof nonconformance.The assessment can and should be made both
internally and externally.
Internal
Supervisors,managers,or quality assuranceofficers are the primary agents
for performinginternalassessmentof the quality assurancesystem.A plan should
be developedto review various aspectsof the programon a frequencythat corre-
spondsto the importanceof the particularaspect.Somequality assuranceactivi-
ties may only needto be reviewedon a quarterly basis,whereasothersmay need
attention weekly. The assessorshould use observationof activities along with
reviews of documentationto determine the degree of conformance.Training
32 KLESTA & BARTZ
needsmay be determinedfrom the internal assessment

function. Good managers
not only know where improvementsare needed,but also will take the necessary
stepsto correctthe situation.
External
Audits performedby regulatoryagencies,certifying organizations,or qual-

ity assuranceunits, as describedin Good LaboratoryPractices(GLPs) (Code of
FederalRegulations),are the most commonsourcesof externalaudits.The audit
processshouldbe as nondisruptiveas possible.The auditorsshouldbe trainedin
auditing techniques,communicationskills, and the samplingand analyticalactiv-
ities taking place.The written report should be preparedas soon as possibleand
shouldbe usedto implementcorrectiveaction. Two typesof auditsare generally
conducted:(i) A performanceaudit reviews quality control data, benchsheets,
logbooks,and proficiency testing results.The determinationof the precisionand
accuracyof the generateddatais the principal goal of a performanceaudit. It may
be considereda quantitative assessmentof quality. (ii) A system audit reviews
training records,quality manuals,SOPs,methodsmanuals,samplingprocedures,
internal assessment of QA programimplementation,and correctiveaction prac-
tices. It may be considereda qualitative assessment of quality. In either case,the
auditor should use a checklistto be certainthat all areasare addressed.
PROFICIENCYTESTING
Proficiency testing schemesplay an important role in a quality assurance
system.Samplesof known concentrationare submittedto a group of laboratories.
The samplesare "blind" in as much as the laboratoriesdo not know the true value
for the sample.Someprogramsmay include "doubleblind" samples.In this case,
the laboratoriesdo not known that the sampleis a proficiency sample.The mate-
rials are submittedunder the pretenseof being a routine sample.In either case,
the resultsare usedto determinethe correctnessof the laboratory'sdataand may
be used to certify the laboratory for future work. The "true value" of the profi-
ciencysamplecanbe determinedin a variety of ways.The concentrationof a syn-
thetic samplecan be determinedby weights and volumes. All of the nonoutlier
data receivedfrom the laboratoriescan be used to determinethe "true value."
This is commonly referredto as the consensusmethod.A small group of previ-
ously qualified laboratoriesmay be usedin a preliminary round to determinethe
"true value" that will be usedfor subsequentrounds.Reportsfrom the organizers
of the proficiency testing scheme shouldbe usedas a feedbackmechanismto the
laboratorypersonnel.
QUALITY CONTROL
Analytical chemistrywithout QC is guesswork.For the words"quality con-
trol," one may substituteother words such as calibration, contaminationcontrol,
stability, precision,or accuracywhich are all part of QC.
Quality control is the componentof a quality assuranceprogramthat is pri-

marily basedon statisticalevaluationof the data resulting from certain control
samples.Control samplesincludeduplicateandspikedroutine samplesandqual-
ity control checksamples.The statisticalrequirementsfor control datamay be set
accordingto the needsof the datausersor may be baseduponthe "best" the ana-
lytical methodandinstrumentmeasurement systemcanattain.Generallythe por-
tions of the quality assuranceprogramthat are underthe control of the analysts
at the benchor instrumentconstitutequality control.
A sufficient frequencyof control samplesneedsto be establishedto allow
evaluationof imprecision and inaccuracyfrom both sampling and laboratory
sources.For example,samplingrelatedimprecisionsand inaccuraciesmay arise
becauseof within horizon variability or spatial variability within a soil type or
becauseof contamination.Laboratoryrelatedimprecisionsand inaccuraciesmay
be related,for example,to multiple calibrations,analysts,and sampleprepara-
tions or changesin the measurement systemsuchas instrumentaldrift within an
analyticalrun.
Quality control samplesalsoaretools that canserveasa routinechecksthat
analytical resultsare consistentover time. The quality control data that are col-
lected along with routine data over severalagricultural growing seasons,over
severalyears,or in independentprojectscan be usedto prove that dataare com-
parableandthereforecanbe usedasonecollectivedataset.Conversely,the qual-
ity control datamay showthat the dataarebiasedwith respectto growing season,
year, or project; however,the quality control dataare then useful in quantifying
the amountof bias and can be usedto normalizethe routine data.
DEFINmONS
Quality control is basedprimarily on the use of statistics. A thorough
review of statisticalconceptsis suggested.Severalstatisticalterms are defmed
hereto aid the discussion.In addition,severalanalyticalquality control termsare
defined.
TestPortion
The test portion is the volume or weight of material that is preparedfor
measuringthe parameterof interest.The amountmust be sufficient to be repre-
sentativeof the sample.In tum, the sampleis assumedto be representativeof the
populationof interest.For solid materials,particle size reductionby crushingor
grinding may be necessaryto allow the useof a smallertestportion weight. Such
mechanicalsamplemanipulationmay alter physicalcharacteristicsthat, in tum,
affect sampleattributessuchas adsorption,mineral structure,or reactivity.
Replicate
The term replicatedescribesmultiple test portions or multiple instrument
measurements on onepreparedtestportion. The datamay be evaluatedby exam-
ining the relative percentdifference,relative standarddeviation,or coefficientof
variation.
34 KLESTA & BARTZ
Duplicates
Duplicatesare specificreplicates.This term usually describestwo separate
test portionsthat are subjectedto the samepreparationprocedureand then to the
samemeasurementprocedure.As for replicates,the data may be evaluatedby
examiningthe relative percentdifference,relative standarddeviation, or coeffi-
cient of variation.
Spiked (or Fortified) Samples

Spikedor fortified samplesare routinesamplesto which known amountsof
analyteare added.The datamay be evaluatedby examiningpercentagerecovery
of the spike addition. The spike recoverydatamay be usedto evaluatethe accu-
racy (and suitability) of the analytical methodfor the samplesof interest.How-
ever, for the following reasons,thesedatamust not be usedto "correct" the rou-
tine data:(i) 'Die spikemay be more easily recoveredthan the naturallyoccurring
analyteof interest.(ii) The spike may be consumed,absorbed,or "fixed" by the
routine sample.(iii) The spike may be otherwiseaffectedby the sampleor solu-
tion matrix.
Spikesalso may be addedto the solution that resultsfrom sampledissolu-
tion or extraction.Thesespike recoverydatamay be usedto evaluatethe accura-
cy and precision of the determinativeportion of the method.As statedearlier,
routine datamust not be correctedfor spike recovery.
Quality Control CheckSamples

Quality control checksamplesare takenfrom materialswith parametersof
known value thatare subjectedto the samepreparationand analyticalprocedures
asthe routinesamples.A certainparameteris measuredrepeatedlyto evaluatethe
control statusof both the analytical preparationand the instrumentalmeasure-
ment system.It is advantageous to use a quality control checksamplethat is of
similar matrix to the routine samples,if such material is available. It also is
advantageous for the parameterof interestto be within the concentration
rangeof
samplesthat are routinely analyzed.If the matrix and concentrationare similar to
that of the routine samples,the analytical precisionand accuracydeterminedfor
the quality control checksamplecanbe attributedto the routine samples,aswell.
LaboratoryControl Samples
A laboratorycontrol sampleis spikedreagentwater or other blank materi-
al that is measuredrepeatedlyto evaluatethe control statusof the calibration of
the instrumentmeasurementsystem.Laboratorycontrol samplesalso are called
continuingcalibrationverification (CCV) samples.The analyticalprecisionesti-
matedfor the laboratory control samplecan be attributed to that concentration
level on the calibrationcurve.
StandardReferenceMaterials
StandardReferenceMaterials(SRMs)areproducedandsold by recognized
standardsorganizationssuch as the National Institute of Standardsand Testing
(NIST). The certified valuesand acceptancecriteria are determinedfor the mate-

rial basedon a variety of analytical methods.Not all matrix types are available
from NIST. Confidencein the accuracyof the analytical procedureis increased
significantly when a laboratory can demonstratesatisfactory performancefor
SRM analysis.
Control Charts
Control chartsare the real-time plots of dataderivedfrom control samples

such as duplicates,spikes,and quality control check samples.Acceptancecrite-
ria are determinedand usedto evaluatethe data.The plots are madeand evaluat-
ed by the analystso that any out-of-control situation may be correctedimmedi-
ately. It is important to note that control chartswhich are not plotted at the time
of analysisare merely historical recordsof performanceand are not viable mech-
anismsfor the control of the analytical process.
Precision
Precision is the closenessof multiple independentmeasurementsto each

other. Precisioncan be expressedby the range,relative standarddeviation,coef-
ficient of variation, or standarddeviationof the measurements.
Precisionis used
to demonstratethe existenceor absenceof randomerror.
Reproducibility
Reproducibility is the measurementof the precisionof analyticaldatagen-

eratedusing the sameanalytical methodat a minimum of two independentlabo-
ratories.
Accuracy
Accuracyis the agreementof a measuredvalue to the true valueof the para-

meterof interest.Accuracy is often expressedas percentagerecoveryor the per-
centagedifferencefrom the certified value of a standardreferencematerial.
Bias
Bias refersto a systematicdifferencebetweenthe determinedvalue and the

true value. Bias is the measurementof systematicerror.
StatisticalProcessControl
Statistical ProcessControl (SPC) is a systemwhich usesstatistical para-

meterssuch as the mean and standarddeviation to calculateacceptancecriteria
which are used for real time control of the process.Statistical processcontrol
doesnot imply accuracy,but must be establishedbefore accuracycan be evalu-
ated.
36 KLESTA & BARTZ
STATISTICS
Quality control samplesare useful tools in day-to-day laboratory opera-

tions. it is importantthat the statistics usedfor the QC dataare easyto calculate
and to understand.Interpretationis facilitated by the use of control charts.Ease
of calculationis aidedby the hand-held,scientific calculatorwhich is usually pre-
programmedto calculatearithmeticmean,variance,and standarddeviation.Sta-
tistics basedon the datacollectedare the "sample"statistics,whereasthe true val-
ues are "population" statistics.Calculationof the samplestatisticsare basedon
(n - 1) degreesof freedom.
Most analytical data for QC samplesare normally distributed (assuming
that the laboratoryis "in control"). The dataare dispersedsymmetricallyabouta
central value (the arithmetic mean) and small deviationsfrom the central value
occurmore frequentlythan large deviations.If plottedas frequencyof occurrence
vs. analyteconcentration,the dataappearas a bell-shapedcurve.Within onestan-
dard deviation,66.6%of the datawill occur.Within two standarddeviations,95%
of the data will occur. Finally, within three deviations,99.7% of the data will
occur.This is the basisfor establishingcontrol limits equalto plus-or-minusthree
standarddeviations.
The mean,variance,and standarddeviation are the statisticsupon which
other evaluationsof the sampledataare based.
Control Charts
A control chartfor standardreferencematerials,QC checksamples,or lab-

oratory control samplesusesthe arithmetic mean as the estimateof the central
value. Initially, control limits are defined as a percentageof the mean, usually
± 10% of the arithmeticmeanof a seriesof measurements. However,after at least
sevenmeasurements havebeenmade,statisticalcontrol limits can be established.
The warning limits are set at plus-or-minustwo standarddeviations from the
mean(and should contain 95% of the valuesthat are derived from the QC sam-
ple), the controllimits areset at plus-or-minusthreestandarddeviationsfrom the
mean(and shouldcontain99.7% of the valuesfor the QC sample).
Five percentageof the datafor the QC sampleswill fall outsidethe warn-
ing limits, and lessthan 1% of the datawill fall outsidethe control limits. An out-
of-control situation is consideredto exist if two consecutivevaluesfall outside
the warning limits (Taylor, 1987). Because99.7% of the data should fall within
three standarddeviationsof the mean, a point outside the control limit is most
probably out-of-control, and correctiveaction is warranted.For example,if the
out-of-control value is for the standardreferencematerial or QC check sample,
the entire analytical batch should be rerun. This may require reanalysisagainst
new calibration standardsor could require taking new test portions through the
entire preparationprocedure.However, if the out-of-control result is for a CCV
sample,only the routine sampleswithin the interval bracketedby the out-of-con-
trol CCV and the previousin-control CCV needto be rerun. Usually this situa-
tion is a result of instrumentaldrift or someother time-dependentcharacteristic.
Four consecutivepoints outsidethe one standarddeviation limits also is a cause

for concern.
A systematictrend in QC data also indicates an out-of-control situation.
Suchtrendsmay be shownby a seriesof sevenvaluesthat occursaboveor below
the meanor by patternsthat appearin the data,which may relateto variablessuch
as room temperature,time of day, or the analyst.
As additional data are obtained, warning and control limits need to be
updatedon a periodic basis.Dependingupon the amountof data generated,this
updatingmay occur weekly, monthly, yearly, or after a certainnumberof values
are obtained.Examinationof the data will indicate if the control limits are ade-
quate.If dataconsistentlyfall within one standarddeviationof the mean,the con-
trollimits are too wide to be useful in "controlling" the analytical system.Alter-
nately, if more than 5% of the datafall outsidetwo standarddeviationsfrom the
mean, either the control limits do not adequatelyaddressthe variability of the
analyticalsystemand needto be updatedor the systemis severelyout-of-control.
When the control limits are updated,all data that have been accumulated
should be used for estimatingthe statistics: mean and standarddeviation. This
can best be accomplishedby pooling the data. A statisticaltext can be consulted
as a sourcefor the appropriateequations.
When control chartsare maintainedand evaluatedat the time of analysis,
correctiveactionsmay be taken immediately.This will result in significant time
savings,becauseroutine samplesare not analyzedwhen the measurement system
is out-of-control. Somesamplesmay be subjectto a holding time during which
the parameterof interestmust be measured.Plotting and evaluatingcontrol charts
in real time will allow for any reruns to be done before the samplesare invali-
datedbecauseof holding times.
Outliers
An outlier is something that does not belong in the population that is
describedby the accumulatedstatisticaldata. An outlier may refer to a sample,a
test portion, an analytical measurement,or a grouping of data from one analyti-
cal run or from a laboratoryin a proficiency program.
A suspectedoutlier may be identified on a control chart as a result that is
out-of-control. When all replicate measurementsare ranked in order of magni-
tude, a suspectedoutlier may appearas an extraneousvalue. Often the outlier is
due to a transcriptionor calculationerror, and the value can be corrected.In other
cases,contaminationor analyte-lossmay be suspected,especially when a QC
checksampleindicatespossiblecontaminationor loss. Thoseoutlier valuescan
be eliminatedfrom the dataset for cause.
Severalstatistical tests have been developedto evaluatedata for outliers,
including Dixon, Grubbs,Cochran,and Youdentests.A statisticaltext (Barnett&
Lewis, 1978) can be consultedas a sourcefor the appropriatetest for identifying
outliers in a dataset.
As a word of caution, data should not be eliminated capriciously; there
mustbe eitheran attributablecauseor a statisticalbasisfor the eliminationof out-
liers from a data set. After outliers are removed,the descriptive statistics(i.e.,
mean,variance,and standarddeviation) are determined.
38 KLESTA & BARTZ
PRECISION
Precision,as describedearlier, is an importantcharacteristicthat is usedto

demonstratethat the processis in control. The range,relative standarddeviation,
coefficient of variation, or standarddeviation conveysthe degreeof agreement
amongthe independentmeasurements.
Precisionof the Analysis
To control the analytical precision or, conversely,to limit the analytical

imprecision,laboratorycontrol samplesand QC check samplesare used.These
samplesare chosenat concentrationsnear the midpoint of the calibration range.
Laboratory control samplesare usedfor instrumentalanalysesthat are particu-
larly susceptibleto instrumentaldrift. For all analyses,at least one QC check
sampleshouldbe includedwith eachanalyticalbatch(run).
The laboratory control sampleis analyzedon a certain interval, such as
every 10th measurement, to evaluatethe calibrationof the instrument.The result
is comparedto control limits. If the result is acceptable,the analysiscontinues;if
the result is not acceptable,the instrumentis recalibratedand all samplessince
the last in-control result must be reanalyzed.The standarddeviationof datafrom
the laboratorycontrol samplesis a measureof the precisionof the instrumental
measurementsystem.
The quality control check samplemay be a SRM or an in-housestandard.
The parameterof interestmust be stable over time in the quality control check
sample.The quality control checksampleshouldbe of similar matrix to the rou-
tine samples,and the concentrationshould be within the samerange as for the
routine samples.If the in-housestandardis a solution, its sourceshould be dif-
ferent from that of the calibration standards.The quality control check sample
containsa "known" amountof the analyte of interest: for the SRM, valuesfor
mean and standarddeviation are listed on a certificate of analysis. For the in-
house standard, the mean analyte concentrationand standard deviation are
derivedfrom repetitive analysis,usually conductedin comparisonto an SRM (if
an SRM is available).The quality control checksampleis subjectedto the same
analyticalpreparationprocedureas the routinesamples.If the analyticalresultfor
the quality control checksamplefalls within two standarddeviationsof the mean,
the quality control check sample is in control. In other words, the analytical
preparationand the instrumentalmeasurementsystemare consideredto be per-
forming with acceptableprecision.If the quality control checksampleis out-of-
control, various problemsare suspectedincluding instrumentalmalfunction or
analytecontaminationor loss.Mer appropriatecorrectiveaction, suchas instru-
ment recalibrationor repreparationof the entire batch of samples,reanalysisis
conducted.
Precisionmay vary with analyteconcentration.In assessingoverall analyt-
ical precision fora large study, SRMs or in-housestandardsthat representsever-
al analyteconcentrations(low, medium,high) may be analyzedwith eachbatch.
The resultingdataareusedto derive thestandarddeviationat eachconcentration.
EvaluatingSourcesof Imprecision
The conceptof precisionmay be appliedto samplereplicatesderivedfrom

the routine samples,such as sampling (field) duplicates,preparationduplicates
(splits), analytical duplicates,and measurementduplicates.From such sample
pairs, imprecisionsdue to sampling,preparation(suchas sampledrying, sieving,
and splitting), analytical preparation, and instrument measurementcan be
assessed.Note that the analytical results for the sampling duplicatesalso will
contain componentsof imprecision due to samplesplitting and the laboratory
analysis(Le., preparationand analysis),whereasthe preparationduplicatesalso
will containa componentof imprecisiondue to the laboratoryanalysis.For stud-
ies that require multiple laboratoriesor multiple seasons,betweenlaboratoryor
betweenseasonprecisionis assessed by using split samplesor replicateanalyses
of SRMs. Precisionmay vary with analyteconcentration;therefore,it is prudent
to chooseanalyticalduplicateswithout concernfor analytecQncentration.
Initial evaluationsof the sampling, preparation,analytical and measure-
ment duplicatescan be madeby using the relative standarddeviation(RSD)
s
RSD = -=- x 100
X
wheres = standarddeviation andj{ = mean

BecauseRSD is dependentupon the meananalyteconcentration,the RSD
for duplicateswith low concentrationsof the analyteof interestmay be extreme-
ly high values. The RSD for duplicatescontaining the analyte of interest at or
abovethe quantitationlimit will stabilize at a value of 20% or lower, depending
on the type of duplicateand the precisionof the analytical method.For sampling
duplicates,any RSD less than 20% may representacceptableroutine data. Usu-
ally preparationduplicateshave an RSD of approximately10 to 15%, and ana-
lytical and measurement duplicates, approximately5 to 10%.
If at leastone duplicatepair is includedin eachsamplingevent,in eachday
of samplepreparation,in each analytical batch, and in eachinstrumentrun, the
imprecisionassociatedwith eachactivity can be assessed over the courseof the
entire project. Usually, analytical duplicatesare included at a frequencyof 5 or
10%, that is,either1 in 20 or 1 in 10, with a minimum of one per analyticalbatch.
Plots of RSD vs. increasingmeananalyteconcentrationcan servetwo pur-
posesin evaluatingroutine sampledata. Pointsthat fall abovethe curve indicate
suspectdata that should be examinedfor possibleerrors, such as transcription,
miscalculation,or samplemisidentification.Points abovethe curve may identify
outliers. In addition,suchplots can illustrate the methodquantitationlimit for the
routine samples.
Within (Le., within the samplingevent,within the daily samplepreparation
procedure,within the analytical batch, within the instrument run) and overall
(i.e., over all samplingevents,over all analytical batches, and over all instrument
runs) imprecisions canbe determinedfrom statisticalanalysisof study data.
40 KLESTA & BARTZ
Statistically,the total varianceequalsthe sum of the variancesattributable

to eachsource
2 -- S2Sampling + S2Prep + S2Analysis + .,. + S2n

STotal
Repeatabilityand Reproducibility
For purposesof collaborativestudy of analytical methods,AOAC Interna-

tional describesrepeatabilityand reproducibility as terms that relate specifically
to variability within and amonglaboratories. Repeatability (sr) is the expression
of "within" laboratoryprecisiononly. Reproducibility(SR) describesthe precision
of data that occurs "among laboratories"and includes the "within laboratory"
component.For purposesof understanding,a third term is specified asthe impre-
cision amonglaboratories(sd.
Theserelate to one anotheras follows
The InternationalStandardsOrganization(ISO) definestwo terms: repeata-

bility value (r) and reproducibility value (R). Theseterms predict the 95% prob-
ability of agreementbetweentwo measuredvaluesof identical test materialover
the shott-termand the long-term, respectively.The terms repeatabilityvalue (r)
and reproducibility value (R) are calculatedas follows
r = (2...J2)sr
R = (2...J2)SR
Taylor (1987) usestheseconceptsto describethe short-termand long-term

standarddeviations.The short-termstandarddeviationis usually smallerthan the
long-termstandarddeviationor, in other words, the measurementsystemis usu-
ally more preciseover short periodsof time than over long periodsof time. This
is becausesomesourcesof variability may not vary over short intervals of time.
For example,the samecalibration curve may be usedfor samplemeasurements
or the same calibration standardsmay be used to derive the calibration curve.
Samplesmay be preparedwith the samelots of reagents.The long-termstandard
deviationis subjectto a greateramountof variability from the identified sources.
ACCURACY
Accuracy is the agreementof a measuredvalue to the true or theoretical

value of the parameterof interest.
True Value
The true value is the meanof the populationof interest.This value cannot
be known, but can be estimatedby the arithmetic mean derived from samples.
The bestestimateof the true value is madeby the samplemeanof measurements

that are free of systematicerror, that is, measurementsthat are unbiased.Both
preciseand imprecisemeasurement systemscan yield good estimatesof the true
value. However, imprecisemeasurementsystems(e.g., an analyte with a large
standarddeviation)needa larger numberof replicatesto yield a good estimateof
the true value.The more precisea measurementsystemis, the fewer replicates
are neededto yield a good estimateof the true value.
Bias
Bias exists when a measurementsystemderives a central value higher or

lower than the true value. Bias may exist betweenor amongdata setsthat have
beengeneratedby two or more analyticalmethods,by two or more analysts,over
two or more seasons,or by two or more analytical laboratories.Bias may exist,
evenwhen the datafro~ the independentsourceshavebeenshownto be in con-
trol. Bias also may exist becauseof different sampling or sample preparation
practices.Bias should be evaluatedbefore data sets from different sourcesare
combined.
Bias can be assessed only for datathat havebeenderivedfrom a measure-
ment systemdemonstratingstatisticalcontrol, as describedin the section"Con-
trol Charts." The effect of bias on sampleresults may be negativeor positive,
additive or multiplicative, constantor concentration-related, or some combina-
tion of all possibilities.Data shouldbe correctedfor bias only if the origin of the
bias is understoodand the cumulativeeffect of all biaseson the datacan be deter-
mined.
Bias may be attributedto a systematicerror in the measurementsystemor
to an analyticalmethodthat producesbiaseddata.For example,an incompletedi-
gestionor an extractioninefficiency causedby a temperatureor time effect would
introducemethodbias. Examplesof measurement systembias are calibrationer-
ror, dilution error, contaminationof blanksor samples,and loss of the analyteof
interest.The common(althoughincorrect)practiceof reportingthe "besttwo out
of three" replicateanalyses introduces bias, as well.
For someprojects,sourcesof biascan be identified and eliminatedor min-
imized at the time of samplecollection and analysis.An exampleis contamina-
tion. Adequate use of blanks in the project design (sampling blanks and trip
blanks) and in the analytical laboratory(methodor reagentblanks) can identify
possiblesourcesof contamination.If contaminationis shown to be a problem,
proceduralcontrolscan be instituted to eliminate or minimize identified sources
of contamination.Data are generatedand reportedto documentthe analytelevels
found in the various blank samples.Laboratorydatashould not be correctedfor
methodblanks. Likewise, analytical instrumentsshould not be "zeroed" on the
method blank. More information is conveyedto the data user by reporting the
blank and routine data"as analyzed"ratherthan "corrected."
Other sourcesof bias may be identified through the use of various quality
control measures.For example,daily balancecalibration checkswill minimize
bias due to weighing. A checkon the precisionof delivery from a fixed or vari-
able volume pipet may identify the potential for bias in samplealiquoting. The
42 KLESTA & BARTZ
analysisof serial dilutions may identify sample dilution errors. An instrument

performancecheck will limit potential for bias from an instrumentalmeasure-
ment system.Control limits for the slope of the calibration curve for an instru-
ment may identify bias introduced through calibration error. Quality control
checksampleswill indicatepotentialbias from a variety of sourcesin the overall
measurementsystem. When such control measuresare part of the day-to-day
operationof a laboratory,corrective actioncan be taken when any check is out-
of-control. Therefore,the potentialfor producingbiaseddata is minimized.
Anotherway to identify bias is by examiningpercentagerecoveryof spiked
samples.If percentagerecovery is high, e.g.,80 to 120%, the analytical method
is probably suitablefor the samplesof interest.If the recoveryfalls within estab-
lished control limits, the measurementprocessis within statistical control. No
matterwhat value is obtainedfor recovery,the result shouldbe reportedwith the
routine data. When recoveriesare variable or do not fall within establishedcon-
trol limits, corrective action is warranted.However, for the following reasons,
recoverydata must not be usedto correct the routine data: (i) The spike may be
more easily recoveredthan the naturally occurring analyte of interest. (ii) The
spike may be consumedor "fixed" by the routine sample.(iii) The spike may oth-
erwise be affectedby the sampleor solution matrix. Recoveryfrom spiked sam-
ples is the least useful way in which to evaluatesamplebias. Note that recovery
from laboratory control samples,i.e., spiked blanks, describescalibration bias
and doesnot estimatesamplebias.
Someanalytical methodsproducebiaseddata. This bias may be evaluated
by analysisof SRMs. Another alternative is to comparedata derived from one
methodto data derived from another,independentmethod.A statistically signif-
icant difference betweenthe analytical result from the method under evaluation
and the known analytevalue for the SRM, or the value obtainedby using the ref-
erencemethod, can be attributed to bias. The bias should be examinedfor the
rangeof concentrationof the analyteand in various samplematrices.(Consider
that the constituentsof mineral and organic soils occur in such variation that
"soil" may describemultiple matrices.)Methodbiasmay be relatedto the amount
of somereactiveconstituentin the samplematrix rather than to the analytecon-
centration.Often, if the biasedmethod producesmore precisedata, the biased
methodmay be preferred.Anotherconsiderationmay be the cost and time to per-
form the analysis.If the biased methodis more cost and time efficient, the biased
methodmay be preferred.
In summary,biaseddata may be obtainedfrom unbiasedanalytical meth-
ods. Many of the sourcesof bias can be identified andeliminatedor, at least,min-
imized. Real correctiveaction is preferredto correctingthe datafor bias. In some
situations,an analyticalmethodthat is known to producebiaseddatamay be pre-
ferred for use. For approachesto the examinationof interlaboratorybias, refer to
the section"Blind Duplicatesand Split Samples."
CONTAMINATION
Contaminationof the samplemay occurduring sampling,shipment,prepa-
ration, storage,or laboratory analysis.The types of blank samplesthat can be
usedto examinethesesourcesof contaminationwere consideredin the section

"Bias."
Crossoveror "memory effect" is a condition that may occurduring analy-
sis and will influencethe instrumentresponse.Theseeffectsmust be minimized,
becauseit is extremelydifficult to quantify them and apply an appropriatecor-
rection factor. For example,when samplepreparationsare introducedinto instru-
mentssuchasultraviolet/visible(UV -Vis) spectrophotometers, atomicabsorption
spectrophotometers, and inductively coupledplasmaspectrometers,the samples
may contactvarioussurfaces,suchas plastic,glass,quartz,andmetal.Thesesur-
facesabsorbor adsorbpart of the samplematerial.Whena samplecontainsa high
amountof an analyte,measurements on subsequentsamplesmay be influenced
by the releaseof the absorbed/adsorbed analyteover an extendedlength of time.
An analystmay test for the susceptibilityof an instrumentsystemfor demon-
strating memory effects by analyzingthe calibrationstandardsin randomorder
ratherthan in the orderof increasingconcentration,Whenevera high concentra-
tion samplehasbeenintroducedinto an instrument,the calibrationblank can be
run after the sampleas a checkfor memory effects. To rectify the problem,the
systemcanbe purgedwith deionizedwater,dilute acid, or the methodblank until
a subsequentsampleis introduced.Someautomatedinstrumentsystemsinclude
a rinse betweensamples,and often the length of time for the rinse can be con-
trolled by the instrumentsoftware.
Gaschromatographs may exhibit the sameconditionswhen sampleswhich
contain high boiling compoundsare not sufficiently purged from the system
beforethe subsequentsampleis introduced.Elimination of the problemis simi-
lar to the proceduredescribedabove in that a purge operationshould be intro-
ducedbetweensamples.
BLIND DUPLICATES AND SPLIT SAMPLES
In the section"EvaluatingSourcesof Imprecision,"the useof samplerepli-

catesto evaluatesourcesof imprecisionwas presented.Two additionaltypesof
replicatesamplesare usedto examinelaboratoryperformance.Within a labora-
tory, blind duplicatesare submittedas a further assessment of within laboratory
precision.Theseintralaboratorysamplesalso can be used to evaluatethe ana-
lysts' ability to handle"difficult" matrices.Betweenor amonglaboratories,split
samplesare analyzedto estimateinterlaboratoryprecisionand bias.In both situ-
ations,the dataalso shouldbe usedas a basisfor correctiveaction.
The use of split samplesis influencedby the analytical "situation" that is
underevaluation:
ScenarioOne
In this situation, analytical data are generatedfor a specific project. The

split samplesare part of the projectdesign.Either the project involves the useof
severallaboratories,or onelaboratoryanalyzessamplesoverseveralyearsor sea-
sons.Ultimately, the datafrom all laboratoriesor all seasonsare to be combined
for use.
44 KLESTA & BARTZ
Blind Duplicates
For meaningfulstatisticsto be obtainedfor intralaboratoryprecision,each
laboratory(or onelaboratoryin eachseasonof the project)analyzesat leastseven
pairs of blind duplicatesfor eachanalyteat eachconcentrationrangeof interest.
The incorporationof theseadditionalblind duplicatesis unnecessaryif the dupli-
catesdescribedin the section"EvaluatingSourcesof Imprecision"(i) are chosen
randomlyor are designatedby the project management;(ii) are not given prefer-
ential treatmentby the analyst, that is, the duplicate samplesare preparedand
analyzedin the same manneras are routine samples;and (iii) are sufficient in
numberto derive meaningfulstatisticsfor within laboratoryprecision.
Split Samples
For the assessment of interlaboratoryprecisionand bias, splits from a sta-
ble, homogeneoussampleare analyzedby eachlaboratory(or by one laboratory
during eachseason)for eachanalyteat eachconcentrationof interest.Again, at
leastsevensplits shouldbe providedto eachlaboratory(or to one laboratorydur-
ing eachseason)for analysisof eachanalytein eachconcentrationrangeof inter-
est. Statisticsare usedto evaluateestimatesof precisionand bias betweenlabo-
ratories.
If interlaboratorybias is negligible (Le., thereare no significant differences
betweenanalytemeans)and precisionis not significantly different betweenlab-
oratories,the data are comparable.If there are significant differencesbetween
analyte means,but the estimatesof within laboratory precision are not signifi-
cantly different, the statisticaldata from the split samplesmay be usedto derive
correctionfactors to normalizethe routine dataprior to further evaluation.If data
generatedfrom the split samplescan be correlatedto SRM performancedata
from the laboratoriesduring the sametime frame, accuracycan be evaluatedfor
all laboratories(or eachseason)in addition to the relative interlaboratorybias.
ScenarioTwo
In this situation,datafrom multiple laboratoriesare usedto makedecisions,

such as fertilizer recommendationsor whetherto excavatecontaminatedsoil or
not. Usually theselaboratoriesare commercialor "production" laboratories.Ulti-
mately the managementand the customerwant assurancethat the datafrom dif-
ferent sources canbe usedin combinationand will result in the samerecommen-
dation.
Blind Duplicates
Blind duplicatesare submittedby the laboratory managementor quality
assurancepersonnelon aperiodic basisfor eachanalyteof interest.The identity
of the sampleis not known by the analyst,and the analystis not given any infor-
mation aboutthe expectedanalyteconcentration;thesesamplesare referredto as
"double blinds." The submitterexaminesthe resultingdatain comparisonto the
original, routine data and evaluatesthe results accordingto the expectationfor
within laboratory precision. When data for any analyte do not meet the accep-
tancecriteria, possibleerrorsin areassuchas calculation,weighing, diluting, and

calibration are investigated.Reanalysisor the submissionof anotherblind dupli-
catemay occurif attributablecausescannotbe identified. This is an effective way
to identify parts of the analytical processwhich may need corrective action or
additionaloversight.
Split Samples
Split samplesfor "production" laboratoriesmay be run in conjunctionwith
a referencelaboratoryor may be from a round robin.
When a referencelaboratoryis used,the originating laboratoryprovidesa
split sampleto the referencelaboratory. The referencelaboratory must use the
sameanalytical procedurethat was used to generatethe original, routine data.
The datafrom the originating and referencelaboratoriesare evaluatedaccording
to the objectivefor interlaboratoryprecision.Data which do not meetcriteria are
investigatedin both the originating and the referencelaboratoriesfor attributable
errors. This is anotheropportunity for identifying parts of the analytical process
which may needcorrectiveaction or additionaloversight.If a group of "produc-
tion" laboratoriesusesthe samereferencelaboratory,interpretationscan be made
regardingthe interlaboratorybias or comparability of data among the "produc-
tion" laboratories.Trendsin data can be observedeven if interlaboratorybias is
not rigorously determined.
If a proficiency testing program(round robin) is employedfor identifying
interlaboratorybias, splits from one homogeneousmaterialare sentfrom the ref-
ereeto the various participatinglaboratories.All participatinglaboratoriesmay
not use the sameanalytical procedurefor the analysesunlessit is a requirement
of the program.The data are reportedto the refereewithin a certain time frame.
Then the refereeassemblesthe data, performsstatistical evaluationof the data,
and issuesa report to the participatinglaboratories.In this way, the proficiency
of the laboratory is assessedagainstthe consensusstatistics.If the data are pre-
sentedin a graphicdisplay, the relative interlaboratorybias is illustrated.
CALIBRATION
The purposeof calibration is to eliminate or minimize bias in the overall

measurementsystem. The measurementof analyte concentrationis dependent
upon an assemblageof samplingand measurementprocesses,all of which need
to be calibrated.Often, only the calibration of the detection/instrumentsystem
receivesattention, and the support components,that measurevariablessuch as
temperature,volume, mass,particle size, and flow rates,are overlooked.These
supportcomponentsneedto be calibratedto assurethat their accuracyis within
acceptablelimits.
In order to producedefensibledata,the calibrationof all componentsmust
be traceableto an acceptablestandardor physicalconstant.For example,a ther-
mometermay be calibratedagainstan NIST standardthermometer,or the ther-
mometermay be calibratedagainstthe physicalconstantsof an ice bath and boil-
ing water (taking into account barometric pressureand solute content of the
46 KLESTA & BARTZ
water). An analytical balancemay be calibratedby a service technicianagainst

weightsthat were calibratedby NIST or by a stateweights-and-measures labora-
tory in comparisonto NIST-calibrated weights. In the calibration of physical
standards,the responsibilityfor traceabilitybelongsto the personor organization
that performsthe calibration.
Calibrationof an instrumentsystemmust be relatedto standards.The stan-
dardsmay be preparedby the analystfrom materialswhosepurity or concentra-
tion was verified by a manufactureragainstSRMs from NIST. In this example,
only the certified value of the SRM is traceablet~ NIST; all other links in the
traceabilitychain are the responsibilityof the manufacturerand the analyst.In the
laboratory, recordsfor calibration standardsmust documentsourceof material
and details of preparationand also must include traceability for physical mea-
surements,such as massand volume, that were madeduring the preparation.
Rangeof Analysis
Becauseof inherentcharacteristics,someanalytical methodsmay be suit-

able for a specific analyteconcentrationrangeonly. Other methodsare applica-
ble over a wide range,and their usefulnessis dependentupon the instrumentation
that is employedfor the detectionof the analyte.Most detection/instrumentsys-
tems rely on an indirect comparisonbetweencalibrationstandardsand samples.
The most commoncalibration protocol is to use a certain numberof cali-
bration standards(usually at least three plus a calibration blank) to establisha
"response"at eachconcentration.Eachresponseis plotted vs. the corresponding
concentration,and a straightline is drawn or fitted to the datausing regressionor
leastsquarestechniques.A plot on graph paperor using computersoftwarewill
show if the data are linear and will identify potential outliers in the calibration
data. Without the aid of complicated,computer-assisted protocols to transform
dataor to fit quadraticor polynomial curvesto calibrationstandards,a linear cal-
ibration curve is the easiestto constructand use.
A calibration curve has lower and upper limits to linearity. The interval
betweenthe lowest standardand the origin (0,0) of the plot must not be assumed
to be a straightline. Also, the curve abovethe highestcalibrationstandardshould
not be extrapolatedas a linear curve. In generalterms,calibrationstandardsmust
bracketthe expectedconcentrationsof the analytein the routine samples,or the
samplesmust be diluted so that the resulting concentrationfalls within the cali-
bration range.
Thereare several characteristics of analyticaldataderivedfrom calibration
curvesthat impact the resultsthat are reported.Theseare methoddetectionlimit,
limit of quantitation,and limit of linearity.
Method DetectionLimit
The methoddetectionlimit (MDL) is basedon the ability of the methodto

determinethe concentrationof an analytein a samplematrix. The MDL is calcu-
lated as three times the standarddeviation (so) of at least sevenreplicate mea-
surementsof methodblanks(if an instrumentalresponsecan be measured)or low
level samples(at approximately3 to 5 times the estimatedMOL). To assessthe

potential variability associatedwith different analysts,different days of prepara-
tion, and different instrumentcalibrations,the sevenreplicatemeasurements (six
degreesof freedom)should be madeon different days or shifts.
MDL = 3so
BecauseMOLs are basedon the standarddeviation which is not an addi-

tive statistic, MOLscannotbe averaged.However, data that are obtainedover a
period of time can be pooled to obtain an MOL. Often a low-concentrationsam-
ple is includedin eachanalyticalrun as an MOL checksample.The resultingdata
can be usedto determinewhetherthe statedMOL is maintainedover time.
Limit of Quantitation
The limit of quantitation(LOO) is the lowest level at which analyticalmea-

surementbecomesmeaningful in quantitatinga result. Analytical results below
this limit are reportedas "less than" values. Although the LOO has been arbi-
trarily definedby an AmericanChemicalSocietycommitteeas 10 times the stan-
dard deviationof the blank, the datausershouldexaminethe datato decideif this
definition is justified. This value is probably suitablefor spikedreagentwater,but
is not suitable for more complex matrices.It is more likely that, for soils, the
appropriateLOO is a higher mUltiple of the standarddeviation. An empirical
value for the LOO can be derived by examiningthe inflection point in the curve
of RSO vs. increasing analyte concentration for analytical duplicates, as
describedin the section"EvaluatingSourcesof Imprecision."
Limit of Linearity
The limit of linearity (LOL) is the upper level of reliable measurement.

Beyondthe LOL, the calibrationcurve is no longerlinear, that is, a changein con-
centrationno longer resultsin a consistentchangein the instrumentresponse.
Linear VersusQuadraticCalibrationCurves
Linear curvesare most commonly usedfor laboratoryanalysis.A straight-

line relationshipbetweenthe concentrationand the responseis easily understood
and allows direct calculationof the result. A correlationcoefficient can be calcu-
lated to determinethe "straightness"of the curve. It is commonlyacceptedthat a
calibrationcurve shouldhave no lessthan threestandardsdistributedfrom low to
high concentration.Criteria can be establishedto evaluatethe acceptabilityof the
curve before it is used.Mter determiningthe linearity of a curve by using multi-
ple standards,curveswith very high correlationcoefficientscan be determinedon
a routine basiswith as few as two standards.
Nonlinear calibration curves can be used effectively to analyze samples.
The shapeof the curve must be reproducibleboth in curvatureand magnitude.A
nonlinearcurve can be usedto increasethe rangeof analysis.In atomic absorp-
48 KLESTA & BARTZ
tion spectroscopy,for example,a lesssensitivewavelengthfor an elementcan be

chosento eliminate the needfor serial dilutions. It is possibleto choosea wave-
length less influenced by interferenceswhich has a nonlinearcurve for an ele-
ment to improve the accuracyof the results.
The numberof calibrationstandardsto usefor a nonlinearcurve is inverse-
ly proportionalto the linearity.When a nonlinearcurve is to be usedfor analysis,
the numberof calibrationstandardsmust be increasedsignificantly.
Plotting nonlinearcurvesmanually is an acceptableprocedure.Day-to-day
comparisonsof curvatureand magnitudecan be madereadily.
Computersoftware used on analytical instrumentsallows for the use of
nonlinearcalibrationcurves.Thesecomputersand softwarepackageswhich per-
form curve-fitting operationscan be used effectively. Quadratic equationsor
polynomial equationscan be used to define the calibration curve. The analyst
must havesufficient experienceto choosethe correctequationfor the curve. Care
should be taken to assurethat the correct equationis usedfor the curve and that
the systemgeneratingthe curve is reproducible.
REFERENCES
American Society for Testing and Materials. 1991. Annual book of ASTM standards.ASTM,
Philadelphia,PA.
American Public Health Association. 1992. Standardmethodsfor the examination of water and
wastewater.18th ed. Am. Public Health Assoc., Am. Water Works Assoc., Water Environ.
Fed.,Washington,DC.
Associationof Official Analytical Chemists-International.1990. Official methodsof analysis. 15th
ed. AOAC-Int., Arlington, VA.
Barnett,v., and T. Lewis. 1978. Outliers in statisticaldata.JohnWiley & Sons,Chichester,England.
Cochran,W.G. 1947. Someconsequences when the assumptionsfor analysisof varianceare not sat-
isfied. Biometrics3:22-38.
Code of FederalRegulations.Good laboratory practice for nonclinicallaboratorystudies.Title 21,
Part 58. U.S. Gov. Print. Office, Washington,DC.
Code of FederalRegulations.Good laboratory practicestandards.Title 40, Parts 160 and 792. U.S.
Gov. Print. Office, Washington,DC.
Dixon, W.J., and EJ. Massey,Jr. 1969. Introductionto statisticalanalysis.3rd ed. McGraw-Hili Book
Co., New York.
Grubbs,EE. 1969. Proceduresfor detectingoutlying observationsin samples.p. 1-21. In Techno-
metrics. Vol. 11.
Klute, A. 1986. Methodsof soil analysis.2nd ed. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Taylor, J.K. 1987. Quality assuranceof chemicalmeasurements. Lewis Publ., Inc., Chelsea,MI.
U.S. EnvironmentalProtectionAgency. 1986. Test methodsfor evaluatingsolid waste.SW-846.3rd
ed. U.S. Gov. Print. Office, Washington,DC.
Youden, w.I., and E.H. Steiner. 1975. Statistical manual of the associationof official analytical
chemists.AOAC, Washington,DC.
Published 1996
Chapter 3
Dissolution for Total Elemental Analysis
L. R. HOSSNER,Texas A&M University, College Station, Texas
Analytical methodsfor total elementalanalysesrequire a homogeneoussolution

as the test medium. Elementalanalysesof nonhomogeneous soils and rocks are
commonlyconductedafter bringing the sampleinto solution using either: (i) acid
digestion,or (ii) fusion agents(Bennett & Hawley, 1965; Groves, 1951; Hille-
brand et aI., 1953; Jackson,1958; Jeffery, 1970; Maxwell, 1968; Washington,
1930). Dissolution in acids is usually preferredto fusion decompositionbecause
of the lower concentrationof extraneousmaterial in the final solution and less
interferencein the determinationof the solution concentrationof elements,par-
ticularly if atomic absorptionspectrophotometryor inductively coupled plasma
emissionspectrophometryis used.Someof the advantagesand disadvantages of
the dissolutionmethodsdiscussedin this chapterare given in Table 3-1.
Stronginorganicacidsor mixturesof inorganicacids,suchas HCI, HN03,
HCI04 and H2S04, have been used for the decompositionof certain solids.
Intermediatestrength H3P04 and weak HF exhibit complexing propertiesthat
enhancedecomposition.Selectedpropertiesof mineral acids that are commonly
usedin dissolutionof soil and geologicsamplesare presentedin Table 3-2.
Fusionagentsare normally employedfor decompositionof substances that
are insolublein acids.The decomposingeffect of fusion agentsis mainly by high
fusion temperature.Fusion agentscommonly usedfor soil and mineral analyses
are anhydroussodiumcarbonate,Na2C03,andsodiumperoxide,Na202,although
borates,hydroxides,disulfates,fluorides, oxides andS are usedfor certainappli-
cations(Table 3-3). Many of the fluxes have specific applicationsbut, in gener-
al, the effectivenessof a flux in attacking silicate rocks increasesin the order
Na2C03< NaOH < Na202(Potts, 1987).
Commercial sourcesof Na2C03 and Na202 are usually pure enough for
determination of major and minor soil constituents. Sodium bicarbonate,
NaHC03, may be usedas the fusion agentsince it is availablein very pure form
and is convertedto carbonateat 300°C. The melting point of Na2C03 is 851°C
and the melting point of Na202is 460°C.
The exact methodof analysiswill usually dependon the natureof the test
sampleand the accuracyand precisionof the requiredanalysis.High purity acids
can be obtainedmore readily than high purity fusion agents.
Book Seriesno. 5.
49
50 HOSSNER
Table 3-1. Someadvantagesand disadvantages

of dissolution methodsfor total elementalanalysis.
Dissolution
method Advantages Disadvantages
Acid Inexpensive,semirapid, high-purity acids Loss of volatiles (e.g., As, B, Sb, Si) from
available,low salt matrix in final solu- opencrucible, HF will attackglass,
tion, can removeSi as SiF4 in open corrosiveand potentially explosive
crucible (HCl04) acids
Fusion Corrosiveand potentially explosiveacids High salt matrix in final solution, high
not used,Teflon-lined digestionbombs potential for contaminantsin fusion
not required salts,cannotanalyzefor crucible ele-
ment, relatively slow, loss of vola-
tiles (e.g., As, Hg, Se,Tl), crucibles
(Pt) are expensive
Microwave Rapid, high-purity acidsavailable,low Equipmentexpense,loss of volatiles
salt matrix in final solution from open containers,corrosiveacids,
low recoveryof Cr and Ti hasbeen
reported
Table 3-2. Selectedpropertiesof mineral acids(Potts, 1987; Fletcher,1981).
Reagentgrade
Specific Boiling
Acid Formula Acid Molarity gravity temperature Azeotropet
% °C %
Hydrochloric HCl 36 12 1.18 110 20.2
Hydrofluoric HF 48 29 1.15 112 38.3
Nitric HN03 70 15 1.42 120 68.0
Perchloric HClO4 70 12 1.67 203 72.4
Phosphoric H3P04 85 15 1.70 213 :j:
Sulfuric HZS04 98 18 1.84 338 98.3
t A constantboiling mixture with water is achievedat this concentrationof acid.
:j: Decomposesto HP03 at this temperature.
Table 3-3. Saltsthat are commonly usedas fluxes for fusions of soils for total analysis(Lide, 1991;
Potts, 1978).
Salt Formula Melting point Crucible for fusion
°C
Sodium carbonate NazC03 851 Pt
Sodium hydroxide NaOH 318 Zr
Sodium peroxide NazOz 460 Zr
Potassiumcarbonate K ZC03 891 Zr
Potassiumfluoride KF 846 Zr
Lithium metaborate LiB0 3 845 Pt
Recentinnovationsin acid dissolutionof rocks and soils have resultedin

the introduction of closedvesselsand heatingwith microwave radiation where
dissolutionis achievedat relatively low temperatures(150-220°C)and pressures
in polytetrafluoroethylene(Teflon) cruciblesor bottles.The importantadvantages
of these proceduresinclude shorter dissolution time, smaller volume of acid,
decompositionof resistantsolids, and retentionof volatile compounds.
TOTAL ELEMENTAL ANALYSIS DISSOLUTION 51
SOURCESOF ERRORIN DISSOLUTION OF SOLIDS
The composition of the final solution for analysis reflects all the errors
associatedwith samplingerror, heterogeneityof the test elementin the solid ana-
lyzed, chemical changes and contaminationof the sampleduring crushing and
screeningto the requiredgrain size, and the decompositionprocessitself.
Dissolution of solids by acid treatmentor fusion always introducesa dan-
ger of contaminationby impurities in the reagentsor by leachingof contaminants
from the walls of the reactionvessel.Additional sourcesof error include incom-
plete dissolution of the solid, lossesof volatile components,formation of spar-
ingly soluble compounds,formation of soluble complex compounds,and reac-
tions with the vesselwall.
PRINCIPLES
Digestionwith Aqua Regiaand Hydrofluoric Acid
A Teflon decompositionvesselwith a sealedcover allows rapid decompo-

sition of silicate minerals in aqua regia and HF without losses of SiF4 gas
(Bernas,1968).The primary problemin dissolutionof a samplein HF is insuring
the rapid and completedissolutionof reactionproducts(Bailey & Wood, 1974).
The methodhas been adaptedfor the determinationof 18 elements(Buckley &
Cranston, 1971) and employs a 250-mL polypropylene bottle and saturated
H3B03 solution to prevent volatilization losses of the SiF4 gas (Sridhar &
Jackson,1974; Jackson,1974).
Boric acid is addedimmediatelyafter openingthe sealedvesselto promote
the transformationof insolublecomplexfluorides into solublefluoroborates.The
addition of H3B03 effects the dissolution of precipitatedmetal hexafluorates,
therebycompletingthe decompostionof the silicates.The H3B03 reactswith HF
to form tetrafluoboricacid (HBF4) in a two-stepexothermicreaction(Ryss,1956)
H3B03 + 3HF = HBF30H + 2H20
HBF30H + HF = HBF4 + H20
The presence of an excessof H3B03 suppresses the hydrolytic decomposi-

tion ofHBF4 to hydroxyfluoborateions [BF30H-, BF2(OH)i, BF(OH)3], H3B03,
and HF for a period of at least2 h (Bernas,1968). No contaminationeffectswill
result from the useof volumetric and other laboratoryglasswareduring this peri-
od if polypropylenevesselsare not available.
The stablematrix system(HBF4-H3BOT ionic constituentsof silicates)pro-
vides a suitablesalt-freesingle matrix that greatly diminishesthe chemicalion-
ization, matrix, and instrumental interferencesfor atomic absorptionspectrome-
try determinations.One of the principle advantagesof this techniqueis that ele-
mental analysismay be madefrom a single samplesolution and with the use of
only four to six combinedstandardsolutions(Lim & Jackson,1982).
52 ROSSNER
Digestionwith Hydrofluoric, Perchloricand Sulfuric Acids
The useof HF for silicate decompositionavoids the introductionof appre-

ciable quantitiesof extraneousions, particularly the alkali ions, and, therefore,is
desirablefor the determinationof a broad range of elements(Jackson,1958;
Bennett& Hawley, 1965).Decompositionof silicatemineralsis effectedby reac-
tion of HF with Si to form gaseousSiF4, Most of the cations form fluorides
which are convertedto sulfatesas the HF and HCI04 volatilize. Perchloricacid
is a powerful oxidizer and dehydrating agent and all common perchlorates,
exceptpotassiumperchlorate,are readily soluble.Hillebrandet al. (1953) report-
ed that HCI04 is not as effective as H2S04 in the expulsionof HF, but that if the
solutionis evaporatedto drynesswith HCI04, the HF is effectively removed.The
HCI04 is sometimespreferredto H2S04 as the residualacid becauseit produces
less interferencein spectrometricdeterminations(Pratt, 1965). Pretreatmentof
sampleshigh in organic matter with HN03 will preventexplosive reactionsof
the organic materialwith HCI04 .
Fusion with Alkali Carbonate
Fusion is employedfor decompositionof substancesthat are insoluble in

acids. Samplesfused in Na2C03 can be used for a broad range of elemental
analysis.The elementsare convertedinto a form that is completelysoluble in
HCI. The rate of decompositionof solids dependslargely on the surfaceareaof
the substanceto be dissolved.A limiting factor is the high viscosity of melts
comparedto solutions,the low rate of diffusion of liberatedions from the reac-
tion interface,and the formation of insoluble productsat the reaction interface.
Strongacid dissolutionof fused silicatesproceedsmore readily for silicateswith
a low ratio of silica to metalsand thosesilicateswith a higher concentrationof
the more basicmetals(Kolthoff & Sandell,1952).
Fusion with carbonatesresults in partial volatilization of As and Se and
completevolatilization of TI and Hg (Bock & Jacob,1964; Sulcek& Povondra,
1991). A large excessof Na2C03must be usedto ensurea completedissolution
of the fused samplein acid. A large Na2C03to sampleratio also is desireable so
that both the temperatureand time of fusion can be kept to a minimum (Bennett
& Hawley, 1965).The Na2C03to sampleratio should be 4:1 by weight for most
materials(Washington,1930) but in soils dominatedby Fe and AI the amountof
Na2C03 should be increasedto six times the weight of the sample (Lim &
Jackson,1982). The finely ground mineral sampleand the fusion agentmust be
thoroughly mixed and fused at a temperaturehigh enoughto ensurecomplete
dissolution of the melt in HCI. Lim and Jackson(1982) recommend1000°C,
Iron, Mn, Li, Sn, Pb, Sb and As tend to alloy with Pt and can causeseriousdam-
age to the fusion crucible at high temperatures.Pretreatmentof sampleshigh in
Fe oxide (>40%) andMn oxide (> 1%) with H20 2 and HCI to reduceMn in Mn02
(Jackson,1958), and then with aqua regia to dissolve as much of the oxides as
possible, obviates this problem (Kanehiro & Sherman,1965; see section on
"Method for Pretreatmentof Soils High in Ferric Oxide and Manganese
Dioxide").
Fusionwith SodiumPeroxide
Sodium peroxide, Na202' (melting point 460°C) is a strongly alkaline

fusion agentthat simultaneouslyactsas a strongoxidant. Resistantcompoundsof
Zr, Ti and Cr that are difficult to decomposeby Na2C03aloneare readily attacked
by an excessof Na202or a Na20z-Na2C03mixture. Fusionwith Na202 ensures
efficient decomposition of resistant borates, silicates, phosphates,sulfates,
borides, carbides, nitrides and halides (Donaldson, 1979; Donaldson, 1980;
Bhargavaet aI., 1980). Sodium peroxidealone,or with modifying admixturesof
alkali carbonatesor hydroxides,is often usedto decomposerefractory materials
suchas chromiteand zirconium oxide ceramics,bauxites,chromites,zircon, tan-
talite-columbites,beryl, ilmenite sand, and fired clays (Sulcek & Povondra,
1991).
Sodium peroxide melts without dangerof explosion, however, it can be
dangerouswhen its melt is in contact with substancesthat are easily oxidized
such as various forms of C, elementalS, and AI or other metal powders(Sulcek
& Povondra,1991).
Porcelain,Ag, Fe, Ni, Pt and corundumcruciblesare significantly corrod-
ed by Na202 at temperaturesabove 400°C. Zirconium and glassy C crucibles
resist molten Na202' Decompositionwith mixed fusion agentsof peroxide-car-
bonatewill suppressthe corrosiveeffect of the peroxidealoneon the crucible and
will decreasethe fusion temperature.Use of a reducingflame or fusion in an Or
free atmosphereof an electricfurnacewill reducelossesof the zirconiumcrucible
during fusion operations.Glassy carbon crucibles are suitable for fusion with
Na202up to 700°Cbut are rapidly oxidized at highertemperatures.The massloss
from a glassy carbon or zirconium crucible at a fusion temperatureof 500 to
600°C is about4 to 8 mg per fusion with Na202 (Sulcek& Povondra,1991).
Microwave Digestion
Decompositionof soils with acids heatedin a microwaveoven resultsin a

rapid, accelerateddissolution procedure.The sourceof microwaveradiation is a
magnetron,and its power can be regulatedfrom 1 to 750 W. Microwaves are a
form of electromagneticenergy.This form of energyhas alternatingelectric and
magneticfields (or waves)which travel at the velocity of light, about 300000 km
S-1 (Gilman, 1988).A frequencyof 2450 MHZ (wavelengthof 12.2 cm) hasbeen
usedfor heatingof aqueoussolutions.Domesticmicrowaveovensoperateon this
frequency and can be utilized directly for dissolutionof solids in aqueoussolu-
tion. Microwave energyis below the visible light and infrared regionsof the elec-
tromagneticspectrumand above radio waves.If a material absorbsmicrowave
energyit will increasein temperature.
Aqueous acid solutions canbe heated to the required temperaturein a
microwavefield within a few minutes. Microwave heatingis many times more
efficient than classicalheatingtechniques.Microwave radiation acts as a source
of intenseenergyto rapidly heatthe sample,however,a chemicalreactionis nec-
essaryfor completedissolution in the acid mixture. Microwave heatingis inter-
54 HOSSNER
nal and externalasopposedto conventionalheatingwhich is only external.Better

contactbetweenthe particlesand acids is the key to rapid solubility. Local inter-
nal heatingtaking place on individual particles can result in their rupture, thus
exposingfresh surfaceto acid attack.Thermalconvectivecurrentstend to agitate
the mixture and exposefresh surfacesto fresh solution(Nadkarni,1984a;1984b).
Pressurizedcontainersenhancechemicaldissolutionby allowing higher sample-
solution temperatures.They also decreasethe amountof acid neededand reduce
or eliminatethe loss of volatile elements.
Aqueoussolutionsof acidsabsorbmicrowaveradiationlessefficiently than
water. The efficiency of absorptionof microwave energyby acids decreasesin
the following order (Kingston & Jassie,1986)
The amount of energy absorbedincreaseswith incre<!sing dilution of the

acid. For absorptionof microwaveenergy,the dielectric constantof the solvent
must behigh. The dielectric constantof water decreaseswith increasingtemper-
ature.
Quartzglassand plasticsare suitablefor microwavedigestionbecause they
are transparentto microwave radiation. Digestion can be in open vessels,how-
ever, acid vaporsthat are formed must be exhaustedfrom the microwaveoven to
protectthe magnetronand the body of the ovenfrom acid corrosion.Good results
have beenobtainedwith closedvesselsof translucentTeflon (perfluoroallcoxy)
bottles. The lifetime of the bottles dependson the conditions in the microwave
oven and the compositionof the solvent. Direct measurements indicate that the
bottle cap fails when the pressureinside the vesselexceeds1.1 MPa. This occurs
within a temperatureinterval of 200 to 260°C (Kingston & Jassie,1986).
Decompositionvesselsthat contains no metallic parts are commercially
availablefor microwavedigestion.The internal Teflon vesselwith an O-ring is
placedin a polymeric resin mantle. The temperaturein the internal Teflon vessel
shouldnot exceed250°C and the pressureshould be lower than 8.3 MPa.
SafetyNotes
Proceduresdescribedin this chaptercommonlyrequirestrongacids,raised

to high temperatures,to effect dissolutionof soil constituents.Caremustbe taken
when dealingwith thesematerialsand an understandingof their propertiesis use-
ful. All of the acidscan causeburnsto skin and producecausticand toxic fumes.
These materials should never be dispensedby mouth pipette. Full protective
clothing shouldbe donnedbeforeworking with theseacids. Hydrofluoric, HCl04
and H 2S04 have additional propertieswhich should be fully appreciatedprior to
their use in the laboratory.
Hydrofluoric acid (48% HF) is one of the most hazardousmineral acids
usedin the laboratory.Hydrofluoric acid appearsto attackCa in the body and to
form a complex with bone. All manipulationsshould be carried out in a fume
hood. A face visor, gloves,and full laboratorycoat should be consideredas min-
imum protectiveclothing.
TOTAL ELEMENTAL ANALYSIS DISSOLUTION ss
Perchloricacid (70% HCI04) is one of the strongestmineral acidsand is a

powerful oxidizing agentwhen concentratedand hot. Hot concentratedperchlo-
ric acid will react explosivelywith organics.Nitric acid pretreatment can
reduce
the potentialfor explosiveoxidation by oxidizing the more reactivecomponents
of the sampleat a lower temperature.Inorganicperchloratesare normally soluble
and stable while moist or in aqueoussolution, however, some of these salts
becomespontaneouslyflammable in the anhydrousform. A few milliliters of
H2S04 canbe addedto the digestionmix to preventthe samplefrom going to dry-
ness.Digestion should take place only in fume hoodsconstructedof the proper
materials(high densitypolypropyleneor stainlesssteel)and fitted with an accept-
able washdownsystemso the entire fume hood exhaustductwork can be sprayed
with water to preventbuildup of anhydrousperchloricdeposits.
Sulfuric acid (98% H2S04) is not commonlyusedfor the digestionof soils
due to the low solubility of some inorganic sulfates.The anhydrousacid has a
high boiling point (338°C) and oxidizing properties.Cold concentratedH2S04
has an extremelyhigh affinity for water. A greatdeal of heat is evolvedwith the
two are mixed. When diluting, the concentratedacid alwaysmust be addedslow-
ly to excesswater. Water should never be addedto concentratedsulfuric acid.
METHOD FOR PREPARATIONOF SAMPLES
SpecialApparatus
1. Agate or mullite mortar and pestle

2. Stainlesssteel screen«74 11m, 200 mesh)
Procedure
This procedureis adaptedfrom the methodreportedby Jackson(1958) and

Robinson(1945).Air dry about50 to 100 g of representativesamplethat hasbeen
lightly crushedand passedthrough a 250- to 125-llm (8- to 16-mesh)stainless
steel screen.Reducethe samplesize to 10 to 15 g by crushing,sieving, mixing,
and partitioning (by the "quarter" system).Grind the samplein an agateor mul-
lite mortar until the entire quantity passesthrougha 74-llm (200-mesh)stainless
steelor Nylon screen.
Comments
Extreme care in the collection and handling of samplesfor total analysis

shouldbe exercised.Field collectionfollowed by propersubsamplingof a repre-
sentativesampleis essential.Care should be taken in collection, crushing,sub-
sampling andscreeningof samplesto insurethat contaminationis not introduced
during theseprocedures.Contaminationduring screeningof samplesis possible
with metal screens.Thompsonand Bauleston(1970) reportedB, Co, Fe, Mn, Ni
and Zn contaminationof sampleswith stainlesssteel screensand B, Co, Fe, Mn,
56 HOSSNER
Ni, Zn, Cu and Sn contaminationwith brassscreens.There was no contamina-

tion if a Nylon screenwas used. Nylon screenscan be obtainedfrom SPEX
Industries,Inc., Edison, NJ.
METHOD FOR DIGESTION WITH AQUA REGIA AND

HYDROFLUORIC ACID IN A CLOSED VESSEL
SpecialApparatus
1. Widemouth polypropylenebottle, 250-mL capacity, with tight-fitting

screwcap
2. Bottle shaker
3. Electric hot plate
Reagents
1. Aqua regia. Justbeforeuse,mix 3 mL of 12 M (36%) hydrochloricacid

(HCI) and 1 mL of 15 M (70%) nitric acid (HN03)
2. Hydrofluoric acid (HF), 29 M (48%)
3. Boric acid (H3B03), saturatedsolution
Procedure
This procedureis adaptedfrom the methodreportedby Jackson(1974)and

SridharandJackson(1974). Placea 0.1- to O.5-g samplein a 250-mL widemouth
polypropylenebottle. Add 1 mL of aquaregia along the inside of the bottle by
meansof a pipette.Add 10 mL of HF by meansof a plastic pipette,and cap the
bottle immediately.Double-wrapthe threadswith Teflon tapeto insurea tight fit
of the bottle cap. Shakethe bottle in a shakerfor a period of 2 to 8 h, depending
on the nature of the sample,to dissolve the sample.A white insoluble residue
(metal fluorides), which may be presentafter this treatment,is easily dissolved
later by the H3B03 treatment.If there isan indication(color) of undissolvedsam-
ple after 2 h, heatthe bottle for 30 min on the hot plate,with the temperatureset
at 75 to 100°C. More heatingtime may be necessaryfor unusualsamples.After
the sampleis decomposed,cool the bottle (if it has beenheated)to room tem-
perature.
Add 100 mL of saturatedH3B03 solution to the decomposedsampleby
meansof a graduatedcylinder. Immediatelycap the bottle to preventthe escape
of SiF4 gas. Dissolution of insoluble metal fluorides may take anywherefrom
minutesto severalhours, dependingon the particle size of the original sample.
Becausethe addition of H3B03 brings about an exothermicreaction,the bottle
and contentsmust be cooled to 25°C. Placethe bottle on the weighing pan of a
balance,and dilute the contents withwater to give a tared200-g weight (+/- 0.1
g) of extract. Store the samplesolution in the 250-mL polypropylenebottle for
subsequentdeterminationof elementalcomposition.
Comments
The acid matrix will keep the samplesolution and standardsolutionsfrom

colloidal precipitationof silica and other salts for a period of 4 wk (Bernas,
1968). The decomposedsampleis well suitedto atomic absorptionanalysis.The
use of the gravimetric rather than the volumetric method in elementalanalysis
eliminatesthe need to transfer the decomposedsamplesand standardsto volu-
metric laboratoryglasswareand thereforesaves timeand excludespotentialcon-
taminationeffects.
METHOD FOR DIGESTION WITH HYDROFLUORIC, SULFURIC,

AND PERCHLORICACIDS
SpecialApparatus
1. Platinumcrucible, 30 mL with lid or Teflon beaker,50 mL with lid
Reagents
1. Hydrofluoric acid (HF), 29 M, (48%)

2. Sulfuric acid (H 2S04), 18 M, (98%)
3. Perchloricacid (HCI04), 12 M, (70%)
4. Hydrochloric acid (HCl), 6 M
5. Nitric acid (HN03), 15 M, (70%)
Procedure
This procedureis adaptedfrom the method reported by Jackson(1958).

Placea 0.1- to 0.5-g sampleof finely groundsoil «74 11m) in a 30-mL platinum
crucible.Wet the soil with a few dropsof H2S04 and add5 mL of HF and 0.5 mL
of HCI04• With organicsoils, add 3 mL of HN03 and 1 mL of HCI04 to the cru-
cible. Heat the soil-acid mixture on a hot plate until fumesof HCI04 appear.Cool
the crucible and then add 5 mL of HE Placethe crucible in a sandbath,and cover
aboutnine-tenthsof the crucible top with a platinum lid. Heat the crucible to 200
to 225°C and evaporatethe contentsto dryness.Cool the crucible and add2 mL
of water and a few drops of HCI04. Replacethe crucible in the sand bath and
evaporatethe contentsto dryness.If organicmatterstainsare still presenton the
sidesor lid of the crucible, direct the flamed of a Meker burneron the sidesand
lid until the organicmatteris oxidized. A faint red heatis sufficient. Removethe
crucible,andwhen it is cool, add 5 mL of 6 M HCI and about5 mL of water. Heat
the crucible on a hot plate or over a burner until the solution boils gently. If the
sampledoesnot dissolvecompletely,evaporatethe solution to drynessand repeat
the procedure,startingwith 1 drop of H2S04, 5 mL of HF and 0.5 mL of HCI04.
When the residuecompletelydissolvesin HCI, transferthe sampleto a 50-
mL volumetric flask and dilute the contentsto volume.
58 HOSSNER
Comments
A relatively inexpensive50-mL Teflon beakercan substitutefor the plat-

inum crucible. The Teflon beakeris heat resistantto 260°C and is inert to all
materialsexceptmolten metals.The procedureis the same exceptthat the con-
tentsof the beakerare digestedon an electric hot plate.
MEmOD FOR PRETREATMENTOF SOILS HIGH IN FERRIC

OXIDE AND MANGANESE DIOXIDE
SpecialApparatus
1. Platinumcrucible, 30 mL with cover
Reagents
1. Hydrogenperoxide(H20 2), 30%

2. Hydrochloric acid (HCI), 12 M, (36%)
3. Hydrochloric acid (HCI), 6 M
5. Nitric acid (HN03, (15 M), (70%)
Procedure
This procedureis adaptedfrom the methodreportedby Jackson(1958) and

Kanchiro and Sherman(1965). Pretreatsoils to be usedfor Na2C03fusion that
contain>40% Fe203,1 % Mn02' or both, in the following manner.Placean accu-
rately weighedquantity (0.5 to 1 g) of ignited soil (Lim & Jackson,1982) in a
150-mL beaker.To dissolveMn02, add 10 mL of 2 M HCI and 30 mL of H20 2.
Digest first at room temperatureand then on a low temperaturehot plate and
evaporatethe contentsto dryness.Add 10 mL of 15 M HN03 and 20 mL of 12
M HCI. Digest the sampleon a hot plate at low heatfor a few hours,and evapo-
rate the contentsto dryness.After cooling the beaker,dissolvethe solublemate-
rials with about 20 mL of 6 M HCI using a stirring rod to aid solution. Transfer
the contentsof the beaker quantitatively to a low-ash filter paper. Wash the
beakerand paperseveraltimes with 2 M HCI and a few more times with water.
Savethe filtrate and washingsfor silica and other elementalanalyses.
Placethe filter paperplus residuein a platinumcrucible, oven dry the cru-
cible and contentsat about100°C,and ignite them with a Meker burner.Startthe
ignition with a low flame on the side of the crucible, and gradually increasethe
temperatureat the bottom of the crucible until it is the color of cherry red.
Continueignition until all the C hasbeenburnedaway. Adjust the cover so that
the crucible is aboutone-fifth open at the start and aboutone-halfor more open
at the end of ignition. After cooling thecrucible, usethe residuefor the Na2C03
or Na202fusion.
TOTAL ELEMENTAL ANALYSIS DISSOLUTION S9
METHOD FOR FUSION WITH SODIUM CARBONATE
SpecialApparatus
1. Platinumcrucible, 30 mL with cover
Reagents
1. Sodiumcarbonate(Na2C03)'anhydrous
3. Hydrochloric acid (HCI), 12 M, (36%)
Procedure
Add 4 to 5 g of anhydrousNa2C03to a 30-mL platinum crucible contain-

ing 0.5 to 1.0 g of soil or to the residueproducedin the section on"Procedures"
under "Method for Pretreatmentof Soils High in Ferric Oxide and Manganese
Dioxide." Thoroughlymix the Na2C03with the soil usinga spatulaor a glassrod
with a fire-polishedtip. Place1 g of Na2C03on top of the mixture.
Placethe coveredcrucible at a slight angleon a triangle, and warm it over
a Meker burner.Do not let the reducingportion of the flame comein contactwith
the crucible, and do not let the flame envelopthe crucible.Heat the crucible with
a low flame for the first 10 min. Cautiouslylift the cover to let air in through a
slit after the initial dangerof spatteringhas passed.Gradually increasethe heat
to almosta full flame of the burner.
Increasethe Meker burnerflame intensity until the bottom of the crucible
is the color of cherry red (about900°C). Maintain this temperaturefor 10 to 20
min. Lift the coverperiodically to provide anoxidizing environment.During the
latter stagesof fusion, adjustthe coverso that the crucible top is aboutone-fourth
open.Increasethe heatuntil the crucible bottom is the color of bright cherry red
(aboutlOOO°C), and continueheatingfor 5 to 10 min. If thereare any patchesof
fused materialsticking to the sidesof the crucible, heat theseplacesto causeall
the material to run togetherin one mass.At the end of this period, there should
be no effervescence,and the molten mass should be quiescent.Remove the
cover, and heatthe crucible a few minutesmore to finish the fusion.
Removethe flame, and after a secondor so, graspthe crucible with tongs,
androtateit in sucha way that the fusion will solidify alongthe sides.This action
facilitates removal of the solidified melt. Formation of bubbles or miniature
cracksduring the first momentsof cooling indicatesincompletedecomposition.
In such an event, remelt the cake, and heat the crucible until it is the color of
bright cherry red for another15 min. If the sampleis high in silica, add water to
the thoroughly cold crucible until the cake is coveredto a depth of at least 0.5
cm.
Heat the crucible cautiouslywith a low flame to loosen the cake. Avoid
boiling the water. Transferthe loosenedcake to a Pyrex beaker(or to a Teflon
60 HOSSNER
beaker). Wash the crucible with water to transfer any loose particles to the
beaker.Add approximately5 mL of 6 M HCI to the crucible,heatcautiouslywith
a low flame or an electrichot plate to disintegratethe remainingcake,and trans-
fer the crucible contentsto the beaker.Use a policemanand a few dropsof 6 M
HCI to transferall materialsadheringto the coverinto the beaker.Someanalysts
preferthe techniqueof flexing a warm platinum crucible as a way to removethe
cakefor analysis.
In the caseof an obstinatesample,place the cold crucible on its side in a
250-mL beaker,and add enough water to cover the crucible. Gently heat the
beakerto loosenthe cake. Cover the beakerwith a watch glass,and add20 mL
of 12 M HCI throughthe lip of the beaker,addinga small portion at a time to pre-
vent any loss through violent effervescence.Use a flattenedstirring rod to help
loosenthe cakeanddisintegratethe lumps,taking carenot to scratchthe crucible.
Heat the beakerat low heatto hastendisintegrationof the cake. Lift the crucible
by meansof a stirring rod, and washoff adheringmaterialsinto the beaker.
If the fusion has beenconductedproperly, no hard gritty particlesshould
be present,and the liquid, except for gelatinousparticles of silica, should be
clear. If the samplewas pretreatedfor removal of Fe and Mn oxides before the
Na2C03 fusion, add the filtrate and washings saved from the section
"Procedures"under"Method for Pretreatmentof Soils High in Ferric Oxide and
~anganese Dioxide" to this acid solution. Transferthe combineds~lution to a
200-mL volumetric flask and bring to volume with water.
Comments
The use of a Meker burner is necessaryfor obtaining a temperature

between900 and 1200°C.This burneris constructedso that a very hot oxidizing
flame is possible. A fusion carried out at the color of bright cherry red heat
(about 1000°C) is adequatefor sampleshigh in silicate content,however,with
soils low in silica and high in free oxides, the sample should be ignited to
1200°C, as indicatedby an orangecolor of the crucible bottom. A longer igni-
tion period also is requiredfor thesesoils.
Soils that are high in Mn02 are digested(see section on "Method for
Pretreatmentof Soils High in Ferric Oxide and ManganeseDioxide") beforethe
Na2C03fusion or the formation of manganatecan be expectedduring fusion. A
bluish-green-coloredfusion cake is an indication of the presenceof manganate.
This oxidized form of Mn will attackPt in acid chloride solution becauseof the
releaseof free Cl. To prevent damageto the crucible, the manganatemay be
reducedby the addition of 1 mL of 95% ethanolto the crucible beforethe addi-
tion of HCl.
The liquid in the beakercontainssilica in solution as solublesilicic acid or
as insoluble particlesand all the other soil constituentsin solution as chlorides.
The liquid can be evaporatedto drynessto renderthe silica insoluble and there-
by separatedfrom the rest of the elements.The silica becomessomewhatsolu-
ble as basic salts if dehydration takes place above 130°C (Jackson, 1958).
Dehydrationof silica after the addition of a few milliliters of HCI04 provides
approximatelya lO-fold decreasein the amount of silica dissolvedduring the
washingand also permitsdehydrationup to 200°C(Smith, 1940;Jackson,1958).

The filtrate left after separationof dehydratedsilica contains the other con-
stituents.The filter papercarrying the dehydratedSi02 may be oven dried, ignit-
ed in a platinum crucible and weighed.Since evaporationof the acid solution is
carriedout at a fairly low temperature,the amountof glassfrom the Pyrexbeaker
going into solution is almost negligible, and the Teflon is inert.
METHOD FOR SODIUM PEROXIDE FUSION
SpecialApparatus
1. Zirconium crucible, 30 mL with cover

2. Teflon beaker,400 mL with cover
Reagents
1. Sodiumperoxide(Na202)
2. Hydrochloric acid (HCl), 12 M
Procedure
Thoroughly mix a 0.4- to 1.0-g sample,or the residuefrom the sectionon

"Procedures"under"Method for Pretreatmentof Soils High in Ferric Oxide and
ManganeseDioxide" with a 5- to IS-fold excessof Na202 in a 30- mL zirconi-
um crucible. Cover the mixture with a I-g layer of Na202' Cautiously fuse the
mixture using a Meker burner,or to the dark red heatof a Meker burner,until the
contentsof the crucible are the color of cherry red and clear. Keep the contents
in a molten state for approximately 30 s to insure complete decomposition.
Allow the melt to cool, then transfer the crucible to a covered400-mL beaker
containingapproximately50 mL of water and 10 mL of concentratedHCl. When
the melt hasdissolved, removethe crucible and beakercover, rinsing them thor-
oughly with water. Cover the beakerand evaporatethe solution to approximate-
ly 50 mL. The solution shouldbe kept almostcompletelycoveredduring the ini-
tial evaporationto avoid loss by spray. Cool to room temperaturethen transfer
the contentsof the beakerto a 200-mL volumetric flask and dilute to volume
with water.
Comments
Sodiumperoxidefusion and its efficiency can be modified by additionsof

alkali carbonates,hydroxides, and borates. Sodium peroxide is often used to
decomposerefractory and resistantmaterials such as chromites,zircon, beryl,
and fired clays. Admixtures of carbonate,hydroxide, or boron oxidewill sup-
press the corrosive effect of the peroxide alone on the crucible material and
decreasethe fusion temperature.Sodium peroxide alone or with modifying
admixtureshasbeenrecommendedfor fusion of resistantberyl, chromiumores,
62 HOSSNER
fired clays,Ti concentrates,iron oresand zirconiumsands.Titanium and chromi-

um oresthat were difficult to decomposeby Na2C03alonewere readily attacked
by an eightfold excessof a 3 + 1 Na202-NaKC03mixture (Schinkel, 1984).
Chromiteswere decomposedby fusion at the temperatureof liquefaction of the
mixture with a 10- to 15-fold excessof the fusion agent(Donaldson,1980).
Fusion also can be accomplishedby meansof a muffle furnace. Placethe
crucible and its contents,coveredwith a lid, in a cold muffle furnace. The tem-
peratureis slowly increasedto attain 650°C. The liquid is then swirled to permit
decompositionof any aggregated particles and the heatingcontinuedfor another
5 min. The contentsare cooled, extractedwith a minimum amountof water and
acidified with HCI or H2S04 to completedissolution.
Vitreous carboncruciblesavoid contaminatingthe subsequentsolution but
are expensiveand have a limited life. Zirconium cruciblesare much less expen-
sive, can be usedfor Na202fusion and have a wide rangeof usefulness.
METHOD FOR DECOMPOSmONWITH MICROWAVE DIGESTION
SpecialApparatus
1. Microwave oven with 650 W output and polypropyleneturntable

2. Polypropylenebottle, 250 mL with polypropyl screw-oncap
Reagents
1. Aqua regia. Just before use, mix one part of 15 M (70%) nitric acid
(HN03) with threepartsof 12 M (36%) hydrochloricacid (HCI)
2. Hydrofluoric acid (HF), 29 M, (48%)
3. Boric acid (H3B03), 1.5% solution
Procedure
Weigh 0.1 g of sample,groundto <74 ~m, into a tared250-mL translucent

Teflon PFA bottle. Add 5 mL of aquaregia and 3 mL HE Cap the bottle with a
polypropyl screw-oncap double-sealedwith Teflon tape to insure a tight seal.
Carouseltraysprovidedwith the microwaveovencommonlyhold 8 to 12 bottles.
Bottles containingwater or reagentblanks must be included to maintain a con-
stantvolume of solution in the oven during eachrun if fewer samplesare decom-
posed. Place the bottles in the microwave oven in an annular array on the
polypropylene tray so that each sample will receive the same amount of
microwaveradiation. The melting point of Teflon PFA is 250°C. Heat at 650 W
microwave output for 30 s. Removethe bottles, retighten the caps, and return
them to the oven. Heat once more for 2 min at the 650-W setting. Removethe
samplefrom the oven, cool in an ice waterbath. When the contentsof the bottles
are cool, open, and add1.5% H3B03 solution to make the contentsof the bottle
to 100 mL by weighing. Recapthe bottles. Heat for 10 to 30 min on a hot plate
with the temperatureset at 75 to lOO°C or until the solution is clear, and then
cool.
Comments
The magnetronmustbe protectedagainstcorrosiveacid vaporsandagainst

unconsumedor reflected microwave energy, therefore,a beakerwith water is
commonly placed in the oven (Sulcek & Povondra,1991). Minerals such as
chromite,rutile, corundum,cassiterite,and zircon have beenfound resistantto
microwavedigestionso recoveryof Cr and Ti is low. If the samplesgo to dry-
nessin opencontainers,substantiallossesof Si as SiF4 will result. Boric acid is
addedafter the dissolutionto neutralizethe HF in solutionby forming tetrafluo-
boric acid. Without this stepthe solutioncannotbe usedin many inductively cou-
pled plasmaemissionspectrophotometers, sinceit will attackthe quartztorch of
the instrument.Lossesof Cr(III) andPb which occurfrom opensystemshavenot
occurredin closedTeflon vessels.Biological materialsandoil shaleshavesome
organicresisuesleft after microwavedigestion,while coal samplesappearto be
essentiallyunchanged(Nadkami,1984a,b).
Ventillation systemsmustbe devisedto evacuateacid fumesfrom the oven
cavity when open digestionsystemsare usedor to protect the oven from acid
fumes that may escapefrom closedsystems.Ventilation has been achievedby
variablespeedfans or compressedair to evacuatefumes from the cavity of the
microwaveoven into a fume hood after metal parts inside the oven cavity have
beencoveredwith plastictape(Klock & Lamothe,1986).Samplecontainerscan
be loadedinto a largerpolyethylenecontainer,suchas a vacuumdesiccatorwith
the vent open and a tube leadingto an iced Na2C03trap, to collect acid fumes
(Mahanet aI., 1987).
Closedsystemshave been developedfor digestion of samplesat higher
pressuresand temperaturesby a number of companies including CEM
Corporation, Matthews, North Carolina; Parr Instrument Company, Moline,
Illinois; and Savillex Teflon-PFAvesselsfrom the Savillex Corporation,Minne-
tonka, Minnesota.
REFERENCES
Bailey, N.T., and SJ. Wood. 1974. A comparisonof two rapid methodsfor the analysisof copper
smeltingslagsby atomic absorptionspectrometry.Anal. Chim. Acta 69:19-25.
Bennett,H., and W.O. Hawley. 1965. Methodsof silicate analysis.Acad. Press,Inc., New York.
Bernas,B. 1968.A new methodfor decompositionand comprehensiveanalysisof silicatesby atom-
ic absorptionspectrometry.Ana!. Chern.40:1682-1686.
Bhargava,O.P., M. Omitro and W.O. Hines. 1980. Universaltechniquefor rapid dissolutionof mate-
rials encounteredin the steelindustry and its applicationsin the photometricdeterminationof
arsenic,phosphorus,titanium and vanadiumin iron ores.Talanta27:263-267.
Bock, R., and D. Jacob.1964. Die bestimmungvon selenspuren.Fres.A. Anal Chern.200:81439.
Buckley, D.E., and R.E. Cranston.1971. Atomic absorptionanalysesof 18 elementsfrom a single
decompositionof aluminosilicate.Chern.Oeo!. 7:273-284.
Donaldson,E.M. 1979. Determinationof bismuth in ores, concentratesand non-ferrousalloys by
atomicabsorptionspectrophotometry after separationby diethyldithiocarbamate extractionor
iron collection.Talanta26:1119-1123.
64 HOSSNER
Donaldson,E.M. 1980. Detenninationof chromium in ores, rocks, and related materials,iron, steel
and non-ferrousalloys by atomic absorptionspectrophotometryafter separationby tribenzy-
lamine chlorofonnextraction.Talanta27:779-786.
Fletcher,w.K. 1981.Analytical methodsin geochemicalprospecting.Vol. 1. Elsevier Sci.Publ. Co.,
New York.
Gilman, L.B. 1988. Generalguidelinesfor microwave sample preparation.CEM Corp., Mathews,
NC.
Groves,AW. 1951. Silicate analysis.2nd ed. GeorgeAllen and Unwin, Ltd., London.
Hillebrand, W.F., G.E.F. Lundell, H.A Bright, and J.1. Hoffman. 1953. Applied inorganic analysis
with special referenceto the analysisof metals, mineralsand rocks. 2nd ed. John Wiley &
Sons,Inc., New York.
Jackson,M.L. 1958. Soil chemicalanalysis.Prentice-Hall,Inc., EnglewoodCliffs, NJ.
Jackson,M.L. 1974. Soil chemical analysis-advancedcourse. 2nd ed. Published by the author,
Madison,Wisconsin.
Jeffery, P.G. 1970. Chemicalmethodsof rock analysis.PergamonPress,Inc., Elmsford, NY.
Kanehiro,Y., and G.D. Shennan.1965. Fusionwith sodiumcarbonatefor total elementalanalysis.p.
952-958.In C.A Black et al. (ed.) Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA
and SSSA,Madison,WI.
Kingston, H.M., and L.B. Jassie.1986. Microwave energy for acid decompositionat elevatedtem-
peraturesand pressuresusing biological and botanicalsamples.Anal. Chern.58:2534-2541.
Klock, P.R., and P.J. Lamothe.1986. Detenninationof ammoniumin a buddingtonitesampleby ion-
chromatography. Talanta 33:495-498.
Kolthoff, I.M., and E.B. Sandell. 1952. Textbook of quantitative inorganic analysis. 3rd ed. The
Macmillan Co., New York.
Lide, D.R. 1991. Handbookof Chemistryand Physics.CRC Press,Inc., Boca Raton, FL.
Lim, C.-H., and M.L. Jackson.1982. Dissolution for total elementalanalysis.p. 1-12. In AL. Page
et al. (ed.) Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Mahan, K.I., T.A. Foderaro,T.L. Garza,R.M. Martinez, G.A Maroney, M.R. Trivisonno, and E.M.
Willging. 1987.Microwave digestiontechniquesin the sequentialextractionof Ca, Fe, Fr. Mn,
Pb, and Zn in sediments.Anal. Chern.59:938-945.
Maxwell, J.A 1968. Rock and mineral analysis.Interscience,New York.
Nadkami,R.A 1984a.A handbookof silicate rock analysis.B1ackie& Sons,Ltd., London.
Nadkami,R.A 1984b.Applicationsof microwaveoven sampledissolutionin analysis.Anal. Chern.
56:2233-2237.
Potts,PJ. 1987. A handbookof silicate rock analysis.B1ackie& Sons,Ltd., London.
Pratt, P.F. 1965. Digestion with hydrofluoric and perchloric acids for total potassiumand sodium. p.
1019-1021.In C.A Black et al. (ed.) Methodsof soil analysis.Part2. Agron. Monogr. 9. ASA
Robinson,W.O. 1945. The fusion analysisof soils. Soil Sci. 59:7-11.
Ryss, I.G. 1956.The chemistryof fluorine and its compounds.(In English.) U.S. At. Energy Comm.
TranslationSer. AEC-tr-3927. U.S. At. Energy Comm. Tech. Infonn. Serv., Oak Ridge, TN.
Schinkel, H. 1984. Detenninationof Ca, Mg, Sr, K, Na, Li, Fe, Mn, Cr, Ni, Cu, Co, Zn and Cd by
flame AAS. A universalmethodfor the analysisof waters,coals,ashes,ores, rocks,building
materials,metals,and similar samples.Fres. Z. Anal Chern. 317:10-26.
Smith, G.F. 1940. Perchloricacid. 4th ed. G.F. Smith Chern. Co., Columbus,OH.
Sridhar,K., and M.L. Jackson.1974. Layer chargedecreaseby tetrahedralcation removaland silicon
incorporationduring natural weatheringof phlogopite to saponite.Soil Sci. Soc. Am. Proc.
38:847-850.
Sulcek,Z., and P. Povondra.1991. Methodsof decompositionin inorganicanalysis.CRC Press,Inc.,
Boca Raton, FL.
Thompson,G., and D.C. Bauleston.1970. Samplingcontaminationfor grinding and sieving deter-
mined by emissionspectroscopy.Appl. Spectrosc.24:210-219.
Washington,H.S. 1930.The chemicalanalysisof rocks. 4th ed. JohnWiley & Sons,Inc., New York.
Published 1996
Chapter4
Atomic Absorption and Flame

Emission Spectrometry
ROBERTJ. WRIGHT
TOMASZ STUCZYNSKI, USDA-ARS, Beltsville, Maryland
AND
Soil scientistshave used spectrochemicalmethodsto determineelementalcon-

tentsof soil digests,soil extractsand plant digestsfor many years.Early investi-
gationswere concernedwith measurementof essentialplant nutrients to deter-
mine their soil chemistry and to make appropriatefertility recommendations.
Flame emissionspectrometrywas successfullyusedto make analytical determi-
nationsof K, Ca and Mg. With the developmentof flame atomic absorptionspec-
troscopyin 1955 and its subsequentwidespreaduse in the 1960sanalyticalmeth-
ods were extendedto micronutrientsand nonessentialelementsthat can affect
crop growth and animal health. More recently soil scientistshave becomecon-
cernedwith the fate of introducedtraceelementsin the soil environmentfrom the
standpointof water quality, food safetyand humanhealth.Over the last 20 yr the
introduction and developmentof furnace atomic absorptionsystemshas greatly
increasedour ability to determine trace levels of both plant essential and
nonessentialelementsin the soil environment.Various accessories allow analysis
of samplesin gas, liquid, slurry and solid forms. Modem computer-controlled
furnace atomic absorptionsystemswith multilamp turrets and autosamplerscan
be setupfor unattendedmeasurementof a variety of elementsat the micrograms
per liter level.
PRINCIPLES
Atomic spectroscopyis basedon energyabsorbedor emittedduring transi-

tions betweenelectronicenergylevelsof an atom. Atoms of a given elementhave
a characteristicset of allowed electronic energy levels. Radiation absorbedor
emitted as a result of transitionsbetweentheseelectroniclevels can be usedfor
qualitative and quantitative analysis. Quantitative analysis by atomic spec-
troscopydependson measurementof radiation intensity and the assumptionthat
radiationemittedor absorbedis proportionalto atomic concentration.A compar-
Book Seriesno. 5.
6S
WRIGHT & STUCZYNSKI
ison of relative intensity valuesfor standardsand samplesis usedto derive ele-

mentalconcentrations.
The lowest energy electronicconfigurationof an atom is known as the
groundstate.Undermostconditionsthe greatmajority of the atomsof an element
are in the ground state.If energy is suppliedto an atom by a Sourcesuch as a
flame, outer-shellelectronsare promotedto a higher energyexcited state.The
excitedstateis unstableandatomsexist in this configurationfor only a very short
time, typically 10-8 s. When an outer-shellelectronreturnsto a more stablecon-
figuration or the ground state,light with energyrepresentedby hef)., is emitted
where h is Planck'sconstant,e is the speedof light and A. is the wavelength.
Transitionsfrom and to the groundstateare known as resonancetransitionsand
give rise to spectrallines known as resonancelines. Eachelementhasa charac-
teristic set of excitedstatessuchthat a specific quantity of energymust be sup-
plied to promotean atom to an excitedstateand light of a definite wavelengthis
emittedwhen the excitedelectronreturnsto a lower energystate.Thereforeeach
elementhasa characteristicsetof resonancelines for excitationand decay.
Three commonly usedanalytical atomic spectroscopytechniquesdepend
on measurementof energyabsorbedduring excitationor energyemittedduring
decay.Atomic absorption,atomicemissionandatomicfluorescenceareschemat-
ically representedin Fig. 4-1. Atomic emissionspectroscopyis basedon mea-
surementof the intensityof light emittedwhenan excitedatomreturnsto a lower
energystate.Excitedstateatomsareproducedby collisionsof ground-stateatoms
with electrons,other atomsor moleculesin flames, plasmasand electrical dis-
charges.An external light source provides atomic excitation in both atomic
absorption and atomic fluorescence spectroscopy.Atomic absorption spec-
troscopyconsistsof passinglight of a specificwavelengththat can be absorbed
by the elementof interestthroughan atomicvaporof the elementand measuring
attenuationof light intensityasa resultof absorption.Sincemostatomsare in the
ground-stateelectronicconfiguration,transitionsto the first excitedstateusually
havethe largesttransitionprobability andareoften usedasthe basisfor the atom-
ic absorptionmeasurement. Atomic vaporis producedby supplyingthermalener-
gy from a flame or an electrically heatedgraphite tube. Atomic fluorescence
spectroscopymeasuresthe intensityof light emittedasan excitedatomreturnsto
a lower energystateor the groundstate.The light sourcein atomicfluorescence
is positionedso that only light emittedfrom excitedatomsfalls on the detector.
Atomic fluorescencespectroscopywill be describedin detail in Chapter7 (Kara-
thanasis& Hajek, 1996).
Comparisonof Techniques
Atomic absorptionand atomic emissionspectroscopytechniquesare more

widely used than atomic fluorescencespectroscopy.Current atomic emission
techniquesgenerallydependon an AI plasmaratherthan a flame to atomizeand
excite the elementof interest.The plasma,consistingof positively chargedAI
ions and free electrons,is energizedwith high-frequencyelectromagneticfields
or with direct current (Ingle & Crouch, 1988). Today most analytical methods
basedon atomic emissionuse radio frequency inductively coupled AI plasma
ATOMIC ABSORPTION& FLAME EMISSION SPECTROMETRY 67
Emission
hC/A
Optics
Detector
Readout
Atomic Vapor at Temperature T
-n°
y . . . ·~,-_~_~_:e_i~_Sto_r
Absorption Light
Attenuation ,--_ _ _ _--,
~~;:e - _ _....I
Readout
Atomic Vapor
Fluorescence
-+ hC/A
Optics
Detector
Readout
*
Light
Source
Fig. 4-1. Schematicrepresentationof atomic emission,atomic absorptionand atomic fluorescence.
(rcP) systems.The analyst must choosethe most appropriatesystemfor a par-

ticular determinationfrom among flame atomic absorption, graphite furnace
atomic absorptionor rcp atomic emissiontechniques.
Flame atomic absorption (FAA), graphite furnace atomic absorption
(GFAA) and rcp atomic emissiontechniqueseach have advantagesand disad-
vantageswhen comparedacross a range of operationalcharacteristics(Table
4-1). Flameatomic absorptionand rcp generallyhavesimilar detectionlimits for
most elements.However, the much higher temperaturesof the plasma (up to
10 OOO°C) comparedto a flame (s;2900°C)makesrcp more effective in detect-
ing lower concentrationsof refractory elementssuch as W, Zr, Ta, Hf and rare
earth elements.Elementswith resonancelines in the very far ultraviolet (UV)
(below 190 nm) such as C, N, 0, F, S, CI, Br, rand P are not readily determined
by FAA. Flame atomic absorptiongenerallyhas betterdetectionlimits than rcp
for group r metalsand selectedvolatile elementsincluding Na, K, Pb, Zn and Cd.
On a relative concentrationbasisdetectionlimits obtainedusing GFAA are gen-
erally 10 to 100 times betterthan FAA or rcp for most elements(Slavin, 1986).
Graphite furnace atomic absorption is not used for refractory elements,rare
earths and the previously mentionedelementswith resonancelines below 190
nm.
68 WRIGHT & STUCZYNSKI
Table 4-1. Comparisonof atomic emissionand atomic absorptiontechniques.

Factor ICP FAA GFAA
Detectionlimits Best for refractory ele- Similar to ICP, but bet- Generally10-100times
ments,rare earthsand ter for group I metals more sensitivethan
elementswith reso- and selectedvolatile FAA or ICP when the
nancelines below 190 elementsNa, K, Pb, elementcan be atom-
nm such as P and S Zn,Cd ized
Linear concentration 1Q4-106 103 102
range
Within run precision 0.3-2% RSD 0.1-1% RSD 0.5-5% RSD
Interferences
Chemical Lowest Intermediate Highest
Spectral Highest Lowest Intermediate
Ionization Lowest Highest Intermediate
Speedof standardi- Most rapid for 6 or Most rapid for lessthan Slow
zation and analy- more elementsper 6 elementsper sample
sis sample
Cost
Initial Highest Lowest Intermediate
Operating Highest Lowest Intermediate
Other factors Doesnot use combusti- Usescombustiblegases Doesnot usecombusti-
ble gasesand can be and cannotbe left ble gasesand can be
usedfor unattended unattended usedfor unattended
overnight operation overnightoperation
Becauseof limited self absorption(light emitted by an excited atom of an

elementis absorbedby anotheratom of that element)ICP has a concentration
rangefor linear operationof approximatelylOS. This is an importantcharacteris-
tic for automatedanalysis.By contrast,the concentrationrangefor linear opera-
tion is approximately103 for FAA and 102 for GFAA. Within-run precision is
generallybestfor FAA with typical relative standarddeviations(RSD) of 0.1 to
1.0% and worst for GFAA where the within-run precision is in the 0.5 to 5%
range(Slavin, 1986).
A number of different chemical and physical effects in the vapor phase
cause interferencesin atomic absorption and atomic emission spectroscopy.
Chemicalinterferenceslimit formation of free atomsin the vaporphase(Frechet
aI., 1980). This type of interferenceis virtually eliminatedwith ICP where high
plasmatemperaturesreadily break chemicalbondsto createfree atoms.Spectral
interferencesoccur when atomic or molecular speciesother than the element
beinganalyzedabsorbor emit energyat the wavelengthof interest.Spectralinter-
ferencesare generallylessof a problemwith FAA than GFAA or ICP. Ionization
interferencesoccur in hot flames wherethermalexcitationcan causeelectronsto
be completely removed from an atom, thereby lowering the popUlation of
ground-stateatoms available for light absorption. Ionization interferencesare
essentiallyeliminated in ICP since the high population of free electronsin the
plasmasuppressesionization. Interferencesand methodsof correction for FAA
and GFAA will be discussedin detail in a later sectionof this chapter.
Speedof sampleanalysisis an important factor if a laboratorydetermines
a variety of elementsin a large numberof samples.Modem FAA systemshave
becomehighly automatedand can perform simultaneousmultielementanalyses
(Farah& Sneddon,1993). Flame atomic absorptionsystemsgive the most rapid
resultsif thereare only a few elementsto analyze.In a routine analysissituation

sequentialICP becomesfaster than FAA when there are six or more elementsto
analyzein a sample.Graphitefurnaceatomic absorptionis a very slow technique
comparedto FAA and ICP.
The initial purchaseprice is highestfor ICP systemsand lowest for FAA.
Operatingcostsincluding gases,powerand consumableitemsare highestfor ICP
systemsand lowest for FAA. Inductively coupled argon plasma emissionand
GFAA havethe advantagethat they do not usecombustiblegasesand can be pro-
grammedfor overnight,unattendedoperation.
The needsof a particular laboratorywill dictate the instrumentof choice
from amongFAA, GFAA and ICP. Flameatomic absorptionis a well-established
versatiletechnique,it has a low initial cost, systemsare automatedand easy to
operate,and interferencesare known andcanbe controlled.Flameatomic absorp-
tion would be the instrumentof choicewherecost is a major consideration,sam-
ple load will not be large and extremely low detectionlimits are not required.
Inductively coupled argon plasmaemissioncan provide faster analysisof sam-
ples where multielementdeterminationsare required.Inductively coupledargon
plasma emissionhas a greaterconcentrationworking rangeso samplesmay not
haveto be diluted or concentratedto analyzedifferent elementsin the samesam-
ple. Inductively coupledargon plasmaemissioncan eliminate chemicalinterfer-
encesandprovideanalysisof certainnonmetalsand refractoryelementsnot read-
ily determinedby FAA and GFAA. If determinationof trace metalsat low con-
centrationsis the most importantconsideration,GFAA is the systemof choice.
The remainderof the chapterwill be devotedto atomic absorptionspec-
troscopy.Detailedinformationconcerningflame atomicemissionspectroscopyis
available from other sources(Hieftje et aI., 1976; Alkemade & Joseph,1979;
Ingle & Crouch,1988)and ICP is covered inChapter5 (Soltanpouret aI., 1996).
A detailedexplanationof equipmentcomponentswill not be given sincethat has
beeneffectively treatedin a numberof review articles(Varma, 1986; Marshall et
aI., 1987). However, an overview of the equipmentwill be given, sampleanaly-
sis problems will be identified and methods to overcome these problems
addressed.
INSTRUMENTATION
All atomic absorptionspectrometers havea numberof componentsin com-

mon including a light source,a device to produceground-stateatoms,a mono-
chromatorto isolate the spectralline of interest,a detectorand a signal proces-
sor/readoutdevice. Thesecomponentsare illustrated schematicallyin Fig. 4--2.
The light sourceproducesresonancelines that can be absorbedby atomsof the
elementof interest.An atomic vapor of ground-stateatomsis generatedthermal-
ly in a flame or furnace.Absorption occurswhen light of the appropriatewave-
length is passedthroughthe atomic vapor, thus reducingthe intensity of the inci-
dent beam. The monochromatorfocuses light of the desired wavelengthon a
detectorwhich producesa current proportional to incident light intensity. The
currentfrom the detectoris processedand convertedto a readoutin concentration
C=J. . .;;/. ~ noD

Monochromator
Light Chopper Detector Signal Processor/
Source Atomization Readout
Device
Fig. 4-2. Diagramof the basiccomponentsof a single beamatomic absorptionspectrometer.A typ-

ical double-beaminstrumentalso would include mirrors to split the beam into sampleand refer-
encecomponentsprior to the atomizationsourceand to recombineand focus the split beamson
the monochromatorentranceslit.
units which may be displayed and/or stored for further processing.Although

simultaneousmultielementatomic absorptioninstrumentswith high-powercon-
tinuum light sourcesare available(O'Haver & Messman,1986), the discussion
in this chapterwill focus on line sourceinstruments.
Light Sources
A light sourcemust emit high-intensity light at discretewavelengthsthat

can be absorbedby the atomsof the elementof interest.The light sourceshould
producea steady,low noisesignal with minimum interferenceand a long operat-
ing life. Hollow cathodelamps (HCL) and electrodelessdischargelamps (EDL)
are the light sourcesgenerallyusedin atomic absorptionspectroscopy.The HCL
consistsof a sealedglass tube filled with Ar or Ne, a cathodeconstructedfrom
the elementof interestand an anode(Beaty, 1988).An electricalpotentialapplied
betweenthe cathodeand anodecausesionization of the fill gas. Fill gas ions are
acceleratedtoward the cathode,striking the surfaceand dislodging atomsof the
analyteelementwhich are then excitedby collisions with fill gas ions. As analyte
atomsundergode-excitationthey emit light of the properwavelengthfor absorp-
tion by atomsof the sameelementin the flame or furnace.Hollow cathodelamps
may be single or multielementbasedon the material used to constructthe cath-
ode. Multielement lampsgenerallyhave a shorteroperatinglife, lower emission
intensity and a greaterpossibility for interferencethan single elementlamps.
Hollow cathodelamps are generallyusedas the light sourcefor most ele-
ments,but with volatile elements,short lamp life and low light intensity may be
a problem. Electrodelessdischargelamps can be used to overcometheseprob-
lems and to provide betterprecisionand lower detectionlimits for elementssuch
as As, Sb, Bi, Pb, Se, Te and Sn (Barnett et aI., 1976). The elementor a salt of
the elementis placedinside a quartz bulb filled with inert gas. Atoms of the ele-
ment are vaporized and excited by application of a radio frequency field.
Electrodelessdischargelamps have a longer life and produce higher intensity
light than HCl, but they are more expensive.
Atomization Systems
Atomization occurswhen thermal energyfrom a flame or furnace is used

to breakchemicalbondsto form the free ground-stateatomsnecessaryfor atom-
ATOMIC ABSORPTION & FLAME EMISSION SPECTROMETRY 71
ic absorption.A flame is the most commonly usedatomizationdevice.The gen-

eral componentsof a flame systeminclude a nebulizer,an impact device,a mix-
ing chamberand burnerhead.A nebulizer,which may be pneumaticor ultrason-
ic, convertssamplesolution to mist droplets.A fine aerosolof samplein oxidant
gasis createdby sprayingmist dropletsthroughor againstan impact devicesuch
as a flow spoiler or beadto removelarge droplets. Fine dropletsof sampleand
oxidantgasare mixed with fuel in a chamber.The resultingmixture is introduced
to a burnerheadwherecombustion occurs and an atomic vapor of the elementto
be analyzedis produced.The processis not highly efficient since only a small
part of the samplegets into the flame and only a small fraction of that is in the
properatomic statefor absorption.However,nebulizationconditionsand impact
devicescan be adjusted,and gasesand burnersselectedto give the best possible
signal with minimum interference.
The two most commonoxidant-fuel mixtures usedin FAA are air-acety-
lene and nitrous oxide-acetylene.Air-acetylenemixturesgive a flame tempera-
ture of approximately23()()OC andcanbe usedfor mostelementsthat do not form
refractoryoxides. Metal-oxygenbondswith dissociationenergiesin excessof 5
eV are not readily broken in air-acetyleneflames resulting in chemicalinterfer-
encesand lower sensitivity (Frech et aI., 1980). Nitrous oxide-acetyleneflames
havea maximumtemperatureof 29()()OCthus allowing determinationof elements
such as AI, Si, Ti and Cr which form refractory oxides. The higher temperature
of nitrous oxide-acetyleneflames reduces chemical interferences,but may
depletethe atomic populationthrough excessionization.
A numberof different burner headsare availableto accommodatevarious
sampleconditionsand gas mixtures (Fernandezet aI., 1980a).Burnersare nor-
mally madefrom Ti to provide heatandcorrosionresistance.Premixburnerswith
a 10-cmslot are typically usedwith air-acetyleneflames. This burnerprovidesa
long path for absorbanceand can be usedwithout clogging with solutionscon-
taining low levels of total solids. Nitrous oxide burnershavea shorterslot (5 cm)
and usea highergasflow rate to minimize C buildup which is commonwith high
temperatureflames. Otherburnersare availablefor specialsituationssuchashigh
solids burner headswith a wider slot to accommodatehigh-concentrationsolu-
tions.
The sensitivityof FAA is limited becauseonly a small fraction of the sam-
ple gets into the flame, certain elementsform refractory compoundsin the pres-
ence of O2 which are not readily convertedto atoms, and those atoms that are
formed only remain in the light path for a short time. Other atomizationtech-
niqueshavebeendevelopedwhich increasesensitivity by more efficiently intro-
ducing atoms of the analyte element into the light path and allowing them to
remaintherefor a longer time. Cold vapor and hydride generationtechniquesare
specializedmethodsthat canbe usedto increasesensitivity for selectedelements.
Theseatomizationmethodswill be discussedin a later section of this chapter.
However,the most commonlyusedmethodto achievegreatersensitivity in atom-
ic absorptionis electrothermalatomization.
The most commontype of electrothermalatomizeris the graphitetube fur-
nace(L'vov, 1978;Carnrick& Slavin, 1988).The basiccomponentsof a graphite
furnace system include an atomizer, a power supply and a programmer.The
atomizerassemblygenerallyconsistsof a graphitetube placedhorizontally in the

light path of the spectrometer.The assemblyis housedin a water-cooled jacket
with inert gasflow internal and externalto the graphitetube. The inert gas, usu-
ally Ar, servesto protect the furnace from oxidation and provides a meansto
purge constituentsfrom the furnace with a controllable internal gas flow. An
autosampleror a manualpiston pipetteis usedto dispensethe sampleon the wall
of the graphitetube or onto a graphite platformwithin the tube. The tube is heat-
ed by applying an electricalpotentialto graphitecontactswhich hold the tube in
place. Upon heating,an atomic vapor of the elementto be analyzedis createdin
a confinedspace.The flow of inert gasthrough the graphitetube can be stopped
and a longer residencetime for absorptionof light can be achievedfor atomic
vapor than in a flame or plasma.Thus graphite furnace atomizationresults in
greatersensitivity and lower detectionlimits than atomizationusing a flame or
plasma.Becauseof thesefactors, much smaller samplescan be used in GFAA
than in FAA. Samplevolume in GFAA is less than 100/lL, with 20 IlL being a
typical samplevolume.
In GFAA a sequenceof heatingstepsis usedto dry, pyrolyze and atomize
the sample.The drying step removessolvent and typically involves heating to
100 to 120°C for aqueoussolvents.A pyrolysis or ashingstep is conductedat a
temperaturethat will volatilize matrix components,but retain the analyte.Internal
inert gas flow through the graphite tube is maintainedduring the drying and
pyrolysis stepsto·removevolatilized solvent and matrix components.The atom-
ization stepis carriedout at a temperaturethat will allow productionof an atom-
ic vapor of the analyteelement.The internal gasflow is stoppedduring atomiza-
tion to increasethe residencetime of the atomic vapor in the furnace. A cool-
down stepis sometimesinsertedprior to the atomizationstep.This allows a max-
imum heating rate to be employedand extendsthe isothermalzone within the
tube. A higher temperatureclean-outstep may be insertedafter the atomization
step to removeany residualanalytebefore the next sampleis injected. The user
develops a specific temperatureprogram for each analyte element using the
instrumentsupplier'ssuggestedconditionsas a guide. Theseprogramscan con-
trol internal inert gas flow, method of standardization,modifier addition and
spectrometercontrol functions in addition to regulatingthe heatingsequence.
Optical System
The optical systemof an atomic absorptionspectrometerconsistsof a light

source,a monochromatorand a detector.Hollow cathodelamps and EDL light
sourceswere discussedin a previoussection.Thesesourcesproduceresonance
lines that can be absorbedby atoms of the analyte elementin the vapor" phase.
Analyte atomsin a furnaceor flame can be excitedandwill emit light of the same
wavelengthas the light from the source.Therefore,if light absorptionfrom only
the sourceis to be measuredthe instrumentmust correct for light emissionby
analyteatoms.This is accomplishedby mechanicallyor electronicallyinterrupt-
ing light from the source.A rotating chopperinstalled betweenthe light source
and the atomizermechanicallymodulatesthe light source.Electronicmodulation
is achievedby pulsing the currentto the light source.
ATOMIC ABSORPTION & FlAME EMISSION SPECTROMETRY 73
Atomic absorptionspectrometersmay have a single-beamor a double-

beam configuration. In a single-beaminstrument light from the source passes
through the atomizerand to the detector.In a double-beaminstrumentlight from
the sourceis split into a samplebeam and a referencebeam which is directed
around the atomizer.The referencebeam allows correction of variation in the
light sourceand system electronics. Absorbanceis takenas the ratio of the sam-
ple beam to the referencebeam thus allowing electronic cancellation of any
changesin signal other than thosedue to absorptionin the flame. Double-beam
instrumentshave beenconfiguredfor dual-channeland sequentialmultielement
operations.They offer some advantagesin speedof operationand can extend
source lamp life by reducing or eliminating warm-up time. Many of the past
advantagesof double-beaminstrumentshave been eliminatedby improved light
sources,greaterelectronicstability and improved electroniccorrectionmethods
employedin modemsingle-beaminstruments.
In addition to the spectrallil1e of interest a lot of unwantedlight of other
wavelengthsis present.A devicecalleda monochromatoris usedto disperselight
of different wavelengthsand focus the desired spectral line onto the detector.
Light dispersionis generally accomplishedby using a grating. A grating is a
reflective surface, scored either mechanicallyor holographically with parallel
grooves,that can be designedfor different wavelengthregions.The ability of a
monochromatorto separateclosely spacedwavelengthsof light is given by
"resolving power" (A/ll.')...,), where ll.')..., is the smallestwavelengthdifference that
can be separatedinto two lines at wavelength')...,. The "resolving power" of grat-
ings is controlled by width of the entranceslit which allows light to fall on the
grating, density of lines on the grating, angle of the lines on the grating, surface
area of the grating and size of the exit slit to the detector.The best resolution
occurswhen gratingshave a high line density and sufficient surfaceareato cap-
ture all incident light coming through the entranceslit. Since gratings are most
effective in a given wavelengthrange, instrumentswill frequently be equipped
with two gratings to cover the wavelengthrange (189-851 nm) used in atomic
absorptionspectroscopy.
Light isolatedby the monochromatorpassesthrough the exit slit and falls
on the detector.The most common type of detectoris the photomultiplier tube.
This device can convertvery low levels of light to large output currentswith an
intensity proportionalto the strengthof the incident light. Current from the pho-
tomultiplier is amplified and processedto determinelight intensity before(10) and
after (I) passingthrough the atomic vapor in the atomizer. Light absorbanceby
the analyteelementis characterizedby absorbance(A) which is relatedto 10 and
I by the expression:A = log loll.
In FAA a constantsignal should be obtainedas long as solution is aspirat-
ed into the flame. The pulse height of the signal can be electronicallyconverted
to intensity. Furnaceatomizationdoesnot result in a normally distributedsignal,
therefore,peakheight doesnot give a reliable measureof intensity (Fernandezet
aI., 1980b).Furnaceatomizationresultsin increasinglight absorptionas the atom
population in the furnace increases.When atom diffusion from the furnace
exceedsatom generation,light absorptiondrops. Generally integratedpeak area
must be determinedfor the entire atomizationperiod to get an appropriatesignal
with furnace atomization. Calibration with analyte element standardsallows

direct readoutof concentrationvaluesevenwhen calibrationcurvesare nonlin-
ear. The dataalso canbe storedfor further calculations,manipulationandreport
generation.
SENSITIVITY AND DETECTION LIMITS
A number oftermsincluding sensitivity,characteristicconcentration,char-

acteristicmassand detectionlimit are usedto describeinstrumentperformance
relative to the determinationof a particularanalyteelement.Atomic absorption
sensitivityis defmedasthe concentrationof an elementrequiredto producea 1%
absorptionreading (0.0044absorbance)under optimum operating conditions
(Ingle & Crouch,1988).Sensitivity canbe representedby the expression
concentrationof standardx 0.0044

sensitivity =--m-e-as-u-re-=d:-"a-=-b-so-r-=-b-a-nc-e---
The conceptof sensitivity is basedon concentrationin FAA becausesolution is

aspirateduntil some fmal equilibrium signal is attained. Sensitivity may be
referredto as characteristicconcentrationand expressedas milligrams per liter
or microgramsper liter of elementrequiredto producea 1% absorptionsignal.
Graphite furnace atomic absorption uses the entire sample, measuring an
absoluteamountof analyte.The term "characteristicmass"is usedto represent
sensitivity. Characteristicmassis defined as the amountof analyte elementin
picogramsrequired to produce a 1% absorptionreading (0.0044 absorbance)
(Slavin & Camrick,1985).A furnacesamplevolumeof 20 ilL is commonlyused
to compareflame and furnacesensitivitiesfor a given element.
Sensitivity is a function of the slope of the line relating concentrationto
absorbanceand is relatedto instrumentoperatingconditions.Sensitivity can be
calculatedfrom the equationlisted previouslyif concentrationvs. absorbanceis
linear. When the relationshipis nonlinear,sensitivityis definedat every point on
the curve by dc/dx where x is the measuredsignal and c is concentrationor
amountof elementto be determined.Sensitivity in FAA is largely dependenton
path lengthfor light absorption,optimizationof slit widths and efficiency of the
nebulization:-atomization process.In GFAA sensitivitydependson completeness
of atomizationandresidencetime of free atomsin the graphitetube.A variety of
factors can influence sensitivity including: atomizationtemperature,shapeand
dimensionsof the graphitetube,useof a platformwithin the tube,tubewall qual-
ity and matrix modification to avoid interferences(Slavin & Carnrick, 1985).
Knowledgeof sensitivityfor a particularelementcanhelp the operator determine
if instrumentalconditionsare optimized. Knowing sensitivity also can help the
operatorpick a concentrationrangethat will produceoptimum absorbancemea-
surements.
Sensitivity gives the concentrationor amountof an elementthat will pro-
duce a given absorbancesignal, but it doesnot indicatewhetherthat signal can
be measuredagainstthe baselinenoise of the instrument.The term detection
limit gives the lowest concentrationor amountof an elementthat can be detect-

ed with 95% probability. Detection limit is defined as the concentrationor
amountof an elementthat producesan absorptionsignal two or threetimes high-
er than the standarddeviation in noisefluctuation of the backgroundsignal under
experimentalconditions.Measurementsat the detectionlimit have a high uncer-
tainty, therefore,to obtain reasonableprecision, concentrationsat least 5 to 10
times the detectionlimit shouldbe employedin routine analyticalwork.
Detection limits for a numberof elementsof interestin soils are given in
Table 4-2. A comparisonof detectionlimits amongdifferent techniquesis sub-
ject to substantialuncertaintybecausemany factors influencethe detectionlimit.
Detectionlimits for a given elementmay vary considerablywith instrument,sam-
ple matrix and operatingconditions.The detectionlimits in Table 4-2 are based
on concentrationandwere calculatedfor GFAA by assuminga samplevolume of
20 ilL. The values in Table 4-2 should be viewed as approximationsand only
usedto make comparisonsin a generalsense.Someauthorshave suggestedthat
detectionlimits are similar if they comparewithin a factor of four (Parsonset ai.,
1983).
Table 4-2. Detectionlimits in microgramsper liter for selectedelementsdeterminedby flame atom-

ic absorption,graphitefurnaceatomic absorptionand ICP atomic emission-.t
Element FAA:j: GFAA§ Iep
AI 30 0.1 20
As 200 0.5 20
B 2000 10 4
Ba 20 0.25 1.3
Ca 1 0.25 0.5
Cd 1 0.01 2.5
Co 10 0.3 6
Cr 5 0.03 5
Cu 2 0.05 2
Fe 6 0.25 3
Hg 500 5 50
K 2 0.05 200
Li 8 0.25 190
Mg 0.3 0.004 0.15
Mn 2 0.01 1.4
Mo 30 0.5 5
Na 0.5 0.01 29
Ni 8 0.5 10
P 20000 15
Pb 8 0.1 50
S 1000
Se 200 0.5 50
Si 20 1 12
Sr 4 0.5 0.5
Ti 20 1 3.8
V 25 1 5
Zn 1 0.01 1.8
t Detectionlimits in Table 4-2 are from Boumanset aI., 1987; Ingle and Crouch, 1988; Date, 1991.
:j: Air-acetyleneor nitrous oxide-acetyleneflame.
§ Basedon a samplevolume of 20 .uL.
Detection limits are quite low for most elementsand in generalall three
methodswould be adequatefor many routine soil analysisoperations.Detection
limits are generally much lower for GFAA systems thanFAA or ICP atomic
emission systems.Graphite furnace atomic absorptioncan be used when low
detection limits are required or when sample size is small. Several elements
including C, N, 0, F, S, CI, Br, I and P possessresonancelines below 190 nm and
cannotbe effectively determinedby atomic absorption(Slavin, 1986). Refractory
elementssuch as W, Zr, Ta, Hf and rare earth elementsare not normally deter-
mined by atomic absorptionbecauseof poor detectionlimits. Detectionlimits are
high for FAA determinationof elementssuch as Hg, As and Se. Cold vapor and
hydride generationatomizationtechniquesfor theseelementswill be discussedin
a later sectionof this chapter.Flame atomic absorptionand ICP atomic emission
have similar detectionlimits for many of the other elementscommonly deter-
mined in soils, althoughFAA doesoffer lower detectionlimits for elementssuch
as K, Na, P~ and Cd.
INTERFERENCES
Interferencesthat reducethe accuracyof the atomic absorptionmeasure-

ment can occur at various stagesin the analysis.Theseinterferencesmay result
from physical property differences between samplesand standards,chemical
reactionsthat limit formation of atomic vapor and emissionor absorptionof light
at the wavelengthof interest by speciesother than the analyte element.These
interferences can be grouped into nonspectral and spectral categories.
Nonspectralinterferencesinfluenceformation of the atomic vapor while spectral
interferencesproduceabsorptionor emissionof light at the resonanceline of the
analyteelement.Theseinterferencesand methodsfor their correctionwill be dis-
cussedin the following sections.
NonspectralInterferences
Physical property differencesbetweensampleand standardsolutions can

influence the nebulization/atomizationprocess in FAA. Differences in acid
strength,organics present,or the amount of dissolved solids changessolution
propertiessuch as viscosity and surface tension resulting in different rates of
introductionof standardsand samplesinto the flame. This type of physical inter-
ferenceis known as matrix interference andcan suppress or enhancethe signal of
the analytesolution relative to the standard.High concentrationsof acidsor dis-
solved solids generallysuppressthe analytesignal as a result of increasedsolu-
tion viscosity which reducesaspirationrate. Depositson the nebulizerand burn-
er may lead to high noiselevels and occlusionof solution by large particlesmay
retardvaporization.The presenceof organicsolventswill enhancesamplenebu-
lization efficiency and analytesignal relative to standards.Thesetypes of inter-
ferencescan be controlledby matchingas closely as possiblethe standardmatrix
to that of the sample.The methodof standardaddition also may be usedto com-
pensatefor this type of interference(Varma, 1986). Aliquots of standardare
addeddirectly to the sampleand all solutionsare diluted to the samevolume thus

automaticallyachievingmatrix matching.This techniqueworks only in the linear
portion of the calibrationcurve and doesnot accountfor spectralinterferencesor
chemicalinterferences.
Free atomsof the analyteelementmust be producedfor atomic absorption
to occur. Molecular forms of the analyteelementmust be dissociatedat the atom-
ization temperatureof the flame or furnace. If the samplecontainsa constituent
that forms a thermally stablecompoundwith the analyte,andthis compounddoes
not dissociateat the atomization temperature,a chemical interference exists.
Chemical interferencesare the most prevalent interferencesfound in atomic
absorptionspectrometryand the most difficult to correct.
A typical exampleof a chemicalinterferencein soil analysisby FAA would
be P interferencewith Ca determination(Frech et aI., 1980). Calcium and P in
solution react during evaporationto form calcium phosphatewhich is converted
to stablecalcium pyrophosphateat air-acetyleneflame temperatures.Formation
of this stablecompoundreducesfree Ca atom populationin the flame. This type
of interferencecan be overcomeby adding to samplesolution and standardsan
excessof an element that can form a stable compound with the interfering
species.Elementslike La and Sr form stable bonds with oxyanionsand can be
used to eliminate interferencessuch as those of phosphatewith Ca. Chelation
methodsalso can be usedto reduceanion chemicalinterferenceswith the deter-
mination of elementssuchas Ca, Mg, Co, Cu, Cr and Mn. In this approachcom-
plexation of thesemetalswith chelatorssuch as ethylenediaminetetraacetic acid
(EDTA), ascorbicacid, or citric acid can prevent formation of metal-{)xyanion
complexeswhich could result in formation of refractory oxidesat flame temper-
atures(Tominaga& Umezaki, 1982).
Higher flame temperaturescan reducechemical interferences by supplying
additionalthermal energyto dissociatestablecompoundsof the analyteelement.
Nitrous oxide-acetyleneflame temperaturesare severalhundreddegreescenti-
gradehotterthan thosein an air-acetyleneflame. Nitrous oxide-acetyleneflames
can reduce chemical interferencesby causing dissociation of refractory com-
poundslike calcium pyrophosphate.Higher atomizationtemperatures,however,
pose the risk of anothertype of interference.At high flame temperaturesone or
more electronsmay be completely removedfrom the atom (ionization) creating
an ionic form of the analyteelementthus reducingthe populationof ground-state
atoms available to absorb light. This ionization interferenceis most commonly
observedwith alkali and alkaline earth elementswhich have low ionization
potentials.Ionization interferencecan be controlled by addition of an easily ion-
ized elementsuch as K, Na, Rb or Cs to both samplesand standards.Easily ion-
ized elementscreatean excessof free electronsin the flame, thus suppressing
ionization of the analyte. Lanthanumcan have a dual function in flame applica-
tions, serving as an ionization buffer for elementslike AI, Ca, Mg and Si and
reducinganion interferencesby forming refractory compoundswith oxyanions.
Stepsmust be takento deal with prevalentchemicalinterferencesin GFAA.
Severaldesign and operationalparametersof the graphite furnace can be con-
trolled to minimize nonspectralinterferences.The surfaceof the graphitefurnace
contributes to nonspectral interferencesthrough carbide formation (Styris &
Redfield, 1993). Use of pyrolytically coated graphite tubes decreasesanalyte

reactionwith the tubesurface,reducescarbideformation and resultsin decreased
chemicalinterferences(Slavin et aI., 1982).
In early designgraphitefurnacesthe samplewasplaceon the inside wall of
the graphitetube. As the furnace was heatedto the atomizationtemperaturethe
walls of the tube were hotter than the internal inert gas atmosphere.Analyte
atomswere releasedinto an inert gas atmospherethat was cooler than the atom-
ization temperature,resultingin formation of molecularspeciesthat could inhib-
it subsequentatomizationof the analyte(Slavin & Carnrick, 1985).This problem
was solved by placing the sampleon a solid pyrolytic graphite platform in the
bottom of the graphitetube. As the graphitetube is heated,the temperatureof the
platform lags behindthe wall temperature.When the platform reachesthe atom-
ization temperature,analyteatomsare releasedinto an inert gasatmospherealso
at the atomizationtemperature.In this environmentrecombinationof the analyte
with interfering speciesis suppn;ssed,molecular dissociationis enhancedand
precisionand sensitivity are increased.The platform also offers the advantageof
extendinggraphitetube life sincecorrosivematerialsare not placedin direct con-
tact with tube walls. A numberof different typesof platformsare currently used
in GFAA (Ajayi et aI., 1989).
Releaseof analyteatomsinto a constanttemperaturefurnace environment
is an important considerationin elimination of chemical interferences.One
approachto the creationof a uniform environmentis rapid heatingduring atom-
ization, known as maximum power atomization.Mter pyrolysis, the furnace is
allowed to cool to ambienttemperature,followed by rapid heatingto the atom-
ization temperature.The walls of the tube and the atmosphereare heatedmuch
more rapidly than the platform allowing analyteatomsto be volatilized into a uni-
form temperatureatmosphere.This approachutilizing a cool-downstepfollowed
by maximum power atomizationhas beenshownto dramaticallyreducenonspe-
cific interference(Carnrick & Slavin, 1989).
SpectralInterferences
Spectral interferences occur when atoms, moleculesor particulatesin the

flame or furnaceemit, absorbor scatterlight at the absorbancewavelengthof the
analyte.Spectralinterferencesmust be removedor a correctionapplied to get a
true absorbancereading.There are two commontypes of spectralinterferences,
emission interference and background absorption. Emission interference can
occur when atomsor moleculesin the flame or furnace emit light at the wave-
length of the analyte resonanceline. More commonly, emission interference
occurs as a result of light emissionfrom the flame and "black body" radiation
from the hot graphitetube or platform. Interferencesfrom light emissionfrom the
flame and "black body" radiation increasewith temperature."Black body" radi-
ation is emitted as a continuum over a wide wavelengthrange (280-780 nm),
with a maximumat approximately580 nm. Therefore,elementswith lower wave-
length absorptionlines such as Zn at 213.9 nm are not influenced by emission
interferences,while elementslike Ba with absorptionat 553.6 nm are quite sus-
ceptible.Emissioninterferencescan be controlledby adjustingslit widths so light
ATOMIC ABSORPrION & FLAME EMISSION SPECTROMETRY 79
2 ng Se + 20 I1g Ni in water
0.20
Q)
o 0.15
c
o
o0.10
.0
~
0.05
o '--
200 600 1000
Pyrolysis Temperature eC)
Fig. 4-3. Stabilizationof Se using a Ni (N03)2 matrix modifier. The presenceof Ni (N03h reduces
the volatility of Se and allows samplematrix to be removedby pyrolysis without volatile loss of
Se (modified from Beaty, 1988).
emittedfrom the flame or furnacedoesnot get to the monochromator.Control of

atomization temperaturealso can influence emission interferences.Using the
lowest temperaturepossiblefor efficient atomizationwill reduceemissioninter-
ferencesfrom the flame or furnace.
Backgroundabsorption,the most commontype of spectralinterference,is
quite prevalent in GFAA. Background absorption results from broad-band
absorptionof light by undissociatedmolecularspeciesand scatteringof light over
a wide wavelengthrangeby particulates(Beaty, 1988).Sincebackgroundabsorp-
tion coversa substantialwavelengthrange,there is a high probability of overlap
with analytewavelengths.Backgroundabsorptioncan be decreasedby reducing
samplesize or changingto a different absorptionwavelengthif adequatesensi-
tivity can be maintained.In general,the problem only can be solved by removal
of interfering matrix componentsor optical backgroundcorrection techniques.
Very few samplematrices,especially in GFAA can be analyzedwithout back-
groundcorrectionand/or matrix modification.
Removal of interfering matrix constituentsprior to atomization offers a
direct approachto control of spectralinterferencesassociatedwith background
absorption.Graphitefurnace atomic absorptiontemperatureprogramscontain a
pyrolysis step to remove matrix components.The effective removal of matrix
componentswithout loss of analyte will dependon relative volatility of matrix
and analyte.Chemicalsubstancesknown as matrix modifiers can be addedto the
sampleand standardsto changethe thermochemicalbehaviorof the analyte,the
matrix or both. Matrix modifiers can be used to convert the matrix to a more
volatile form or the analyte to a less volatile form (Fig. 4-3). Either approach
would allow selection of a pyrolysis temperaturethat would result in matrix
removal and analyte retention. Matrix modifiers that stabilize the analyte until
atomization temperaturesare reachedcan help reduce chemical interferences.
Therefore,matrix modifiers can servea dual role by enhancinganalytedissocia-
tion at atomization temperaturesand contributing to removal of matrix con-
stituentsthat can causebackgroundabsorption.Chemical modifiers also can be
usedto modify reactivity and structureof the atomizersurface,facilitate better

sample-atomizersurfacecontactand influencegas phasecomposition(Tsalev et
aI.,1990).
Many volatile analyteelementsincluding Cd, Pb, Se,As, Hg andTe require
addition of matrix modifiers to preventvolatile loss of analyteduring pyrolysis
and to increasevolatility of matrix constituents(Ming & Quan, 1987). A variety
of inorganic salts, organics and gases have been used as matrix modifiers.
Inorganicsaltsare the most commonlyusedmatrix modifiers with ammoniumor
nitrate salts consideredto be preferable.Mixed modifiers such as Pd(N03)z +
Mg(N03)2 are frequently used with complicatedsamples(Tsalev et aI., 1990).
Selectionof an appropriatemodifier is difficult becausemodifier effectiveness
dependson a varietyof factors including analytechemistry,matrix composition,
modifier chemistry,modifier physicalform, atomizerconstruction andefficiency,
and atomizationtemperature.
Becauseof these complexities, a universal chemical modifier does not
exist. However,Pd hasexcellentmatrix modificationcharacteristicsunderreduc-
ing conditions.A mixed modifier consistingof Pd(N03)2 + Mg(N03)2 has been
usedto effectively stabilize a numberof analyte elements(Hinds et aI., 1988).
Matrix modifiers also can createproblemsif precautionsare not taken to select
the properchemicalmodifier. Modifiers may containunacceptablelevelsof trace
elementsor may contaminatethe furnace, thereby producingmemory effects in
subsequentanalyses.This problem canbe overcomeby selectinga modifier that
is rarely determinedby furnace atomic absorption(Pd, Mg, Ce, La, Ta, W, Zr).
Modifiers also may contribute to backgroundabsorption,and have a corrosive
action on the graphitefurnaceor reducesensitivity. A useful overview of matrix
modification can be found in a review article by Tsalev et al. (1990).
Sinceit is not possibleto eliminateall backgroundabsorption, background
correction techniquesare necessary,especiallywith GFAA. Three major back-
ground correctiontechniqueswill be discussedin this section: continuumback-
ground correction (CBC), Zeeman backgroundcorrection (ZBC), and Smith-
Hieftje backgroundcorrection(SHBC). Continuum backgroundcorrectionsys-
tems have been usedfor many years and are still widely used today. The tech-
nique is basedon selectiveabsorptionof a discretewavelengthof light by the
analytecomparedto broad-bandbackgroundabsorptionexhibited by interfering
molecularspecies.
A deuteriumarc source,which has broad band emissionacrossthe wave-
length rangefrom approximately180 to 400 nm, is the mostcommonlyusedcon-
tinuum source(Slavin & Carnrick, 1988).Deuteriumarc emissiongenerallycov-
ers the wavelengthrangewhere backgroundcorrection is most needed.A tung-
sten-halidelamp can be used to extend backgroundcorrection into the visible
range.Light from the deuteriumarc and HCL are alternativelypassedthroughthe
atomizationchamber.Light from the HCL is attenuatedby the analyteand mol-
ecular speciescausingbackgroundabsorption,while continuum sourcelight is
absorbedonly by molecularinterferences.The differencebetweenthe two signals
is taken as absorptiondue to the analyte. Continuum backgroundcorrection is
fairly effective with FAA systems,but suffers some limitations with GFAA.
Continuumbackgroundcorrectioncannotaccuratelydeal with high background
First Excited -....,.--":::- - - -

State
285.2 nm - +
0' 1t 0'
Ground
State
No Magnetic Field 10 kG Magnetic Field
Fig. 4-4. Simple Zeemanenergylevel splitting of Mg in a magneticfield. The 1t componentremains

at the wavelengthof the resonanceline while the cr componentsare shifted to shorterand longer
wavelengths(modified from Sneddon,1989).
absorption and fine structure in backgroundabsorption. In addition, difficult

alignmentand balancingproblemsoccur sincecorrectedand uncorrectedsignals
come from two separatesources(Sneddon,1989).
Zeemanbackgroundcorrectionis basedon magneticfield alterationof the
energy levels of an atom (Fig. 4--4). In the simplest casethe absorptionline is
split int.o three lines. One line (7t component)has reducedintensity and rt?mains
at the original wavelength,polarizedin the planeof the magneticfield. Two lines
(0 components)eachwith 25% of the original intensity are polarizedin a plane
perpendicularto the magneticfield and displacedfrom the original wavelength
(Slavin & Camrick, 1988). A numberof different instrumentconfigurationsare
possiblewith ZBC. The magneticfield can be constant ormodulated,the magnet
can be located at the light source (direct Zeeman) or the atomizer (inverse
Zeeman), andthe magneticfield can be perpendicular(transversesystem)or par-
allel (longitudinal system) to the optical axis. A typical modem system could
containan electromagneton the atomizerto producea magneticfield perpendic-
ular to the optical axis and a fixed polarizerto excludelight from the lamp polar-
ized parallel to the magneticfield. With the magneticfield off, the total absorp-
tion of the analyteplus backgroundabsorptionis measured.When the magnetic
field is on, Zeemansplitting occurs withthe 0 componentsof the analyteideally
split far enoughfrom the resonanceline that they do not absorbperpendicularly
polarized light transmittedby the optical system.Therefore,with the magnetic
field on, only backgroundabsorptionoccurs.The correctedsignal is obtainedby
subtractingabsorbancewith the magneton, from absorbancewith the magnetoff.
Zeemanbackgroundcorrectionsystemshaveseveraladvantagesover CBC
systemsespeciallywith regard to GFAA operation.Zeemanbackgroundcorrec-
tion can more effectively deal with higher levels of backgroundabsorptionfound
in GFAA. Thesesystemscan correctfor spectralinterferencesthat differ from the
analyteline by 0.02 nm, and therefore,they are less affectedby structuredback-
groundabsorption.There is only one light sourceand measurements are madeat
the analyte wavelengthin ZBC. This configuration avoids sourcematchingand
optical alignment problems found with CBC. Zeeman backgroundcorrection,
however,is not free from problems.As analyteconcentrationincreases,broaden-
~ 0.60
C
CO
..c
....
o
If) 0.40
..c
«
"C
U 0.20
o I
100
I
200
I
300
Lamp Peak Current (rnA)
Fig. 4-5. Influenceof hollow cathodelamp currenton apparentabsorbanceof a 1 mgIL Cd solution.
Broadeningof the analytesourceline at high lamp current resultsin reducedanalyte absorption
and is the basisfor Smith-Hieftje backgroundcorrection(modified from Smith and Hieftje, 1983).
ing of analytecr componentsoccursin the magneticfield allowing absorptionof

perpendicularlypolarizedlight. Therefore,somesignalfrom the sampleis includ-
ed in background, thus resulting in overcorrection (Wenrich et aI., 1989).
Sensitivity also can be reducedfor elementsthat havecomplexZeemansplitting
patterns(all elementsof interestexceptMg, Ca, Sr, Ba, Cd and Zn) since some
analyteabsorptionmay be includedin the background(Slavin & Carnrick, 1988).
Polarizerscontribute to a significant loss of light. Finally, ZBC is basedon the
assumptionthat backgroundabsorptionis unaffectedby magneticfield strength
and light sourcepolarization. Previousinvestigationshave indicatedthat this is
not always the case(Massmann,1982).
Smith-Hieftje backgroundcorrection is basedon inducing a self-reversal
(self-absorption)effect in the HCL (Smith & Hieftje, 1983). Under normal low
current conditionsthe lamp producesa distinct spectralline. At high lamp cur-
rent, emitted light is self-absorbedby atomsin the lamp. Self-absorptionwill be
a maximum at the centerof the resonanceline. This broadenedsourceline will
result in greatly reducedanalyte absorption(Fig. 4-5), but will not influence
broad-bandbackgroundabsorption.The SHBC techniqueis performedby first
measuringanalyteplus backgroundabsorptionat low lamp current. The lamp is
then pulsed to a much higher current where sourceline broadeningoccurs and
absorptionin the atomizeris primarily due to backgroundabsorption.Times for
low and high current cycles are preciselycontrolled to give an equal amountof
light in eachphaseof the cycle. The backgroundcorrectedanalytesignal is taken
as the differencebetweenlow and high currentsignals.
Smith-Hieftjebackgroundcorrectioncoversboth visible and UV regionsof
the spectrumand can be usedwith flame and furnace systems.It can deal with
high backgroundabsorption while correcting for structured background.The
techniqueuses a single light source, eliminating source balancingand optical
alignment problemsthat occur with CBC. Smith-Hieftje backgroundcorrection
does not require ancillary equipmentsuch as magnetsand polarizers,thus these
systemsare generally less expensivethan CBC or ZBC systems.Smith-Hieftje
backgroundcorrectionsystemshave a limited analytical rangeand reducedsen-
Table 4-3. Comparisonof continuumsource,Zeemanand Smith-Hieftje backgroundcorrectionsys-

temsfor line-sourceatomic absorption.
Background
correction Advantages Disadvantages
Continuumsource Correctsfor broadband background Cannotcorrect high backgroundab-
absorption sorption
Highestsensitivity Cannotcorrect structuredbackground
Reasonablylow cost Difficult to match line sourceand
continuumsourceintensities
Optical alignmentproblems
Baselinedrift problems
Zeeman Correctsfor high backgroundabsorp- Reducedsensitivity of someele-
tion ments
Correctsspectralinterferencesif Analyte signal may be included in
lines are 0.02 nm apart backgroundcorrection
Correctsfor structuredbackground Backgroundabsorptionmay be af-
absorption fected by magneticfield and light
Doesnot require matchingof two line polarization
sources Polarizerscontributeto light loss
Avoids optical alignmentproblems Cost
Smith-Hieftje Correctsfor high backgroundabsorp- Cannotprovide backgroundcorrec-
tion tion for elementslike Mo and Ti
Correctsfor structuredbackground Reducedsensitivity comparedto
absorption continuumsource
Allows correctionin the visible and Analyte signal may be included in
UVregions backgroundcorrection
Doesnot require matchingof two Specialhollow cathodelampsare
light sources required
Avoids optical alignmentproblems
Doesnot require ancillary equipment
such as magnetsand polarizers
sitivity relative to deuteriumarc CBC. This is a result of analyteabsorptiondur-

ing the high currentpulsewhich leadsto an overestimationof background(Sotera
& Kahn, 1982). A long decaytime after the high currentpulsemay place restric-
tions on the speedof analyte and backgroundsignal collection during GFAA
(Slavin & Camrick, 1988). Finally, every HCL cannot produce a useableself-
reversaleffect for backgroundcorrection. Therefore,SHBC systemscannot be
effectively used for certain refractory elementssuch as Mo and Ti (Sneddon,
1989).
A summary of the advantagesand disadvantagesassociatedwith CBC,
ZBC and SHBC systemsfor line-source atomic absorptionspectrometryare
shown in Table 4-3. Continuumbackgroundcorrectionwith a deuteriumarc is
the most widely usedsystemfor FAA applications.It providesa higher sensitiv-
ity and a greateranalytical rangethan ZBC or SHBC systemswhen background
correction is not a serious problem. Continuum backgroundcorrection cannot
adequatelydealwith high backgroundabsorptionfound in GFAA, ZBC or SHBC
systemsmust be used.Since SHBC systemscannotprovide backgroundcorrec-
tion for someelements,ZBC would seemto be the preferredsystemfor GFAA.
SAMPLE PREPARATION
Methodof samplepreparationwill strongly influencethe atomicabsorption

techniqueselectedfor analysis.The choiceof a suitableinstrumentalmethodwill
dependon chemicalform and expectedconcentrationof elements tobe analyzed
and interferencesassociatedwith the samplematrix. Generalconsiderationsasso-
ciatedwith the determinationof analyteelementsin soil digests,soil extractsand
soil solutionswill be discussedin this section.
Soil Digests
Total soil analysisis generallyperformedby dissolutionin somecombina-

tion of mineral acidssuchas HN03, H2S04, HCI04 and HF or lesscommonlyby
fusion with Na2C03,Na202or Na or Li saltsof B (Cresseret aI., 1988; McGrath
& Cunliffe, 1985). Thesetechniquescan leave acidic or basic matricesthat can
cause analysis problems. However, matrix problems can be reduced in open
digestionsystems(nonpressurized)by heatingto drynessto removestrongacids
followed by dissolutionof the residuein an appropriatereagent.The analystcan
control the dilution factor (ratio of final volume of solution to samplesize) allow-
ing elementspresentin low concentrationsto be analyzedin a low dilution fac-
tor solution followed by dilution to bring elementspresentat higher concentra-
tions into an appropriaterange for analysis. Using an open system digestion
approach,total elementalconcentrationsof Ca, Mg, K, Na, Fe, AI, Mn, Si, Ti, Zn,
Pb, Ni, Cr, Cd and many other metallic soil constituentscan be determinedby
FAA. Digest residuesare frequently redissolvedin dilute HCl. Certain elements
such as Pb, Cu and Cd form thermally stablechlorideswhich only partially dis-
sociateat flame atomizationtemperatures,thus sensitivity is considerablysup-
pressed(Parsonset aI., 1975). This is not a major problemsince total concentra-
tions of theseelementsare generallyhigh enoughfor determinationsto be made
by FAA evenif a loss of sensitivity occurs.If analytelevels in the digest are too
low for direct analysisby FAA a preconcentrationstepusing ion exchangeor sol-
vent extractioncan be used.A methyl isobutyl ketone amine synergisticiodide
complex(MAGIC) extractionsystemwas found to be effective for analysisof 18
elementsby FAA (Clark & Viets, 1981). Preconcentrationstepsoffer the addi-
tional benefit of matrix removal, resulting in reducedinterferences.Flow injec-
tion systems(Tyson, 1991)can be coupledto FAA to preconcentratethe analyte.
Hydride generationtechniquesalso can be used to enhancedeterminationof
hydride-formingelements(Dedina, 1988).
Graphite furnace atomic absorptioncan be used to determineelemental
concentrationsof digestconstituentsthat are presentat levels too low for detec-
tion by FAA. A preconcentrationstep will not be neededfrom a detectionlimit
standpoint with GFAA, but may be helpful to remove matrix components.
Chemicalinterferencesand backgroundabsorptionwill presentproblemsin the
determinationof analyteelementsin soil digests.However,thesedifficulties can
be overcomewith appropriatechemical modifiers and ZBC (Nham & Brodie,
1989). Dilute HN03 (approximately1%) is the preferredmatrix for most analyt-
ical determinations.More concentratedHN03 (:dO%) may improve recoveryof
certain elements,but will hastendeteriorationof graphitetubes.The appearance

of the analyte elementas a chloride in the samplematrix will lower sensitivity.
The mechanismof chloride interferencein GFAA analysisis complex,involving
occlusionof the analytein chloride matrix microcrystalsfollowed by expulsion
during rapid expansionof gaseousproductsof decompositionduring atomization
(Styris & Redfield, 1993).Stablegaseousanalyte-chloridespeciesmay form dur-
ing the atomizationstep and diffuse from the atomizer prior to atomization.
Matrix modification and removal of chloride during the pyrolysis step will be a
necessarycomponentof GFAA analysis.
Dissolutionof soil samplesin closedmicrowavedigestionsystems(Warren
et aI., 1990) presentssomeproblemsfor subsequentatomic absorptionanalysis.
Samplesize is generallylimited to 0.5 g becauseof pressurebuild-up in the sys-
tem during digestion. In addition, acids used in the digestion processare not
removedby evaporationto drynessas in open digestionsystems.Dilution of the
digest is required to lower acid concentrationsthat would decre,asenebulization
efficiency, reduce sensitivity and possibly damagethe equipment.Assuming a
sample size of 0.5 g, a dilution factor of at least 50 will be required for a
microwave digest. This will not be a problem for most elementsof interest in
soils since dilution will be required to bring their concentrationsinto a linear
absorbancerange. However, for certain soil constituentslike Cd, Co, Sc and V,
this level of dilution will make FAA analysis impractical. While microwave
digestion methodsoffer many advantagesin terms of speed,'lower reagentuse,
and reducedcontaminationof samplesand the laboratory atmosphere;they do
createproblemswith regard to analysisof selectedelementsin total digestsby
atomic absorption.Open digestionsystemsoffer advantagesrelative to selection
of samplesize and dilution factor, and ability to remove strong acids from the
matrix.
Soil Solutionsand Soil Extracts
Soil solutions, soil leachatesand soil extracts are routinely analyzedfor

nutrientsand traceelementsusing atomic absorptionspectrometry.Analysts may
be concernedwith the phytoavailability of nutrientsor the movementand food
chain availability of trace elementsfrom municipal, industrial or animal wastes
addedto soils. Soil solutionsand water extractswould, in general,have the low-
est levels of analyteelements.Most major constituentscould still be determined
by FAA. Equilibrium levels of trace elementsin soil solution would, in general,
be quite low and dependenton a numberof factors including total concentration
of the elementpresent,soil pH, sorption characteristicsof the soil and the pres-
enceof complexingagents.Preconcentrationtechniquessuchas ion exchangeor
solventextractionwould be requiredfor FAA analysisor GFAA could be used.
A toxicity characteristicleachingprocedure(TCLP) is used to determine
the mobility of organics and inorganics (Ag,As, Ba, Cd, Cr, Hg, Pb and Se)
addedto soils with variouswastematerials(USEPA, 1992). If traceelementsare
presentat regulatorylevels (mg/L), FAA can generallybe usedto determineall
elementsin the leachateexceptHg which requiresa specialized coldvapor atom-
ization technique. A recent study demonstratedthat simultaneousFAA with
SHBC provided a rapid and efficient methodfor the determinationof all TCLP
elements except Hg at their milligram per liter regulatory levels (Farah&
Sneddon,1993).
Elementconcentrationsin most soil extractsare at high enoughlevels to
allow utilization of FAA. Normal precautionsincluding use of ionization buffers
and releasingagentsare neededfor FAA determinationof exchangeablecations
in 1 M NH40Ac (pH 7) extracts.Soil diethylenetriaminepentaacetic acid (DTPA)
extracts(Lindsay & Norvell, 1978) can be analyzedfor Fe, Mn, Cu, Zn and Cd
by FAA althoughpreconcentrationor GFAA may be requiredfor Cd analysisin
some soils. Diethylenetriaminepentaacetic acid in the extract also limits forma-
tion of refractory componentswhich would reducesensitivity of the FAA deter-
mination. Sequentialextraction techniquesfor fractionation of trace elements
suchas Cd, Co, Cu, Ni, Pb, Zn, Fe and Mn (Tessieret aI., 1979) may requiretrace
elementpreconcentrationor GFAA analysisin somefractions.Addition of appro-
priate matrix modifiers and ZBC would be requiredto overcomeinterferencesin
GFAA determinations.
SPECIALIZED SAMPLE INTRODUCTION METHODS
Hydride Generation
Hydride generationis a useful techniqueto improvethe sensitivityof detec-

tion of elementssuch as As, Sb, Bi, Ge, Pb, Se, Te and Sn which form volatile
hydrides(Dedina, 1988). The techniqueinvolves conversionof the analyte to a
gaseoushydride, transportof the hydride to the atomizerand dissociationof the
molecularhydride speciesfor analysisby FAA or GFAA. Sensitivity is dramati-
cally improved relative to direct FAA analysisbecauserelatively large samples
(10-50mL) canbe used,preconcentrationtechniquescanbe employed,the entire
sample is introduced to the atomizer, atomic vapor residencetime for light
absorption is increasedand separationof the analyte from the sample matrix
reducesinterferences(Beaty, 1988).
Sodiumborohydrideis the reducingagentgenerallyusedto release analyte
hydrides from an acidified sample solution. Efficiency of releaseis generally
greaterthan 95% for most hydride-formingelementsand greaterthan 80% for
Pb, the hydride most difficult to generate(Dedina, 1988). The hydridesgenerat-
ed can be collectedor directly transportedto the atomizer.Flow injection meth-
ods have been developedwhich allow automation of the hydride generation
processand preconcentrationof the analyte(Chan& Sadana,1992).Atomization
can occur in devicessuch as a tube atomizer in a Hr 0 2 flame or directly in a
graphitefurnace(Dedina,1988). Dissociationof the hydride molecularspeciesis
generally consideredto occur by reaction with H radicals in the flame and by
thermal dissociationin GFAA (Welz & Schubert-Jacobs, 1986).
Although spectralinterferencesare greatly reducedin hydride generation,
interferencescan still occurduring hydride release,during transportto the atom-
izer and in the atomizer.Theseinterferencesare generallyassociatedwith biva-
lent transition metals and hydride-forming elements other than the analyte
(Petrick & Krivan, 1987). Methodsinvolving adjustmentof acid strengthin the

sample solution and utilization of exchangeresins have been used to remove
interferencesand allow effective determinationof hydride-formingelementsin
soils andenvironmentalsamples(Hersheyet aI., 1988; Chan& Sadana,1992). In
some soil analysis situations hydride generationmay not be necessaryfor the
determinationof elementslike Se. Direct GFAA analysiswith chemical modi-
fiers and ZBC canprovide comparablesensitivitiesto hydride generation(Nham
& Brodie, 1989).
Cold Vapor Method
Mercury can exist as an atomic vapor at room temperature,therefore,heat

is not requiredto causemoleculardissociation.A cold vapor technique,similar
in someaspectsto hydride generation,is usedfor atomic absorptiondetermina-
tion of Hg. Samplesolution is placedin a reactionvesselandtreatedwith a strong
reducing agent such as SnCI2, KBH4 or NaBH4 (Cresseret aI., 1988). The Hg
atomicvaporproducedis transportedto a sample cellin the light pathof the spec-
trometer for the absorptionmeasurement.The systemcan be designedfor one
passanalysis,Hg can be recycledthrough the reactionvesselor preconcentrated
prior to analysis.This techniqueis quite sensitivecomparedto conventionalFAA
becauselargesamplevolumes(50 mL) canbe used,efficiency of Hg releasefrom
solution is high, atom residencetime in the light path is long and little, if any,
backgroundinterferenceshould be present. Detection limits of approximately
0.02 ~gIL make the cold vapor methodthe most sensitiveand reliable technique
for determinationof low concentrationsof Hg by atomic absorption (Beaty,
1988).
Solid Samples
Thereis growing interestin direct determinationby GFAA of elementcon-

centrationsin solid sampleswithout prior dissolution(Brown et aI., 1987; Hinds
& Jackson,1987;Cresseret aI., 1988; Hoeniget aI., 1989;Carbonellet aI., 1992).
Direct introduction of solid samplescould theoretically reducesampleprepara-
tion time, reducerisk of analyte loss during digestionand eliminate contamina-
tion from reagents(Brown et aI., 1987).The samplecan be introducedas a small
solid microsampleor as a slurry of the homogenizedsample (Cresseret aI.,
1988). Solid sampleshave been analyzed in flames and furnaces using cups,
probes,boatsand direct placementon the graphiteplatform (Brown et aI., 1987;
Hoenig et aI., 1989). Since samplesizes are generally less than 5 mg, sample
inhomogeneitybecomesa seriousproblem.
Introductionof solid samplesin a slurry form seemsto be preferablesince
an autosamplercan be used(Hoeniget aI., 1989).Sampleinhomogeneityand sta-
bility of the suspensionare still major problems.Hoenig et ai. (1989) were able
to determineCu, Cd,Pb, Co, Cr and Ni at the milligram per kilogram level in soil
and sedimentslurries using GFAA. Significant chemical and spectral interfer-
enceswill occur, but in somesituationsthey can be overcomewith matrix mod-
ification and ZBe. Difficulties in finding appropriatereferencestandards,and
matching sample and standardmatrices, present calibration problems. Direct

solid sampleintroduction can be usedwhen only a small amount of sampleis
available,digestionis difficult or time consuming,interferencescan be overcome
and a reducedlevel of accuracyand precisionis acceptable.
Flow Injection
Flow injection sampleintroduction systemshave the potential to greatly

enhanceperformanceof conventionalatomic absorptionspectrometers.These
systemshave a variety of configurations,but generally include one or more
pumpsto managecarrierstreamflow, manifold componentsfor manipulationand
chemical modification of the sample and electronically controlled valves for
injection of the sample(Tyson, 1991). Flow injection systemsautomatethe sam-
ple preparationprocessby diluting or preconcentrating the analyte,addingmatrix
modifiers, removing interfering matrix. constituentsand transportingthe analyte
directly to the atomizer. On-line flow injection separationproceduresbasedon
liquid-liquid extraction,dialysis, ion exchange,precipitation,gas diffusion and
electrochemicalprinciples have been developedfor atomic absorptionsystems
(Fang, 1993). Trace metals, presentat concentrationsbelow the detectionlimit
for GFAA, have been isolated from sample matrices and preconcentratedby
organic extraction to detectablelevels in a flow injection system(Bengtsson&
Johansson,1984). Flow injection systemshave been coupled with automated
hydride generationto preconcentrateand determineAs and Se in soil and sedi-
ment samples(Chan & Sadana,1992). A flow injection systemhas beendevel-
opedto performdilutions and to add releasingagentsand ionization buffersin the
automateddeterminationof basic cationsby FAA (Cresseret aI., 1988).
Flow injection systemshave severaladvantageswhen comparedto batch
operationsincluding: samplingfrequencycan be increasedto 500/h with little or
no loss of precision,lower consumptionof sampleand reagent,betterprecision,
reducedrisk of contaminationand automatedoperation(Fang, 1991). On-line
columnseparationand preconcentrationof analyteusingvariousresinsandsolid-
phaseextractantsseemsto be the most promising current use of flow injection
systems.At the present time these techniqueswould be most appropriately
appliedto aqueoussoil extractsand soil solutions.A comprehensivetreatmentof
flow injection separationand preconcentrationsystemsis given by Fang(1993).
Although commercially available manifolds to carry out many separationand
preconcentrationoperationsstill needto be developed,flow injection is likely to
be a major componentof future automatedatomic absorptionsystemsfor soil
analysis.
REFERENCES
Ajayi, 0.0., D. Littlejohn, and C.B. Boss. 1989. Influence of graphitefurnacetube designon vapour
temperaturesand chemicalinterferencesin ETA-AAS. Talanta36:805--810.
Alkemade, c., and T. Joseph.1979. Fundamentalsof analytical flame spectroscopy.Adam Hilger
Ltd., Bristo!'
Barnett, W.B., J.W. Vollmer, and S.M. DeNuzzo. 1976. The application of electrodeless discharge
lampsin atomic absorption.At. Absorp. News!. 15:33-41.
ATOMIC ABS0RYI10N & FLAME EMISSION SPECTROMETRY 89
Beaty,R.D. 1988. Concepts,instrumentationand techniquesin atomicabsorptionspectrophotometry.

Perkin-ElmerCorp., Norwalk, cr.
Bengtsson,M., and G. Iohansson.1984. Preconcentrationand matrix isolation of heavy metals
through a two-stagesolventextractionin a flow system.Anal. Chim. Acta 158:147-156.
Boumans,P.W., I.M. Yrakking, and I.IAM. Yrakking. 1987. Detectionlimits of about 350 promi-
nent lines of 65 elementsobservedin 50 and 27 MHz inductively coupled plasmas(lCP):
Effects of source characteristics,noise and spectral bandwith. Spectrochim. Acta
42B:553-579.
Brown, A.A., M. Lee, G. Kullemer, and A. Rosopulo.1987. Soil samplingin graphitefurnaceatom-
ic absorptionspectrometry.FreseniusZ. Anal. Chern.328:354-358.
v.,
Carbonell, A. Morales-Rubio,A. Salvador,M. de La Guardia,I.L. Burguera,and M. Burguera.
1992. Atomic absorptionspectrometricanalysis of solids with on-line microwave-assisted
digestion.1. Anal. At. Spectrom.7:1085-1089.
Carnrick, G.R., and W. Slavin. 1988. Modem graphitefurnaceAA. Am. Lab. 20(10):88-95.
Carnrick, G.R., and W. Slavin. 1989. Modem graphitefurnaceAA. Am. Lab. 21(2):90-95.
Chan,C.C.Y., and R.S. Sadana.1992. Determinationof arsenicand seleniumin environmentalsam-
ples by flow-injection hydride generationatomic absorptionspectrometry.Anal. Chim. Acta
270:231-238.
Clark, R.I., andI.G. Viets. 1981.Multielementextractionsystemfor the determinationof 18 traceele-
mentsin geochemicalsamples.Anal. Chern.53:61-65.
Cresser,M.S., L. Ebdon,and I.R. Dean.1988.Atomic spectroscopyupdate-environmental analysis.
1. Anal. At. Spectrom.3:1-28.
Date, A.R. 1991. Inductively coupledplasmamassspectrometry.Spectrochim.Acta Rev. 14:3-32.
Dedina,I. 1988. Evaluationof hydride generationand atomizationfor AAS. Prog.Analyt. Spectrosc.
11:251-360.
Fang,Z. 1991.Flow-injectionon-line column preconcentrationin atomic spectrometry.Spectrochim.
Acta Rev. 14:235-259.
Fang,Z. 1993. Flow injection separationand preconcentration.YCH, New York.
Farah,K.S., and1. Sneddon.1993.The useof simultaneousflame AAS for the determinationof inor-
ganicsincludedin the TCLP. Am. Environ. Lab 5:1,6,8.
Fernandez,F.1., B. Lumas,and M.M. Beaty. 1980a.A new burnersystemfor atomicabsorptionspec-
trophotometry.Atom. Spectrosc.1:55-57.
Fernandez,F.1., M.M. Beaty,and W.B. Barnett. 1980b.Use of the L'vov platform for furnaceatomic
absorptionapplications.At. Spectrosc.1:16-21.
Frech,W., I.A. Persson,and A. Cedergren.1980. Chemicalreactionsin atom reservoirsusedin atom
absorptionspectroscopy.Prog. Anal.At. Spectrosc.5:279-297.
Hershey,J.W., T.S. Oostdykand P.N. Keliker. 1988. Determinationof arsenicand seleniumin envi-
ronmentaland agricultural samplesby hydride generationatomic absorptionspectrometry.J.
Assoc. Off. Anal. Chern. 71:1090-1093.
Hieftje, G.M., T.R. Copeland,and D.R. De-Olivares.1976. Flame emission,atomic absorptionand
atomic fluorescencespectrometry.Anal. Chern.48:142R-158R.
Hinds, M.W., and K.W.Iackson.1987.Lead atomisationfrom soil by slurry introductionelectrother-
mal atomisationatomic absorptionspectrometry.Part 1. Effects of matrix componentson the
absorbanceversustime profile. J. Anal. At. Spectrom.2:441-445.
Hinds, M.W., M. Katyal, and K.w. Jackson.1988. Effectivenessof palladium plus manganeseas a
matrix modifier for the determinationof lead in solutionsand soil slurries by electrothermal
atomisationatomic absorptionspectrometry.J. Anal. At. Spectrom.3:83-87.
Hoenig, M., P. Regnier,and R. Wollast. 1989. Automatedtrace metal analysisof slurried solid sam-
ples by graphite furnace atomic absorptionspectrometrywith application to sedimentsand
suspendedmattercollectedin natural waters.J. Anal. At. Spectrom.4:631-634.
Ingle, 1.0., and S.R. Crouch. 1988. Spectrochemicalanalysis.PrenticeHall, EnglewoodCliffs, NJ.
Lindsay, W.L., and W.A. Norvell. 1978. Developmentof a DTPA test for zinc, iron, manganeseand
copper.Soil Sci. Soc. Am. J. 42:421-428.
L'vov, B.Y. 1978. Electrothermalatomization-theway toward absolutemethodsof atomic absorp-
tion analysis.Spectrochim.Acta 33B:153-193.
Marshall,1., S.1. Haswell,and S.1. Hill. 1987.Atomic spectrometryupdate-instrumentation. J. Anal.
At. Spectrom.2:79R-115R.
Massmann,H. 1982. The origin of systematicerrorsof backgroundmeasurements in Zeemanatom-
ic absorptionspectrometry.Talanta29:1051-1060.
McGrath, S.P.,and C.H. Cunliffe. 1985. A simplified methodfor the extractionof the metalsFe, Zn,
Cu, Ni, Cd, Pb, Cr, Co andMn from soils andsewagesludges.1. Sci. FoodAgric. 36:794-798.
Ming, N.Z., and S.x. Quan. 1987. The reductionand elimination of matrix interferencesin graphite
furnaceatomic absorptionspectrometry.Spectrochim.Acta 42B:937-949.
Nham,T.T., and K.G. Brodie. 1989.Evaluationof digestionproceduresfor the determinationof sele-
nium in soil comparingvapourgenerationand graphitefurnaceatomic absorptionspectrome-
try with Zeeman-effectbackgroundcorrection.J. Anal. At. Spectrom.4:697-700.
O'Haver, T.C., and J.D. Messman.1986. Continuum sourceatomic absorptionspectrometry.Prog.
Analyst. Spectrosc.9:483-503.
Parsons,M.L., B.W. Smith, and G.E. Bentley. 1975. Handbookof flame spectroscopy.PlenumPress,
New York.
Parsons,M.L., S. Major, and AR. Forster. 1983. Trace elementdeterminationby atomic spectro-
scopicmethods-state of the art. Appl. Spectrosc.37:411-418.
Petrick, K., and V. Krivan. 1987. Interferencesof hydride forming elementsand of mercury in the
determinationof antimony, arsenic,seleniumand tin by hydride-generationAAS. Fresenius
Z. Anal. Chern.327:338-342.
Rhoades,J.D. 1996. Salinity: Electrical conductivity and total dissolvedsolids. p. 417-435.In D.L.
Sparkset al. (ed.) Methodsof soil analysis. Part 3. Chemical methods.SSSA Book Ser. 5.
SSSAand ASA, Madison,WI.
Slavin, W. 1986. Flames,furnaces,plasmas,how do we choose?Anal. Chern. 58:589A-597A
Slavin,w., G.R. Carnrick, and D.C. Manning. 1982. Graphite-tubeeffects on perchloric acid inter-
ferenceson aluminum and thallium in the stabilized-temperatureplatform furnace. Anal.
Chim. Acta 138:103-110.
Slavin,w., and G.R. Carnrick. 1985. A survey of applicationsof the stabilizedtemperatureplatform
furnaceand Zeemancorrection.At. Spectrosc.6:157-160.
Slavin,w., and G.R. Carnrick. 1988. Backgroundcorrectionin atomic absorptionspectroscopy.CRC
Crit. Rev. Anal. Chern. 19:95-134.
Smith, S.B., Jr., and G.M. Hieftje. 1983. A new backgroundcorrectionmethodfor atomic absorption
spectrometry.Appl. Spectrosc.37:419-424.
Sneddon,J. 1989. Backgroundcorrectiontechniquesin atomic spectroscopy.Spectroscopy2:38-45.
Sotera,IJ., and H.L. Kahn. 1982. Backgroundcorrectionin AAS. Am. Lab. 14(11):93-99.
Soltanpour,P.N., G.W. Johnson,S.W. Workman, J.B. Jones,Jr., and R.O. Miller. 1996. Inductively
coupledplasmaemissionspectrometryand inductively coupledplasma-massspectrometry.
p. 91-139. In D.L. Sparkset al. (ed.) Methods of soil analysis. Part 3. Chemical methods.
SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Styris, D.L., and D.A Redfield. 1993. Perspectiveson mechanismsof electrothermalatomization.
Spectrochim.Acta Rev. 15:71-123.
Tessier,A, P.G.C.Campbell,and M. Bisson. 1979. Sequentialextractionprocedurefor the speciation
of particulatetrace metals.Anal. Chern.51:844-851.
Tominaga,M., and Y. Umezaki. 1982. Comparisonof ascorbicacid and relatedcompoundsas inter-
ference suppressorsin electrothermalatomic absorption spectrometry.Anal. Chim. Acta
139:279-285.
Tsalev,D.L., V.1. Siaveykova,and P.B. Mandjukov. 1990. Chemicalmodification in graphite-furnace
atomic absorptionspectrometry.Spectrochim.Acta Rev. 13:225-274.
Tyson, I.F. 1991. Flow injection atomic spectrometry.Spectrochim.Acta Rev. 14:169-233.
U.S. EnvironmentalProtectionAgency. 1992.Method 1311 toxicity characteristicleachingprocedure
(TCLP). 40CFR, Part 216, Appendix 2. Office Fed. Reg., Natl. Arch. Records Admin.,
Washington,DC.
Varma, A 1986. Handbookof atomic absorptionanalysis.CRC Press,Inc., Boca Raton, FL.
Warren, c.J., B. Xing, and MJ. Dudas. 1990. Simple microwavedigestion techniquefor elemental
analysisof mineral soil samples.Can. J. Soil Sci. 70:617-620.
Welz, B., and M. Schubert-Jacobs. 1986. Investigationson atomizationmechanismsin hydride-gen-
erationatomic absorptionspectrometry.FreseniusZ. Anal. Chern.324:832-838.
Wenrich, R., W. Frech, and E. Lundberg. 1989. Spectralinterferencesin graphite furnace Zeeman-
effect atomic absorptionspectrometry.Spectrochim.Acta. 44B:239-246.
Zhe-Ming, N., and S. Xiao-Quan. 1987. The reduction and elimination of matrix interferencesin
graphitefurnaceatomic absorptionspectrometry.Spectrochim.Acta 42B:937-949.
Published 1996
Chapter 5
Inductively Coupled Plasma Emission

Spectrometry and Inductively Coupled
Plasma-Mass Spectrometry
PARVIZ N. SOLTANPOUR,Colorado State University, Fort Collins,

Colorado
GREG W. JOHNSON,Matheson Gas Products, Longmont, Colorado
STEPHENM. WORKMAN,Analytical Technologies, Inc., Fort Collins,

Colorado
J. BENTON JONES,JR.,Macro-Micro Analytical Services, Athens, Georgia
ROBERT O. MILLER, University of California, Davis, California
The application of inductively coupled plasma-atomicemission spectroscopy

(ICP-AES) to the analysisof soil and biological materialswas reviewedin 1982
for this book series(Soltanpour etaI., 1982). Over the last 14 yr many studiesin
soil sciencehave incorporateddata obtainedon ICP-AES instrumentation,and
many advancesin the ICP-AES field have beenmade. In addition, inductively
coupledplasma-massspectrometry(ICP-MS) has becomecommercially avail-
able. The purposeof this chapteris to review basic ICP principles; highlight
some of the applications of ICP-AES in the study of soils; and present an
overviewof someof the instrumentalfeatures,capabilities,and limitations of the
ICP-MS for applicationto soil sciencework.
Relatively new developmentsin ICP-AES include: suspensionnebuliza-
tion analysisof clays (Laird et aI., 1991); interfacing ICP spectrometerswith
flow injection analyzersfor automaticdilution, calibration, separation,concen-
tration, standard additions and many other operations (Greenfield, 1983;
LaFemiereet aI., 1985); interfacing ICP-AES with liquid chromatographsfor
concentrationandspeciationof elements(Roychowdhury& Koropchack,1990);
high salt neulizersto preventcloggingof nebulizers(Legere& Burgener,1985);
successfuluseof concentrationand elimination or reductionof spectralinterfer-
ence techniquessuch as chelation/solventextraction (Huang & Wai, 1986;
Book Seriesno. 5.
91
92 SOLTANPOURET AL.
Bradford & Bakhtar,1991); useof computerprogramssuch as orthogonalpoly-

nomials(Hassan& Loux, 1989),simplexoptimization(Belchamberet aI., 1986),
and that recommendedby Taylor and Schutyser,1986,for optimizationof spec-
trometeroperatingconditionsand automaticcorrectionfor spectralinterferences;
and compilation of ICP emissionlines (McLaren & Berman, 1985; Boumans,
1984; Parsonset aI., 1980).
The ICP-MS techniquehas beendevelopedover the last 20 yr, particular-
ly by Houk et ai. (1981) who showedsuprathermalionization in an ICP Ar plas-
ma. Within about the last 10 yr it has becomepossibleto apply the method to
routine analytical concentrationdeterminations.Several review articles docu-
ment the ICP-MS developmentalmilestones (Beauchemin,1989; Hieftje &
Vickers, 1989; Douglas, 1989; Houk & Thompson,1988; Houk, 1986; Gray,
1985; Douglas & Houk, 1985). Between 1986 and 1988, ICP-MS enjoyed a
surgeof popularity. According to Cresseret ai. (1988), the late AR. Date attrib-
uted the successof ICP-MS to spectralsimplicity, very high sensitivity, and iso-
tope ratio capability. Cresserattributes the followingquoteto Date (1986),"ICP-
MS is the greatestthing to happento atomic spectroscopysince choppedlight."
Each year since 1986, papersdealing with ICP-AES and ICP-MS analysisof
environmentalsamplesincluding soil sampleshave beenreviewedby Malcolm
S. Cresser,and coworkers(Cresseret aI., 1986, 1988, 1989, 1990, 1991, 1992;
~bdon et aI., 1987). Another sourceof information for the ICP-MS analysisof
geologicaland inorganic materialsis the biennial review publication appearing
in Analytical Chemistry (Jackson et aI., 1989, 1991). The ICP Information
Newsletter publishersan ICP bibliography eachJanuary(Barnes,1992) and-
like the Cresserreview-abstractspaperson ICP-MS presentedat national and
internationalconferences.
An inorganic mass spectrometry,ICP-MS, and x-ray fluorescencespec-
trometry review has beenpublishedyearly since 1988 (Ure et aI., 1988; Bacon
et aI., 1989, 1990,1991). A text on applicationsof ICP-MS was editedby Date
and Gray (1989), and a compilation of 21 selectedpapersfrom the Second
InternationalConferenceon PlasmaSourceMass Spectrometryheld at Durham
University, 24 to 28 Sept. 1990,was editedby Holland and Eatonand published
as a bound volume (1991). A comprehensivebook on ICP-MS and its applica-
tions has beenwritten by Jarvis (1992).
Isotopesof 71 naturally occurring elementscan be monitoredusing con-
ventionalpositive ion, solution nebulizationICP-MS. Accuraciesof theconcen-
trations estimatedusing thesemeasurements at the Division of Agriculture and
Natural Resources(DANR) Analytical Lab at the University of California,
Davis, using internal standardcorrections,are typically within 2.5% of the true
concentrationsin favorable cases.For about 70% of theseelements,more than
one stable isotopeoccursin nature.Thus, analysescan be done for them using
isotope ratios and/or isotope dilution. Isotope ratios show precision of 0.1 to
0.3% (Gregoire, 1989). Concentrationscalculatedusing the method of isotope
dilution (Fasset& Paulsen,1989) are generallywithin 1% of their true concen-
trations;a higher accuracyand precisionthan ICP-MS analysesdonewithout the
use of stable isotope addition (Viczian et aI., 1990; Van Heuzen et aI., 1989,
Garbarino & Taylor, 1987; McLaren et aI., 1987; Dolan et aI., 1990).
ICP EMISSION SPECTROMETRY 93
Concentrationsfor 13 other nuclei that are not naturally occurring also can be
estimatedusingthe ICP-MS (Brown et aI., 1988;Igarashiet aI., 1990; Kim et aI.,
1989a,b,1991).
While elementalcoverageand detectionlimits under relatively ideal con-
ditions are excellent, there are some problem areas in ICP-MS that must be
investigatedbeforedecidingwhetheror not the ICP-MS technique"will work for
you (Hieftje, 1992)." While mostof the following problemshavebeenovercome
or circumventedto meet analytical needsin selectedinstances,the statements
that follow are generallyvalid for a generic,normal resolution(i.e., peakwidths
between0.5 and 1.0 dalton), normal aqueousaerosolgenerationICP-MS:
1. Although the ICP-MS has beenfound to be satisfactoryfor soils work,
comparedto ICP-AES the accuracyand precisionof ICP-MS data are approxi-
mately three times worse. However,for concentrationsdeterminedfrom isotope
dilution/ratio measurements,precision and accuracy is somewhat better
(Gregoire,1989; Dolan et aI., 1990; Taylor et aI., 1991).
2. Isobaricoverlaps(spectralinterferences)occurwith someregularity for
elementsbetweenapproximately28 to 80 daltons,and do occur throughoutthe
massrange.They are a result of a commonunit masssharedby more than one
element,doubly chargedions overlappingan isotopewith half the unit massof
the doubly chargedspecies(Vaughan& Horlick, 1986), elementaloxide, ele-
mental hydride, and/or elementalhydroxide ions overlappingisotopesof other
elements(Vaughan& Horlick, 1986; Munro et aI., 1986; Date et aI., 1987; Gray,
1986), and backgroundspectral problems (Vaughan & Horlick, 1986; Gray,
1986; Tan & Horlick, 1986). The isobaric interferencesinvolving O2 can be
eliminatedusing techniques such as electrothermalvaporization(ETV) atomiz-
ers or Laserablationsampleaerosolproduction(Gregoire,1989).
3. Ion responseis significantly suppressed by matrix elements.The thresh-
old matrix elementvaluesare low comparedto emissionsuppressionsnotedfor
ICP-AES. Nonspectroscopic interferencesresult from excessivedissolvedsolids
in the test solutions. For a numberof reasons,the analyteion arrival rate at the
detector (i.e., analyte response) is suppressedunder these circumstances
(Beaucheminet aI., 1987; Olivares & Houk, 1986; Douglas & Kerr, 1988;
Gregoire, 1987a,b;Hieftje, 1992). Although the onsetof suppressionis usually
observedin the neighborhoodof 100 to 500 mg L-l at the DANR Analytical Lab,
Gregoire (1989) indicated somewhathigher levels using the same instrument
model/manufacturer (i.e., Perkin-ElmerSCIEX 250).
4. The ICP, generatedin Ar with normal aqueoussolution nebulization,
may be unableto producemeasurableamountsof positive ions for someanalytes
that could be of interest,e.g.,F, CI, and/orS. However,the halogenscan be deter-
mined in the negativeion mode(Hieftje et aI., 1988; Chisum, 1992). Sulfur can
be detectedif the water is removedfrom the sample prior to injection of dry
aerosol into the ICP and/or a high resolution ICP-MS is available (Douhitt,
1994a,b).Some water vapor can be removedfrom the sampleaerosolusing a
cooled spray chamber(Hutton & Eaton, 1987). Water can be completely sepa-
rated from the S using an electrothermalatomizer(Gregoire, 1989) or partially
removed using nebulization-desolvationequipment (Veillon & Margoshes,
1968).
94 SOLTANPOURET AI...
Fig. 5-1. Magneticfields (H) and eddy currents(shaded)

generated by high-frequency currents (I) flowing
throughcoil (reproducedfrom Fassel& Knisely, 1974,
with permission obtained from the publishers of
H
Analytical Chemistry).
5. The cost of instrumentation,operation,and maintenancefor ICP-MS are

generallyhigher than thosefor ICP-AES, leadingto higher cost per analytecon-
centrationdetermination.We calculate that the cost per analyte concentration
determinationfor an off-the-shelfICP-MS is about two and one-halftimes that
of a state-of-the-artautomatedsequentialscanningICP-AES instrument,using
the samedepreciationschedulefor both instruments:Gregoire(1989) points out,
however,that the relative cost of analysisusing ICP-MS is low relative to other
methodscapableof producingdataon individual isotopes.Similarly, the sample
throughputis abouta factor of five greaterfor ICP-MS than obtainableby these
other isotopemethods.
6. Finally, while multielementcapability existsfor the ICP-MS, true simul-
taneousmultielementanalysisdoesnot (Hieftje, 1992). For an ICP-AES simul-
taneousmultielementsystem,addingmore analytesdoesnot require longermea-
surementtimes per sampleto preservedetectionlimits. For the ICP-MS howev-
er, addingadditionalanalyticalisotopesrequiresa longer analysistime per sam-
ple to avoid detectionlimit and/orprecisiondegradation.
INDUCTIVELY COUPLED PLASMA-ATOMIC EMISSION

SPECTROMETRYAND INDUCTIVELY COUPLED
PLASMA-MASS SPECTROMETRY
Instrumentation
Inductively CoupledPlasmaGeneration
The ICP is producedby passinginitially ionized Ar gas through a quartz
torch locatedinside a Cu coil connectedto a radio frequency(RF) generator.The
RF generatorprovides upto 3 kW forward power (in most commercialunits) at
a frequencyof 27.1 MHZ. "The high-frequencycurrentsflowing in the Cu coil
generateoscillating magnetic fields whose lines of force are axially oriented
inside the quartztube and follow elliptical closedpathsoutsidethe coil," Fassel,
1977. The Cu coil-torch assemblyis shownschematicallyin Fig. 5-1 (Fassel&
OPTICAL EMISSION SPECTROMETRY
E
E
...J
5 TEMPERATURE (K)
u (± IO%)
a
<{
25 6000
0
...J 20 6200
w
>
0
15 6500
CD 6800
<{ 8000 (ESTIMATE)
..... 10000
:J:
(!)
iii
:J:
SAMPLE AEROSOL
Fig. 5-2. Temperaturesin the plasmaas measuredby the spectroscopicslope method(reproduced

from Fassel,1977,in Pure and Applied Chemistry, with permissionobtainedfrom PergamonPress
Ltd., Oxford).
Kniseley, 1974). Electronsand ions passingthrough the oscillating electromag-

netic field flow at high accelerationratesin closedannularpathsvarying in their
direction and strength, resulting in electron accelerationon each half cycle.
Collisions betweenacceleratedelectronsand ions, and ensuingunionizedAI gas
causefurther ionization.The collisionscauseohmic heatingand,when measured
spectroscopically,give thermal temperaturesranging from 6000 to 10 ooooK
(Fig. 5-2; Fassel,1977).
The quartztorch hasthreeconcentricchannels.The outerchannelconducts
AI gasat about 15 to 17 L min-I to the plasmato sustainthe plasmaand to iso-
late the quartz tube from high temperatures.The innermostchannelis for intro-
duction of sampleinto the plasma.The middle channelconductsthe auxiliary AI
gasat about 1 L min-I and is usedin ICP-AES only when startingthe plasmaor
for organic samples,and is routinely used for all types of samplesfor ICP-MS
(Fig. 5-3; Fassel& Knisely, 1974). The ICP has an annular, or donut, shape
when it is viewed from above.The hole has a lower temperaturethan the donut
and, once the flow of vaporizedsamplecarried in a flow of nebulizer argon is
established,offers less resistanceto the sampleinjection. The sampleis injected
into the plasmaby using Ar carriergasat a rate of about 1 L min- 1 for ICP-AES
work. For ICP-MS work the aerosolflow is approximately1.5 L min-I.
Propertiesof Inductively CoupledPlasma

The ICP generated,as discussedabove,hasuniquephysicalpropertiesthat
make it an excellent sourcefor elementalvaporization/atomization/ionization/
excitation. For ICP-AES, the aerosoldropletscontaining the analyte are desol-
vated, the analyte salts/oxidesare vaporized,and the analyte is atomizedat the
high temperatureregion of the plasmanearthe Cu coil (Fig. 5-2). An initial radi-
ation zone (IRZ) has beendefined by Koirtyohannet al. (1980) as the zone that
96 SOLTANPOURET AL.
Fig. 5-3. Typical quartz torch and inductively coupled

plasmaconfiguration. Flow A is auxillary flow used
mf m ARGON PLASMA
@ @ SUPPORT FLOW
for organic samples (reproduced from Fassel &
Knisely, 1974, with permission obtained from the r
AEROSOL CARR IER
publishersof Analytical Chemistry). ARGON FLOW
beginsin the sampleaerosolchannelinside the load coil for ICP-AES. The IRZ
extendsupwardto 1 or 2 mm abovethe load coil, taking on the appearance of an
amber"bullet" during nebulizationof soil, plant and water samples.The CaO
moleculeson the surfaceof the "bullet" emit an ambercolored radiation which
changesto a deepblue- or purple-coloredradiationfurther downstreamas emis-
sion from Ca atomsand ions dominate.The blue/purpleregion is termedthe nor-
mal analytical zone (NAZ), and is the region in which the analyte emissionis
observedby the spectrometer.Color photographsillustrating the appearanceof
the IRZ and NAZ while nebulizing an elevatedconcentrationof Y into an ICP
have recently beenpublishedfor ICP-AES (Winge et ai., 1988), which clearly
define thesecritical regions.The NAZ is 15 to 20 mm abovethe coil, or about
14 to 19 mm abovethe tip of the IRZ, in an environmentrelatively low in back-
groundemission.
The backgroundemissionconsistsof Ar lines and someweak band emis-
sion from OH, NO, and CN moleculespresentin the plasma(Ward, 1978a).By
the time the decompositionproductsof the samplereachthe NAZ, they havehad
a residencetime of about2 ms at spectroscopicallymeasuredtemperaturesrang-
ing from about 8000 to 5000 K (Fassel,1977). The residencetime and temper-
0
ature experiencedby samplesintroduced into the plasmasare about twice as

large as those in the hottestflames, e.g., N20-C2H2. The high temperatureand
residencetime combination,at the sampleaerosolflow ratestypical for the ICP-
AES, lead to completesamplevaporizationand atomizationin contrastto flames
that require releasingagentsfor refractory compounds(Larson et ai., 1975).
Oncethe free compounds,atoms,and ions are formed in ICP-AES, they are in a
chemically inert environmentin contrast to highly reactive combustionflame
environments.Ionization interferencesare generally negligible in ICP-AES.
Self-absorption(a phenomenonresponsiblefor the flattening of the standard
curve at high analyteconcentrations)is practically absent,which leadsto a wide
linear dynamic analytical rangeof three to five decades.No samplingor skim-
mer conesare used in the ICP-AES, and thereforecontaminationfrom them is
absent.
For ICP-MS, the vaporizationand atomizationbegin at approximatelythe

samelocation relativeto the load coil as do theseprocessesin the ICP-AES, in
a relatively hot region of the plasmanear the Cu coil (Fig. 5-2). Comparedto
ICP-AES, however,the flow of sampleand/or auxiliary argon are increased,so
that the IRZ extendsfurther from the downstreampart of the load coil. Sample
aerosoland/orauxiliary Ar flow ratesusedfor ICP-MS work mustbe higher than
usedfor ICP-AES to obtain an analytically usefulpopulationof ions (Winge et
aI., 1991),while keepingthe samplingconea safedistancefrom the load coil to
preventarcing betweenthe coneand the load coil. The IRZ extendswell beyond
the downstreamside of the load coil. The water dropletsproducedin a conven-
tional concentricnebulizer,althoughapparentlyextremelyfew in numbercom-
paredto the total numberof aerosoldropletsproduced,can survive the rigorous
desolvationlatomizationconditionsgeneratedby the ICP (Winge et aI., 1991).
Although the downstreamside of the load coil-to-IRZ tip distancevaries from
one lab to another,.itis generallybetween10 and 20 mm. Unlike ICP-AES, this
leavesmuch of the analyte vaporizationand atomizationto be done in regions
beyondthe hottestpartsof the ICP in the ICP-MS case.The samplingcone ori-
fice definesthe NAZ in the ICP-MS (Fig. 5-4), and can be another0- to 10 mm
downstreamfrom the tip of the IRZ. In the DANR Analytical Lab, the IRZ
extendsapproximately19 mm downstreamfrom the spectrometerside of the
load coil and the.samplerconeorifice is positioned0 to 3 mm downstreamfrom
the IRZ tips which resultsin placementof the NAZ a total of between19 to 22
mm from the nearestsurfaceof the load coil. Most of the particle beamis sucked
throughthe samplingconeinto the intermediatevacuumregion of a differential-
ly pumpedaperture.The tip of a secondcone,called the skimmer, is immersed
in what is termeda barrelshock(Gray, 1989)that resultsfrom supersonicexpan-
sion of the plasmagas as it passesfrom atmosphericpressurethrough the sam-
pling cone orifice into a vacuumof about 1 torr. The kinetic temperatureof the
gaseousparticlesat the tip of the skimmerconehasbeenmeasuredto be 2200 K 0
(Lim et aI., 1989). Although the position of the sampler with respectto the
extendedIRZ of the ICP is adjustedbeforeeachanalytical run to achievemaxi-
mum ion arrival rate at the detector,it also is samplingaerosolthat has under-
gonesolutevaporizationand atomizationreactionsoutsidethe hottestregionsof
the ICP. This is thought to contributeto the appearanceof more molecularions
in the massspectraand higher susceptibilityto nonspectroscopic matrix effects
than if the aerosolflow rate and/orauxiliary Ar flow rate could besloweddown
enoughto put the IRZ back to within 1 or 2 mm of the downstreamside of the
load coil. However,this is not possiblebecauseof the arcingthat occursbetween
the load coil and the metallic samplingcone in instancesin which the cone is
placedtoo closespatially to the load coil.
In summary,the NAZ is much closerto the tip of the IRZ in ICP-MS (~1O
mm) than the NAZ is to the tip of the IRZ in ICP-AES (14-19 mm). The closer
proximity usedfor the ICP-MS measurements increasesthe concentrationof ions
to a level at which they are analytically useful; but, to preventarcingbetweenthe
load coil and the metal samplercone, high flow rates for the nebulizerand/or
auxiliary gasesmust be usedto push the IRZ tip away from the load coil. Thus,
analytesolutevaporizationand atomizationoccurin a region of the plasmawith
98 SOLTANPOURET AL.
A. rep Spati al Configurati on for Ato'llic Emission Spectroscopy
1001"= =",'" " ,
Load Coil
B. rcp Spatial Configuration for Mass Spectrometry
Key: PressuresPI' p,. and p,: ConesS, and S,:
p,=760 torr S,=SamplingCon,
p,=1.2 torr S,=SkimmerCom
p,=10.7 to 10" torr
Fig. 5-4. Spatial nomenclaturefor ICP (adaptedfrom Koirtyohann, 1980), where IRZ denotesthe
Initial RadiationZone. In agronomicwork, it likely will appearred to pink in color due to emis-
sion of CaO and CaOH molecules.NAZ denotesNormal Analytical Zone,the locationof the NAZ
dependingon whetheranalytical measurements are being perfonnedfor AES (A) or MS (B).
cooler excitation and gas temperaturesthan the ionization temperatures.

Undoubtedly,the requirement forhigh ion densityat a distancewell downstream
from maximum gas and excitation temperaturespromote formation of metal
oxide ions and nonspectroscopicmatrix element supressioneffects that are
observedin the ICP-MS. A number of modifications that will be mentioned
below, most involving variations on the usual sampleintroduction techniques,
have beenfound to significantly reducetheseproblems.
SampleIntroduction Systems
Nebulizers.Nebulizersare devices used for the injection of the sample
into the plasmas.Thereare threegeneraltypesof nebulizers-pneumatic nebuliz-
ers, Babingtonstyle nebulizers,and ultrasonicnebulizers(USN's) (Thompson&
Walsh, 1983).The pneumatictype usesthe Venturei effect to draw samplesolu-
tions into the spraychamber.The Babingtonrequiresa pump to deliver the solu-

tion to a pinhole orifice from which Ar gas is emergingat high velocity. The
USN also requiresa pump to deliver the solution, this time to a vibrating plate.
Thereare two commontypesof pneumaticnebulizers:crossflow and con-
centric. For the crossflow, as the solution emergesfrom the rigid capillary tube
carrying the samplesolution, anothertube positionedat a right angle blastsAr
past it to shearoff fine aerosol particles. The cross-flow nebulizersare often
made of highly corrosion-resistantcapillary metal tubes, e.g., Pt-Ir alloy. One
capillary carriesAr at approximately1 L min-1 and the other, samplesolution.
The orientationof the tips is fixed by the manufacturer,and may include a sap-
phire edge at the tip of the solution tube to producea fine, uniform mist out of
approximately 10% of the solution drawn in. The cross-flow systemsin the
authors'laboratorieshave held up to the most demandingapplicationsfor 2 or 3
yr with no sign of degradation.
The concentricflow (Meinhardt type) glassnebulizersare routinely used
at the DANR Analytical Lab for both ICP-AES and ICP-MS work. Theseare
madeentirely of glassin a "T" type configuration.The main barrel of the nebu-
lizer consistsof a fine glasstube taperedto capillary size. The capillary portion,
measuring2.5 cm. in length, carries the samplesolution and is through a tube
joined in a "T" shapeto this barrel. The Ar pressurecan be anywherefrom 124
kPa to 345 kPa (18-50 psi), resulting in flow rates of betweenapproximately
0.75 to 1.5 L min- 1. The openendsof the argontube and the capillary tube meet
at a taper, and a fine mist is producedas the Ar (flowing concentricallyaround
the capillary) shearsoff small fragmentsof water droplets at the capillary tip.
Thesenebulizersare very steadyand, like the cross-flow type, produceaerosol
from about 10% of the solution going through the tip.
The cross-flow and concentricnebulizerseasily clog with high salt solu-
tions. Soltanpouret a1. (1979a)treated1 M NH4HC03-O.005 M DTPA (diethyl-
enetriaminepentaacetic acid) soil extractswith 0.5 M HN03 to overcomeclog-
ging. However, the Colorado State University Soil Testing Laboratory
(CSUSTL) presentlyusesa Legere1 teflon nebulizer(Legere& Burgener,1985)
attached to a peristaltic pump; eliminating the need for acid pre-treatment.
Wolcott and Butler (1979) designeda pneumaticnebulizer that could aspirate
solutions containing up to 36% suspendedsolids. To overcomedifferencesin
surfacetension,density, and viscosity, the analystcan use a peristaltic pump to
introducesamplesolutionsinto the nebulizer(Beasecker& Williams, 1978). For
concentricnebulizers,care must be taken to eliminate small insoluble particles
from test solutionsthat would otherwiseclog the capillary. If a particle becomes
lodged in the capillary or betweenthe capillary and the taperedtip, then great
caremust be exercisedwhile removingthe blockageto avoid breakingthe frag-
ile glasstubing. One methodis to carefully removethe nebulizerfrom the Ar and
sampledelivery tubes and squirt acetonefrom the nebulizer tip into the barrel
while tapping with a finger, and then force Ar through the tip into the barrel
while continuingto tap2.
1 Distributedby Burtec InstrumentCorporation,Post Office Box 235, Delmar, NY 12054.

2 Petrie,K., PrecisionGlassblowingof Colorado,Englewood,CO.
100 SOLTANPOURET AL.
In Babingtonnebulizers(Suddendorf& Boyer, 1978), aerosolis produced

by pumpingsamplesolution into a V-groove and rupturing the cohesiveness of
the liquid streamas it flows over a small hole in the groove from which Ar gas
is flowing. Glassfrit nebulizersand the like (e.g., the Hildebrandnebulizeil) use
the sameprinciple as the Babingtonexceptwith a multitude of orifices. These
nebulizerscan be usedfor high saltsolutions.Since no constrictingorifices are
neededto produce aerosol, they are relatively clog free. For pneumaticand
Babingtonnebulizers,larger dropletssettle out in the spray chamberand drain
off, leaving the finer aerosoldropletssuspendedin the flow streamof argonfor
transportto the plasma.
In USN's, transducersare used to produce the sample aerosol. USN's
improve the detectionlimit of ICP spectrometersby one-to two ordersof mag-
nitudecomparedwith pneumaticenbulizers(Olsonet aI., 1977).A three-to four-
order of magnitudeimprovementin ICP-MS detection limits has been noted
using the USN with a high resolution, double focusing, ICP-MS instrument
(Tsumura& Yamasaki,1991). The USN's are operatedwith a sampleaerosol
desolvation system that follows aerosol production by the transducer.The
aerosoldesolvationsystemis a heating assemblyfollowed by a condensercol-
umn. Thus, factors involved in improvedanalyticalperformanceof the ICP-MS
with use of the USN observedin the latter reference(1991) are: (i) improved
sample transport to the plasma, (ii) reduction in water vapor present in the
aerosolintroducedto the plasma(Hutton & Eaton, 1987), (iii) reductionin the
amount of O2 and OH- presentas reactive gasesin the differentially pumped
interface(Gregoire,1989; Lim et aI., 1989; Veillon & Marghoshes,1968), and
(iv) reducedbackgroundas a result of reducedO2 and OH- levels in the spec-
trum (Gregoire, 1989). Coupled with the high resolution characteristicof the
doublefocusing MS, detectionlimits achievedby Tsumuraand Yamasaki(1991)
are in the low partsper quadrillion range.
Pneumaticand Babingtonnebulizersare the most commonly encountered
waysof enablinga sampleto be injectedinto an ICP, and are usedwith both ICP-
AES and ICP-MS instrumentation.Some of the alternatemethodsof sample
aerosol production and/or sample injection which also are applicable to both
ICP-AES and ICP-MS techniquesare (i) hydride generators(Workman &
Soltanpour,1980; Thompsonet aI., 1978a,b;Ek et aI., 1991), (ii) LASER abla-
tion (Denoyer,1991; Hager, 1989; Abell, 1991; Denoyeret aI., 1991; Pearceet
aI., 1992), (iii) high performanceliquid chromatography(HPLC), including ion
chromatography(Braverman,1992),(iv) liquid-liquid solventextraction(Plantz
et aI., 1989; Serfasset aI., 1986), (v) flow injection (FI) analysis(Thompson&
Houk, 1986; Dean et aI., 1988), (vi) ETV (Gregoire, 1989; Denoyer & Stroh,
1992; Denoyeret aI., 1991), (vii) aerosoldesolvation(Veillon and Margoshes.
1968), (viii) DIN4, (ix) direct insertion devices(Gervais& Salin, 1991), and (x)
ultrasonicnebulizersystems(Olson et aI., 1977). Hydride generators(see sec-
tions "Hydride and Mercury Vapor Generators"and"Determinationof Ultratrace
Levels of Arsenic, Selenium,and Mercury by Use of Hydride-Mercury Vapor
3 LeemanLabs Inc., Lowell, MA.

4 CETAC Technologies,Inc., Omaha,Nebraska.
Generator"),direct injection (see "Direct Injection Nebulizers"), LASER abla-

tion systems(see "LASER Sampling of Solids") and flow injection principles
(see"SpecialApparatus")are discussedbelow, othersystemsare describedin the
abovereferences.
Hydride and Mercury Vapor Generator. Certain elements, when

reducedby NaBH4' form gasesthat can be directly introducedinto the plasma.
Among these elements,As, Sb, Bi, Se and Te are reduced to form volatile
hydride gases,while Hg is reducedto its volatile elementalform. This methodof
sampleintroductiongreatly improvesthe detectionlimits of theseelementscom-
pared with pneumaticnebulizationdue to an improvementin sample delivery
and a decreasein matrix effect. Thompsonet al. (1978 a,b) simultaneouslydeter-
minedAs, Sb, Bi, Se, andTe by useof ICP-AES and a hydridegenerator.Studies
at the CSUSTL have indicatedthat by reducingAs and Se to their hydridesand
Hg to its vapor form and introducing these gasesinto the ICP, they could be
quantitatively detectedat 1.0, 0.5, and 0.5 )1g L-t, respectively(Workman &
Soltanpour,1980). Recently, Ek et al. (1991) have used an analogoussystem
with ICP-MS instrumentationto improve Se detectionlimits to 0.05)1g L-I.
LASER Samplingof Solids.Many solid samplesare difficult or time con-
suming to put into solution, e.g., soils and ceramics.Sometimesthe elemental
compositionof grain features and small inclusions in the solid are of greater
interestthan the overall composition, e.g.,minerals.To savetime in samplepre-
treatment and to permit feature analysis, surface sampling methods using a
LASER havebeendeveloped(Denoyer,1991; Hager,1989;Abell, 1991; Denoy-
er et al., 1991b;Pearceet al., 1992). LASER ablationcan be usedin conjunction
with ICP-AES,but mostly the ablatedaerosolis injectedinto an ICP and the ions
producedare subsequentlydetectedusing a massspectrometer.At least two of
the currentmanufacturersof ICP-MS instrumentationmarketa LASER ablation
accessory5.The accessoryis equippedwith a NdYAG (Nd-Y-Al-Garnet) LASER
and an ablation stand. The ablation stand has an X-Y-Z translationalspecimen
stagethat is moved either with joystick control or rasteredunder computercon-
trol. Vendor softwaresupportstime-resolveddata acquisition and semiquantita-
tive analytical reports. LASER repetition ratesare adjustablefrom a single shot
to hundredsof bursts per second.Beamscan be used in a defocused mode to
cover approximately1 mm of surfacearea, or sharply focusedto less than 0.02
mm (Pearceet al., 1992). The time durationsand numberof repeatingshotscan
be selectedby the operator.The amountof energyper pulse is variable. There is
a thresholdenergyrequiredto fire the LASER. The upperlimit on repetition rate
and energyper pulse is set either by the limitations of the LASER output or the
window material degradationthreshold.A typical pulse can be as short as a few
nanoseconds (Q-switched) and delivers approximately0.1 J of energy.
In operation,the sampleargonto the ICP is momentarilyinterruptedwhile
the ablation stagecover is removed,the samplespecimenplaced on the teflon
ablation stage,and the ablation cover replaced.The sampleAr flow is resumed,
5 FisonsInstrumentsInc., Danvers,MA 01923; Perkin-ElmerSciex, Norwalk, cr 06859-0215.

and the portion of the sampleto be ablatedis locatedwithin the ocular of a light
microscope.The specimenis focusedusing the X-Y-Z movementof the sample
stage.The computeris notified of impending analysis,and the LASER fired.
Preablationtimes,LASER repetitionratesandLASER powerper pulseare some
of the more importantvariables.
The detectionlimits of metalsin the solid are usually lessthan 1 mg kg-1.
The dry sampleaerosolproducedby the laserburstsis free of manyof the recom-
bination polyatomicions that would ordinarily accompanya major element(M)
in a nebulizedsample(e.g., MO+, MW, MOH+, seeDate et al., 1987). Argide
polyatomic ion species,e.g., MAr+, may persisthowever. Sampleanalysisrate
can be rapid, but dependson the analyticalobjectivesandthe variability between
samples.The accuracyof the analysesare highly dependenton the availability of
certified materialsof compositionsimilar to the sample.At ultralow concentra-
tions, memory effects must be taken into consideration.For example,assumea
Au nuggetis to be ablatedto determineapproximateelementalcomposition.On
the next samplean elementalassayis requestedon a metallic inclusion in a piece
of quartzfor Au content.To reducethe Au backgroundbetweenthe two samples,
the entire ICP-MS systemshould be shut down to permit thoroughcleaningof
the sampleand skimmercones,the ICP torch, and aerosolcarrier line from the
LASER stand,andthe interior of the glasssamplestagecover.Cleaningthe glass
sample stage cover is probably the most critical becausethe interior of the
LASER ablationwindow becomescoatedwith a metallic film of elementalcom-
positiongenerallyrepresentativeof the ablatedsample,andreablationof the film
can occurduring ablationon subsequentsamples.Thus,for the analysisproblem
at hand, the total analysistime can be a few minutesor a few hours depending
on whetherthe quartzpiececan be run aheadof the Au nugget,and, more gen-
erally, what the detectionlimit and accuracyrequirementsare.
Direct Injection Nebulizers.Direct Injection Nebulizers(DIN) provide
for the direct injection of microvolumeaqueous liquidsamplesinto the baseof
torch plasmausing fused silica capillary tube and a high pressureHPLC type
pump. It can be utilized on either ICP-AES or ICP-MS instrumentation.Direct
injection nebulizersis uniquely suitedfor the determinationof nebulizermemo-
ry-pronemassisotopesof B, Hg, I, C, S, and Br or wheresamplevolume is lim-
ited. Studiesof Smith et al. (1991) haveshownit capableof detectingas little as
1 nglg of B in biological materialsusing ICP-MS. Powell and Boomer (1995)
have shown that the techniqueunder optimal conditions accuratelycapableof
detectingCr at the 30 ngIL concentrationrangefor multiple Cr(lll) and Cr(VI)
species.In addition, the techniqueprovidesthe fast samplewash-outand high
samplethrough-put.
Spectrometers
Atomic Emission Spectrometry.Atoms of elementsin a samplewhen
excited emit light of characteristicwavelengthswith an intensity directly pro-
portional to the elementconcentration.The light is focusedon the entranceslit
of the spectrometerto illuminate the diffraction grating. The diffraction grating
separateslight into its componentwavelengthsof lines (spectrum).The spectral
line of an analytepassesthroughthe apertureof an exit slit and strikesa photo-
multiplier tube. Photomultiplier tubes produce signals directly proportional to

the intensity of the spectralline. The signal is fed to the readoutsystem,which
displaysintensities,concentrations,or both. Readout systems are computercon-
trolled. The computerstoresthe intensitiesof standardsand usesthesedata to
calculatethe concentrationsof unknowns.Systemsare availablethat checkcali-
bration curve accuracyperiodically, so that if the quality control (QC) limits are
exceeded, thesystem automatically updatesthe calibration6• If the system is
equippedwith tandemnebulizers7, one nebulizercould be shutdown by the com-
puter if indications are that it is clogged or another irrecoverableerror has
occurred,leaving the second nebulization systemto finish running the samples.
If the sensitivity is degradedbeyondprescribedlimits, or if the run is finished,
there are commercially availablesystemsthat shut down the ICP generator,Ar
flow, and other systemfunctions automatically.
Two types of spectrometersare commonly used(Slavin, 1971, p. 53-80):
(i) direct-readingpolychromators(direct readers),and (ii) scanningmonochro-
mators.Somesystemsare equippedwith both spectrometers.
Direct readersare designedto reduce the possibility of unwantedlight
reachingthe photomultiplier tubes.The refractor platesusedfor fine alignment
of the spectrallines also are filters that exclude stray light and/or radiation in
unwantedordersof diffraction. The photomultipliertube-exitslit assembliesare
protected bylight shield and the internal surfacesof the spectrometerare black-
enedto reducereflections.
Scanningmonochromatorsuse a variety of techniquesto make a wide
range of useful analytical wavelengthsaccessible.Fixed or movable gratings,
single or multiple detectors,and movableentranceand exit slits are a few of the
options available amonga variety of manufacturers.The scanningis computer
controlled,fast and accurate.
Direct readershave the advantageof being faster if concentrationsare
beingdeterminedon more than a few elementsper sample,and a smallersample
volume is required in thesecircumstancescomparedwith scanningmonochro-
mators. Thedisadvantageof direct readersis their fixed wavelengths.In contrast,
scanningmonochromatorsallow the analyst to scan the entire spectrum and
choosethe most useful line (see "Selection of Wavelength"). For laboratories
engagedin both routine and researchactivities, a spectrometerwith both a scan-
ning monochromatorand a polychromatoris the bestsystem.
The manufacturersof spectrometersusually provide the requiredsoftware
(computer programs) for the operation of the spectrometer.These programs
enablethe computerto do many tasksautomatically.Throughcomputerprogram
commands,modemspectrometersare able to perform standardization,normal-
ization of standardsolution readings,correction of the interelementalspectral
interferences,printing data,etc. When a spectrometer ispurchased,suchfactors
as computer size, available software, speedof the printer, automatic interele-
mental spectral interference corrections, and other computer-relatedfactors
shouldbe consideredin addition to optical systemfactors.
6 (f. Zalinski, personalcommunication)

7 (G. Coleman,personalcommunication)
Inductively CoupledPlasma-MassSpectrometry.Thereare at leastfive

manufacturersof ICP-MS instruments.Threeproducequadrupolespectrometers,
one manufacturerproducesa high-resolutiondouble-focusingmassspectrome-
ter, and one manufacturerproducesboth quadrupoleand double-focusingmod-
els. The normal peakwidths on quadrupolesare typically one-halfto 1 dalton
acrossthe mass range: 6 to 250 daltons. This is sufficient to separateto the
responsebaselineisotopesdiffering in one atomic massunit that may differ up
to a factor of 2 x 107 in concentrationin the sample.The high resolutiondouble-
focusingelectrostatic-magnetic sectormassspectrometers are capableof achiev-
ing a resolutionof 50 000. Note that the resolution,R, is definedas being equal
to M, the massof interest, divided by 11M, the peak width at 5% of the peak
height, or R =M/I1M. The resolutionof the double-focusingsystemcan be used
to avoid isobaricoverlapsthat occur on unit resolutionunits in many instances.
However,the analyticalsensivityis inverselyproportionalto the resolution, and
the cost of a double-focusingunit is about three times the cost of a quadrupole
equippedunit. All manufacturersmake extensiveuse of computersfor instru-
ment control and dataprocessing.
Among the commonfeaturesof commerciallyavailableICP-MS systems
are the following: (i) An ICP is usedas the ionization device; (ii) The ions are
sampledat atmosphericpressureand detectedat high vacuum,requiring a dif-
ferentially pumpedinterfaceat an intermediate.vacuum;typically 1 torr (1/760
atmosphere);(iii) The pressureis very low inside the spectrometerthat produces
the mass-tochargeseparation,typically 10-4 to 10-7 torr; (iv) The systemsare
highly automated,with computersusedfor instrumentcontrol and data process-
ing; (v) The systemsall have rapid sequentialmultiisotope capability, and are
able to quantitatively analyze isotopes for more than 70 elements; (vi)
Measurementsare sequentialin nature.Spectrometersdo not as yet exhibit true
simultaneous multielement capability; (vii) Detectionlimits are in the low part-
per-trillion range(ng L-1) for genericICP-MS units and low part-per-quadrillion
range(pg L-1, see,for example,Tsumura& Yamasaki,1991)for many elements
using high-resolutionand ultrasonicnebulization,but degradeas a result of sev-
eral factors including the numberof elementsin the analytical suite, the com-
plexity of the samplecomposition,and the amountof dissolvedsolids in the ana-
lytical testsolutions.Severalafter-manufacture,add-onaccessories are available
for ICP-MS and ICP-AES, i.e., ultrasonicnebulizers(USN), DIN, high-perfor-
manceliquid chromatographic (HPLC)systems,Fl· accessory,hydride genera-
tion equipment,ETV accessory,and LASER ablationsolid samplingequipment.
Analytical Capabilities
Selectionof Wavelength
The numberand wavelengthsof spectrallines generatedby de-excitation
of atomsdependson their electronconfiguration.The theory and explanationof
atomic emissionmakefor exciting readingbut are beyondthe scopeof this dis-
cussion. Those wishing to explore spectral theory more thoroughly can read
Boumans'(1966) book on the subject.
For the analystusingthe spectrometrictechnique,line selectionbecomesa

matterof finding the most useful line, sufficiently intenseto be easily detected
with a minimum of spectralinterferencefrom other spectrallines and back-
ground.Line selectioncanbe a difficult processrequiringcarefulexaminationof
the spectrum.In someinstances,the most useful lines may lie outsidethe spec-
tral rangeof the spectrometeror fall in areasof high background.If an analyst
wants to measurethe concentrationof an elementsuchas S that emits the most
suitableradiationfor analytical purposesin the ultraviolet (UV), then that ana-
lyst shouldhave a vacuumor Ar-purgedspectrometeravailableto him because
much of the radiationin the UV spectrum,including UV emissionlines of S, is
efficiently absorbedby 02.
For some elements,only one or two useful lines are available, whereas
otherelementsoffer severaluseful lines.
Winge et at. (1979) determinedthe relative intensitiesof atomic and ionic
lines of elements excitedin ICP. This informationis partially reproducedin Table
5-1.
Table 5-1. Prominentlines of the elementsemitted by the ICP. The list is arrangedalphabetically
with the lines of eachelementlisted in order of decreasingI..JIb ratio (adaptedfrom Winge et aI.,
1979).
Estimated
*
detection
Element S of It Wavelength I..JIb Concentration§ Iimi~ Comments#
nm l1g/mL
Ag I 328.068 38.0 10.0 0.007
I 338.289 23.0 10.0 0.01
II 243.779 2.5 10.0 0.1
II 224.641 2.3 10.0 0.1
II 241.318 1.5 10.0 0.2
II 211.383 0.9 10.0 0.3
II 232.505 0.7 10.0 0.4
II 224.874 0.6 10.0 0.5
II 233.137 0.5 10.0 0.6
AI 309.271 13.0 10.0 0.02 OH band,NRtt
309.284 13.0 10.0 0.02 OHband,NR
396.152 10.5 10.0 0.03
237.335 10.0 10.0 0.03 NR
237.312 10.0 10.0 0.03 NR
226.922 9.0 10.0 0.03 NR
226.910 9.0 10.0 0.03 NR
308.215 6.6 10.0 0.04 OHband
394.401 6.3 10.0 0.05
236.705 5.8 10.0 0.05
226.346 5.0 10.0 0.06
221.006 4.8 10.0 0.06
257.510 4.0 10.0 0.08
As 193.696 56.0 100.0 0.05
197.197 39.0 100.0 0.08
228.812 36.0 100.0 0.08
200.334 25.0 100.0 0.1
189.042 22.0 100.0 0.1
(continuedon next page)
106 SOLTANPOUR ET AL.
Table 5-1. Continued.

Estimated
detection
Element S of It Wavelength IJ1b* Concentration§ limit1l CommentS#
nm llg!mL
234.984 21.0 100.0 0.1
198.970 16.0 100.0 0.2
200.919 6.1 100.0 0.5
278.022 5.7 100.0 0.5
199.048 5.5 100.0 0.5
B 249.773 63.0 10.0 0.005
249.678 53.0 10.0 0.006
208.959 30.0 10.0 0.01
208.893 25.0 10.0 0.01
Ba II 455.403 230.0 10.0 0.001
II 493.408 130.0 10.0 0.002
II 233.527 75.0 10.0 0.004
II 230.424 73.0 10.0 0.004
II 413.066 9.1 10.0 0.03
II 234.758 7.8 10.0 0.04
II 389.178 5.2 10.0 0.06 H 388.905
II 489.997 3.7 10.0 0.08
II 225.473 2.0 10.0 0.2
II 452.493 1.9 10.0 0.2
Be II 313.042 110.0 1.0 0.0003 OHband'
I 234.861 96.0 1.0 0.0003
II 313.107 41.0 1.0 0.0007 OHband
I 249.473 8.0 1.0 0.004 Group NR**
I 265.045 6.4 1.0 0.005 Group NR
I 217.510 2.5 1.0 0.01 NR
I 217.499 2.5 1.0 0.01 NR
I 332.134 1.4 1.0 0.02 Group NR
I 205.590 0.7 1.0 0.04 NR
I 205.601 0.7 1.0 0.04 NR
Bi I 223.061 87.0 100.0 0.03
I 306.772 40.0 100.0 0.08 OHband
I 222.825 36.0 100.0 0.09
I 206.170 35.0 100.0 0.09
I 195.389 14.0 100.0 0.2
I 227.658 12.0 100.0 0.3
II 190.241 10.0 100.0 0.3
I 213.363 10.0 100.0 0.3
I 289.798 9.0 100.0 0.3
I 211.026 7.8 100.0 0.4
Ca II 393.366 89.0 0.5 0.0002
II 396.847 30.0 0.5 0.0005 H 397.007
II 317.933 1.5 0.5 0.01 OHband
I 422.673 1.5 0.5 0.01
Cd II 214.438 120.0 10.0 0.003
I 228.802 110.0 10.0 0.003
II 226.502 89.0 10.0 0.003
I 361.051 1.3 10.0 0.2
I 326.106 0.9 10.0 0.3
I 346.620 0.7 10.0 0.4
I 231.284 0.5 10.0 0.6


Estimated
detection
Element S of It Wavelength l,/lb* Concentration§ Iimit~ Comments#
nm llg/mL
479.992 0.5 10.0 0.6
Co II 238.890 50.0 10.0 0.006
II 228.616 43.0 10.0 0.007
II 237.862 31.0 10.0 0.01
II 230.786 31.0 10.0 0.01
II 236.379 27.0 10.0 0.01
II 231.160 23.0 10.0 0.01
II 238.346 21.0 10.0 0.01
II 231.405 18.0 10.0 0.02
II 235.342 17.0 10.0 0.02
II 238.636 14.0 10.0 0.02
II 234.426 14.0 10.0 0.02
II 231.498 13.0 10.0 0.02
II 234.739 13.0 10.0 0.02
Cr II 205.552 49.0 10.0 0.006
II 206.149 42.0 10.0 0.007
II 267.716 42.0 10.0 0.007
II 283.563 42.0 10.0 0.007
II 284.325 35.0 10.0 0.009
II 206.542 31.0 10.0 0.01
II 276.654 22.2 10.0 0.01
II 284.984 21.0 10.0 0.01
II 285.568 16.0 10.0 0.02
II 276.259 15.0 10.0 0.02
II 286.257 15.0 10.0 0.02
II 266.602 14.0 10.0 0.02
II 286.511 14.0 10.0 0.02
II 286.674 13.0 10.0 0.02
I 357.869 13.0 10.0 0.02
Cu I 324.754 56.0 10.0 0.005 OHband
II 224.700 39.0 10.0 0.008
I 219.958 31.0 10.0 0.01
I 327.396 31.0 10.0 0.01
II 213.598 25.0 10.0 0.01
I 223.008 23.0 10.0 0.01
I 222.778 19.0 10.0 0.02
II 221.810 17.0 10.0 0.02
II 219.226 17.0 10.0 0.02
I 217.894 17.0 10.0 0.02
I 221.458 13.0 10.0 0.02
Fe II 238.204 65.0 10.0 0.005
II 239.562 59.0 10.0 0.005
II 259.940 48.0 10.0 0.006
II 234.349 29.0 10.0 0.01
II 240.488 27.0 10.0 0.01
II 259.837 24.0 10.0 0.01
II 261.187 24.0 10.0 0.01
II 234.810 23.0 10.0 0.01 NR
II 234.830 23.0 10.0 0.01 NR
II 258.588 20.0 10.0 0.02
II 238.863 20.0 10.0 0.02

108 SOLTANPOUR ET AL

Estimated
detection
Element S of It Wavelength IJ1bt. Concentration§ limit~ Comments#
nm JlglmL
II 263.105 19.0 10.0 0.02 NR
II 263.132 19.0 10.0 0.02 NR
II 274.932 19.0 10.0 0.02
II 275.574 16.0 10.0 0.02
II 233.280 15.0 10.0 0.02
II 273.955 15.0 10.0 0.02
Ga I 294.364 64.0 100.0 0.05
I 417.206 45.0 100.0 0.07
I 287.424 38.0 100.0 0.08
I 403.298 27.0 100.0 0.01
I 250.017 16.0 100.0 0.2
II 209.134 11.0 100.0 0.3
I 245.007 10.0 100.0 0.3
I 294.418 9.4 100.0 0.3
I 271.965 5.7 100.0 0.5
I 233.828 3.9 100.0 0.8
I 265.987 3.6 100.0 0.8
Hg II 194.227 120.0 100.0 0.03
I 253.652 49.0 100.0 0.06
I 296.728 1.7 100.0 1.8
I 435.835 1.1 100.0 2.7
I 265.204 0.7 100.0 4.3
I 302.150 0.6 100.0 5.0
I 365.483 0.3 100.0 10.0
In II 230.606 47.0 100.0 0.06
I 325.609 25.0 100.0 0.1
I 303.936 20.0 100.0 0.2
I 451.131 16.0 100.0 0.2
I 410.176 6.4 100.0 0.5 H 410.174
I 271.026 5.4 100.0 0.6
I 325.856 5.0 100.0 0.6
II 207.926 4.2 100.0 0.7
I 256.015 4.2 100.0 0.7
I 293.263 2.0 100.0 1.5
II 197.745 1.7 100.0 1.8
I 175.388 1.6 100.0 1.9
K 404.721 0.7 1000.0 42.9
404.414 NM 1000.0 NM AR404.442
Li 460.286 3.5 100.0 0.9
323.263 2.8 100.0 1.1 OHband
274.118 1.9 100.0 1.6
497.170 1.4 100.0 2.1
256.231 0.7 100.0 4.3
413.262 0.4 100.0 7.5 NR
I 413.256 0.4 100.0 7.5 NR
Mg II 279.553 195.0 1.0 0.0002
II 280.270 100.0 1.0 0.0003
I 285.213 19.0 1.0 0.002
II 279.806 2.0 1.0 0.02
I 202.582 1.3 1.0 0.02
II 279.079 1.0 1.0 0.03

Estimated
*
detection
Element S of It Wavelength IJlb Concentration§ Iimit~ Comments#
nm llg/mL
I 383.826 0.9 1.0 0.03
I 383.231 0.7 1.0 0.04
I 277.983 0.6 1.0 0.05
II 293.654 0.5 1.0 0.06
Mn II 257.610 220.0 10.0 0.001
II 259.373 190.0 10.0 0.002
II 260.569 145.0 10.0 0.002
II 294.920 39.0 10.0 0.008
II 293.930 29.0 10.0 0.01
I 279.482 24.0 10.0 0.01
II 293.306 22.0 10.0 0.01
I 279.827 18.0 10.0 0.02
I 280.106 14.0 10.0 0.02
I 403.076 6.8 10.0 0.04
II 344.199 6.6 10.0 0.05
I 403.307 6.3 10.0 0.05
II 191.510 5.8 10.0 0.05
Mo II 202.030 38.0 10.0 0.008
II 203.844 24.0 10.0 0.01
II 204.598 24.0 10.0 0.01
II 281.615 21.0 10.0 0.01
II 201.511 16.0 10.0 0.02
II 284.823 15.0 10.0 0.02
II 277.540 12.0 10.0 0.03
II 287.151 11.0 10.0 0.03
II 268.414 10.0 10.0 0.03
II 263.876 8.0 10.0 0.04
II 292.339 8.0 10.0 0.04
Na I 588.995 101.0 100.0 0.03 AR588.859
I 589.592 43.0 100.0 0.Q7
I 330.237 1.6 100.0 1.9
I 330.298 0.7 100.0 4.3
I 285.301 1.1 1000.0 27.3 NR
I 285.281 1.1 1000.0 27.3
II 288.114 0.6 1000.0 50.0
Nb II 309.418 83.0 100.0 0.04 OHband
II 316.340 75.0 100.0 0.04 OHband
II 313.079 60.0 100.0 0.05 OHband
II 269.706 43.0 100.0 0.Q7
II 322.548 42.0 100.0 0.Q7 OHband
II 319.498 41.0 100.0 0.Q7 OHband
II 295.088 40.0 100.0 0.08
II 292.781 40.0 100.0 0.08
II 271.662 34.0 100.0 0.09
II 288.318 31.0 100.0 0.1
II 210.942 31.0 100.0 0.1
II 272.198 30.0 100.0 0.1
II 287.539 28.0 100.0 0.1
Ni II 221.647 29.0 10.0 0.01
I 232.003 20.0 10.0 0.02
II 231.604 19.0 10.0 0.02

Estimated
*
detection
Element S of It Wavelength IJlb Concentration§ limit'!! Comments#
nm llglmL
II 216.556 17.0 10.0 0.02
II 217.467 13.0 10.0 0.02
II 230.300 13.0 10.0 0.02
II 227.021 12.0 10.0 0.03
II 225.386 12.0 10.0 0.03
I 234.554 9.5 10.0 0.03
II 239.452 7.8 10.0 0.04
I 352.454 6.6 10.0 0.05
I 341.476 6.2 10.0 0.05
P I 213.618 39.0 100.0 0.08
I 214.914 39.0 100.0 0.08
I 253.565 11.0 100.0 0.3
I 213.547 8.5 100.0 0.4
I 203.349 7.4 100.0 0.4
I 215.408 7.2 100.0 0.4
I 255.328 5.2 100.0 0.6
I 202.347 3.8 100.0 0.8
I 215.294 3.4 100.0 0.9
I 253.401 3.0 100.0 1.0
Pb II 220.353 70.0 100.0 0.04
I 216.999 33.0 100.0 0.09
I 261.418 23.0 100.0 0.1
I 283.306 21.0 100.0 0.1
I 280.199 19.0 100.0 0.2
405.783 11.0 100.0 0.3
224.688 3.0 100.0 0.3
368.348 8.6 100.0 0.3
266.316 7.7 100.0 0.4
239.379 6.3 100.0 0.5
363.958 5.2 100.0 0.6
247.638 5.1 100.0 0.6
S 180.669 30.0 100.0 0.1 Vac.line§§
181.979 30.0 100.0 0.1 Vac. line
Sb 206.833 91.0 100.0 0.03
217.581 68.0 100.0 0.04
231.147 49.0 100.0 0.06
252.852 28.0 100.0 0.1
259.805 28.0 100.0 0.1 NR
259.809 28.0 100.0 0.1 NR
217.919 19.0 100.0 0.2
195.039 18.0 100.0 0.2
213.969 16.0 100.0 0.2
204.957 15.0 100.0 0.2
214.486 12.0 100.0 0.3
209.841 8.7 100.0 0.3
203.977 6.6 100.0 0.5
220.845 6.5 100.0 0.5
287.792 4.7 100.0 0.6
Se 196.026 40.0 100.0 0.08
203.985 26.0 100.0 0.1
206.279 10.0 100.0 0.3

Estimated
*
detection
Element S of It Wavelength IJlb Concentration§ Iimit~ CommentS#
nm I!glmL
207.479 1.9 100.0 1.6
199.511 0.6 100.0 5.0
Si 251.611 250.0 100.0 0.01
212.412 180.0 100.0 0.02
288.158 110.0 100.0 0.03
250.690 100.0 100.0 0.03
252.851 95.0 100.0 0.03
251.432 79.0 100.0 0.04
252.411 75.0 100.0 0.04
221.667 72.0 100.0 0.04
251.920 61.0 100.0 0.05
198.899 50.0 100.0 0.06
221.089 47.0 100.0 0.06
243.515 36.0 100.0 0.08
190.134 23.0 100.0 0.1
220.798 23.0 100.0 0.1
205.813 23.0 100.0 0.1
Sn II 189.989 120.0 100.0 0.03
I 235.484 31.0 100.0 0.1
I 242.949 31.0 100.0 0.1
I 283.999 27.0 100.0 0.1
I 226.891 25.0 100.0 0.1
I 224.605 25.0 100.0 0.1
I 242.170 19.0 100.0 0.2
I 270.651 18.0 100.0 0.2
I 220.965 16.0 100.0 0.2
I 286.333 14.0 100.0 0.2
I 317.505 14.0 100.0 0.2 OH band
Sr II 407.771 72.0 1.0 0.0004
II 421.552 39.0 1.0 0.0008
II 216.596 36.0 10.0 0.008
II 215.284 29.0 10.0 0.01
II 346.446 13.0 10.0 0.02
II 338.071 8.8 10.0 0.03
II 430.545 4.8 10.0 0.06
I 460.733 4.4 10.0 0.07
II 232.235 2.9 10.0 0.1
II 416.180 2.4 10.0 0.1
Te 214.281 73.0 100.0 0.04
225.902 17.0 100.0 0.2
238.578 17.0 100.0 0.2
I 214.725 14.0 100.0 0.2
I 200.202 12.0 100.0 0.3
I 238.326 11.0 100.0 0.3
I 208.116 11.0 100.0 0.3
I 199.418 6.3 100.0 0.5
I 225.548 2.7 100.0 1.1
I 226.555 2.7 100.0 1.2
Ti II 334.941 79.0 10.0 0.0004
II 336.121 57.0 10.0 0.0005
II 323.452 56.0 10.0 0.005 OHband

Estimated
detection
Element S of It Wavelength 1r/1b+ Concentration§ Iimit1l CommentS#
nm ~g!mL
II 337.480 45.0 10.0 0.007

II 334.904 40.0 10.0 0.008
II 308.802 39.0 10.0 0.008 OHband
II 307.864 37.0 10.0 0.008 OHband
II 338.376 37.0 10.0 0.008
II 323.657 30.0 10.0 0.01 OHband
II 323.904 29.0 10.0 0.01 OHband
II 368.520 26.0 10.0 0.01
TI II 190.864 74.0 100.0 0.04
I 276.787 25.0 100.0 0.1
I 351.924 15.0 100.0 0.2
I 377.572 13.0 100.0 0.2
I 237.969 7.0 100.0 0.4
I 291.832 2.9 100.0 1.0
I 223.785 2.2 100.0 1.4
I 352.943 1.7 100.0 1.8
I 258.014 1.7 100.0 1.8
V II 309.311 60.0 10.0 0.005 OHband
II 310.230 47.0 10.0 0.006 OHband
II 292.402 40.0 10.0 0.008
II 290.882 34.0 10.0 0.009
II 311.071 30.0 10.0 0.01 OHband
II 289.332 29.0 10.0 0.01
II 268.796 29.0 10.0 0.01
II 311.838 25.0 10.0 0.01 OHband
II 214.009 20.0 10.0 0.02
II 312.528 20.0 10.0 0.02 OHband
II 327.612 19.0 10.0 0.02
II 292.464 18.0 10.0 0.02
II 270.094 17.0 10.0 0.02
Zn I 213.856 170.0 10.0 0.002
II 202.548 75.0 10.0 0.004
II 206.200 51.0 10.0 0.006
I 334.502 2.2 10.0 0.1
I 330.259 1.3 10.0 0.2
I 481.053 1.3 10.0 0.4
I 472.216 0.7 10.0 0.4
I 328.233 0.6 10.0 0.5
I 334.557 0.4 10.0 0.8
I 280.106 0.4 10.0 0.8 NR
I 280.087 0.4 10.0 0.8 NR
Zr II 343.823 42.0 10.0 0.007
II 339.198 39.0 10.0 0.008
II 257.139 31.0 10.0 0.01
II 349.621 30.0 10.0 0.01
II 357.247 30.0 10.0 0.01
II 327.305 25.0 10.0 0.01
II 256.887 22.0 10.0 0.01
II 327.926 21.0 10.0 0.02
II 267.863 20.0 10.0 0.02


Estimated
detection
Element S of It Wavelength IJIb+ Concentration§ limitll CommentS#
nm llg!mL
II 272.261 16.0 10.0 0.02
II 273.486 14.0 10.0 0.02
II 274.256 14.0 10.0 0.02
II 270.013 12.0 10.0 0.03
II 350.567 12.0 10.0 0.03
II 355.660 12.0 10.0 0.03
II 348.115 12.0 10.0 0.03
II 256.764 11.0 10.0 0.03
II 272.649 11.0 10.0 0.03
II 330.628 11.0 10.0 0.03
II 316.597 11.0 10.0 0.03 OH band
II 318.286 11.0 10.0 0.03 OHband
II 328.471 10.0 10.0 0.03
II 274.586 10.0 10.0 0.03
II 275.221 10.0 10.0 0.03
II 357.685 10.0 10.0 0.03
t S of I = stateof ionization. SymbolsI, II, and III indicatethat the spectrallines originatefrom the
neutral atom, singly ionized, and doubly ionized states,respectively.
+ InlIb =ratio of net analyte intensity to backgroundintensity.
§ Concentration= concentrationof the single element analyte solution used for the wavelength
scansfrom which the prominentlines were determined.
11 Detectionlimits estimatedfrom the IJIb ratios using the formula: DL =O.03C/(IJIb) where C is
concentrationof analyte.
# Includes interferenceinformation when a componentof the backgroundspectrumoverlapsan
analyte line (e.g., the Ba 389.178-nmline is locatedon the H 338.905-nmline) or when an ana-
Iyte line is located in complex molecular band system (e.g., the OH 306.36-nmsystem)where
bandcomponentsmay causespectralinterferences.The notationof molecularbandsdoesnot pre-
clude the use of analytewavelengthswithin the band region.
tt NR = not resolved.This descriptionindicatescomponentsof an unresolvedpair of lines.
+:!: Group NR indicatescomponentsof an unresolvedgroup (three or more lines). Only the wave-
length of the strongestline is listed.
§§ Vacuum lines for S require either a vacuum or an Ar purged spectrometer,see "Selection of
Wavelength."
Selection of Isotope
Ideally, the most abundantisotope is selectedfor an element becauseit
usually will producethe strongestsignal to backgroundratio. Thus, the analyst
obtains the lowest detection limit, the best accuracy,and the best sensitivity.
However, the analytical isotope selection processcan be complicatedby the
presenceof isobaric interferencefrom atomic and molecular ions originating
from the plasma,the solvent, and the reagentsusedto preparethe test solutions
as well as from nonanalytesampleelementalconstituents(Vaughan& Horlick,
1986; Munro et ai., 1986; Date et ai., 1987; Gray, 1986; Tan & Horlick, 1986).
In the isotopeselectionprocessthe following are considered:analytical isotope
abundance,background isobaricspecies,and isobaric speciesresulting from
samplesolution composition.Relative abundanceof the isotopesof all natural
elementsare given by Date and Gray (1989). Tables by Holden and Walker
(1972) and Weastand Astle (1979) containnot only the naturally occurringiso-
114 SOLTANPOURET AL
topesbut thoseproducedas a result of man'sactivities as well. These areuseful

when usedwith the relative abundancesin accuratelycalculatingatomic masses
of the elements, and
by themselvesfor thoseconsideringhigh resolution,double-
focusing ICP-MS.
DetectionLimits (Inductively CoupledPlasma-AtomicEmission

Spectrometry)
Detection limit is defined as the analyte concentrationequivalentto two
times the standard deviation of the background beneath the analyte line.
However,concentrationsfive timesthe detectionlimits aregenerallyrequiredfor
quantitativemeasurements. Hence,the latter is referredto as "quantitativedetec-
tion limit."
From the definition, it is obvious that detection limits are a function of
excitation sourceand samplematrix as they affect the intensity of background
radiation. The detectionlimits are also affected by sampledelivery efficiency.
Detectionlimits are further affectedby operationalparametersof plasmaspec-
trometers;suchas power, height of observation,and the flow rate of samplecar-
rier gas. The optimum operationalparametersare often different for different
groups of elements,and therefore, one should select compromiseoperational
parametersfor simultaneousmultielementanalysis.
Winge et at (1979) calculatedthe ICP detection limits of different ele-
ments (Table 5-1), assumingthat the standarddeviation of the backgroundis
approximately1% of the backgroundsignal level and that the detectionlimit is
threetimes the standarddeviationof the background.The formula usedfor these
calculationsis given in the footnote underTable 5-1. Thesevaluescan be used
as first approximationin the absenceof measuredvaluesfor detectionlimits.
Table 5-2 gives the measuredICP detection limits of some elementsin
pure water, in 10% HCl, and in a solution containingmajor elementsof a typical
arid region soil digest (assuminga dilution factor of 50). The pure water values
were obtainedfrom literature(Robin, 1979).Othervalueswere determinedat the
CSUSTL. The blank or 0 concentrationsolutions were analyzed 10 times to
determinethe standarddeviation of the backgroundsignal at the wavelengthof
the elementsof interest.Thesestandarddeviationswere multiplied by two and
changedto their apparentconcentrationequivalentsusing appropriatecalibration
curves.Thesevaluesare called detectionlimits by definition.
The detectionlimits in 10% HCl solution are given for thoseelementsfor
which the CSUSTL ICP spectrometerhas channels.The simulatedsoil digest
was preparedin a 10% HCl solution containingAI, Fe, K, Na, Ca, Mg, Ti, Mn,
and P; therefore,the detectionlimits for theseelementsare not given in the third
column of Table 5-2.
It should be emphasizedthat the two last columnsof Table 5-2 represent
detectionlimits that are more realistic and could be obtainedunderoutline con-
ditions. The detectionlimits close to thosein 10% HCI are probably obtainable
for soil waterextracts,NH4HC03-DTPA extracts,and otherextractsof low back-
ground.However,for total soil digests,the last columnof Table5-2 is more real-
istic. It seemsdetectionlimits get poorer by one order of magnitudefrom soil
extractsto total soil digests.
Table 5-2. Detectionlimits in an ideal solution (pure water), 10% HCl, and in a simulatedarid soil
digest.
ICP
Element Ideal solutiont 10% HCl:j: Simulatedsoil digest§
l1g!mL
Ag 0.004
AI 0.0002 0.03
As 0.04 0.02 0.7
Au 0.04
B 0.0007 0.002 0.03
Ba 0.00002 0.001 0.001
Be 0.0004
Bi 0.05
Ca 0.00002 0.002
Cd 0.002 0.001 0.006
Ce 0.0007
Co 0.003 0.01 0.02
Cr 0.0003 0.003 0.01
Cu 0.0001 0.003 0.01
Fe 0.0003 0.003
Ga 0.0006
Hf 0.01
Hg 0.001 0.02 0.2
In 0.03
K 0.1
La 0.00005
Li 0.0003
Mg 0.00001 0.0004
Mn 0.00006 0.001
Mo 0.0002 0.004 0.02
Na 0.0002 0.01
Nb 0.00007
Ni 0.0004 0.005 0.009
p 0.02 0.03
Pb 0.002 0.03 0.07
Pt 0.08
Sb 0.2
Se 0.02 0.2
Si 0.01
Sn 0.03
Sr 0.00002 0.0006 0.007
Ta 0.002
Ti 0.00007 0.002
U 0.03
V 0.0002
W 0.0007
Zn 0.002 0.002 0.008
Zr 0.0004
t Data from Robin (1979).
:j: Data from CSUSTL.
§ Data from CSUSTL. Simulatedsoil digest contained10% HCl and the following elementsat the
given concentrationsin microgramsper milliliter: AI = 1500, Fe = 500, K = 400, Ca = 200, Na =
200, Mg = 100,Ti = 60, Mn = 20, P = 15.
116 SOLTANPOURET AI...
Table 5-2 revealsthe deteriorationof the detectionlimits due to interele-

ment spectralinterferences.For example,in the caseof As, a detectionlimit of
0.7 ppm is shown.This detectionlimit will precludedeterminationof As in total
soil digestsusingdirect nebulization.In this case,As is separatedfrom major soil
constituents by the hydride generation technique (see "Determination of
Ultratrace Levels of Arsenic, Selenium, and Mercury by Use of Hydride-
Mercury Vapor Generator").When interelemental interferenceis not assevereas
in the caseof As, other correctiontechniquescan be used(see"Correction for
Interferences(Inductively CoupledPlasma-AtomicEmissionSpectrometry),,).
DetectionLimits (Inductively CoupledPlasma-MassSpectrometry)

Detection limits are one to three orders of magnitudelower by ICP-MS
than by ICP-AES for most elementsmeasurableby both techniques.However,
thereare a few key analytesof interestin soil and plant analysisthat exhibit bet-
ter detectionlimits by ICP-AES than by ICP-MS; including S and Ca. A list of
ideal detectionlimits has beencompiledby Date and Gray (1989). Someof the
detection limits listed by them can be lowered if necessaryby adopting strict
cleanlinessproceduresto reduce analyte contaminationin reagentsand glass-
ware. Detectionlimits also can be improved by using massspectrometerscapa-
ble of resolution equal to or better than 3500 (Tsumura & Yamasaki, 1991;
Bradshawet aI., 1989).In addition, detectionlimits could be reducedby increas-
ing the duty cycle on the analytical mass.This could be done by using time of
flight mass spectrometersinstead of quadrupolemass spectrometers(Hieftje,
1992).
Net analyticalsignal for genericICP-MS, in units of analyteresponseper
unit analyteconcentration,decreaserapidly with increasingmatrix elementcon-
centrationandcommencebeingdegradedat lower matrix elementconcentrations
in ICP-MS than ICP-AES (Beauchemin,1989; Houk & Thompson,1988; Houk,
1986;Gregoire,1989;Beaucheminet aI., 1987;Douglas& Kerr, 1988;Gregoire,
1987a,b).Thus, comparisonof detectionlimits for the two methodsis accurate
for describinganalysisof test solutionswith total dissolvedsolids up to approx-
imately 100 to 500 mg L-1, the rangedependingon severalfactors including the
mass(es)and ionization potential(s) of the matrix elements(Gregoire, 1989).
This fact, coupledwith the outstandingdetectionlimits exhibitedby the ICP-MS,
makesit a morenaturalchoicefor a chromatographicdetectorthan the ICP-AES.
With continuing interest in chemicalspeciation,numerouspapersand publica-
tions have appearedin the area of ICP-MS involving ion exchange,HPLC,
and/orliquid-liquid solventextractionprior to detection.
Comparisonof detection limits betweenICP-MS and ICP-AES must be
doneon an analysisby analysisbasis.If digestionof solid materialsis involved,
the detectionlimits of ICP-MS and ICP-AES could be about the samebecause
the ICP-AEScantolerate10 to 100 times more dissolvedsolids than the ICP-MS
before the analytical sensitivity becomesadverselyaffected.Many of the appa-
ratus mentioned in "Special Apparatus (Inductively Coupled Plasma-Mass
Spectrometry"are designedto allow the concentrationof matrix elementin the
samplesolutionsto be increasedwhile maintainingthe analyticalresponse.
Interferences(Inductively CoupledPlasma-AtomicEmission
Spectrometry)
Solute Vaporation.In emissionspectrometry,refractory compoundssuch
as calcium phosphatesor calcium aluminatesare vaporized in the excitation
sourcesand henceinterferewith analysis.For example,Johnson(1979) showed
that AI suppressesthe Ca signal in DCP emission spectrometry.The solute
vaporizationinterferenceis negligible in ICP (Larson et aI., 1975).
Ionization. When atomic or ionic speciesof an elementin a plasmaemit

their characteristicline radiation,any shift in the ratio betweenthesetwo species
causesa shift in the intensity of the atomic and ionic lines. Johnsonet ai. (1979b)
reportedthe enhancementeffect of Cs and Li on K, Na, Ba, AI, Cr, etc. in DCP.
This enhancement effect is negligible in ICP when recommendedparametersfor
power input, observationheight, and carrier gasflow rate are used(Larsonet aI.,
1975).
Unwanted Radiation. Unwanted radiation refers to that radiation other
than the analyteradiation reachingthe analytedetector.In any emissionsystem,
the analytesignal consistsof the wantedanalyteradiationand the unwantedradi-
ation. The latter may be divided into the following categories(Ward & Myers,
1979):
1. Sourcebackground.
2. Extractingsolution background.
3. Stray radiation in the spectrometer.
4. Spectralline or band interference.
Sourcebackgroundrefers to the radiation originating, for example,from
Ar. This backgroundradiationis very stablein the caseof Ar plasmas.Extracting
solutionbackgroundrefers tothe continuumoriginatingfrom the extractingsolu-
tion.
Stray light radiationin somedirect readerscreatesthe most seriouserror in
determinationof trace metals (U.S. Dep. CommerceNati. Tech. Inform. Serv.,
1977). Three main sourcesof stray light are: (i) grating scatter;(ii) reflections
and scatterin the secondaryoptics, i.e., the region betweenthe exitslits and pho-
tomultipliers; and (iii) generalscatterfrom reflectionsby internal surfacesof the
direct readers.
The grating scatteris due to the grating imperfectionsand has been dis-
cussedby Larson et ai. (1976). The degreeof stray light interferenceof the lat-
ter two types dependson the engineeringdesignof the direct readersand could
be reducedsignificantly by the use of nonreflectivecoatings,light traps,baffles,
and general optical design (U.S. Dep. CommerceNati. Tech. Inform. Serv.,
1977).
The spectralline or band interferencesarise when there are spectralover-
laps betweenthe analyte and matrix elementspecies.In some instances,matrix
element speciesmay elevate the intensity of the backgroundcontinuum. The
method used to correct for the spectral overlaps, backgroundelevations, and
stray light interferences is discussed in "Corrections for Interferences
(Inductively CoupledPlasma-AtomicEmissionSpectrometry)".
Correction for Interferences(Inductively Coupled Plasma-Atomic

EmissionSpectrometry).To correct for solute vaporizationeffect, the analyst
should add a releasingagent to both sample and standards.For example, to
reducethe effect of AI on Ca, one may add Sr to both the sampleand standard
solutions.The Sr will combinewith AI and reduceits effect on Ca. To suppress
ionization interference,an easily ionizable elementsuch as Cs or Li is usually
addedto standardand sample solutions.Thesesamplepretreatmentsare not nec-
essaryfor ICP-AES.
To correctfor sourceand extractingsolution background,the analystwill
zero the spectrometerwith the blank solution madeup of the extractingsolution
(blank correction).
The interferencesdue to stray light, backgroundelevation, and spectral
overlapscould be correctedfor if the blank and the samplesolutionswere iden-
tical in compositionexceptfor the analyte.This ideal solutionis beyondthe prac-
tical realm, especiallywhen a multielementanalysisis desired.However, if the
samplesare rather uniform in major interfering species,thesecould be addedto
the blank and the standardsto compensatefor their interference.But addition of
major interfering speciesto the samplesolutionspreventsanalystsfrom deter-
mining these elementswith other elementssimultaneously.This dilemma is
resolvedby using a schemeknown as interelemental spectral interference cor-
rection, which is discussedbelow.
Interelementalinterferenceis observedwhenthe analytedetector(channel)
receivessignalsfrom the interfering elements.
When the soil water extracts, NH4HC03-DTPA extracts, dilute acid
extracts,and other extractswith low concentrationsof interfering elementare
analyzed by ICP, the degree of interelementalinterference is usually small.
However,a soil analysisfor total elementalcontentresultsin high concentrations
of interferingelementsandcorrespondinglylargeinterelementalinterferences.In
the latter case,one should determinesignificant interelementalinterferencesin
samplesolutionsand correctfor them.
To determinethe degreeof interelementalinterference,the spectrometer
shouldbe standardized,a pure solutionof interfering(affecting) elementsshould
be aspirated,and the apparentconcentrationof the affectedelementsdetermined.
When the sampleis analyzed,the concentrationof the interfering elementsis
determinedand the necessarycorrectionsmadeon the apparentconcentrationof
the affected elements.Computerprogramsare available for the automaticcor-
rection of the interelementalinterference(Dahlquist& Knoll, 1978).
The following exampleis given to show the useof the interelementalinter-
ferencecorrectionmethod.A syntheticsolutioncontaining1.0 mg L -1 of Pb read
3.66 mg L-1 of Pb when AI, Fe, K, Ca, Na, Mg, Ti, Mn, and P were addedto the
Pb solution as 1,500-,500-,400-,200-,200-, 100-,60-,20-,and 15-mgL-1 con-
centrations,respectively.When pure solutionsof the aboveelementsat the above
concentrationswere aspiratedinto the plasma,the apparentconcentrationsof Pb
were 2.62,0.164,and 0.038 for AI, Fe, and Ti solutions,respectively.Other ele-
mentsdid not produceany noticeableunwantedradiation at the Pb wavelength
(220.3 nm). Subtractingthe above interferencesfrom 3.66 gave a Pb value of
0.84, which is much closerto the true value of 1.0 mg L-1 than the uncorrected
value of 3.66. In this case,the spectrometerwas not restandardizedbefore the

high backgroundPb solution was read. This may explain the reasonfor obtain-
ing 0.84 value insteadof 1.0 for Pb. However,when a solution containingall the
above elementconcentrationsexcept Pb was aspirated,it gave an apparentPb
concentreationof 2.80, which is almost identical to the sum of the apparentPb
concentrationsin AI, Fe, and Ti solutions.The aboveexampleshowsthe validi-
ty of interelementalinterference correction methoddiscussedabove.
Some precautionsto be observedin interelementalspectral interference
correctionare discussedin the remainderof this section.Care must be taken to
ensurethat an adequaterinsebetweenthe introductionof eachinterferentialsolu-
tion is performed.Purechemicalssuchas SpecPure(Curtis MathesonScientific,
Inc. ChernPure Brand, Houston,TX) reagentsshould be usedfor determination
of interelementalinterferences.If the chemicalsare not pure, an impurity of ana-
lyte in thesechemicalswill createrather large errors in the results. For soil HF-
HCI04 digests, the interferenceof major soil constituentson other elements
should be determined. Soil extracts also should be examined for possible
interelementalinterferences.Theseinterferencesare specific for a given instru-
ment dependingon the wavelengthused for each elementand effective use of
baffles and black interiors to reducestray light interferences.
In the instrument (Jarrell-Ash Model 975 ICP AtomComp) used at the
CSUSTL, it was found tha~ the following elementssignificantly interfere with
sometraceelementsin HF-HCI04 soil digests:AI, Fe, Mg, Mn, andTi. However,
one should be on guard against interferencefrom other elementsthat may be
found in large quantitiesin contaminatedsoils. The interelementalspectralinter-
ferencesfound in the ICP-AES systemmentionedaboveare shownin Table5-3.
In the ColoradoStateUniversity instrument,Ca interferencewith other elements
is very low in contrastwith publishedreportson anotherinstrument(U.S. Dep.
CommerceNatl. Tech. Inform. Serv., 1977).
The order of elementalcorrectionis important (Marciello & Ward, 1978).
As a generalrule, interelementcorrectionshould be programmedinto the com-
puter in the following order:
1. Major matrix componentsas interfering elements.
2. In order of magnitudeof interferenceeffect for minor matrix compo-
nents.
In correcting for interelementalinterference,one should be aware of the
fact that inferenceper unit concentrationof the interfering elementmay not be
linear. This has beenshown to be the casein the particular instrumentusedfor
the Ca interferenceon As, Se, Pb, and Sn (U.S. Dep. CommerceNatl. Tech.
Inform. Serv., 1977). In this event, curvesshould be plotted showing the appar-
ent concentrationof the affectedelementas a function of the concentrationof the
affecting elements.Thesecurvesshould then be usedto correctfor interelemen-
tal interferences.Computerprogramsand computersinterfacedwith the direct-
readerscapableof performing thesetasks wit make the interelementalinterfer-
encecorrectionseasierand much faster.
Another schemefor interferencecorrection,called backgroundcorrection
by some,is by the useof a devicecalled a spectrumshifter. The spectrumshifter
N
...<=
Table 5-3. Examplesof someinterelementalspectralinterferencesobservedin an ICP spectrometerat the CSUSTL.

Wavelength(nm)
Affecting 324.7 206.2x 3 213/6 x 2 202.0 214.4 228.6 267.7 407.7 249.7 455.4 220.3 253.6 193.6 196.0
element Concentration Cu Zn Ni Mo Cd Co Cr Sr B Ba Pb Hg As Se
llg/mL Apparentconcentrationof affectedelementsllg/mL
K 400t
Ca 50 0.003
200t 0.002 0.D15 0.024
500 0.004 0.039 0.079
Mg 50 0.007 0.009 0.005 0.004 0.006 0.024 0.054
lOOt 0.012 0.012 0.005 0.008 0.010 0.039 0.043 0.087
200 0.024 0.038 0.008 0..013 0.004 0.066 0.083 0.168
Na 200t
P 15t 0.030
Ti 60t 0.026 0.003 0.111 0.008 0.028 0.038 0.197
Mn 20t 0.002 0.003 0.010 0.D18
Fe 100 0.005 0.006 0.011 0.010 0.008 0.016 0.004 0.285 0.064 1.36 0.053 0.083
500t 0.014 0.021 0.048 0.044 0.046 0.072 0.024 1.32 0.164 6.96 0.250 0.564
1000 0.025 0.040 0.093 0.082 0.088 0.140 0.046 2.51 0.004 0.302 13.1 0.494 1.07
AI 500 0.088 0.002 0.253 0.080 0.008 0.053 0.006 0.006 0.866 0.019 5.96 1.46
rJl
1000 0.002 0.176 0.013 0.517 0.164 0.008 0.016 0.106 0.020 0.012 1.76 0.030 12.2 2.90 0
1500t 0.002 0.261 0.016 0.760 0.242 0.018 0.024 0.156 0.020 0.D18 2.62 0.054 18.0 4.28 ~
High backgroundsolutiont 0.046 0.295 0.080 0.807 0.288 0.200 0.074 0.172 1.26 0.023 2.80 6.84 18.2 4.98
~
t Indicateshigh backgroundsolution was madefrom the elementsand concentrationsmarkedby t. All solutionswere 10% in HCI. -=0
~
:=
~
>
r
measuresthe backgroundradiationcloseto the analyteexit slit. This radiation is

assumedto originatefrom nonanalytesourcesand to increasethe radiationat the
analytewavelength.The averagebackgroundradiationsignal is subtractedfrom
the signalobservedat the analytewavelengthto correctfor the interference.This
correction method may be used when the analyst is not aware of all the con-
stituentsof the sampleor dealswith samplesof varying matrixes.This correc-
tion methodis obviously not valid when the backgroundradiationreceivedat the
analytewavelengthis grossly different from the one measuredby the spectrum
shifter.
Interferences(Inductively CoupledPlasma/MassSpectrometry)
In a concise explanation of interference effects in ICP-MS, Conrad
Gregoire (1989) has divided the subjectof interferenceeffects in the ICP-MS
into three areas.Theseare isobaric interference,nonspectroscopicinterference,
and massdiscrimination.The isobaricproblemsweresubdividedfurther into two
categories:(i) Molecular ion interferencesincluding oxide ions, and (ii) Spectral
interferencesdue to other elements.The nonspectroscopicsuppressioneffects
were discussedin the contextof spacechargeand ionizationsuppressioneffects.
Massdiscriminationeffectswereviewed in two categories:(i) Instrumentalmass
discrimination,and (ii) Matrix dependentmassdiscrimination.To the discussion
of nonspectroscopic interferencewill be addedsuppressionsdue to solids depo-
sition on the samplerand skimmerconesand suppressionsdue to solutevapor-
ization.
SolidsDepositionon Samplerand SkimmerCones.Depositionof solids
on the skimmerand samplerconescancauseunwantedchangesin the analytical
response,i.e., reductionsin the quantity (ion arrival rate at the detectorper unit
analyte concentrationin the test solutions) (Douglas & Kerr, 1988). At the
DANR Analytical Lab, long runs involving analytical measurementson plant
digestspreparedusing a microwavebomb technique(Sah & Miller, 1992) have
beenmadeusing the ICP-MS instrumentation.During a run, coatingsof calcium
sulfate and oxide on the samplerand skimmer conesoccur. Unlessthe dilution
factors and sample nebulization times of the test solutions are carefully con-
trolled, the solid depositionson the samplerandskimmerconescan result in seri-
oussuppressionof analyticalresponse.The coatingof the skimmerconenearthe
tip, to the point of obstructingthe orifice, causesthe most seriousdecreasein
analyticalresponse;which persistsindependentof matrix elementconcentration
in the test solution for the durationof the analyticalrun. The suppressioncan be
at onceeliminatedby cleaningthe skimmer cone.
Non-SpectroscopicInterferences(Inductively Coupled Plasma-Mass
Spectrometry).Nonspectroscopicinterferenceis the general term adoptedin
ICP-MS for describingreduction in analytical responsewith increasingmatrix
elementconcentrationin the test solution. Nonspectroscopicinterferenceis a
complex issue,with severalfactors contributing to the suppression(s)observed.
To successfullyuse ICP-MS for soils analysis,the major elementconcentration
of the solutionsmust be anticipatedto vary widely. Among the severalfactors
discussedbelow are: (i) Solutevaporization,(ii) Ionization suppression,and (iii)

Space-charge.
Solute vaporization interference occursin instancesin which the solute
doesnot havesufficient time and/orthe sourcedoesnot havesufficient energyto
dissociatethe solute before the analytical speciesmoves into and through the
region of observation.Typical manifestationsof the interferenceare suppression
and/orincreasedvariability of the analytesignal as a function of increasedcon-
comitantconcentration.For example,Johnsonet al. (1979a)showedthat AI sup-
pressesthe Ca signal in a direct current plasma.Winge et al. (1991) published
high-speedphotographsproviding evidence that speciesgenerally associated
with low spectroscopictemperaturescan persistthrough the central channelof
the ICP to enterthe samplingconeof the ICP-MS, and discussedsimilar reports
of unvaporizedsolventdoing the same.Ionization interferencescan be notedfor
ICP-MS. Partially ionized elementsin an ICP, e.g., Au and B, are susceptibleto
ion suppressionfrom fully ionized interferants(smaller effective analyte ion
molar ratio/interferantion) and lesseffective in causingion suppressionfor fully
ionized analytes(larger analyteion molar ratio/interferention) (Gregoire,1989).
In ICP-MS measurements, ionization interferencescausesuppressionof analyti-
cal response(Houk & Thompson,1988; Tan & Horlick, 1986).
Easily ionized matrix elementinterferencehasbeenrecognizedin ICP-MS
data. For a given matrix elemention interference,the analyticalresponsefor the
lighter atomic massisotopesare suppressedmore than the heavierones. For a
given analyte,lighter matrix elementssuppressthe responseless than the heav-
ier ones.Thesetrendsare consistentwith what would be expectedfrom a space-
chargeeffect (Gregoire,1989; Hieftje, 1992).Tracingthe courseof the ions from
the point of ion productionin the plasma,the ions that move throughthe sampler
and skimmer, through the ion optics, and through the quadrupole,are reflected
away from the deflectorand acceleratedonto the detector.As the particle beam
electronsdiffuse out of the beammore quickly than the positive ions-aprocess
termedambipolardiffusion (Ahearn,1972).As a consequence, coulombicrepul-
sion spreadsthe ion beam.The larger ions stay on coursebetterthan the lighter
ones. Equivalently, the trajectoriesof the lighter ions are affectedmore by the
ions of the heavier mass isotopesthan the trajectoriesof the heavier ions are
affectedby the lighter massisotopes.
In practice, it is difficult to separatethe nonspectroscopicinterferences.
Thesecan be compensatedfor by matrix matching,but that seriouslylimits the
rangeof matrix elementconcentrationvariability betweentest solutionswithin
the run. Another approachthat hasbeenusedsuccessfullyby many is the inter-
nal standard calibration method, discussedin "Methods of Correction for
Interferencesand StandardizationProcedures(Inductively Coupled Plasma-
Mass Spectrometry)."
Mass Discrimination. Gregoire (1989) defines mass discrimination as
bias in ion transmissionto the detectorthe magnitudeof which is dependent
upon the massof the analytical isotope.Further, the effects can be divided into
two categoriesdependingupon the origin of the massbias. The first categoryis
referred to as instrumentalmassdiscrimination effects which are the result of
massdiscriminationoccurring at the interface(sampleand skimmercones),ion
lenses,quadrupolemassfilter and detector.Correction factors for instrumental

massdiscriminationare normally found by comparingmeasuredisotoperatios to
the known isotoperatio for a substanceof known or certified isotopic composi-
tion, and applying the correction to the samplesrun during the same time.
Instrumentalmassdiscriminationcan range from 50% per dalton for light ele-
mentsto 2% per dalton for heavy elements.
The other type of massdiscriminationresultsfrom the presenceof matrix
elements,has beenreportedonly twice, and effects only Li and B. Briefly, the
magnitudeof the effect is dependentupon five factors:
1. Absolute massof the analyte.
2. Degreeof ionization of the analyte.
3. Difference in massbetweenthe two isotopes.
4. Mass of the matrix elements.
5. Degreeof ionization of the matrix elements.
The above dependentfactors involved with this effect are very similar to the
space-chargeinterpretationof analyte responsesuppression,so for all practical
purposesmatrix dependentmassdiscriminationcan be considereda special case
of the more general space-chargeeffect. The interestedreader is referred to
Gregoire(1989) for more details.
Unwanted Ions (Inductively Coupled Plasma-MassSpectroscopy).
Unwantedions refer to that part of the ion beamother than the analyteion part
reachingthe detector.In a massspectrometer system, particularly one operated
with peak widths in the neighborhoodof one dalton, the analytesignal may be
accompaniedby unwantedions. Theseunwantedions occur as a consequence of
several factors (Gray, 1985; Date & Gray, 1989; Gregoire, 1989; Vaughan &
Horlick, 1986; Munro et aI., 1986; Date et aI., 1987; Gray, 1989; Tan & Horlick,
1986), and include the following:
1. Elemental ions (NM+) of the same unit mass as the analyte isotope
(NN).
2. Elementalhydride (N{N-IMIHV) molecular ions that are of the same
unit massas the analyteisotope(NA +).
3. Elementoxide (N{N-16M160+) molecularions that are of the sameunit
massas the analyteisotope(NA +).
4. Element hydroxide (N{N-17M1601HV) molecular ions that are of the
sameunit massas the analyteisotope(NA +).
5. Elemental(2NM2+) ions that are doubly chargedand twice the unit mass
of the analyte isotope (NA +). Also, for unit resolution spectrometers,
elemental«2N-l)M2+) and (2N+IlM2+) ions must be includedas ions that
will overlap,albeit partially with the analytical mass(NA +).
6. Elemental Argide (N{N-36M36Ar}+' N{N-38M38Ar}+' N{N-40M 40 ArV)
molecular ions that are of the same unit mass as the analyte isotope
(NN).
7. Elemental Hydrogen Argide (N{N-37MIW6Ar}+' N{N-39MIH38Ar}+'
N{N-41MIH40ArV) ions that are of the sameunit massas the analyteiso-
tope (NA+).
Methods of Correctionfor Interferencesand StandardizationProce-

dures (Inductively Coupled Plasma-MassSpectrometry).The presenceof
both isobaric and nonspectralmatrix dependentinterferencesrequires special
attention. Severalpapersdescribein more or less detail the procedureused to
correctisobaricoverlap(Dateet aI., 1987;Tsumura& Yamasaki,1991; Evans&
Ebdon, 1989; Lam & McLaren, 1990). Methods for correcting for analyte
responsechangesas a function of time and matrix elementconcentrationsin test
solutionshave also beendescribedand evaluated(Doherty, 1989; Thompson&
Houk, 1987; Johnsonet aI., 1992a,b).Softwareissuedby the instrumentmanu-
facturerscorrectsfor isobaricinterference,allows periodic updateof calibration
data to correctfor drift, and includesinternal standardcorrectioncalculationsto
correctfor analyteresponsechangesas a function of matrix elementconcentra-
tion in the test solutions. In addition, severalinstrumentadd-onsare available
that tend to reducethe isobaricand/or nonspectroscopic matrix dependentinter-
ferences.
In a correction scheme routinely used at the DANR Analytical Lab
(Johnsonet aI., 1992a,b), from two to four levels of analyte concentration
(including 0 ).lg L-1) are preparedin triplicate as calibration standardsolutions.
Two- thirds of theseare spikedwith an appropriateconcentrationof Y, e.g., 100
and 200 mg L-1. The Y is purchasedas the oxide at 99.999% purity and dis-
solvedusingnitric acid. Singleelementconcentrationsof the major concomitants
generallyrepresentativeof the concentrationsanticipatedto be found in the sam-
ples are preparedin a similar manner,i.e., with and without Y. The calibration
solutions are run at the beginning of the analytical run sequenceand approxi-
mately every 20 to 30 samplesthereafter, and again at the end of the run
sequence.Three internal standardsare usedto cover the instrumentmassrange.
Theseare usually Be, In, and Bi, but can be varied as necessarydependingon
whetheror not any of theseare analytes.Data are written on the hard disk of the
computerin a format standardizedby the manufacturer(Perkin Elmer Sciex,500
ICP-MS hardware,5000ICP-MS software).The file is copiedonto a floppy disc,
transportedto an off-line computer,and processedwith a BASIC program for
importationinto the QuattroPro spreadsheet by Borland.
Concentrationsare estimatedby a simple four-stepprocedure:
1. Zero).lg L -1 responsecalculated.
2. Isobaricinterferencescorrected.
3. Analytical responsecorrectedfor nonspectroscopic
interferences.
4. Concentrationscomputed.
In the absenceof spectralinterference,accuracyof measuredvs. true con-
centration estimatesfor the standardconcentrationsread as samplesdepend
somewhaton the numberof internal standardsapplied to the data. In step three
above,as many as threeinternal standardshavebeenappliedto datafor one ana-
lytical isotopein eithera sequentialor simultaneousfashionusingregressionsta-
tistics. Note that interactionterms can be, and are includedwith the simultane-
ous regressionmodel. Accuraciesof measuredconcentrationsimprove from
±15% to betterthan ±1.5% of the correspondingtrue concentrationsfrom one to
three iterationsusing Bi, Be, and In as internal standards.Accuracy is about the
samewith sequentialvs. simultaneous,but the simultaneousis easierto apply.

Trends for improvementin analytical sensitivities(Skogerboe& Grant, 1970)
were about the sameas notedfor accuracies.While the isobaricoverlapcorrec-
tion procedureis still being evaluated,it appearsthat the calibration- subtrac-
tion approach,supplemented by assumptionsconcerningthe naturaldistribution
of the interfering isotopeswhereconvenientand/or necessary,can yield correc-
tions within ±2.5% of the true correction.The accuracyof the final resultsthen
dependson the severity of the specific interference(Johnsonet aI., 1979b).
PracticalApplications
Grinding Soil Samples
Soil samplesshouldbe air dried as close to the time of samplingas possi-
ble (drying and grinding of soil samplesare not recommendedfor Mn and Cr).
Soil samplesmay containclods or large aggregates.Many laboratoriesuse auto-
matic grinders to crush soil. Recent studies haveshown that the amount of
extractablemicronutrientsfrom soils is affectedlargely by the degreeof grind-
ing (Soltanpouret aI., 1976, 1979a);therefore,grinding variablessuch as force
and time shouldbe standardized.When soils are groundfor extractablemicroele-
ments, care must be taken to avoid excessivegrinding. It is important to use
grinders that do not contaminatethe soil. At the CSUSTL, a high-densityalu-
minum oxide auger made by the Coors Porcelain Co., Golden, Colorado, is
attachedto a Nasco-AspIinautomaticgrinder. The grinder is equippedwith a 2-
mm stainlesssteel sieve. This grinder minimizes the degreeof soil contamina-
tion with trace elements.In soil analysis,it is a standardprocedureto passthe
soil through a 2-mm sieve after mild grinding. Then all analytical results are
basedon a 2-mm soil. For total elementalanalysis,the 2-mm soil may be further
groundso that all of it passesthrougha O.16-mm(lOO-mesh)polyvinyl chloride
(PVC) sieve.
ObtainingSoil Extracts
For simultaneousmultielement determination,obviously single-element
extraction solutions are not useful. Therefore, Soltanpourand Schwab (1977)
developeda 1M NH 4HCO T O.005M DTPA (AB-DTPA) solution for simultane-
ousextractionof P, K, Zn, Fe, Cu, Mn andnitratesfrom soils. This testwas mod-
ified by Soltanpourand Workman(1979) to omit C black, which sometimescon-
taminatedthe sampleand adsorbedmetal chelates.The above test is routinely
usedby the CSUSTL to assesssoil fertility of the Coloradofarms. After extrac-
tion, ICP-AES is usedto simultaneouslyanalyzetheseextractsfor P, K, Zn, Fe,
Cu, and Mn. Experiencehas shownthat AB-DTPA solution should be acidified
to get rid of the carbonate-bicarbonate
matrix in order to preventclogging of the
capillary tip (Soltanpouret aI., 1979b). However, use of high salt nebulizers
(Legere & Burgener, 1985) has obviated the use of acid pretreatment
(Soltanpour,1991).
Soil water extractsand DTPA extracts(Lindsay & Norvell, 1978) can be
analyzedby ICP-AES. We are analyzingthe soil saturationextractssimultane-
ously for Ca, Mg, Na, and K and then calculatingthe Na absorptionratio. Plant
digestsalso are analyzedby ICP-AES.
The CSUSTL and many environmentallaboratoriesin the western U.S.
analyzemine overburdenand mine spoil materialsby useof AB-DTPA and ICP-
AES for P, Zn, Cu, Mn, B, Cd, As, Se, Mo, Pb, Ni, andotherelementsfor screen-
ing these materials for their potential toxicity to plants and their consumers
(Soltanpour,1991). The AB-DTPA extract is low in Ca, Mg, AI, Fe, Mn and Ti,
which causeinterelementalinterference,and thereforeis well adaptedto ICP-
AES analysis.Obviously water extractsare ideal for ICP-AES determinations,
but someelementsin waterextractsare found in very low concentrationsthat are
below the ICP-AES detectionlimits.
Jones(1977) determinedCa, K, Mg, and P simultaneouslyin the double-
acid extractsof Georgiasoils.
The type of vesseland shakerand the speedof the shakermay affect the
amountof someextractableelements,but their effect is small comparedwith the
grinding variables(Soltanpouret aI., 1976).
For obtainingsaturationextracts,refer to Chapter14 (Rhoades,1996). For
obtainingdouble-acidextractions,refer to Chapter19 (Helmke & Sparks,1996).
For obtainingMehlich 3 extracts(a modification of double-acidextractfor mul-
tielementextraction)refer to Chapter19 (Helmke & Sparks,1996).
Special Equipment
1. Reciprocalshaker.
Reagents
1. Ammonium bicarbonate(NH4HC03).
2. Diethylenetriaminepentaacetic
acid (DTPA).
3. Ammonium hydroxide (N~OH).
5. Ammonium bicarbonate-diethylenetriaminepentaacetic
acid solution
(AB-DTPA).
To make aIM NH4HCOr -O.OO5 M DTPA solution, add 1.87 g of DTPA
to 800 mL of distilled-deionizedwater (DDW). Add approximately2 mL of 1:1
NH40H to facilitate dissolution and to preventeffervescence.Shakeuntil most
of the DTPA is dissolved.Then add 79.06 g of NH4HC03, and stir gently until
dissolved.Adjust the pH to 7.6 with NH40H. Dilute the solution to 1.0 L with
DDW. This solution is unstablewith regardto pH. If the solution is storedunder
about3 cm of mineral oil, the pH remainsstable.However,it is preferableto use
a fresh solution.
Procedure.Weigh 10 g of a 2-mm soil into a 125-mL conical flask. Add
20 ml of AB-DTPA solution. Shakeon the reciprocalshakerfor 15 min at 180
cycles/minwith flasks kept open. Filter the extractsthroughWhatmanno. 42 fil-
ter paperor its equivalent.Take a 2-mL aliquot of the extract,and add0.25 mL
of concentratedHN03• Shake for 10 min to get rid of carbonate-bicarbonate
matrix to preventclogging of the capillary tip in cross-flowor concentricnebu-
lizers. This solution is now ready for simultaneousmultielementdetermination.
With high-saltnebulizers(Babingtontype) the acid pretreatmentis not required.
Digestionof OrganicMatter and Dissolutionof Silicates

for Total ElementalAnalysis
Digestion(oxidation)of organicmatteranddissolutionof silicatesare nec-
essarystepsin bringing all elementsinto solution. Theseprocessesare referred
to as digestionin this chapter.Certainly, it is advantageousto use methodsthat
yield themselvesto multisample rather than single sample digestion. Another
important considerationin choosing a method is preservationof the easily
volatilized elementsin the sample.Fusion and other high temperaturemethods
of digestionlead to the loss of volatile substancessuch as As, Se, Sb, and Hg.
The methoddescribedbelow avoidsthe loss of As, Se, and Sb in the presenceof
silicates(Bajo, 1978); therefore,we recommendthis method. Research is under
way on the use of other methodssuchas the use of cappedpolyethylenebottles
for HF digestionof siliceousmaterial(Odegard,1979; Langmyhr& Paus,1968;
see Chapter3, Hossner,1996). An all-Teflon bomb (Bernas, 1968; Lechler &
Leininger, 1979) hasbeenusedfor analysisof siliceousmaterial.
Special Equipment
1. Stainlesssteel perchlorichood.
2. Hot plate.
3. Teflon beaker,100 mL.
4. Teflon watch cover.
Reagents
1. Perchloricacid (HCI04).
2. Nitric acid (HN03).
3. Hydrofluoric acid (HF).
4. Hydrochloric acid (HCl).
Procedure.Weigh 1.0 g of the lOO-mesh(0.15-mm) soil into a 100-mL
Teflon beaker.Add 10 mL of HN03 and 10 mL of HCI04• Cover with a Teflon
watch cover,and heatat 200°C for I h. Removethe cover and continueheating
until the volume is 2 to 3 mL. Cool the sample,add 5 mL of HCI04 and 10 mL
of HF, coverwith a Teflon watch cover, and heatovernightat 200°C. Overnight
digestion is for convenience,but one may terminate digestion as soon as all
siliceousmaterial has beendissolved.Removethe cover, and continueheating
until the volume is 2 to 3 mL. Cool the digest, add 10 mL of 50% HCI, cover,
and heatat 100°C for 30 min. Removefrom the hot plate, and allow it to cool.
Transferthe solution quantitativelyinto a 50-mL volumetric flask, and bring to
volume. The solution is then ready for ICP determination.
Comments.The reasona 100-mesh(0.15-mm)soil sampleinsteadof a 2-
mm sampleis usedin digestionis to speed upthe breakdownof silicates.At the
CSUSTL, 2-mm soil sampleshave beendigestedwith no difficulty when soils
were digestedovernightin the presenceof HF-HCI04•
The HCI04 hood should be washedperiodically to remove perchlorates
and to avoid dangerof explosion.Do not let HCl04 solution get dry, anhydrous
HCl04 is explosive.
Analysis of Soil Extractsand Digests

Special Apparatus
1. An inductively coupledplasmaatomic emissionspectrometer.
SpecialApparatus(Inductively CoupledPlasma-MassSpectrometry).
There are several different after manufactureadd-ons available for sample
aerosolproductionand/orintroductionfor ICP-MS systemsfrom different man-
ufacturers. These include USN, DIN, HPLC systems(Braverman, 1992), FI
accessory(Thompson& Houk, 1986; Deanet aI., 1988; Denoyer& Stroh, 1992;
Denoyeret aI., 1991a),hydride generationequipment(Denoyer& Stroh, 1992a),
ETV accessory(Denoyer et aI., 1991a), and LASER ablation solid sampling
equipment(Denoyer, 1991; Hager, 1989; Abell, 1991; Denoyer et aI., 1991b;
Pearceet aI., 1992). Theseaccessoriesalso are available for use on ICP-AES
instrumentation.
One of the most versatilepiecesof equipmentin this list is the FI accesso-
ry (Denoyer& Stroh, 1992).Thereare many possiblephysicaland chemicalpro-
ceduresthat can be combinedwith the ICP-MS via a flow injection apparatus,
including on-line dilution, isotopedilution, standardadditions,hydride genera-
tion, cation exchange,anion exchange,and ETV. The flow injection apparatus
introduces a discrete sample aliquot into a flowing carrier stream for direct
analysisof solutions.Samplevolumesfor a typical analyticalmeasurement range
from 50 to 500 pL, comparedto 1 to 3 mL for continuoussolution aspiration.
Full massscanscan be performedon a quadrupolein about 100 ms,or 10 scans
per second.The high scanningspeedof the quadrupoleallows transientsignals
generatedby flow injection to be capturedand measured.Relativestandarddevi-
ationsare 2 to 5% on replicateinjections.Rinse-outtimes areconsiderablyshort-
er for FI-equippedsystemsthan for intermittent nebulization. An impressive
advantageof FI is that it allows analysisof solutionscontainingapproximately
50 times more matrix element concentrationthan conventional nebulization
equipmentfor comparablereductionsin analytical responsedue to nonspectro-
scopicmatrix dependentinterferences;putting the acceptablelevel of dissolved
solids in the test solutions in the 0.5 to 2.5 g per 100-mL range. Use of the FI
apparatusalso reportedlypreventscloggingof samplerandskimmerconeswhile
reducingthe amountof solids depositedon them per samplesolution. Fourfold
increasesin samplethroughputper unit havebeenreportedusing FI (Deanet aI.,
1988).
Preparationof Stock StandardSolutions.Any emissionspectrometric
methodcomparesthe emissionsignalsfrom the samplesolutionswith that of the
standard solutions to determine the composition of the sample solution.
Thereforeit is imperativethat extreme carebe takenin preparingstandardsolu-
tions. It is recommendedthat ready-to-usestandardsolutionsof highestpurity be
purchased.In some developingcountriestheseready-to-usestandardsmay not
be available.Therefore,in the following paragraphs wehavedescribeda method
for preparationof standards.All acids used for dissolution should be of high
purity, such as Hi-Pure or Ultrex grade. Water usedfor dilution should be dis-
tilled-deionized.Table 5-4 adaptedfrom Ward (1978b)can be usedfor prepara-
tion of solutionscontaining1000 mg L-l of an element.
Table 5-4. Primary standardsolution preparation(adaptedfrom Ward, 1978).t

Element Compound Weight Solvent
g
AI AI 1.0000 6MHCI
AlCI 3 ° 6HzO 8.9481 1MHCI
Sb Sb 1.0000 Aqua regia
SbCI3 1.8736 1MHCI
As As 1.0000 4MHN03
Asz0 3 1.3203 4MHCI
Ba BaClz* 1.1516 Water
BaC03* 1.4369 0.05MHN0 3
BaN03 1.9029 Water
Be Be 1.0000 0.5MHCI
BeO* 2.7753 0.5MHCI
Bi Bi 1.0000 4MHN0 3
Bi z0 3 1.1149 4MHN03
Bi (N03)3 ° 5HzO 2.3211 1MHN03
B B 1.0000 4MHN03
H3B03 5.7195 Water
Cd Cd 1.0000 4MHN0 3
CdO 1.1423 4MHN03
Ca CaC03 2.4972 0.5MHN0 3
Ca (N03)z ° 4HzO* 5.8920 Water
Cr Cr 1.0000 4MHCI
CrCI3 (6H20) 5.1244 Water
Co Co 1.000 4MHCI
CoClz ° 6 HzO 4.0373 Water
Cu Cu 1.0000 4MHN0 3
CuO 1.2518 4MHN0 3
In In 1.0000 Aqua regia
Fe Fe 1.0000 4MHCI
Fez03 1.4297 4MHCI
Pb Pb 1.0000 4MHN03
PbO 1.0772 4MHN03
Pb (N03)z 2.6758 Water
Li Li zC03 5.8241 1MHCI
LiCI 6.1092 Water
Mg MgO 1.6581 0.5MHCI
MgCLz ° 6HzO* 8.3625 Water
Mn Mn 1.0000 4MHN0 3
MnOz 1.5825 4MHN0 3
Hg HgClz 1.3535 Water + 1 g (NH4)zSzOg
Mo Mo 1.0000 Aqua regia
Mo03 1.5003 Aqua regia
Ni Ni 1.0000 4MHCI
NiO 1.2725 4MHCI
NiClzo 6HzO 4.0489 Water
Nb NbzOs 1.4305 Minimum quantity of HF, add 1 M HCI
P NaHzP04 3.8735 Water
NH4HzP04 3.7137 Water
K KCI 1.9067 Water
K ZC03 1.7673 1MHCI
Se SeOz 1.4053 Water
Si NazSi03 ° 9HzO* 10.1190 Water
Ag Ag l.OOOO 4MHN0 3
AgzO 1.0742 4MHN03


Element Compound Weight Solvent
Na NaCI 2.5421 Water
Na2C03 2.3051 IMHCI
Sr SrC03 1.6849 IMHN0 3
Sr(N03h 2.4152 Water
Te Te02 1.2508 4MHCI
TI Tl 20 3 1.1174 4MHCI
TICI 1.1735 Water
Sn Sn 1.0000 4MHCI
SnCI20 2H2O 1.9010 4MHCI
Ti Ti 1.0000 4MHCI
V V 1.0000 4MHN0 3
Zn Zn 1.0000 4MHN0 3
ZnO 1.2448 4MHN0 3
Zn (N03h ° 6H2O 4.5506 Water
t Use 100 to 150 mL of solventto dissolveandbring to 1 L volume to give a concentrationof 1000
mg L-l of element.
:j: Not Specpurematerials.
StandardizationProcedures(Inductively Coupled Plasma-Atomic

EmissionSpectrometry).In calibrating the ICP spectrometer,one should con-
siderthe concentrationrangeto be used,the interelementinterferencecorrection,
and the stability of the standards.Mixtures of chemicalsthat causeprecipitation
should be avoided. McQuakeret al (1979) deviseda calibration schemefor 30
elementsthat satisfiesthe needsof analystsin soil, water, tissue,and particulate
matter analysis.In soil analysis,one set of secondarystandardsis requiredfor
eachextractingsolution and anothersetfor the total soil digest.In making a mul-
tielementstandardsolution, one should avoid makingsolutionscontaininghigh
concentrationsof affecting and low concentrationsof affected elements(see
Table 5-3).
Interelementaleffectsare measuredby using single elementsolutionspre-
paredwith Specpurechemicals.Appropriate computersoftware is used to cor-
rect for interelementaleffects. In caseof nonlinearinterference,computersoft-
ware should be able to storecorrectioncurvesfor interelementalcorrections.
Secondarystandardsshouldbe madein sucha way that standardsolutions
match the sample solutions in concentrationof acids. For AB-DTPA extracts,
standardsshould be made in 1 M NH4HCOr O.005 M DTPA solution that has
beenneutralizedwith concentratedHN03 (Soltanpour& Workman, 1981). For
HN03-HCI04-HF digests, standardsolutions should be made to contain 5%
(vol/vol) HCl04 and 10% (vol/vol) HCl.
Comments.The operationalparametersthat have beenusedwith the ICP
spectrometer(Jarrell-Ash750 AtomComp); Jarrell-AshCorp., Waltham,MA) at
the CSUSTL are given here to aid in interpretationof data given in Tables5-2
and5-3: sampleflow rate, 0.7 mLimin; Ar pressure,550 kPa (80 Ib/in.2); aerosol
carrier Ar flow rate, 1 Llmin; Ar plasmasupportflow rate, 17 Llmin; capillary
usedin crossflow nebulizer,tungstencarbide(o.d. = 0.99 mm, i.d. = 0.25 mm);
height of observationabove coil (NAZ), 15 mm; incident power, 1.25 kW;
reflectedpower, <3 W.
NaBH4
Solution
To
plasma
Sample
60ml coarse
fritted disc
Buchner funnel
Drain
Argon
Pumps l2Imin
Fig. 5-5. The hydride and mercury vapor generator.
It should be noted that the operationalprocedureof different ICP spec-

trometerscannotbe given in a chaptersuch as this. When a particularICP spec-
trometeris purchased,usually the manufacturerswill train the analystsin opera-
tional proceduresand in solving any operationalproblemsthat may arise.
Determinationof UltratraceLevels of Arsenic, Selenium,and Mercury

by Use of Hydride-MercuryVapor Generator
The hydride-mercuryvaporgeneratoris usedto determineultratracelevels
of As, Se, and Hg in soil extractsand total soil digests.Pneumaticnebulization
is not useful when levels of theseelementsare lower than 100Jlg/L. In contrast,
the hydride-mercuryvapor generatorwill enable the analyst to quantitatively
measurelevels aslow as 1.0 Jlg/L.
Soil extractsmust be pretreatedbefore introduction into the hydride gen-
erator. The oxidation state of the analyte is critical. Selenium (IV) is readily
reducedto the hydride, but Se(VI) is not. Arsenic (III) is more readily reduced
than As(V). Also, organicconstituentsin the extractsinterferewith hydride gen-
eration andshouldbe destroyed.At present,no pretreatmentprocedurehasbeen
found to simultaneouslypretreat extractsfor Se and As analysis. The Se pre-
treatmentis not effective in reducingAs(V) to As(III), and the As pretreatment
reducesSe to the metal form, which will not form the hydride.
The total digest solutions resulting from the procedure given in
"Procedure"should contain only Se(IV) so that no pretreatmentfor Se analysis
is needed.However, for As determinationsthe digest solution must be pretreat-
ed.
Mercury may be determinedin solutionspretreatedfor Se or As analysis.
It is recommendedthat spike recovery studies be performed on the particular
type of solution to be analyzedto verify the accuracyof the technique.
Special Equipment
1. Hydride-mercuryvapor generator(Fig. 5-5).
2. Hot water bath.
3. Calibrated50-mL digestiontubes.
Reagents
1. Concentrationhydrochloricacid (HCI).
2. Hydrogenperoxide(H 20 2), 30%.
3. Sodium borohydride(NaBH4) solution: in 1 L of DDW first dissolve
0.5 g of sodiumhydroxide(NaOH), then 2.0 g of NaBH4.
4. Potassiumiodide-ascorbicacid solution: in 100 mL of DDW, dissolve
10 g of potassiumiodide (KI) and 1 g of ascorbicacid.
Description of Generator.The system(Fig. 5-5) consistsof three peri-
staltic pumps and a gas-liquid separator.Pump A (head no. 7014, 5.3 mL/min
flow rate) (Curtis MathesonScientific, Inc., Denver, Colo.) pumps the NaBH4
solution continuously. Pump B (head no. 7016, 20 mL/min flow rate) pumps
either 30-s (lO-mL) aliquots of pretreatedsampleor 30 s of 0.5 M HCI rinse
betweensamples.PumpC (headno. 7015, 42 mL/min flow rate) drains the gas-
liquid separator.Argon is forced through the baseof it 60-mL coarse-fritteddisc
Buchnerfunnel to strip the gasesfrom the liquid and to carry them into the plas-
ma. A signal is producedwhenevera detectableamountof analyteis introduced
into the plasma.Ten-secondintegrationsof the maximumsignal are usedto cal-
culateconcentrations.To beginthe integrationat the propertime, a chartrecorder
is useful initially to observethe signal timing. One sampleper minute is usually
analyzedunlessa very high concentrationis encounteredand a long rise period
becomesnecessary.Total Se determinationin metal-contaminatedsoils needs
special attention. Banuelos8 noticed Cu-like precipitates,after which Se deter-
minationswere not reproducible.The NaBH4 might have beenused up by ele-
ments other than Se. Banuelos recommended sampledilution to overcomethe
aboveinterference.
Pretreatmentof Soil Extracts to be Analyzed for Selenium.The fol-
lowing procedureis usedto reduceSe(VI) to Se(IV) and to destroyorganiccon-
stituentsin aqueoussolutionsbeforegeneratinghydrides.
Place15 mL of extractingsolution into a 50-mL digestiontube. Add 1 mL
of fresh 30% H20 2, and heat for 20 min in a boiling water bath. Add 10 mL of
concentratedHCI, and heat again for 20 min. Cool and bring the solution to a
final volume of 25 mL with DDW. The solution is now ready to be analyzedfor
Se using the hydride-mercuryvapor generator.The solutionsare approximately
4.8 M HC!. Standardsshouldcontainthe sameacid concentrations.
Pretreatmentof Soil Extractsor Total Soil Digeststo be Analyzed for
Arsenic.The following procedureis usedto reduceAs(V) to As(III) beforegen-
eratinghydrides.
Add 3 mL of potassiumiodide-ascorbicacid solution to 10 mL of either
soil extracts previously treated using the procedurein "Pretreatmentof Soil
Extractsto be Analyzedfor Selenium"or total soil digestsfrom the procedurein
"Procedure."Wait at least 10 min. The solution is now ready for analysisusing
the hydride-mercuryvapor generator.Standardsshould contain the sameacid
concentrationand be treatedin the sameway as the samples.
8 Banuelos,Gary, USDA-ARS, Fresno,CA.

Quality Cootrol Methods
In ICP-AES and ICP-MS as in any other methodsof analysis, the QC

shouldbe an integral part of the proceduresusedfor analysis.The quality of the
analytical work may be checkedin severaldifferent ways. Use of blind dupli-
cates,check sample,standardreferencematerials,recoveryof addedelements,
and interlaboratorycomparisonsare recommended.
Blind duplicatesare samplesthat are introducedinto the laboratoryby the
laboratorysupervisoror customers.The analystsanalyzethesesampleswithout
any knowledgeof the presenceof blind duplicates.A checksampleis a sample
that has beenanalyzedmany times by different laboratoriesand is available in
large quantities.This sampleis analyzedwith every batch of samplesto see if
any grosscontaminationor error hasoccurred.Standardreferencematerialsare
materials that have certified analytical values, such as National Bureau of
Standard(NBS) samples.Recently,NBS hasbeenchangedto National Institute
of Science and Technology (NIST). Canadian Land Resource Institute,
Agriculture Canada,and the CanadaCenterfor Mineral and EnergyTechnology
haveprepareda few referencesoil sampleswith recommendedvaluesfor sever-
al elements. Oneof the bestways to evaluateanalytical resultsis to have sam-
ples analyzedby severalreputablelaboratoriesand then to comparethe values,
suchasin a proficiencytestingprogram.Two programsavailablearethe Western
StatesLaboratory Proficiency Testing Programand The Soil & Plant analysis
proficiency testingprogram.Another methodof quality control is to add known
amountsof elementsto soil digestsor extractsand to determinethe percentage
recoveryof the addedelement.The bestguardagainstcontaminationof samples
is maintaininga high standardof cleanlinessthroughoutthe laboratory.For a dis-
cussionof QC methods,seeSkogerboeand Koirtyohann (1976) and Chapter2
(Klesta & Bartz, 1996).
SUMMARY
In this chapterwe have discussedICP-AES and ICP-MS. Our experience

and that of the workers who use ICP specrometryfor routine analysis have
shown that ICP is a generallysuperiorsourcein accuracy,precision,detection
limit, freedom from interferences,and dynamic range. Use of automaticsam-
plers,largecomputers,and the neededsoftwarefacilitatesanalysis.Onecanana-
lyze a solution for many elementsin 1 min with ICP-AES and 2 to 3 min with
ICP-MS. Therefore,large volumesof datacan be generatedvery fast. Isaacand
Johnson(1982)indicatethat with ICP-AESone techniciancando the samework
that formerly required four technicians.The ICP-MS requires highly trained
techniciansand is subjectto morepitfalls than ICP-AES.To our knowledge,only
three soil testing laboratoriesuse ICP-MS in contrastwith many that use ICP-
AES in the USA. However, for elementalisotopeabundancedeterminationthe
ICP-MS is the instrumentof choice.Thoughtshouldbe given to the handlingand
processingof the data.Interfacingthe instrumentwith larger computersfor data
handling and analysisis a must if one contemplatesobtaining large volumesof
data. It is recommendedthat usersof ICP-AES and ICP-MS subscribeto the
newslettersavailablefrom the manufacturerof their unit and other sourcesso

that they can be kept abreastof new developmentsin ICP-AES and ICP-MS.
Journalsreferencesand othersignificant sourcesof information are given in the
referencesection.
REFERENCES
Abell, J.D. 1991. Performancebenefitsof optimisationof laser ablation samplingfor ICP-MS. p.
209-217. In G. Holland and AN. Eaton(ed.) Applicationsof plasmasourcemassspectrom-
etry. R. Soc. Chern.Cambridge,England.
Ahearn,A.J. (ed.). 1972.Traceanalysisby sparksourcemassspectrometry.Acad. Press,New York.
Bacon,J.R.,A.T. Ellis, andJ.G.Williams. 1989.Atomic spectrometryupdate-inorganicmassspec-
trometry and x-ray fluorescencespectrometry.J. Anal. At. Spectrom.4:199R.
Bacon,J.R.,AT. Ellis, andJ.G. Williams. 1990.Atomic spectrometryupdate-inorganicmassspec-
Bacon,J.R.,A.T. Ellis, andJ.G.Williams. 1991.Atomic spectrometryupdate-inorganicmassspec-
Bajo, S. 1978. Volatilization of arsenic(III, V) antimony (III, V) and selenium(IV, VI) from mix-
turesof hydrogenfluoride and perchloncacid solution:Application to silicate analysis.Anal.
Chern.50:649-651.
Barnes,R.M. (ed.). 1992.ICP inform. newsletter.Univ. Massachusetts, Amherst,MA.
Beasecker,D.R., and LL Williams. 1978.An improvedsampledelivery systemfor ICAP analysis.
Jarrell-AshPlasmaNewslett.1(3):5-9.
Beauchemin,D. 1989.Early experiencewith inductively coupledplasmamassspectrometry.J. Anal.
At. Spectrom.4:553.
Beauchemin,D., J.w. Mclaren,andS.S.Berman.1987.Studyof the effectsof concomitantelements
in inductively coupledplasmamassspectrometry.Spectrochim.Acta 42B:467.
Belchamber,R.M., D. Betteridge,AP. Wade,AJ. Cruickshank,and P. Davison.1986.Removalof a
matrix effect in ICP-AESmulti-elementanalysisby simplexoptimisation.Spectrochim.Acta
41:503-505.
Bernas,B. 1968.A new methodfor decompositionandcomprehensiveanalysisof silicatesby atom-
ic absorptionspectrometry.Anal. Chern.40:1682-1686.
Boumans,P'W.J.M. 1966.Theory of spectrochemicalexcitation.PlenumPress,New York.
Boumans,P.W.J.M. 1984. Line coincidencetablesfor inductively coupledplasmaatomic emission
spectrometry.2nd ed. Pergamon,New York.
Bradford, G.R., and D. Bakhtar. 1991. Determinationof trace metalsin saline irrigation drainage
waterswith ICP-OES after preconcentrationby chelatiOn/solventextraction. Environ. Sci.
Technol.25:1704--1708.
Bradshaw,N., E.F.H. Hall, and N.E. Sanderson.1989. Inductively coupledplasmaas an ion source
for high-resolutionmassspectrometry.J. Anal. At. Spect.4:801.
Braverman,D.S. 1992. Determinationof rare earth elements by liquid chromatographyseparation
using inductively coupledplasmamassspectrometricdetection.J. Anal. At. Spect.7:43.
Brown, P.G.,T.M. Davidson,and J.A Caruso.1988.Application of He microwaveinducedplasma
massspectrometryto the detectionof high ionizationpotentialgasphasespecies.J. Anal. At.
Spect.3:763.
Chisum,M.E. 1992.Applicationsof negativeion analyseson the ELAN 250 ICP/MS. At. Spectrosc.
12:155.
Cresser,M.S., LC. Ebdon,C.W. McLeod, and J.C. Burridge. 1986.Atomic spectrometryupdate-
environmental analysis. J. Anal. At. Spectrom.1:1R.
Cresser,M.S., LC. Ebdon,andJ.R. Dean.1988.Atomicspectrometryupdate-environmental analy-
sis. J. Anal. At. Spectrom.3:1R.
Cresser,M.S., LC. Ebdon,andJ.R.Dean.1989.Atomic spectrometryupdate-environmental analy-
sis. J. Anal. At. Spectrom. 4:1R.
Cresser,M.S., LC. Ebdon,J. Armstrong,J.R.Dean,M.H. Ramsey,andM. Cave.1990.Atomic spec-
trometry update_nvironmental analysis.J. Anal. At. Spectrom.5:1R.
Cresser,M.S., J. Armstrong, J.R. Dean, P. Watkins, and M. Cave. 1991. Atomic spectrometry
update-environmental analysis.J. Anal. At. Spectrom.6:1R.
Cresser,M.S., J. Armstrong, J.R. Dean, P. Watkins, and M. Cave. 1992. Atomic spectrometry
update-environmental analysis.J. Anal. At. Spectrom.7:1R.
Dahlquist, R.L., and J.w. Knoll. 1978. Inductively coupled plasma-atomicemissionspectrometry.

Analysis of biological materialsand soils for major, trace, and ultra-traceelements.Appl.
Spectrosc.32:1-29.
Date, AR. 1986. ICP-MS: The best thing in analytical chemistrysince choppedlight? p. 20-26. In
Biennial Natl. Atom. Spectrosc.Symp., Bristol, England.July 1986.
Date, AR., Y.Y. Cheungand M.E. Stuart. 1987. The influence of polyatomic ion interferencesin
analysisby inductively coupled plasmasourcemassspectrometry(ICP-MS). Spectrochim.
Acta 42B:3.
Date, AR, and AL. Gray. (ed.). 1989. Applicationsof inductively coupledplasmamassspectrom-
etry. Blackie and Sons,London.
Dean,J.R.,L.c. Ebdon,H.M. Crews,and RC. Massey.1988. Characteristicsof flow injection induc-
tively coupledplasmamassspectrometryfor trace metal analysis.J. Anal. At. Spect.3:349.
Denoyer,E.R. 1991. Analysis of powderedsamplesby laser samplingICP-MS. p. 199-208.In G.
Holland and AN. Eaton (ed.) Applications of plasma source mass spectrometry.R Soc.
Chern.,Cambridge,England.
Denoyer,E., L. Qinghong,A Stroh, U. Volkopf, and R. Thomas.1991a.ExtendingICP-MS capa-
bilities with flow injection analysis.In Elem. Mass Spectrom.Workshop. Am. Soc. Mass
Spectrom.San Francisco,CA November1991.
Denoyer,E.R, KJ. Fredeen,and J.W. Hager. 1991b. Laser solid samplingfor inductively coupled
plasmamassspectrometry.Anal. Chern. 63:445A
Denoyer,E.R., and A Stroh. 1992. ExpandingICP-MS capabilitiesusing flow injection. Am. Lab.
February74.
Doherty, W. 1989. An internal standardizationprocedurefor the determinationof Yttrium and the
rare earthelementsin geologicalmaterialsby inductively coupledplasma-mass spectrometry.
Spectrochim.Acta 44B:263.
Dolan, R., J. Van Loon, D. Templeton,andA Paudyn.1990.Assessmentof ICP-MS for routine mul-
tielement analysis of soil samplesin environmentaltrace element studies. Fres. J. Anal.
Chern. 336:99.
Douglas,DJ., and R.S. Houk. 1985. Inductively-coupledplasmamassspectrometry(ICP-MS). Prog.
Analyt. At. Spectrosc.8:1.
Douglas,DJ., and L.A Kerr. 1988. Study of solids depositionon inductively coupledplasmamass
spectrometrysamplersand skimmers.J. Anal. At. Spect.3:749.
Douglas,DJ. 1989. Somecurrentperspectiveson ICP-MS. Can. J. Spectrosc.34:38.
Douthitt, c.B. 1994a.A new technology in high performanceICP-MS. p. 16. In Proc. 36th Rocky
Mountain Conf. Anal. Chern.,Finnigan MAT, Pap. 2, Denver,CO. 1 August.
Douthitt, C.B. 1994b.High resolutionICP-MS for ultra trace analysisin difficult matrices.p. 16. In
Proc. Rocky Mountain Conf. Anal. Chern.. Finnigan MAT, Pap 6. Denver,CO. 1 August.
Ebdon, L.c., M.S. Cresser,and C.W. Mcleod. 1987. Atomic spectrometryupdate-environmental
analysis.1. Anal. At. Spectrom.2:1R
Ek, P.G.,S.G. Hulden, E. Johansson,and T. Liljefors. 1991. A continuoushydride generationsystem
for ICP-MS using separatelynebulizedinternal standardsolutions.p. 178-198.In G. Holland
and AN. Eaton (ed.) Application of plasmasourcemassspectrometry.R. Soc. Chern.
Evans, E.H., and L.C. Ebdon. 1989. Simple approachto reducing polyatomic ion interferenceson
arsenicand seleniumin inductively coupled plasmamassspectrometry,J. Anal. At. Spect.
4:299.
Fassel,V.A 1977. Current and potential applicationsof inductively coupled plasma(ICP)-atomic
emissionspectroscopy(AES) in the exploration, mining, and processingof materials.Pure
Appl. Chern. 49:1533-1545.
Fassel,V.A., and RN. Kniseley. 1974. Inductively coupledplasmas.Anal. Chern.46:1155A-1164A
Fasset.J.D., and PJ. Paulsen.1989. Isotopedilution massspectrometryfor accurateelementalanaly-
sis. Anal. Chern. 6L643A.
Garbarino,J.R., and H.E. Taylor. 1987. Stable isotope dilution analysisof hydrologic samplesby
inductively coupledplasmamassspectrometry.Anal. Chern. 59:1568.
Gervais, L.S., and E.D. Salin. 1991. Inserted injector tubes for inductively coupled plasmaspec-
trometry. J. Anal. At. Spect.6:493.
Gray, AL. 1985.The ICP as an ion source-origins,achievementsand prospects.Spectrochim.Acta
40B:1525.
Gray, A.L. 1986. Massspectrometrywith an inductively coupledplasmaas an ion source:The influ-
enceon ultratraceanalysisof backgroundand matrix response.Spectrochim.Acta 41B:151.
Gray, AL. 1989. Visual observationof shock waves in an inductively coupled plasmamassspec-
trometry expansionstage.J. Anal. At. Spect.4:371.
Greenfield,S. 1983.Inductively coupledplasma-atomicemissionspectroscopy(ICP-AES)with flow

injection analysis(PIA). Spectrochim.Acta 38:93-105.
Gregoire, D.C. 1987a. The effect of easily ionizable concomitantelementson non-spectroscopic
interferencesin inductively coupledplasma-mass spectrometry. Spectrochim.Acta 42B:895.
Gregoire, D.C. 1987b. Influence of instrument parameterson nonspectroscopicinterferencesin
inductively coupledplasma-mass spectrometry. Appl. Spectrosc.41:897.
Gregoire,D.C. 1989. Application of isotope ratios determinedby ICP-MS to earth sciencestudies.
Prog. Anal. Spectrosc.12:433.
Hager,J.W 1989. Relative elementalresponsesfor laserablation-ICP-MS.Anal. Chern.61:1243.
Hassan,S.M., and N.T. Loux. 1989.Elimination of spectralinterferencesin inductively coupledplas-
ma-atomic emission spectroscopy using orthogonal polynomials. Spectrochim. Acta
45:719-729.
Helmke, P.A., and D.L. Sparks. 1996. Lithium, sodium, potassium, rubidium, and cesium. p.
551-574. In D.L. Sparkset al. (ed.) Methods of soil analysis. Part 3. Chemical methods.
Hieftje, G.M. 1992.Toward the next generationof plasma-sourcemassspectrometers.p. 16. In Proc.
1992 Winter Conf. PlasmaSpectrochem.,San Diego, CA. 11 January.
Hieftje, G.M., and G.H. Vickers. 1989. Developmentsin plasmasource/massspectrometry.Anal.
Chim. Acta 216:1.
Hieftje, G.M., G.H. Vickers, and D.A. Wilson. 1988. Detectionof NegativeIons by ICP-MS. Anal.
Chern.60:1808-1812.
Holden, N.E., and EW Walker. 1972. Chart of the nuclides. Wall chart. 11th ed. Knolls Atomic
Power Lab. General Electric Co. and Naval Reactors, U.S. At. Energy Comm. Educ.
Relations,G.E. Company,Schenectady,NY.
Holland, G., andA.N. Eaton.1991.Applicationsof plasmasourcemassspectrometry.R Soc.Chern.
Hossner,L.R 19%. Dissolution for total elementalanalysis.p. 000-000.In D.L. Sparkset al. (ed.)
Methodsof soil analysis: Chemical methods.Part 3. SSSA Book Ser. 5. ASA, CSSA, and
SSSA,Madison,WI.
Houk. R.S. 1986. Mass spectrometryof inductively coupledplasmas.Anal. Chern.58:97A.
Houk, RS., Hl. Svec,and V.H. Fassel.1981. Mass spectrometric evidencefor suprathermalioniza-
tion in an inductively coupledargonplasma.Appl. Spectrosc.35:380.
Houk, R.S., and J.J. Thompson. 1988. Inductively coupled plasma mass spectrometry. Mass
Spectrom.Rev. 7:425.
Huang,Y.Q., anc C.M. Wai. 1986. Extraction of arsenicfrom soil digestswith dithiocarbanatesfor
ICP-AES analysis.Commun.Soil Sci. Plant Anal. 17:125-133.
Hutton, RC., and A.N. Eaton. 1987. Role of aerosol watervapour loading in inductively coupled
plasmamassspectrometry.J. Anal. At. Spect.2:595.
Igarashi,Y., C.K. Kim, Y. Takaku, K. Shiraishi,M. Yamamoto,and N. Ikeda. 1990. Application of
inductively coupled plasmamassspectrometryto the measurementof long-lived radionu-
c1idesin environmentalsamples.A review. Anal. Sci. 6:157.
Isaac,RA., and WC. Johnson.1982. High speedanalysisof agricultural samplesusing inductively
coupledplasma-atomicemissionspectroscopy.Spectrochim.Acta 38:277-282.
Jackson,L.L., D.M. McKown, J.E. Taggart,Jr., Pl. Lamothe,and EE. Lichte. 1989. Geological and
inorganicmaterials.Anal. Chern. 61:109R
Jackson,L.L., T.L. Fries, J.N. Grossman,B.S.W King, Pl. Lamothe.1991. Geologicaland inorgan-
ic materials.Anal. Chern.63(biennialApplications Reviewsissue):33R-48R
Jarvis, KE. 1992. Handbookof inductively coupledplasmamassspectrometry.Chapmanand Hall,
New York.
Johnson,G.W. 1979. Trace analysiswith a direct currentplasmaechellespectrometersystem.Ph.D.
diss. ColoradoStateUniversity, Fort Collins (Diss. Abstr. 40104:1673-B).
Johnson,G.W., H.E. Taylor, and RK Skogerboe.1979a.Evaluationof solute vaporizationinterfer-
enceeffects in a direct currentplasma.Anal. Chern.51:2403.
Johnson,G.W., H.E. Taylor, and R.K Skogerboe.1979b. Evaluationof spectralinterferencesasso-
ciatedwith a direct currentplasma-multielement atomicemissionspectrometer(DCP-MAES)
system.Appl. Spectrosc.33:451.
Johnson,G.W., RD. Miller, and P.H. Brown. 1992a.ICP-MS for plant analysis.p. 212. In Proc. 1992
Winter Conf. on PlasmaSpectrochem.,San Diego, CA. 10 January.
Johnson,G.W., E.S. Littlefield, and R.O. Miller. 1992b.Botanicaltissueanalysisusingthe USN ICP-
MS technique. p. 49. In Proc. Pacific Conf. Chern. and Spectrosc.,Foster City, CA. 21
October.
Jones,lB., Jr. 1977.Elementalanalysisof soil extractsand plant tissueash by plasmaemissionspec-

troscopy.Commun.Soil Sci. Plant Anal. 8:349-365.
Kim, C.K., Y. Oura, Y. Takaku, H. Nitta, Y. Igarashi, and N.Ikeda. 1989a. Measurementsof
240pu:239pu ratio by fission track methodand inductively coupledplasmamassspectrometry.
1 Radioanal.Nucl. Chern. 136(5):353.
Kim, C.K, Y. Takaku, M. Yamamoto, H. Kawamura, K Siraishi, Y. Igarashi, I. Igarashi, H.
Takayama,and N. Ikeda. 1989b. Determinationof Neptunium - 237 in soil samplesusing
inductively coupledplasmamassspectrometry.Radioisotopes38:153.
Kim, C.K, R. Seki, S. Morita, S.1. Yamasaki,A. Tsumura,Y. Takaku,Y. Igarashi,and M. Yamamoto.
1991.Application of a high resolutioninductively coupledplasmamassspectrometerto mea-
surementof long-lived radionuclides.J. Anal. At. Spect.6:205.
K1esta,E.J.,Jr., and J.K Bartz. 1996. Quality assuranceand quality control. p. 19-48.In D.L. Sparks
et al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand
ASA, Madison,WI.
Koirtyohann,S.R.,IS. Jones,and D.A. Yates. 1980. Nomenclaturesystemfor low-power inductive-
ly coupledplasma.Anal. Chern. 52:1965.
LaFemiere,KE., G.w. Rice, and V.A. Fassel.1985.Flow injection analysiswith inductively coupled
plasma-atomicemissionspectroscopy:Critical comparisonof conventionalpneumatic,ultra-
sonic and direct injection nebulization.SpectrochimicaActa 40:1495-1504.
Laird, D.A., R.H. Dowdy, and R.e. Munter. 1991. Suspensionnebulization analysis of clays by
inductively coupledplasma-atomicemissionspectroscopy.Soil Sci. Soc. Am. J. 55:274-278.
Lam, J.W., and lW. Mclaren. 1990. Use of aerosol processingand nitrogen-argonplasmasfor
reductionof oxide interferencein inductively coupledplasmamassspectrometry.J. Anal. At.
Spect.5:419.
Langmyhr,F.J., and P.E. Paus.1968. The analysisof inorganicsiliceousmaterialsby atomic absorp-
tion spectrophotometryand the hydrofluoric acid decompositiontehcnique.Anal. Chim. Acta
43:506-507.
Larson, G.F., W.Y. Fassel,R.H. Scott, and R.N. Kniseley. 1975. Inductively coupledplasma-optical
emissionanalyticalspectrometry.A study of someinterelementeffects.Anal. Chern.47:238.
Larson,G.F., V.A. Fassel,F.K Winge, and R.N. Kniseley. 1976. Ultratraceanalysesby optical emis-
sion spectroscopy:The stray light problem. Appl. Spectrosc.30:384-391.
Lechler, P.J., and R.K Leininger. 1979. Analysis of black shaleby ICAP spectroscopy.Jarrell-Ash
PlasmaNewslett.2(1):8-10.
Legere, G., and P. Burgener. 1985. Elimination of high-salt interferenceeffects causedby lithium
metaborateusing a new teflon high-saltnebulizer.ICP Inform. Newslett. 11:447-456.
Lim, H.B., R.S. Houk, M.e. Edelson,and KP. Carney.1989. Somefundamentalcharacteristicsof a
reduced-pressure plasmaextractedfrom an inductively coupledplasma.J. Anal. At. Spect.
4:365.
Lindsay, W.L., and W.A. Norvell. 1978. Developmentof a DTPA soil test for zinc, iron, manganese,
and copper.Soil Sci. Soc. Am. J. 42:421-428.
Marciello, I., and A.F. Ward. 1978. Interelementcorrectionsfor spectralline interferences.Jarrell-
Ash PlasmaNewslett. 1(2):12-13.
Mclaren, J.W., and S.S. Berman. 1985. Wavelength selection for trace analysis by ICP-AES.
SpectrochimicaActa 40:217-225.
Mclaren, J.W., D. Beauchemin,and S.S. Berman.1987. Application of inductively coupledplasma
massspectrometryto analysisof marine sediments.Anasl. Chern. 59:610.
McQuaker,N.R., P.D. K1uckner,and G.N. Chang.1979. Calibration of an inductively coupledplas-
ma-atomicemissionspectrometerfor the analysisof environmentalmaterials.Anal. Chern.
51:888-895.
Munro, S., L. Ebdon, and D.J. McWeeny. 1986. Application of inductively coupled plasma mass
spectrometry(ICP-MS) for trace metal determinationin foods. 1 Anal. At. Spec!. 1:211.
Odegard,M. 1979. Determinationof major elementsin geologicalmaterialsby ICAP spectroscopy.
Jarrell-AshPlasmaNewslet!. 2(1):4-7.
Olivares, lA., and R.S. Houk. 1986. Suppressionof analyte signal by various concomitantsalts in
inductively coupledplasmamassspectrometry.Anal. Chern. 58:20--25.
Olson, KW., W.J. Haas,Jr., and Y.A. Fassel.1977. Multielement detectionlimits and samplenebu-
lization efficienciesof an improved ultrasonicnebulizerand a conventionalpneumaticnebu-
lizer in inductively coupledplasma-atomicemissionspectrometry.Anal. Chern.49:632...Q37.
Parsons,M.L., A. Forster, and D. Anderson. 1980. An atlas of spectralinterferencesin ICP spec-
troscopy.Plenum,New York.
Pearce,N.J.G., W.T. Perkins,I. Abell, G.A.T. Duller, and R. Fuge. 1992. Mineral microanalysisby
LASER ablation inductively coupledplasmamassspectrometry.1. Anal. At. Spect.7:53.
Plantz, M.R., I.S. Fritz, F.G. Smith, and R.S. Houk. 1989. Separationof trace metal copmlexesfor
analysisof samplesof high salt contentby inductively coupledplasmamassspectrometry.
Anal. Chern. 61:149.
Powell, M.J., and D.W. Boomer. 1995. Determinationof chromium speciesin environmentalsam-
ples using high-pressureliquid chromatographydirect injection nebulizationand inductively
coupledplasmamassspectrometry.Anal. Chern.67:2474-2478.
Robin, I. 1979. Emissionspectrometrywith the aid of an inductive plasmagenerator,ICP Inform.
Newslett.4(11):495-509.
Roychowdhury,S.B., andI.A Koropchak.1990.Thermosprayenhancedinductively coupledplasma
atomicemissionspectroscopydetectionfor liquid chromatography.Anal. Chern.62:484-489.
Sah, RN., and RO. Miller. 1992. Spontaneousreactionfor acid dissolutionof biological tissuesin
closedvessels.Anal. Chern.64:230.
Serfas,RE., I.1. Thompson,and RS. Houk. 1986. Isotoperatio determinationsby inductively cou-
pled plasma/mass spectrometryfor Zinc bioavailability studies.Anal. Chim. Acta 188:73.
Skogerboe,RK., and S.R Koirtyohann. 1976. Accuracy assurancein the analysisof environmental
samples.Natl. Bur. Stand.,Stand.Publ. 422. u.S. Gov. Print. office, Washington,DC.
Skogerboe,RK., and c.L. Grant. 1970. Commentson the definitions of the terms sensitivity and
detectionlimit. Spectrosc.Leu. 3:215.
Slavin, M. 1971. Emissionspectrochemicalanalysis.Wiley-Intersci., New York. p. 53-80.
Smith, F.G., D.R. Wiedrin and R.S. Houk. 1991. Measurementof boron concentrationsand isotope
ratios in biological samplesby inductively coupled plasmamassspectrometrywith direct
injected nebulization.Anal. Chim. Acta. 248:229-234.
Soltanpour,P.N., A Khan, and W.L. Lindsay. 1976. Factorsaffecting DTPA-extractableZn, Fe, Mn,
and Cu from the soils. Commun.Soil Sci. Plan Anal. 7:797-821.
Soltanpour,P.N., and AP. Schwab.1977. A new soil test for simultaneousextractionof macro-and
micro-nutrientsin alkaline soils. Commun.Soil Sci. Plant Anal. 8:195-207.
Soltanpour,P.N., and S. Workman. 1979. Modification of the N~HCOrDTPA soil test to omit car-
bon black. Commun.Soil Sci. Plant Anal. 10:1411-1420.
Soltanpour,P.N., A. Khan, and A.P. Schwab.1979a.Effect of grinding variableson the NH4HCOr
DTPA soil test values for Fe, Zn, Mn, Cu, P, and K. Commun. Soil Sci. Plant Anal.
10:903-909.
Soitanpour,P.N., S. Workman, and A.P. Schwab.1979b.Use of inductively - coupledplasmaspec-
trometry for the simultaneousdeterminationof macro- and micronutrientsin N~HCOr
DTPA extractsof soils. Soil Sci. Soc. Am. J. 43:75-78.
Soltanpour,P.N., and S. Workman.1981. Soil testingmethodsusedat ColoradoStateUniversity Soil
TestingLaboratoryfor the evaluationof fertility, salinity, sodicity, and traceelementtoxicity.
ColoradoStateUniv. Exp. Stn. Tech. Bull. 142.
Soltanpour,P.N., J.B. Jones,Jr., and S.M. Workman. 1982. Optical emissionspectrometry.p. 29-65.
In AL. Page(ed.) Methods of soil analysis. Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA, Madison,WI.
Soltanpour,P.N. 1991. Determinationof nutrient availability and elementaltoxicity by AB-DTPA
soil test and ICPS. Adv. Soil Sci. 16:165-190.
Suddendorf,R.F., and K.w. Boyer. 1978. Nebulizer for analysisof high salt contentsampleswith
inductively coupledplasmaemissionspectrometry.Anal Chern.50:1769-1771.
Tan, S.H., and G. Horlick. 1986. Backgroundspectralfeaturesin ICP-MS. Appl. Spectrosc.40:445.
Taylor, H.E., J.R. Garbarino, and S.R Koirtyohann. 1991. Flame ionization mass spectrometry:
Isotoperatio determinationsfor potassium.Appl. Spectrosc.45:886.
Taylor, P., and P. Schutyser.1986. Descriptionof a computerprogramfor quantitativespectralanaly-
sis of ICP-AES spectrageneratedwith a high resolutioncomputer-controlledmonocrhoma-
tor. 1985. Spectrochim.Acta 41:81-103.
Thompson,M., B. Pahlavanpour,and S.J. Walton. 1978a.Simultaneousdeterminationof trace con-
centrationsof arsenic,antimony, bismuth, selenium,and tellurium in aqueoussolution by
introductionof the gaseoushydridesinto an inductively coupledplasma sourcefor emission
spectrometry.Part I. Preliminarystudies.Analyst (London) 103:568-579.
Thompson,M., B. Pahlavanpour, and S.J. Walton. 1978b.Simultaneousdeterminationof trace con-
ncentrationsof arsenic,antimony, bismuth, seleniumand tellurium in aqueoussolution by
introduction of the gaseoushydridesinto an inductively coupledplasma sourcefor emission
spectrometry.Part 2. Interferencestudies.Analyst (London) 103:705-713.
Thompson,M., and J.N. Walsh. 1983. A handbookof inductively coupled plasmaspectrometry.
Blackie, London.
Thompson,J.J.,and R.S. Houk. 1986. Inductively coupledplasmamassspectrometricdetectionfor

multielementflow injection analysisandelementalspeciationby reversed-phaseliquid chro-
matography.Anal. Chern.58:2541.
Thompson,J.J.,and R.S. Houk. 1987. Study of internal standardizationin inductively coupledplas-
ma-massspectrometry.Appl. Spectrosc.41:801.
Tsumura,A., andS. Yamasaki.1991.Determinationof ultra-tracelevelsof rare earthelementsin ter-
restrial water by high resolution ICP-MS with an ultrasonic nebulizer. p. 119-129. In G.
Holland and AN. Eaton (ed.) Applications of plasmasource massspectrometry.R. Soc.
Chern.,Cambridge,England.
Ure, AM., AT. Ellis, and J.G. Williams. 1988. Atomic spectrometryupdate-inorganicmassspec-
U.S. Departmentof CommerceNational Technical Information Service. 1977. Evaluation of an
inductively coupledplasma,multichannelspectrometricanalysissystem.Ames Lab., Ames,
Iowa, and Environ. Res.Lab., Athens,GA Anal. Chern.BranchPB-270-918.
Van Heuzen,AA., T. Hoekstra,and B. van Wingerden.1989. Precisionand accuracyattainablewith
isotope dilution analysisapplied to inductively coupledplasmamassspectrometry:Theory
and experiments.J. Anal. At. Spectrom.4:483.
Vaughan,M.A., and G. Horlick. 1986. Oxides, hydroxide and doubly-chargedanalyte speciesin
inductively coupledplasma/mass spectrometry.Appl. Spectrosc.40:434.
Veillon, C., and M. Margoshes.1968. A pneumaticsolution nebulization system producing dry
aerosolfor spectroscopy.Spectrochim.Acta 23B:553.
Viczian, M., A Lasztity, X. Wang and R.M. Barnes.1990.On-line isotopedilution and sampledilu-
tion by flow injection and inductively coupled plasma mass spectrometry.J. Anal. At.
Spectrom.5:125.
Ward, A.F. 1978a. Inductively coupled argon plasmaspectroscopy.Development,technique,and
applications.Am. Lab. 10:79-87.
Ward, AF. 1978b.Stockstandardpreparation.Jarrell-AshPlasmaNewslett. 1(2):14-15.
Ward, AF., and R.B. Myers. 1979. The effect of concomitant speciesupon the analyte signal
observedfrom a modemICAP direct readingspectrometer.ICP Inform. Newslett.4(9):403.
Weast,R.C., and MJ. Astle. (ed.). 1979. CRC handbookof chemistry and physics. 59th ed. CRC
Press,Boca Raton,FL.
Winge, R.K., V.I. Peterson,and V.A. Fassel.1979. Inductively coupled plasma-atomicemission
spectroscopy.Prominentlines. Appl. Spectrosc.33:206-219.
Winge, RK, D.E. Eckels,E.L. DeKalb and V.A. Fassel.1988. Spatiotemporalcharacteristicsof the
inductively coupledplasma.J. Anal. At. Spect.3:849.
Winge, R.K., J.S.Crain, and R.S. Houk. 1991. High speedphotographicstudy of plasmafluctuations
and intact aerosolparticlesor droplets in inductively coupledplasmamassspectrometry.J.
Anal. At. Spect.6:6Ol.
Wolcott, J.F., and C.S. Butler. 1979. A simple nebulizerfor the inductively coupledplasmasource.
ICP Inform. Newslett.4(10):461.
Workman, S.M., and P.N. Soltanpour.1980. Importanceof pre-reducingselenium(VI) to selenium
(IV) and decomposingorganicmatterin soil extractsprior to determinationof seleniumusing
hydride generation.Soil Sci. Soc. Am. J. 44:1331-1333.
Published 1996
Chapter 6
Neutron Activation Analysis
PHILIP A. HELMKE, University o/Wisconsin, Madison, Wisconsin
Neutronactivationanalysis(NAA) is a methodof elementalanalysisutilizing the

propertiesof the atomic nucleus.This feature distinguishesthe techniquefrom
most other methodsof analysiswhich are basedon the behaviorof the elements
resultingfrom their electronconfigurations.Neutronactivation analysisis based
on the principlesof atomic and nuclearstructure,radioactivity, nucleartransfor-
mations,and the interactionof radiationswith matter.
Neutron activation analysis provides a highly sensitive and accurate
methodapplicablefor the analysisof more than 70 elements.About 30 elements
can be readily determinedin soil samples.Most of theseelementscan be deter-
mined by instrumentalneutronactivation analysis(INAA), that is, NAA with no
postirradiationchemical separations.The sensitivity of NAA can be increased
more than 1000-fold by postirradiation radiochemical separationsbut this
approachis labor intensiveand usedonly when justified. The identity of the ele-
mentsdeterminedby INAA rangefrom severalof the major and trace elements
essentialto plantsto many of the trace elementsimportant to geochemicalstud-
ies of soils. Fewer elementsare measurablein plant and animal samplesthan in
soils becausesome elementsare bioaccumulatedat concentrationstoo low to
detectby INAA.
Among the techniquessuitablefor analysisof traceelements,NAA hasthe
important advantageof being relatively free of the effects of contaminationand
reagentblanks. It also is a very efficient methodof analysisfor elements,espe-
cially those with isotopeshaving short half-lives, such as Na, K, CI, and Mn,
where 50 or more samplescan be analyzedin severalhours. The resultsare not
affectedby the chemical stateof the elementbecauseNAA is a nucleartechnique.
Instrumentalneutronactivation analysisalso has the advantageof being nonde-
structive and free of the problemsassociatedwith sampledissolution,an impor-
tant considerationwhen determiningtotal elementconcentrationsin chemically
complex materialssuch as soils. Analyzed samplesare left intact for additional
microscopicor chemicalanalyses,althoughproperconsiderationsmust be made
for the small remnantof radioactivity left in the samples.Helmke and Ney (1992)
is an exampleof where the desirablecharacteristicsof NAA are utilized for the
rapid analysisof Na, K, and CI at trace concentrations.The major disadvantage
Book Seriesno. 5.
141
142 HELMKE
of NAA is that the techniquerequiresaccessto a researchnuclearreactoror other

neutron source.Severalfacilities in the world offer custom analysesat modest
costs.
The principles of NAA were proposed(von Hevesy& Levi, 1936) only 4
yr after the discoveryof neutronsin 1932. The techniquedevelopedrapidly with
the placementof researchnuclearreactorsin university and governmentlabora-
tories in the late 1940sthrough the 1950s. Major improvementsin gamma-ray
detectorsand electronicssince the 1960shave led to the maturationof the tech-
nique.
Neutron activation analysisis a comparisontechnique.Samplesand stan-
dardsare encapsulatedin plasticor silica containersand irradiatedsimultaneous-
ly with neutrons.After irradiation, the radionuclides,usually thoseproducedby
neutroncapture,are radioassayedand the ratio of the decaycorrectedcount rates
areproportionalto the massesof the elementsin the samplesandstandards.Users
should have basic training in analytical skills and knowledgeof nuclearchem-
istry, radiationsafety, and detectioninstruments.Instrumentalneutronactivation
analysis is remarkably free of systematicerrors when practicedwith care and
excellentresultscan be easily obtained.
PRINCIPLES
NeutronInducedReactions
A radioisotopeof each analyte element must be producedby a suitable

nuclearreaction.This is the "activation" process.The most convenientand wide-
ly used activation processis neutron capture. Neutrons are classified by their
kinetic energy.Thermalneutronsare in thermal equilibriumwith the surrounding
atomsand have an energyof about 0.025 eV. They are the most abundantneu-
trons in a nuclear reactor and, fortunately, capture of thermal neutronsby the
atoms inordinary materialsgives many radioisotopeswith propertiessuitablefor
gamma-rayanalysis.
The simplestneutroncapturereactionis the captureof one neutronby a sta-
ble atom to give a new isotope of the same elementwith a mass number one
greaterthan the original atom. An exampleof this reactionfor K is given in the
following equation
[1]
Radiochemistsabbreviatethis equation(where nand y representa neutron and

gammaray, respectively)to
[2]
and the reactionis called a (n, gamma)process.Someisotopesemit gammarays

in this processand thosegammarays are calledprompt gammas. They arisefrom
the excessenergy resulting from the captureof the neutron. They are not nor-
NEUTRON ACTIVATION ANALYSIS 143
mally used for NAA. The nucleususually recoils with sufficient energy during
the emissionof prompt gammasto breakchemicalbonds.The radioactiveatom
produced bythe captureprocesswill often have new or unusualoxidation states.
Some of the product isotopesof neutron irradiation are stable while most are
radioactive.The radioactiveisotopesarepotentially useful for NAA, and assayof
their decayradiationsoccursafter the neutronirradiation.
Other transmutationreactionsmay occurduring irradiation, suchas (n,2n),
(n,d), (n,a), and neutron-inducedfission reactionson atoms of elementswith
atomic numbersgreaterthan 90. Thesereactionsmay provide alternatepathways
for the production of analyte isotopes,and thus provide a potential source of
error. Such nuclearinterferencescannotbe eliminated,but their effects are gen-
erally minimal for most soils and plantsand can be easily characterized.
NeutronCaptureCross-Sections
Definition of Cross-Section
The measureof the probability of a reaction occurring is called its cross
section.For nuclearreactions,the cross-sectionis a measurement of areawith the
units of barns(b). One bam is 10-24 cm2. The value of the crosssectionfor neu-
tron captureis a function of the energyof the neutrons.At low energies,the val-
uesof the cross-sectiondecreasewith increasingneutronenergy.Neutroncapture
cross-sectionvalues for most isotopesare near their maxima for thermal neu-
trons. Resonancemaximaoccurwith neutronshaving energiesof about0.1 to 1.0
MeV. Thesemaximaoccur when the sum of the kinetic and binding energiesof
the neutronsequalsthe energyof an excited stateof the compoundnucleus.
Thin TargetAssumption
Accurateanalysesby neutronactivationdependson the assumptionthat the
numberof radioactiveatomsproducedand thereforethe intensity of the gamma
radiationproducedby the decayof the radioactiveatomsare known functions of
the intensity and durationof the neutronirradiation. Inaccurateresultsoccurif the
samplesand standardsare exposedto different and unknown neutronflux inten-
sities. Most researchreactorsusedfor neutronirradiationshavefacilities to rotate
the sampleduring irradiation to averagethe effects of flux gradientswithin the
reactor.A more furtive sourceof potential error is the casewhere the composi-
tion of the samplesand standardsare suchthat a significant proportionof the neu-
tron flux is absorbedby the outer portions of the sample or standard.This is
called self-shieldingwith the problem being most severewith sampleshaving
high concentrationsof elementswith large cross-sections.
The desiredsituation is that the sampleabsorbs«1% of the impinging neu-
trons. The sampleis then said to be "thin" to the neutronflux. For a samplecom-
posedof i different isotopeswith cross-section(Ji (cm2), each at a density of ni
atomsper cubic centimeter,the ratio of the neutronflux (I) at any distancex (cen-
timeters)into the sampleto the flux of impinging neutrons(fo) must be »0.99, as
calculatedfrom Eq. [3]
144 HELMKE
[3]
The conceptof "thinness"doesnot dependsolely on the linear thicknessof the

samplebut on the productof XLjnjO'j. Samplesof most naturalmaterialsare thin
to neutrons.Samplesthat are not thin may requiredilution by dissolutionbefore
neutron irradiation, although this is generally avoided whenever possible
becauseof potentialproblemsof contaminationand reagentblanks.
The "thinness" concept also can be violated by epithermal neutrons.
Epithermalneutrons,also called fast neutrons,are neutronswith energieshigher
than thermal neutrons.They occur becauseneutronsfrom nuclear fission have
initial energiesseveralordersof magnitudegreaterthan thermal energies.The
energy of the epithermalneutronsare moderatedthrough collisions with light
nuclei, mostly H nuclei in the water surroundingthe reactorcore. The ratio of
thermalto epithermalneutronsvarieswith reactordesignand position within the
reactorcore.
The element most likely to causesoil samplesto violate the thin target
assumptionis Mn, which has a seriesof 200- to 2000-b peaksat 300 to 10 000
eV. Calculationsshouldbe doneusing data orthe neutronenergyspectrumof the
irradiation facility for samplescontainingmore than 10 000 mg kg-1 of Mn and
with samplediametersexceedingabout 5 mm. Other elementswith large cross
sections(Ag, Au, B, Cd, In, rare-earthelements)are presentat low enoughcon-
centrationsin most soil samplesso that the thin target assumptionis valid. The
thin targetassumptionis almost neverviolated for samplesof plants and natural
waters.
Productionand Decayof RadioactiveIsotopes
The rate of productionof radioactiveatomsN (dN/dt) during neutronirra-

diation is proportionalto the neutronflux, $, the numberof targetnuclei (n), and
the cross-sectionfor neutroncapture,0', as given in Eq. [4]
dN/dt = $nO'. [4]
The neutronflux is the numberof neutronspassingthrough an areaof 1 cm2 in

1 s.
If the product isotope is radioactive,some of its atoms decay during the
irradiation period. Radioactivedecayfollows first order kinetics and the loss of
the product radioisotope N is given by Eq. [5]
-dN/dt= AN. [5]
The rate constant,A., has units per time and is equal to In 2 divided by the half-
life of the isotope.In most instances,very few atomscapturemultiple neutrons
and no atomsof the radioactiveisotopeare presentat the start of the irradiation.
The net rate of productionof eachradioactiveisotopeduring irradiation is there-
fore the sum of Eq. [4] and [5]
dN/dl = <l>nO' - AN. [6]
Integrationof Eq. [6] over the time of irradiation, I, gives the total numberof
radioactiveatomsproducedfor eachisotope
N =(<I>nO'/A)[1 - exp(-A/)]. [7]
The sensitivity of the analysisis thus dependenton the magnitudeof the

neutronflux, the value of the cross-section,the numberof targetatoms,and the
decayconstantof the isotopeproduced.The variablesunderthe analyst'Sdirect
control arethe numberof targetatoms,which is proportionalto samplemass,and
the lengthof the irradiation. Within limits, the sensitivityof analysisis increased
by increasingthe samplemass.Longer irradiationsincreasesensitivity only for
isotopesthat do not become"saturated"asdiscussedbelow. The intensityof the
neutronflux varieswith reactordesignand location within the reactor.The flux
for most researchreactorsrangesfrom 1012 to 1014 neutronscm-2 S-1.
It is useful to calculatethe numberof radioactiveatomsat the end of the
analysisto estimatethe sensitivity of the analysis.This cannotbe doneprecise-
ly becausethe valuesfor the cross-sections and the intensity of the neutronflux
are usually not exact.Most isotopesusedanalytically fall into one of two limit-
ing casesof Eq. [7]. In mostcases,the durationof the neutronirradiationis short
comparedto the half-life of the isotopeof interest.The fraction of eachisotope
that decaysduring the irradiation is then usually insignificant. In this case,I is
«/112' and exp(-A/) - 1 - At in Eq. [7], and then
N =<l>nO'/. [8]
This is the situationfor most elementsmeasuredby NAA.

In the casewhere the duration of the irradiation is long comparedto the
half-life of the isotope,which is often the casefor very short-livedisotopes,the
exponentialterm approacheszero and from Eq. [7]
N =<l>nO'/A. [9]
or
AN =<l>nO'. [10]
In this case,the rate of productionequalsthe rate of decay,and the isotopehas

reached"saturation."The sensitivity of analysisin this casecan be improvedby
increasingthe intensity of the neutronflux but not by increasingthe length of
irradiation.
The radioactiveatomscontinueto decayafter irradiation accordingto Eq.
[5] or its integratedform
N =No exp(-At) [11]

146 HELMKE
whereNo is thenumberof radioactiveatomsat someoriginal time, to, andN indi-

catesthe numberat somelater time, t. The quantity t1/2 is known as the half-life
of the isotopeand it can be usedto characterizethe isotopesince it is relatedto
the decayconstant,A, which hasa unique value for eachisotope.
The samplesneedto be radioassayedbeforethe activity of the analyteiso-
topes becomes insignificant, usually within five half-lives. Because many
radionuclidesare producedduring the irradiation of natural samples,the activi-
ties of the variousradioisotopeswill dependupon the lengthof irradiationandthe
lapsedtime from the end of irradiation to radioassay,also known as "cooling
time." Both time periods are chosen to maximize the activity of the analyte
radioisotopesand minimize the activity of potentially interfering radioisotopes.
Samplesare often allowed to cool to decreasethe radioactivity of short-livediso-
topesbefore radioassayof intermediateand long-lived isotopes.In practice this
usually resultsin two irradiations,one short (minutes)and an immediateradioas-
say within hoursand one longer irradiation (hours),and severalradioassaysover
a period of 3 to 40 d.
NuclearDecayand Detectionof Radiation
Typesof NuclearDecay
Radioactiveatomscan decay by five modes:ejection of an electronfrom
the nucleus(~-), ejectionof a positiveelectron(positron,W), ejectionof two pro-
tons and two neutrons(~He, or a), electroncapture(EC), and fission. Most of the
radioisotopesproducedby neutron irradiation of natural samplesdecay by 13-
emission,becausethe addition of a neutronincreasesthe neutron/protonratio of
the nucleus.The emissionof gammarays resultsas the excitednucleusproduced
by the decaygoesto its groundenergystate.Most of the time the excitednucle-
us de-excitesalmost immediately, but some isotopes have metastableenergy
statesand the gammarays are delayed.Thesetransitionsare known as isomeric
transitions.
Gamma-rayspectroscopyis almost always used for radioassayby NAA
becausewhen gammarays interactwith matterthey can lose most or all of their
energyin a singleevent.In contrast,chargedparticles interactwith matterby los-
ing energyin small increments,about34 e V per ion pair produced.
The decayschemesfor most isotopeshave beenstudiedand are compiled
in references(Lederer& Shirley, 1978). The decayschemefor the decayof 42K
[Eq. 12] is shown in Fig. 6-1.
t§K --7 taCa+ ~- + V [12]
The symbol V in Eq. [12] representsan antineutrino.The antineutrinois neces-

sary to satisfy the conditionsof the decay,but it can be ignoredbecauseit hasno
mass,or nearly so, and an insignificantprobability of interactionwith matter,and
thus it can not be easily detected.
As shown in Fig. 6-1, about 18% of all of the decaysof 42K populatethe
1525keV energylevel of 42Ca,while 82% of the decaysgo directly to the ground
12.4 hour
f3 3440
~~K
,...-....
>Q) 0.07% 2423
.::i.
"-" I
>- 0.05% I t 1836
t
C)
0:: I 1525
w 0.18% I I
Z
w 18%
82%
\
Fig. 6-1. Decayschemefor 42K.
state.The 1525 keY level of 42K also is populatedby small contributionsfrom

higher energylevels. The most intensegammaray for the analysisof K is there-
fore the 1525 keY line. The sensitivity of analysiscalculatedfrom Eq. [8] must
thereforebe adjustedupward by a factor of 5.6 (100%/18%)to accountfor the
percentageof all decays that produce the gamma ray used analytically. The
gammarays are called 42K gammarays though they result from the de-excitation
of 42Ca.
The energiesand intensitiesof gammarays emitted by radioisotopespro-
ducedby neutronirradiation are compiledin forms convenientfor NAA (Filby et
aI., 1970; Lis et aI., 1975a,b;Erdtmann & Soyke, 1975a,b).When selectinga
gammaray to be usedanalytically, one must considerthe half-life of the isotope
and its abundance,the intensitiesand energiesof the gammarays, and the poten-
tial spectral interferenceof gammarays from other radioisotopes.The relative
concentrationsof elementsin the sample must be consideredin making these
decisions.Radioisotopesand gammarays suitable for NAA of soils and plants
are given in Table 6-1. Potentialspectraland nuclearinterferencesalso are pre-
sented.The review by Laul (1979) gives a thoroughevaluationand discussionof
interferences.
Detectionof GammaRays
The radiationsemittedby decayingnuclei can only be measuredwhen the
radiationsinteractwith matter.Photons,including gammarays, interactwith mat-
ter by three mechanisms:the photoelectriceffect, Comptoneffect, and pair pro-
duction. The natureof theseinteractionsmust be understoodto properly interpret
gamma-rayspectra.The relative probability of eachtype of interactiondepends
on the energy of the photonsand the compositionof the absorber,which in the
caseof interestis the detector.
The photoelectriceffect involves the total absorptionof the energy of the
photon by an atom in the detector,which causesthe ejection of a K or L shell
electron.The ejectedelectronusually interactswith the other atomsof the detec-
148 HELMKE
Table 6-1. Summaryof radioassayinformation pertainingto neutronactivationanalysisof soils.
Interferences
Energies Spectral
of gamma Count
Nuclide Half-life rays used set Isotopeor reaction§§ Half-life Energy
keY d
24Na 15.0 h 1368.4 3 [24Mg (n,p) 24Na]:j:
1731.9 det [27AI (n,lX) 24Na]:j:
2754.1
38Cl 37.3 min 1642.7 0 [41K(n,lX) 38Cl]:j:
2167.6
42K 12.36h 1524.7 3
47ea 4.54d 1297.0 10 152Eu§ 12.7 yr 1299.2
49ea 8.7 min 3084.1 0
46Sc 83.8d 889.3 10,40
1120.5 182Ta1l 115d 1121.2
51Cr 27.7d 320.1 10,40 177Lu§ 6.74d 321.4
[54Fe (n,lX) 51Cr]:j:
56Mn 155 min 846.7 0
1811.2
59Fe 44.56 d 192.2 40
1099.2
1291.6
6OCO 5.272yr 1173.2 10,40
1332.5
64CU 12.8 h 511.0 0.3# [~+ annihiiation]1I
1345.8
65Cu 5.1 min 1039.0 0
65Ni 152 min 366.5 0#
1481.7
65Zn 243d 1115.5 10,40 152Eu 12.7 yr 1112.2
1~ 72d 1115.3
46Sc 83.9d 1120.5
72Ga 14.12h 629.9 0#
834.0
76As 26.32h 559.1 3
75Se 120d 136.0 40# 181Hf§ 42.5 d 133.1
l3lBa§ 12 d 133.7
181Hf§ 42.5 d 136.3
264.7 40#
82Br 35.34 h 554.3 3,10
698.3
776.5
86Rb 18.82d 1077.2 10,40
87mSr 169.8min 388.5 0#
99Mo 66.7h 181.1 3#
366.4
llOmAgtt 255 d 657.7 40
115Cd 53.5 h 492.3 3#
527.9
122Sbtt 64.34h 564.1 3,10 76As§ 26.4 h 562.8
124Sbtt 60.2d 1691.1 40
1281 25.0 min 443.3 0#
526.4
Interferences
Energies Spectral
of gamma Count
Nuclide Half-life rays used set Isotopeor reaction§§ Half-life Energy
keY d
134es 2.062yr 604.7 40 124Sb§ 60.3 d 602.7
795.8
l3lBa 12.0 d 373.2 10
496.2
140La 40.27 h 328.8 10 23SU fission
487.0 3,10 47Ca§ 4.5 d 489.2
1596.5
141Ce 32.55 d 145.4 40 23SU fission
147Nd 10.98d 531.0 10 IS3Sm§ 46.8 h 531.4
23SU fission
IS3Sm 46.8 h 103.2 3,10 23~p§ 56 h 99.7
23~p§ 56h 103.7
lS2mEu 9.30h 344.2 0
lS2Eu 12.7 yr 121.8 40 7SSe§ 120 d 121.1
778.9 lS4Eu:j::j: 16 yr 123.1
1408.1 l3lBa:j: 12.0 d 124.2
l60Jb 72.1 d 298.6 10,40 233Pa'll 27d 300.1
17SYb 4.19 d 282.5 10 23~p§ 56 h 285.6
396.1 16OJb§ 72d 392.5
133Pa§ 27 d 398.5
l60yb 32.02d 177.0 40
177Lu 6.71 d 208.3 3,10 23~p'll 56h 209.7
181Hf 42.5 d 133.1 40 131Ba 12.0 d 133.7
482.2 10,40
1189.0
182Ta 115d 1221.4 40
1231.0
197Hgtt 65.0h 77.6 3#
191.4
198Autt 64.7 h 411.8 3,10 191Pt§ 3.0d 409.6
233Pa(fh) 6.95 d 311.9 10,40 l6OJb§ 72.1 d 309.6
13~p(U) 56.3 h 228.1 3,10 182ya 115d 229.3
277.6
t de =doubleescape.
:j: Not seriousfor most soil samples.
§ Potentially serious,peaksnot resolvedor marginally resolved,but interfering peak is usually
insignificant in most soil samplesif sampleis radioassayed
at optimum time.
'II Serious,similar size peaksnot resolvedfor nearly all samples,correctivemeasuresnecessary.
# Radiochemicalseparationusually requiredbeforeradioassay.
tt Elementpresentat concentrationsbelow detectionlimit for INM in most soil samples.
:j::j: Not serious;152Eu/154Euratio is constantfor all samples,andthe half-lives are too long to cause
any error.
§§ The symbols,n, p, a, and 11+ representa neutron,proton,alphaparticle,and positron,respective-
ly.
tor, with the net result that all of the gamma-rayenergy is depositedinto the
detector.The hole left by the ejectedelectronis filled by a higherenergyelectron,
accompaniedby the emissionof characteristicx-rays. The photoelectriceffect is
the desiredphenomenafor gamma-rayspectroscopy.
150 HELMKE
During the Compton effect, the incident gammaray transferspart of its

energyto a bound electron.The gammaray emergeswith a lower energy,while
the electronis ejectedfrom the atom. The scatteredgammarays have a continu-
ousenergyspectrum,with their energydependingon the angleof interaction.The
Compton effect is important for gammarays having energiesgreaterthan 0.5
MeV. This phenomenonis the dominantcontributorto the generalbackgroundin
the low-energyregionsof gamma-rayspectra,especiallyif the samplescontain
radioisotopeswith intense,high-energygammarays.
Pair productionresultswhen high-energygammarays interactwith nuclei
to producean electron-positronpair. The minimum energy of the gamma ray
neededfor this phenomenonis 1.02 MeV, which is the energyof the equivalent
rest massof the electron-positronpair. Any extraenergyis carriedaway as kinet-
ic energyof the pair. The positroncan interactwith an electronin the detectorand
both articles are annihilatedto form two photonsof 511 keVeach.Gamma-ray
spectrathereforealmostalwayscontaina peakat 511 keY. Also, single and dou-
ble escapepeaksfor intensegamma rays with energiesabove 1.02 MeV will
occur. The energiesof the escapepeakswill be 511 and 1022 keY lessthan that
of the photopeak.The escapepeaksresult when one or both of the 511 keV anni-
hilation gammarays escapefrom the massof the detector.
EQUIPMENT
NeutronSources
The limited availability of neutron sourcesis the major deterrentto the

more widespreaduse of NAA. Researchnuclearreactorsat university and gov-
ernmentfacilities are most commonly used for irradiations. A completelist of
available research and training reactorscan be obtainedfrom the Departmentof
Energy Research Reactor Database through the FEDIX system (Internet
192.111.228.33).Forty reactorsin the contiguousUSA are currently listed.
The flux of thermal neutronsin reactorscan be as high as 1015 neutrons
cm-2 S-1 but valuesnear1013 neutronscm-2 S-1 are more common.Neutrongen-
eratorsthat utilize the interactionof a radioisotopewith selectedtargetsare avail-
able. Their thermal neutronflux is generallytoo low to be useful for the analysis
of most naturalmaterials,but they are usedfor someindustrial and in vivo analy-
ses.The costsof neutronirradiation vary, but start at about $150/hr(1992 infor-
mation). From 20 to 50 samplesand standardscan be irradiatedsimultaneously
so the cost per sampleis moderate.
Gamma-RayDetectors
A nuclearradiationdetectoris a device that convertsthe energyof nuclear

radiationsinto an electricalsignal having an amplitudeproportionalto the ener-
gy of the radiation. Successfulanalysisby INAA requiresa detectorwith suffi-
cient efficiency and energyresolutionto measurelow count ratesand to resolve
gamma-raypeaksdiffering in energy by only a few kiloelectronvolts. Modem
semiconductordetectorshave almost completely replacedscintillation, such as

thallium dopedsodium iodide crystals [NaI(Tl)], or gas-filled detectors.This is
especiallyso for INAA becausepractical gamma-rayspectroscopyrequiresthe
much superiorenergyresolutionof Ge detectors.Scintillation detectorsare still
sometimesusedfor radiochemicalNAA becauseof their superiorefficiency.
Most modemsemiconductordetectorsfor gamma-rays,known as intrinsic
detectors,are madefrom hyperpureGe. They were introducedin 1976 and now
dominatethe market. The previousgenerationof semiconductordetectorsintro-
ducedin 1965 requiredthe diffusion of Li into part of the crystal to compensate
for the excessp-typeimpuritiesoriginally present.TheseGe(Li) detectorsrequire
a continuoussupply of liquid N to cool the cryostat and crystal becauseof the
mobility of Li at room temperature.Their propertiesare destroyedif they are
without liquid N for evena few minutes.Intrinsic detectorsrequire cooling only
when in useto reduceleakagecurrentsgeneratedby mobile carriersat room tem-
perature.There are still many Ge(Li) detectorsin use.
Interactionof gamma-rayswith the detectorcreateselectricalcarriers,elec-
trons and positive holes, which are collectedby an applied electrical field. The
collectedchargeis proportionalto the energylost by the incident radiation. The
averageenergyneededto producean electron-holepair in Ge at 7rK is 2.98 eY,
which comparesto about300 eV in Na(Tl). This differenceis responsiblefor the
greatly superiorenergyresolutionof Ge detectors.
The resolutionof gamma-rayspectrometers varieswith gamma-rayenergy.
It is usually reportedfor the 122 keY peakof 57CO, the 662 keY peakof 137CS,
and the 1333 keV peakof 6OCo. The resolutionis expressedas the full width-half
maximum (FWHM) in kiloelectronvoltsat the specifiedpeaks.Detectorswith a
resolutionof 1.60 to 1.90 keY at 1333 keY are preferredbecausethey can fully
resolveclosely spacedpeaksof severalimportant elements (e.g., Sc and Zn; and
Br, As, and Sb).
The efficiency of the detectorincreasesas the size of the detectorincreas-
es, especially for high-energygamma rays. Low-energy gamma rays are effi-
ciently stoppedeven by small detectorsor those madeof a low atomic number
material such as Si. Efficiencies rangefrom 10 to 100% relative to a 7.6 by 7.6
cm Na(I) detector.The choice of efficiency is largely dictatedby cost. Detectors
with efficienciesfrom 15 to 25% are adequatefor NAA, especiallysince analyt-
ically useful gamma-raysoccur at energiesbelow 2800 keY, and most are below
2000 keY.
The photopeak/Comptonratio is an indicator of the ability of a detectorto
measurelow-intensity, low-energypeaksin the presenceof higher energyradia-
tion. It is usually measuredas the ratio of the photopeakheight of the 1333 keV
peak of 6OCO to the height of the highest point of the Compton spectrumjust
below the Comptonedge.The peak/Comptonratio increasesas the efficiency and
resolutionof the detectorincreases,with commonvaluesranging from 35 to 55
for detectors with15 to 25% efficiencies.
The cryostat housesthe detectorcrystal and activatedC to provide cryo-
static pumpingto maintain a vacuum.It also containsa preamplifiermatchedto
the characteristicsof the detector.Cryostatsare made in either vertical or hori-
zontal configurations.The cryostatis seatedin a dewarfor liquid N, usually with
152 HELMKE
a capacity of 31 L. Appropriate hardwareis neededto securethe detectorand

sampleholder within the shielding to reproduciblyposition the samplesrelative
to the detector.
The detectorsrequireshieldingagainstbackgroundradiationfrom the nat-
ural U and Th decayseriesand 40K. Commercialshieldsare available,but very
satisfactoryshieldscan be fabricated atlesscost. About 2.0 cm of Pb is adequate
for NAA applications.Lead shielding thicker than 12 em increasesthe back-
ground becauseof the increasedvolume for cosmic ray interactions.Lining the
inside of the lead shield with Cd and Cu sheetswill eliminate the Pb x-rays (75
keV) if they presenta spectralinterference.Plexiglasscan be usedto adsorbthe
Cu x-rays if needed.
MultichannelAnalyzers
Multichannelanalyzersare neededfor pulseheightanalysisandfor record-

ing the gamma-rayspectrum.Other requiredmodulesinclude an amplifier and a
high voltagesourcefor the detector.Electronicmodulesconform to the Nuclear
Instrumentation Module(NIM) standardand are generallyinterchangeablewith
respectto physicalsize and signal characteristics.
Multichannel analyzersare available in single box (hardwired) units or
computer-based configurations.A computer-based system requiresa multichan-
nel buffer with an analogto digital converterand an interfaceto the computer.
Suchsystemsoften can acceptinput simultaneouslyfrom severaldetectors.The
software available with computer-basedsystemsgreatly simplifies data reduc-
tion and management.The multichannel analyzershould have a minimum of
2048 channelsper detectorbut 4096 channelsor larger is preferredfor INAA.
The cost of a completesystemin 1992without shieldingfor the detector(detec-
tor with cryostat,preamplifier anddewar; multichannelanalyzerwith amplifier
and high-voltagepower supply) startsat about $25 000. About another$10 000
is neededfor a computer-basedsystem withsoftware.The cost is considerably
higher if a large detectoris purchased.
GENERAL METHODS
RadiationSafety
The amountsof radioactivity producedduring neutron irradiation of sam-

ples are potentially hazardousand proper precautionsand shielding must be
used.Gram amountsof sampleirradiatedfor severalhourswill requireseight or
more cm of lead shieldingfor severaldaysafter irradiation to limit the doserate
to less than 50 mr h-1 at 30 cm from the surfaceof the container.The intensity
of radiation decreasesquickly as 56Mn (half-life of 155 min), 24Na (half-life of
15 h), and other short-lived radionuclidesdecay. The usual practice is to not
removethe samplesfrom the lead containerusedto transportthe samplesuntil
the samplesare radioassayed.Dose ratesmust be monitoredwith a portablesur-
vey meterand personnelmust wear either film badgesor thermoluminescence
dosimeters.The U.S. Nuclear Regulatory Commission(USNRC, 1983) limits

personnelexposureto 50 mr d-1 and 100 mr wk- 1• Prudenthandlingof the sam-
ples will keeppersonnelexposuresto almostundetectablelevels.
The USNRC regulates the use of radioactive material in the USA.
Individual researchersusually receiveauthorizationto use and possessradionu-
clides through their institutional USNRC license. Fume hoods, lead bricks for
shielding,and a lead containerfor sampletransportmust be available.The insti-
tutional healthphysicistshouldbe consultedto obtain training in radiationsafe-
ty and contaminationcontrol and advice about the adequacyof the facilities for
radionuclides.
Preparationof Samplesfor NeutronIrradiation
Grinding Samples
Samplesof soils and rocks for activationanalysisshouldbe groundto pass
a 150-~m (l00-mesh)plastic sieve to insure a homogenoussample.This size
requirementis sometimesmore severefor activation analysis,especiallywhen
samples<1 mg are analyzed.An automaticgrinderwith a high densityaluminum
oxide mortar and pestle(FisherScientific, Springfield, NJ) resultsin acceptably
low levels of elementcontamination.An agatemortar and pestle is an alterna-
tive. Small samplesof specific mineralscan be analyzedwhole, with the advan-
tage that their integrity is retainedfor later microscopicor other studies.
Plant and other biological samplesshould be oven- or freeze-dried.Wiley
mills and similar units usedfor grinding biological samplespotentially contam-
inate the sampleswith trace amountsof Mn, Co, Fe, Cr, Ni and other elements.
The levels of contaminationare often low enoughto be ignored,but they should
be evaluatedas required.A clean alternativefor specialsamplesis "thrashing"
samplessealedin a Teflon bag, but this works only for samplesthat are brittle.
Insectsand other small samplescan be analyzedwhole.
Standards
Suitablestandardsare previouslycalibratedsoil or rock samplesor labora-
tory preparedsolutions.Four standardsoil sampleswith different compositions
are available from the CanadianCertified ReferenceMaterials Project (CAN-
MET, Energy, Mines and ResourcesCanada,555 Booth St., Ottawa, Canada
KIA OG1). Elementconcentrationsdeterminedby NAA for thesesamplesare
reportedby Koons and Helmke (1978). Gladneyet al. (1985) provide a compi-
lation of all data publishedon thesesamples.United StatesGeological Survey
standardrocks also are suitableas standardsfor soil analyses,especiallySCo-1
(Cody shale). The National Instituteof Standardsand Technology(NIST) has
standardsamplesof orchard leaves[StandardResearchMaterial (SRM) 1571]
and bovine liver (SRM 1577a)that are suitablestandards forbiological samples.
The NIST coal flyash standard(SRM 1633a) is suitable for soils and rocks
(Korotev, 1987).High-purity chemicalsmustbe usedto reduceinterelementcon-
taminationif multielement,laboratorypreparedsolutionsare usedas standards.
Nitrate is the preferredanion for metal standards.
154 HELMKE
Sample Encapsulation for Neutron Irradiation

Samplesand standardsmust be sealedinto individual leakproofcontainers
for irradiation in a nuclearreactor. Heat-sealedpolyethylenetubing or vials are
convenientif the irradiation time is less than about 3 h. Our laboratoryuses65-
mrn (2.5-in.) lengthsof 6.4-mm inner diameter(i.d.) (0.25-in.) polyethylenetub-
ing. One endis heatsealedover a small flame by pinchingthe softenedtubewith
a crucible tong. After taring, the sampleis weighed directly into the tube. The
tube is then heatsealedand labeledwith a waterproofink.
Quartz tubessealedwith an Oz-naturalgas flame are usedfor irradiations
longerthan 3 h, becausepolyethylenebecomesbrittle with long irradiationtimes.
Small diameter(2- or 3-mm i.d.) high purity (99.9999%)quartz tubing is very
convenientfor the analysisof small samples«50 mg) becausethe elementval-
uesfor the tube blank are so small that the samplesdo not needto be transferred
to nonirradiatedcontainersbeforeradioassay.
Considerableinternal pressureis generatedwhen liquid samplesare irradi-
. ated longer that about 1 h becauseof the decompositionof water. Suitablehead
spacemust be allowed in the tubesfor the expansionof gasesand the sealsmust
be secure.Up to 0.5 mL of solution is usedin the polyethylenetubes.Othersam-
ples also may generatesomepressurethat can result in potential loss of fmely
powderedsampleswhen the tubesare opened.This can be eliminatedby cooling
the samplesin liquid N2 beforethey are opened.
Neutron Irradiation
Analystsshouldconsultwith their reactorpersonnelto determinethe con-

ditions of irradiation and the type of outer samplecontainersrequired.The reac-
tor personnelwill assistyou in estimatingthe total amountof radioactivity pro-
ducedand with proper irradiation procedures.Many reactorswill accepta 120-
or 180-mL polyethylenevial that will hold 20 to 50 samples.
Samplesand standardsare irradiated simultaneouslyand it is imperative
that they receiveidenticalneutronfluxes. Most reactorshaveprovisionsfor rotat-
ing the samplecontainersduring irradiation to averagethe effects of small flux
gradients.An alternativeis to monitor the neutronflux. This is done by securing
a short length of Fe wire (about 50 mg) aroundeachtube with Teflon tape (3M
Products,St. Paul, MN) before irradiation. The ratios of the specific activities of
the Fe wire after irradiation are then usedto correctflux heterogeneities.Copper
wire can be usedfor short irradiations.
It is convenientto irradiatethe samplestwice. The first irradiation is short
(10 s-5 min) and is immediately followed by radioassayto determinethe ele-
mentswith short-lived isotopes(such as Na, K, CI, Mn, Dy). A long irradiation
producessuch intenseradioactivitiesfor theseelementsthat radioassayis diffi-
cult or impossible.The secondirradiation for long-lived isotopesis usually 1 to
2 h but can be 6 h or more if the samplesare small or havelow concentrationsof
the elementsof interest.The initial radioassayis done 3 d after the long irradia-
tion so that the intenseradioactivity from the short-livedisotopeshasdecayed.
Radioassay
InstrumentalNeutronActivation Analysis
About 25 elementswith isotopeshaving half-lives >30 min can be easily
and accuratelydeterminedin most soils by INAA (Koons & Helmke, 1978).
Additional elementswith isotopeshaving half-lives <10 min (Al, Ca, Cu, F, Mg,
0, Ti, and V) can be determinedif facilities are availableto radioassaythe sam-
ples within a few secondsor minutesafter irradiation (De Soeteet aI., 1972).
The irradiated samples must be transferred to new containers before
radioassayunlessthe irradiatedcontainersare known to be free of the elements
being determined, includingaccidental contamination.Small, plastic culture
tubeswith a snap-topare convenientcontainersfor radioassay.A plexiglasssam-
ple holder canbe machinedto reproduciblyposition the tubesduring radioassay.
Samplesencapsulatedin high-purity containersfor irradiation can be cleaned
with water and methanoland tapedto small squaresof posterboardthat secure-
ly fit a matching sampleholder. Filtered samplesthat are separatedfor radio-
chemicalneutronactivationcan be heatsealedinto small polyethyleneenvelopes
and mountedonto a similar squareof posterboard.
Within an irradiationset,the volumeof the samplesandstandardsandtheir
position beforethe detectormust be identical. Failure to reproducethe counting
geometryis a common sourceof error, especiallywhen the samplesare a few
centimetersor lessfrom the detectors.The distancefrom the sampleto the detec-
tor shouldbe set so that the most radioactivesampleregistersa deadtime of <30
but usually <10% for the detectorsystem. Dead time occurs becausea small
amountof time is neededto detecteachgammaray andprocessits signal. During
this time the spectrometeris insensitiveto new gammarays. Excessivedead time
leadsto pulsepile up wherethe leadingedgesof successivesignalsoverlap.This
tendsto broadenthe peaksin the spectra.Excessivedead timealso leadsto coin-
cidencelosses,and theseshouldbe corrected ifthe total count rate exceeds5000
countsS-1 (Wyttenbach,1971).
About 5 min is sufficient to radioassaythe short-livedisotopes(Na, K, CI,
Mn, Dy) immediately after irradiation. Of these elements,CI has the shortest
half-life, 37.3 min. The radioassaysshould be completedbefore five half-lives
elapseto have an adequatecount rate. The time and length of the radioassays
after a long irradiation dependon the elementsbeing determined.If most of the
elementslisted in Table 6-1 are being determined.the best resultsare obtained
when the radioassaysare done 3, 10, and 40 d after irradiation. The 3-d radioas-
say requiresabout 1 h to acquiresufficient countsto have reasonablestatistical
uncertainties.The 10- and 40-d radioassayseach require about 6 h. This
sequenceallows mostof the isotopesto be radioassayed at closeto their optimum
times. Also, with this sequence, several isotopesare assayedin more than oneset
so that the quality of eachset of radioassayscan be compared.Most of the same
isotopescan be determinedin two count setsat 7 to 12 d and at 28 to 35 d, but
someof the interferencesare then potentially more significant and careful spec-
tral analysisis required.
156 HELMKE
RadiochemicalNeutronActivation Analysis
Dissolutionof the irradiatedsamplesand subsequentchemicalseparations
greatly improve the sensitivity of analysisand allow additional elementsto be
determined,suchas Cu, Ni, Ga, Sr, Cd, I, Hg, andsomeothersnot listed in Table
6-1. Theseelementsare usually presentin environmentalsamplesat concentra-
tions too low to be accuratelymeasuredby INAA. Chemicalseparationsreduce
the backgrounddue to Comptonscattering,especiallythat from 24Na and 4OK,
and eliminatepotentially overlappingphotopeaks.
Separationof single elementsis not necessary.Elementshaving similar
chemical propertiescan be isolated and radioassayedas a group. Additions of
carriersare neededto provide macroscopicamountsof the elementsfor easeof
handling, but this is an advantagebecausemeasurement of yields eliminatesthe
needfor quantitativeseparations.Simple separation schemes basedon precipita-
tion reactionsand liquid-liquid extractionsare often the quickest and safest.
Specific reagents, such as dimethylglyoxime for Ni or hydroxylamine
hydrochlorideor sodiumdithionite for the reductionof Se or Cu, respectively,to
the elementalform; are very useful and efficient.
The irradiated samplesmust be dissolved in the presenceof carriers.
Fusionwith Na202works well for soil androck samples,unlessvolatile elements
are to be determined.About 10 mg of a nitrate solution of each elementto be
determined,except 2 mg for each rare-earthelement, is added to a 4S-mL
Zirconium crucible as carrier and evaporatedto dryness.Identical amountsof
eachelementgroupedaccordingto the separationschemebeing usedare added
to individual flasks for the receipt of the irradiatedliquid standards.The irradi-
ated containersare openedand the sample poured into the crucible. A small
amountof solid NaCI can be usedto rinse any adheringsamplefrom the con-
tainer. About 10 tolS g of Na202is addedto the crucible and the mixture is fused
over a Meeker burner. After cooling, the fusion cake is dissolvedin water in a
coveredbeakerand acidified until a clear solution is obtained.
Digestionof irradiatedsampleswith nitric acid in a Teflon-linedParrbomb
works well for biological samples.Carriers are added to the Teflon liner and
evaporatedto drynessbeforethe irradiatedsampleis added.The amountof sam-
ple and nitric acid that can be safely useddependson the size of the Parr bomb.
Dissolutionof soils androcks in the Parrbombswith HF is neededif volatile ele-
ments are determined.Soils often need a preliminary treatmentwith HN03 to
destroyorganicmatter.
Analystscan devisetheir own separationschemeor select onefrom those
summarizedby De Soeteet al. (1972) or other sources.Our laboratoryusesthe
methodsof Denechaudet al. (1970), Allen et al. (1970), or Henzleret al. (1974)
with slight modifications for soils and biological samples.The separatedele-
mentsor their compoundsare filtered onto 2- or 3-cm diam. filters. The filters
are heat sealedinto small polyethyleneenvelopesand taped onto a squareof
posterboard for radioassay.The standardsare handledsimilarly. The yields do
not needto be quantitativeand slight contaminationwith nonradioactivemater-
ial is not importantas long as the amountof contaminationis insignificant com-
paredto the amountof carrier.The easiestway to measurethe yield from the sep-
aration is to irradiate the radioassayedsamplesand standardsfor a few seconds

or minutesand then radioassaythe activatedcarriers.Any residualradioactivity
from the initial irradiation mustbe small comparedto the radioactivity of the irra-
diatedcarriers.The carriersalso can be dissolvedand their concentrations deter-
mined by any suitabletechnique,such as atomic absorptionspectrophotometry.
Calculations
Calculationsinvolve determiningthe areaof eachanalytical photopeakin
the spectrumexpressedas a count rate, correctingthe count rate for radioactive
decay,and calculatingthe sampleconcentrationsbasedon the massesof the sam-
ples and standardsand the concentrationsof elementsin the standard.
There are two generalapproachesto determiningthe areaof each photo-
peak-peak-integration methodsand peak-fitting methods.Peak-fitting methods
dependupon fitting the spectraldataby non-linearregressionto a gaussianfunc-
tion modified by suitableexponentialor quadraticfunctions prior to determining
the peakarea.This approachhasadvantagesfor deconvolutingoverlappingpho-
topeaks, especially when the computer model is coupled with a gamma-ray
library that considersthe relative intensity of associatedpeaksand the efficiency
of the detector.Peak-fittingmethodsthat dependon a calibration spectrumhave
the disadvantagethat peak shapeoften dependson count rate, which can vary
with INAA.
Numerical integration methods are less sophisticated.They are usually
basedon fmding the minima in the smoothedspectrumon eitherside of the peak,
summingthe channelsbetweenthe minima, and subtractingthe backgroundas
approximatedby a straightline betweenthe minima. Accurateplacementof the
baselinebetweenclosely spacedpeaks,such as a single channelbetweenpeaks,
requires that the program comparethe smoothedto the real data. A common
approachto this problemfollows. The baselineis identified by finding the mini-
ma on either side of the peak in the five- or three-pointsmoothedspectrathat is
within three times the distance from the peak centroid correspondingto the
FWHM of the peak. If the minima in the five-point smoothedspectrais greater
than one standarddeviation from the actual data, the minima in the three-point
smoothedspectrais comparedto the actualdata.If it also is greaterthan one stan-
dard deviation for the actual date, the value of the actualdata is used.The back-
ground under the peak is the straight line betweenthesevalues.The grosspeak
area is the sum of the countsbetweenand including the backgroundchannels.
The difference is the net peak, which when divided by the live time gives the
count rate. A successfulnumericalintegrationprogramis describedby Lindstrom
and Korotev (1982).
A statistical error can be associatedwith each photopeakarea because
radioactivedecayis a randomprocess.The statisticalerror is determinedby the
error in the grossareaplus that of the baseline.The error in the grossareais the
squareroot of the area.The error in the backgroundis more complicatedbecause
the backgroundis a calculatedvalue.The backgroundareaerror is thesquareroot
of the productof the backgroundareatimes the ratio of the numberof channels
in the peakto the sumofthe numberof smoothedchannelsto determinethe back-
ground.The error in the net peakareais therefore
158 HELMKE
SDnetarea =
backgroundareax peakwidth
grossarea+ width low minimum + width high minimum. [13]
The statistical uncertainty for each photopeakin the standardand samplesare

combinedto give the uncertaintyof the calculatedconcentrationof eachelement.
The count rate for eachisotopein the standardsand samplesmust be cor-
rected forradioactivedecayusing the first-order rate law. Any time canbe select-
ed as t = 0 but a convenienttime is the midpoint of the acquisitionreal time of
the standard.The decaytime is then the differencein the mid-point time of the
samplesand standard.This approachapplieswhen the acquisitiontime is short
compared tothe half-life of the isotope,which is the usualcasefor NAA.
The concentrationof each elementis calculatedfrom the decay corrected
count rate as given below.
count ratesample massstandard

Concsample= x x concstandard [14]
count ratestandard masssample
Valuesfrom multiple determinationsof the sameelementshouldbe comparedfor

consistency.The concentration values determinedfrom multiple gamma-raysof
the same element and/or from multiple radioassaysare averagedto give the
reportedvalue. In caseswhere the statisticaluncertaintiesare very different, the
most precisedeterminationcan be reported.
SystematicErrors
Neutron activation analysisis relatively free of randomerrors. Well-char-
acterized standards in the most common matrices are readily available.
Heterogeneitiesof the neutronflux during irradiation can be monitoredif neces-
sary. Extraterrestrialsamplesand terrestrialsamplesassociatedwith U oresmay
have anomalousisotopic abundances.Neutron-inducedfission productsalso are
potential sourcesof error in sampleshaving high concentrationsof U or Th.
Interfering photopeaksare always a concernin spectroscopy,but nucleardecay
reactionsare well characterizedand scrutiny of the spectrashould detectpoten-
tial problems.Experiencehas shownthat the actual precisionof measurementis
limited by random errors to about 1% even when the statistical uncertainty is
smallerthan this.
REFERENCES
Allen, RO., L.A. Haskin, M.R Anderson,and O. Muller. 1970. Neutron activation analysisfor 39
elementsin small or preciousgeologic samples. J. Radioanal.Chem. 6:115.
Denechaud,E.B., P.A. Helmke, and L.A. Haskin. 1970. Analysis for the rare-earthelementsby neu-
tron activationand Ge(Li) spectrometry.J. Radioanal.Chem.6:97-113.
De Soete,D., R Gijbels, and J. Joste.1972. Neutronactivation analysis.Wiley-Intersci., New York.
Erdtmann,G., and W. Soyka. 1975a.The gamma-raylines of radionuclides,orderedby atomic and
massnumber.Part I. J. Radioanal.Chem.26:375495.
Erdtmann,G., and W. Soyka. 1975b.The gamma-raylines of radionuelides,orderedby atomic and

massnumber.Part II. J. Radioanal.Chern.27:137-286.
Filby, R.H., AI. Davis, K.R. Shah,G.G. Wainscott,WA Haller, andWA Cassatt.1970.Gammaray
energytablesfor neutronactivationanalysis.Nuel. Rad. Ctr. Rep. WSUNRC-97(2),Pullman,
WA. WashingtonStateUniv.
Gladney,E.S., C.E. Bums,andI. Roelandts.1985. 1983compilationof elementalconcentrationdata
for samplesSO-l to SO-4. Geostand.News!. 9:35-68.
Helmke,P.A, andO.M. Ney. 1992.Relationshipsbetweenconcentrationsof sodium,potassium,and
chlorine in unsaltedfoods. J. Agric. Food Chern.40:1547.
Henzler,T.E., R.I. Korda, PA Helmke, M.R. Anderson,M.M. Jimenez,and L.A Haskin. 1974.An
accurateprocedurefor multielementneutronactivationanalysisof traceelementsin biologi-
cal materials.J. Radioanal.Chern.20:649--663.
Koons, R.O.,andP.A Helmke.1978.Neutronactivationanalysisof standardsoils. Soil Sci. Soc. Am.
1. 42:237-240.
Korotev, R.L. 1987. National Bureauof Standardscoal flyash (SRM 1633a)as a multielementstan-
dard'for instrumentalneutronactivationanalysis.J. Radioanal.Chern.110:159-177.
Laul, J.e. 1979. Neutronactivationanalysisof geologicalmaterials.At. Ener. Rev. 173:603--695.
Lederer,C.M., and V.S. Shirley (ed.). 1978.Table of isotopes.7th ed. JohnWiley & Sons,Inc., New
York.
Lindstrom, OJ., and R.L. Korotev. 1982.TEABAGS: Computerprogramsfor instrumentalneutron
activationanalysis.J. Radioanal.Chern.70:439-458.
Lis, S.A, Ph.K. Hopke, and J.L. Fasching.1975a.Gamma-raytablesfor neutron,fast-neutronand
photonactivationanalysis.I. List of all the nuclideswith their associatedgamma-raysin order
of increasingatomic numberand mass.J. Radioana!'Chern.24:125-247.
Lis, S.A, Ph.K. Hopke, and J.L. Fasching.1975b.Gamma-raytablesfor neutron,fast-neutronand
photonactivationanalysis.II. List of all the gamma-raysfor all isotopesin order of increas-
ing energy,.J. Radioana!'Chern.25:303-428.
U.S. NuelearRegulatoryCommission.1983.Standardsfor protectionagainstradiation.p. 20-1 to 20-
26. In TItle 10, Chapter1, Codeof FederalRegulations-Energy.
von Hevesy,G., and H. Levi. 1936.Action of slow neutronson rare earthelements.Gen. Elec. Co.,
Schenectady,NY.
Wyttenbach,A 1971. Coincidencelosses.1. Radioana!.Chern.8:335-343.
Published 1996
Chapter7
Elemental Analysis by X-Ray Fluorescence

Spectroscopy
A. D. KARATHANASIS, University of Kentucky, Lexington, Kentucky
B. F. HAJEK, Auburn University, Auburn, Alabama
X-ray fluorescencespectroscopyor spectrometry(XRFS) is a method of ele-

mental analysis,which assesses the presenceand concentrationof various ele-
mentsin soil and plant materialsby measuringthe characteristicsecondaryradi-
ation emitted from a sample that has been excited with an x-ray source. The
method,which also is known as x-ray emissionspectrographyor x-ray spectro-
chemical analysis, is rapid, reliable, usually nondestructiveand in many cases
quicker and more accurate than conventional chemical analysis techniques.
Modem x-ray spectrometers are capableof detectingmore than 80 elementswith
atomic number>8, often times at concentrationsof a few parts per million and
they usually do not require extensivesamplepreparation.
X-ray fluorescencespectroscopyhas been used for elementalanalysisof
soil, plant and geologicalsamplessincethe 1950s;it is especiallywell-suitedfor
the compositionaldeterminationof major soiland clay components,and the mea-
surementof unusuallylarge concentrationsof certaintrace elementsin soils and
plants. In recent years, increasingenvironmentalconcernsabout soil and water
pollution from agriculturaland industrial chemicalshave triggeredtechnological
advancesin XRFS that allow routinely accurateelementalanalysesof soil, plant,
environmental,geologicaland biological samplesat lower detectionlimits.
Elementalanalysisby XRFS is basedon the principle that primary x-rays
from a target tube causethe elementspresentin the sampleto emit secondary
(fluorescent)x-radiation. Eachelement,when excited,emits x-rays of character-
istic wavelength(inverselyrelatedto the squareof its atomic number)and inten-
sity, which is roughly proportional to the amount of that element present.
Elementsof high atomic number emit radiationsof short wavelengthand high
energy, whereaselementswith low atomic numbersemit radiationswith long
wavelengthand low energy.
The radiation emitted by the sample can be analyzed by two types of
XRFS's. In the wavelength dispersive spectrometer,the emitted radiation is
SegoeRd., Madison,WI 53711.USA. Methods of Soil Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
161
162 KARATHANASIS & HAJEK
directedand diffracted accordingto Bragg'slaw by a single crystal of known d-

spacing,so that only one wavelength(element)is analyzedfor eachangularset-
ting of the crystal. The intensity of the diffracted radiationis measuredby a suit-
able detector (gas-filledproportionalor scintillation counter).In energy-disper-
sive spectrometers,diffraction is not involved. The variouswavelengthsof radi-
ation emittedby the sampleare separatedon the basisof their energyby means
of a solid-statelithium-drifted silicon-detectorand a multichannelanalyzer.The
counterproducespulsesproportionalin height to the energyof the incidentbeam
and the multichannel analyzer sorts out the various pulse heights. Although
wavelength-dispersive spectrometersare more widely usedbecausethey offer
betterresolutionwith low atomic numberelements(Si, Al, Mg, Na) and shorter
counting times, energy-dispersivespectrometersare usually less expensiveand
recentadvancesin solid-statedetectortechnologyhaveimprovedtheir resolution
and led to their successfulutilization for major elementanalysis.
Although elementalanalysisby XRFS was successfullyperformedas early
as 1923 (Von Hevesy,1932), the methoddid not come into generaluse until the
1950s,with the developmentof high-intensity,sealedx-ray tubes,suitableelec-
tronic counters and stable high-voltage power supplies. Since then, steady
improvementsin the standardsof mechanicaland electronic performance, inno-
vative advancesin solid-statedetectortechnologyand microcomputerautoma-
tion to control instrumentaloperatingconditions,and data acquisition,manipu-
lation, andanalysis have made XRFS an easilyaccessibleroutine analyticaltech-
nique rather than a researchinnovation. At presenttime, a relatively inexperi-
encedanalystcan analyzesuccessfullya great variety of specimensusing com-
mercially availableXRFS'sand accessoriesin a very short time.
There are certain considerations,however, that need to be taken into
accountand the analystshouldbe well awareof the limitations of the methodin
order to produceaccurateanalytical results.The major problemsassociatedwith
the elementalanalysisof soil and plant samplesby XRFS are the poor yield and
low energyof radiationsemittedfrom the light elements(abundantin the matrix
of soils especially), the matrix radiation effects produced from interelement
interferencesand particle-sizevariability, and the needto determinelow concen-
trations of important minor constituents.To reduceor eliminate theseeffects, a
variety of choicesin samplepreparation and/or matrix effect correctionsmay be
employed.
This chapterprovides a brief descriptionof the principles of x-ray spec-
troscopy, including sampleexcitation by x- rays, the interactionof x-rays with
matter, the detectionand measurementof secondaryradiations,availability of
instruments,safety measures,and qualitative and quantitative aspectsof the
method.Finally, applicationsof the methodto a variety of samplesare described
in detail, including instrumentalsettings,samplepreparation,and calculationof
results.
Some of the principles of x-ray spectroscopyare sharedby other related
techniquessuch as the electronprobe microanalysis(Castaing,1961; McKinley
et aI., 1966; Reed, 1975; Long, 1977; Heinrich, 1961), x-ray absorptiometry,x-
ray protoelectronspectrometry(Carlson, 1978) and low-energy electron spec-
trometry (Brondle & Baker, 1979).Thesemethodsrequire specialequipmentor
X-RAY FLUORESCENCESPECTROSCOPY 163
adaptationsand special sample preparationand they are not dealt with here.
Broader treatment of XRFS can be found in Birks (1969), Jenkins (1976),
Jenkinset al. (1981, p. 586), Tertian and Claisse(1982), and Williams (1987).
New applications,developments,and methodsare reportedin variousjournals,
including X-Ray Spectrometry, Advances in X-Ray Analysis, Analytical
Chemistry, Applied Spectroscopy, The Analyst, in varioussymposiaproceedings,
and in reportsand newslettersfrom manufacturers.
PRINCIPLES
Natureand Productionof X-Ray Spectra
Propertiesof X-Rays
X-rays are electromagneticradiationwith wavelengthsin the rangeof 0.01
to 5.00 nm, occupyingthe region betweenthe gammaand ultraviolet spectrum.
They are producedwhen incident electrically chargedparticles(electrons,pro-
tons, etc.) or photons with sufficient kinetic energy are deceleratedthrough
inelasticcollisions with atomsof the target material.The energylost in thesecol-
lisions is transferredinto the emitted x-ray photons. Becausethe proportion of
the energy lost by each deceleratedelectron can range from 0 to 12.4 keV, the
x-ray spectrumproducedconsistsof a continuousrange of energiesbetweena
minimum of zero and a maximum which correspondsto the incident electron
energy.Energy and wavelengthare relatedby the fundamental equation
E =hv =he/A [1]
whereE = energy,'A. =wavelength,h = Planck'sconstant(6.626 x 10-34 J), v =

frequency and c = the velocity of the electromagneticradiation. Therefore,the
energyE (keY) of an emittedx-ray photonwith wavelength'A. (nm x 10) is given
by the equation
E = (E j - E f ) = 12.4/'A. [2]
where Ej and Ef representinitial and final energystatesof the electrons,respec-

tively, measuredin kiloelectronvolts.
The x-ray spectrumprovided by electron bombardmentsconsistsof two
parts: the continuousand the characteristicspectrum.The continuousspectrum
always is presentas long as the primary electronradiation is within the rangeof
the x-ray wavelengthband.The characteristicspectrumis only presentwhen the
incident radiation exceedsthe critical excitationenergyrequiredby the elements
presentin the emitting sample. This excitation can be accomplishedeither by
electronbombardmentor by primary x-radiation producedby an x-ray tube as is
the case in XRFS. Becausethe primary x-rays used as excitation sourcesin
XRFS cannotlose their energyin a continuousfashion analogousto the deceler-
ation of electrons,only the characteristicspectrumis produced.
Following suchexcitations,electronsfrom the inner subshellsof the target

atomsare ejectedfrom the atomsor shifted to higher energyouterorbital levels.
At this statethe atom is unstableand regainsits initial groundstateby the trans-
fer of high-energyouter orbital electronsto the temporaryunfilled inner levels.
Theseinward transitionsreleaseenergy,which is then emitted as characteristic
x-ray radiation. Sincethe energiesof the electronsin eachatom are fixed at var-
ious orbital levels, the wavelengthsof the x-rays producedare uniquefor a given
elementand a given electrontransition. Thus, the K spectraare producedfrom
electrontransitionsfrom the L to the K shell giving rise to the Kal and Ka2 lines,
while M to K shell transitionsresult in ~l and K~2 lines. There are about 12
prominentspectrallines resulting from M to L electrontransitions.For general
analyticalwork in XRFS, the Ka, ~, La and L~ spectralines are the most impor-
tant. Different probabilitiesfor electrontransitionsdeterminethe relative inten-
sities of the characteristicspectralines producedfrom a given element,with the
highestenergiesand shortestwavelengthsbeingassociated with the K spectrum.
The principles cited above combinedwith Moseley'srelationshipbetweenthe
atomic number(Z) and the wavelength(A.) of characteristicx-ray spectralines
produced bya given elementserveas the basisfor XRFS.
X-Ray Excitation
Characteristicx-ray spectrallines are producedonly when the i!1cident pri-
mary x-ray beamhasenergygreaterthan the binding energyof the K or L elec-
trons. The wavelengthof the incident radiation must be shorter than a certain
critical wavelength,called the absorptionedge, in order to produce a fluores-
cencespectrum.The absorptionedgefor eachspectrumis just shorterthan the
characteristicwavelength.Alternatively, the characteristicwavelengthsfor each
elementare just longer than the respectiveabsorptionedgesfor that element.A
Cu targettube, for example,producingcharacteristicKa radiationof 0.154nm is
a very efficient exciterof Fe, which hasits K absorptionedgeof 0.174nm. There
is only one absorptionedgefor the K spectrumand three for the L spectra.
The minimum wavelength(Amin) in a continuous spectrum producedby an
x-ray tube is a function of the voltageappliedto the x-ray tube (Amin = 12.35/V).
Substitutingthe absorptionedgefor Amin in this equationallows the calculation
of the minimum voltage in kilovolts necessaryto excite and producethe charac-
teristic spectrumof a particularelement.For K spectraV min rangedform 1.3 kV
for Mg to 116 kV for U, while for L spectrathe range is 0.7 kV for Cr to 21.8
kV for U. In practice,voltagessubstantiallyhigher than V min of particular ele-
mentsare usedto achievegreaterspectralintensities.Acceleratingpotentialsup
to 50 kV are common for obtaining characteristicK x-ray spectraof elements
with Z s 50 and L spectraof elementswith Z > 50.
Tablesof x-ray emissionspectrawavelengthsand absorptionedgesfor dif-
ferent elementscan be found in several textbooks including Jenkinsand DeVries
(1970), White and Johnson(1972), Williams (1987) and others.
Excitation Sources
X-ray tubes,which arethe typical sourcesof x-radiation,usually containa
sourceof electrons(filament, cathode)a high acceleratingvoltagesuppliedby a
generator,and a metal target (anode).High-speedelectronsproducedby the hot

filament and acceleratedby the high voltage of the generatorcollide with the
metal targetand producex-rays. X-rays emitted from the targetescapefrom the
vacuum-tighttube housingvia two or more windows usually madeof beryllium.
X-ray tubesare marketedwith a variety of anodematerials.The most common-
ly used are tungsten(W), gold (Au), platinum (Pt), chromium (Cr), rhodium
(Rh), silver (Ag), molybdenum(Mo), scandium(Sc), and titanium (Ti). Because
of the limited numberof suitabletargetelementsavailablefor x-ray tubes,a tar-
get elementthat will yield effective exciting radiation for a wide range of ele-
mentsis usually selected.Usually an elementof a high melting point (allowing
applicationof high currents)and of high atomic numberis preferredbecausethe
intensity of the primary x-ray radiation is proportional to the atomic number.
However,becausethe most efficient excitationof the x-ray spectraof an element
is achievedby primary x-radiationof wavelengthjust shorterthan the absorption
edge of the element,different target elementsprovide different efficienciesfor
various rangesof x-ray spectra.Tungsten,Pt, and Au for example,with atomic
numbers74, 78,and 79, respectively,are commonlyusedmetal targetsfor exci-
tation of elementswith Z > 24, while chromium, molybdenum and rhodium
anodesare more appropriatefor lighter elements.Sincemost of the kinetic ener-
gy of the electronsstriking the target is convertedto heat, the target is water
cooledto preventits melting.
Obviously, no tube is suitablefor all applications.The choice of an x-ray
tube for a particularanalysismust take into accountthe following considerations:
1. It is not feasible to use a tube with an anodeidentical to the element

sought,especiallyat low concentrations.Characteristicspectrallines
from the tube anodeare scatteredfrom the sampleand producea high
backgroundbeneaththe fluorescentemissions.
2. The characteristicradiation of the anodeshould not interfere with the
analyticalcharacteristiclines of the elementsought.For example,the
Cr-K~ line of Cr tubes interfereswith the Mn-Ka (although the mag-
nitude of this interferencecan be greatly reducedby the use of alu-
minum filters), andthe L spectrumof the tungstentube interfereswith
the Ka lines of Ni, Cu, and Zn.
3. If only one tube is accessible,a generaluse tube with anodesmadeof
heavy metalssuch as Au, Pt or W should be preferred.Theseanodes
withstand high currents that produce a highly energeticcontinuous
spectrumthat excitesthe greatestpart of the fluorescentradiationfrom
most elements.One of thesetubesis desirablefor the analysisof ele-
ments with Z > 24 (Cr). Heavy metal anodes,however, are not as
effective exciters of light elementsbecausethey self-absorba high
proportionof the long-wavelengthspectrum.Another significant por-
tion of this spectrumis absorbedby the beryllium window, which is
especiallythick in thesetubesto withstandheat stressfrom the large
number of electrons scatteredby the anode surface. Rhodium or
molybdenumtubesmay be the compromisein thesesituations,espe-
cially when greaterexcitationof Sr and Rb is desired.
4. Tubeswith lighter anodessuch as chromium or titanium provide bet-

ter analyticalresultsfor light-element(Z < 20) determinationsbecause
the absorptionof the long-wavelengthspectrumby the anodeand the
thinner beryllium window is less severe.The fluorescentintensities
obtainedby thin-window chromium tubesfor light elementssuch as
AI and Si are usually five to six times higher than thoseobtainedwith
heavy metal anodesat the samevoltage.
5. Specialend-windowtubes witha rhodium anodehavebeendeveloped
which enablemore long-wavelengthradiation to be transmittedand
improve the sensitivity for elementsof low atomic numbers.These
tubes have short-anodeto sampledistance,very thin beryllium win-
dows protectedfrom heating,and require less rigorous cooling.
6. Tubes with very thin windows or windowless tubes also have been
designedfor analysisof very light elements(Z = 5-12). Thesetubes
are generally operatedat low voltagesand require special operation
and maintenanceprocedures(Baird, 1970).
A generalizedguide to primary tube selectionand operationfor various
elementsis given in Table 7-l.
Greaterexcitationefficienciescan be achievedby replacingthe x-ray tube
with an electrongun and exciting the sampledirectly (Kimoto et aI., 1966). In
this way relatively low tube currentsand voltagesmay be used, and light ele-
ments and ultra-light elements(with Z = 5-12) can be analyzedsuccessfully
(Poole, 1973; Strasheim& Brandt, 1970). The samplemay be in any form usual
for x-ray spectrographybut must have a conductingsurfaceto remove surface
charges,and the beammust scanonly 1 cm2 of the surface.A refinementof this
techniqueis the electronprobe,in which a beamof electronsas narrow as 1 pm
is focusedon a specimenas a target. This combinesthe methodsof x-ray spec-
trographywith the techniquesof electronoptics (Castaing,1961). Full accounts
of the technique and discussion of its potential are given by Long (1977),
McKinley et al. (1966), and Reed(1975).
Table 7-1. Generalizedguide to primary tube selectionand operation(Williams, 1987).

Analyte elements Target Voltage
K spectra
o (8}-Ti (22) Cr 55 kV
V (23}-Co (27) W 55 kV
Ni (28}-Zn (30) Au 65 kV
Ga (31}-Y (39) Mo 100 kV
Zr (40}-V (92) Au 100kV
L spectra
Mo (42}-Cs (55) Cr 55 kV
Ba (56}-Hf (72) W 55 kV
Ta (73}-Re (75) Au 65 kV
Os (76}-V (92) Mo 100 kV
Interaction of X-Rays with Matter
Absorption
When x-rays encounterany form of matter they are progressively attenu-
atedas a result of a complexseriesof interactionswith atomsof the attenuating
medium.This attenuationis a consequence of absorption,scattering,and/ordif-
fraction phenomenaand its extentis a function of the compositionof the sample.
The intensity (I) of the radiation emergingfrom a homogeneoussubstanceof
thickness(f), and a linear absorptioncoefficient of p, is relatedto the intensity
(10) of the incident x-ray beamby the equation
1=10 exp (-p,f) [3]
By introducingthe density p of the absorbingmedium in this equationwe have
1= 10 exp [- (p,/p) pi] [4]
wherep,lp representsthe massabsorptioncoefficient and pi is the mass/unitarea

of the absorber.
The mass absorption coefficientfor any element or compound usually
increaseswith increasingwavelength,but the relationship is not continuous.
Insteadit is markedby a seriesof abrupt decreasesat eachof the wavelengths
correspondingto the absorptionedgesof the various characteristicspectra(K,
L), and the energy levels at which electronsare expelled. Betweenabsorption
edgesthe massabsorptioncoefficientscan be measuredor calculatedaccording
to the empirical relationship
[5]
where K =constantwith different value at each absorptionedge. Short wave-

lengths,therefore,are highly penetrating(hard), while long wavelengths(soft)
are easily absorbed.Valuesfor massabsorptioncoefficientsfor variouselements
can be found in severaltextbooksincluding White and Johnson(1972), Bertin
(1975), Williams (1987) and others.
For a sampleconsistingof a mixture of n elementsand for a particular
wavelength,the massabsorption coefficientcan be calculatedfrom the weight
fractions Cl> C2, ..• Cn) of its componentsand the correspondingindividual ele-
ment coefficients{(P,/P)l' (pJp)z, (P,/P)m} accordingto the equation
[6]
For example,the massabsorptioncoefficient of quartz for RdKa radiation (A =

0.0163nm) and respectivemass absorptioncoefficientsfor Si =5 and 0 =1 may
be calculatedas follows:
The weight fractions for Si and 0 in quartzare: CSi =28/60 =0.467 and Co =2
x 16/60 =0.533, respectively.Therefore,
(,u/P)Si02 =(0.467 x 5) + (0.533 x 1) '" 2.9. [7]
where 0.467 and 0.533 are the weight fractions for Si (28/60) and0 (16/60 in
quartz), respectively.
Scattering
Although absorptionaccountsfor a major portion of the attenuationof
incident x-rays, somescatteringalso may occur, especiallyin the caseof short
wavelengthx-rays and low atomic number elements.Both characteristicand
continuousradiation from the primary tube can be scatteredby the sampleinto
the spectrometer.Much of it will be coherently scatteredwithout change in
wavelength,while a smaller portion will be incoherentlyscatteredwith small
increasesin wavelength.A spectraltrace of the backgroundin thesecasesmay
show peaksfrom the scatteredlines of the target elementand an asymmetric
broadbandin the regionof white radiation,cuttingoff sharplyon the shortwave-
length side and gently on the other. The massabsorptioncoefficients,u/p report-
ed in the literature usually encompassboth true absorptionand scatteringphe-
nomena.
Enhancement
X-rays interacting with matter undergo not only attenuation but also
enhancementdue to secondaryor tertiary fluorescencex-rays producedby the
matrix that excite elementswith electronsat lower energylevels. For example,
in a sample containing Fe, Zn, and Cu, the FeKa spectraare expectedto be
enhanceddue to secondaryand tertiary fluorescenceof Zn and Cu becausethe
FeK absorptionedge(0.174 nm) is greaterthan thoseof Zn and Cu, and there-
fore both of them are capableof exciting additional FeK radiation. Fortunately,
enhancementeffects are usually much less severethan absorptioneffects but
invariably more complex and much more difficult to correct. Norrish and
Chappell (1977) note that in most minerals and rock analysesthe effects of
enhancement canbe overcomeby judiciouschoiceof calibrationstandardsin the
compositionalrangeof the analyticalsamples.
Dispersion(Diffraction)
X-rays traveling along a common path also may exhibit interference
effects.They may be either reinforcedin intensity if they are in phase witheach
other or reducedin intensity if they are not. This is essentiallya scatteringphe-
nomenonof incident x-rays in all directions by matter. Since the atoms are
arrangedperiodically on alattice of a substance,the rays scatteredby them have
definite phaserelationships.In most directionsof scatteringthe rays are not in
phase(destructiveinterference)and they canceleachother. In a few directions,
however,constructiveinterferencetakesplaceand diffracted beamsare formed.
Diffraction occursonly at thoseparticularanglesof incidenceat which the wave-
length of the x-rays is of the sameorder of magnitudeas the repeat distance
betweenthe scatteringcenters(atoms)accordingto Bragg's law
n'A. = 2d sin e [8]
where 'A. = wavelengthof diffracted rays, d = distancebetweenatom planes,e=

angle of incident beam,and n = order.
In XRFS the diffraction phenomenonhas a very practical application in
that crystalsof known interplanarspacing(d) can be usedto dispersethe radia-
tion emitted by an excited sample,thus permitting the isolation of selectedana-
lytical wavelengthsfrom the rest of the emitted spectrum.This is the principle
on which the wavelength-dispersive spectrometersare based.
Detectionand Measurement
X-rays emitted froman excited sampleare detectedand measuredby spe-
cial detectorswhosefunction is to convertthe energyof individual x-ray photons
into pu~ses of electricalenergy,which canbe countedas a function of time.,There
are different typesof x-ray detectors,all of them dependingbasicallyon the abil-
ity of x-rays to ionize atomsby photoelectricprocesses.The most typically used
include photographicfilm types, gas-filled detectors,scintillation counters,and
semiconductor(solid-state)detectors.The photographicdetectionin which the
intensity of the incident x-rays is basedon the degreeof blackeningof a photo-
graphicfilm are no longer usedin quantitativex-ray spectrometry.
In gas-filled detectors,x-ray photonsenter the countervia an electrically
conductive thin window (1-6 .urn) generally made of metal-coatedMylar,
polypropylene,or polycarbonateand causethe ionization of inert gasessuch as
Xe or Ar under a strong electrical field producedby a high direct current (DC)
voltage applied acrossa cathodeand anode.Heavy gasesare preferredbecause
they have large absorptioncoefficients. Electrons producedby the ionization
processmigrate to the anode, where they produce electric pulses, which are
amplified and fed to electroniccountingcircuits. If the meannumberof ion pairs
producedand the size of the electronicoutput pulseis proportionalto the energy
of radiation, which is usually accomplishedat ionization potential <1500 V, the
detectoris said to operatein the proportionalregion. Detectorsdesigned tooper-
ate under theseconditions are called gas proportionaldetectors.At higher ion-
ization potentials(> 1500V) the proportionalityof responseis lost, and the mag-
nitude of electronscollectedbecomesvirtually independentof the energyof the
detectedx-ray photons.Detectorsof this type are called Geigercountersand are
no longer used in quantitativex-ray spectroscopy.
Proportionaldetectorssensitiveto soft x-rays with wavelengths(0.5-2.0
nm) utilize windows so thin that they allow a small portion of the gas to diffuse
out of the detector.To compensatefor this leakage,a constantstreamof gas is
continuouslysuppliedto the detectorwhich, for this reason,is referredto as gas-
flow proportionalcounter.The time interval following the arrival of a pulsedur-
ing which the detectoris not responsiveto further pulsesis called dead time of
the detection/countingsystemand it is usually in the range of 1 to 2.us. This is
due to the slow rate of migration of the positive ions away from the anode,which
lowers the field strengthin the vicinity of the anode,and therefore,inhibits the
initiation of new pulses and the responseto absorptionof new x-ray photons.
Therefore,the net countingtime during which the detectoris active and able to
record pulsesis somewhatshorter than the actual elapsedtime, by an amount
equivalentto the product of the number of counts recordedand the dead time
betweenpulses. To optimize the operationof the detector at the proportional
region and reducedead time,a quenchgas,suchas P-lO (90% Ar, 10% methane)
is commonlyused,which maintainsa stableflow of positive ions away from the
anode.Also, most modemspectrometersare equippedwith gasdensity stabiliz-
ers which preventgasamplification drifts.
If the energyof the x-ray photonsenteringthe detectoris greaterthan the
absorptionedge energy of the filling gas, characteristicpeakswill be observed
attributedto fluorescenceof the gasatomsin the counter.Theseare calledescape
peaksor pulsesand have energyslightly lower than that of the main peakbeing
analyzed.Sincethe ArK absorptionedgeis 3.20 keY, KKa (3.31 keY) and CaKa
(3.69 keY) usually produce characteristicallystrong escapepeaks in Ar-filled
detectors.These peaks can usuallybe removedby pulse height selection,but
may causeinterferencesin someanalyses.
The scintillation detectorsconsistof a phosphor(usually a sodium iodide
crystal dopedwith Tl), which convertsthe energyof incident x-ray photonsinto
a seriesof light pulses,and a photomultiplierwhich convertsthe light pulse into
voltagepulsesthat can be amplified and countedin a fashion similar to that of a
gas-filled proportionaldetector.Sodium iodide is usedbecauseI is a heavy ele-
ment with high absorptioncoefficient. The deadtime of scintillation countersis
slightly lessthan that of a gasproportionalcounter,but the resolutionwith regard
to photon energyis poorer becausethe processof energytransferis inefficient.
Nevertheless,scintillation countersare often able to handle higher count rates
than gas-filled detectorsand their use is favored in cases wherehigh count rates
are more importantthan detectorresolution.
The optimum wavelengthsensitivity range for scintillation detectorsis
about0.02 to 0.20 nm comparedto the >0.15 nm of the gas-filled detectors.This
makesthe scintillation detectorsmore efficient at radiation wavelengths<0.20
nm and the gas-filled detectorsmore efficient at wavelength>0.20 nm (cutoff
elementsMn or Cr). Therefore,it is not unusualto interchangethe two typesof
detectorsdependingon the wavelengthrangeof interestor combinethem, with
the flow detectormountedin front of the scintillation detectorthroughan auxil-
iary collimator. This doesnot improve the resolutionof the tandemsystem,how-
ever, sinceit is essentiallythat of the scintillation counter.Scintillation detectors
also may produceescapepeak phenomenabut only by very high-energyx-ray
photons.
Both scintillation and proportionalcountersproducesmall pulsesthat can-
not be measureduntil they havebeenamplified. This is achievedby adjustingthe
voltage on the detectorto give maximum gain, or amplification and by using a
linear amplifier. Best resultsare obtainedwith the lowest possiblevoltageon the
detector alongwith a high amplifier gain, i.e., attenuation.Settingsmust be con-
sistentwith maximumamplificationand minimal noisefrom the amplifier. There
are severalother practical problemsassociatedwith the use of scintillation and
gas-flow proportionalcounters,which are discussedlater.
X·RAY FLUORESCENCESPECTROSCOPY 171
Solid-statedetectorsor semiconductorsdevelopedin the 1960s,like gas-

filled detectors,dependon the photoelectricprocessinitiated when an x-ray pho-
ton enters the detection system which producespulses proportional to the
adsorbedx-ray energy.However, this processtakesplace in a solid insteadof a
gaseousmedium and the ionization produces"electron-hole"pairs rather than
"electron-ion" pairs. The most widely used solid-statedetectorin x-ray spec-
troscopyis the lithium-drifted Si detectoror Si(Li).
Pure Si is an intrinsic semiconductorwhich has fully occupiedvalence
bandsseparatedby energygapsfrom their higherenergyconductionbands.The
energyof incident x-ray photonsabsorbedby intrinsic semiconductorsis trans-
ferred by photoelectroninteractionsto valenceelectronswhich are excited and
jump acrossthe bandgap to the conductionband.The producedconductionband
electronsact as negativechargecarrierswhile the electronholesgeneratedin the
valence band act as positive charge carriers. By applying a suitable voltage
acrosssuch a semiconductor,the ion pair chargescan be sweptout and collect-
ed by an electrodeset (anodeand cathode).Since the numberof electron-hole
pairs producedare proportionalto the energyof the incident x-ray photons,the
magnitudeof the chargescollectedalso areproportionalto the x-ray photonener-
gy andthus,the semiconductoris inherentlya true proportionaldetector.To com-
pensatefor B impurities in Si that reduceits efficiency as an intrinsic semicon-
ductor, a certainamountof Li is diffused throughthe Si that producesa detector
with high inherentsensitivity.
The pulsescollectedat the electrodeare amplified and passedto the mul-
tichannelanalyzer,which sortsthe pulseselectronicallyaccordingto their ampli-
tudesinto different channelsand countsthe numberof pulsescollectedin each
channel.Becauseof their superiorenergy resolutionover other counterssolid-
state detectorsoffer practicaladvantagesfor numerousanalytical applications.
They do not require a dispersingcrystal systemor the goniometerto rotate at
specified Bragg diffraction anglesand thereforerequire much less space;they
can sample the complete spectrum of emitted x-ray energiessimultaneously
avoiding the time consumingnecessityfor sequentialscansat different energy
(wavelength)settings; they can be placed much closer to the emitting x-ray
source(sample)without requiring intensity-reducingcollimator, and therefore,
they providesubstantialefficiency increasein x-ray energyconversion.The most
outstandingattribute of thesedetectorswhen used with energy dispersivesys-
tems is the speedof multielementanalyses,which often can be completedwith
adequateprecisionin secondsrather than minutesor even hours requiredby the
wavelengthdispersivesystems.
On the other hand, solid-statedetectorshave some disadvantageswhich
should be consideredin order to ensuresatisfactoryoperation.They mustbe
maintainedconstantlyat low temperature(usually immersedin a dewarof liquid
N2) which requiresregularrefilling to preventexcessiveLi diffusion and serious
degradationof their performance.The detectorcan occasionallybe brought to
room temperaturefor a short period provided the voltage is switched off. The
surfaceof the detectoris very sensitiveto contaminationso that the housingis
kept permanentlyundervacuumwhich requiresthicker windows that reducethe
bestpossibletransmissionand therefore,detectionand resolutionof low energy

x-rays.
Pulse-HeightSelection
All the detectors(gas-filled, scintillation, solid-state)are proportional in
the sensethat they producepulseswith amplitudeproportionalto the energyof
the incidentx-rays. After amplification,thesepulsesare directedto electricalcir-
cuits which are used to distinguishbetweenpulsesof different amplitude,and
therefore,x-rays with different energies(wavelengths).Thesecircuits are called
pulse-heightanalyzersor pulse-heightselectors.
The rangeof pulse heightsthat passesthe selectoris called a window, or
channel,and the lower thresholdis termedthe baseline,or lower level. Both win-
dow and baselinemay be adjusted,and the signal that passescan be fed to a
counting device or to a rate meter. Pulse-heightselectioncan thus improve the
peak/backgroundratio by rejecting the harmonics, much of the instrumental
background"noise," and often but not always,the nearbyand overlappingpeaks
due to otherelements,including an escapepeak.It is particularlynecessarywhen
the scintillation counter is used for detecting relatively soft radiation (A. =
0.15-0.40nm) and when the gas-flow counteris usedfor detectionof radiation
from the light elements(A. > 0.45 nm). Theremay be a problemof shift in pulse
height with the latter. This may be due to: (i) drift in the voltage applied to the
detector;(ii) somevariation in the density of the gascausedby variationsin the
temperatureor pressureof the gas; or (iii) the incident intensity being so great
that photonsare entering the detector beforethe precedingphotonshave been
resolved,i.e., within the deadtime of the counter.
With an energy-dispersivespectrograph,a multichannelanalyzeris used
with at least 1024 channels.This receivesthe amplified output from the silicon
(lithium) detectorand sortsthe pulsesby energyinto the channels.When a suf-
ficiently large count (-UP) has beenaccumulatedin the channelcorresponding
to the highestintensity,the countsin eachchannelare transmittedto the recorder,
oscilloscope,or digital printer so that the intensity over the entire spectrummay
be displayedsimultaneously.Correctionsmay be appliedelectronicallyfor dead
time, background,"pulse pileup," and spectralline interferenceby the computer
associatedwith the spectrograph.Modem computershave sufficient capacityto
make appropriatecorrectionsfor absorptionand enhancementby the sample
matrix.
To monitor the rate at which pulsesare collectedby the detectorsystema
ratemeteror a scaler-timersystemis used.The ratemeteris essentiallyan inte-
grating (averaging)device with analoguevoltage output that varies proportion-
ally with the count rate. The finite time interval over which the averagingof
count rate occursis the time constantof the systemwhich typically rangesfrom
0.1 to 10 s. Ratemeteroutputsare usually directedto a chart-recorderwhich is
convenientfor qualitative or quantitativeanalysis.On a scaler-timersystemthe
amplified detectorpulsesare displayedon a digital device(scaler)and electron-
ically countedby a timer over finite time intervals. The analystcan then select
the counting mode by either fixing the time constant(presenttime mode) and
measurethe counts accumulated,or by fixing the total number of countsto be
accumulated(presetcount mode)and measurethe time interval it takesto collect
them.
Counting Efficiency
The overall efficiency of the counterelectronicssystemin detectinginci-
dent x-ray photonsis the productof absorptionefficiency and detectionefficien-
cy. All countershave a thin window (mica, Be, Mylar) throughwhich the x-rays
passbeforethey reachand are absorbedby the counterchamber.The absorption
efficiency, therefore,can be expressedas
[9]
whereIw = fraction of x-rays absorbed by thewindow, andIe = fraction of x-rays

absorbedby the counter.The detectionefficiency (Eder) on the other hand is (1 -
11), where11 fractional countinglossesoccurreddue to unresolvedpulses(espe-
cially at high counting rates and long-time constants).Therefore, the overall
countingefficiency (E) is given by the equation
[10]
Since Edet for most countersis nearly 100%, E mainly dependson Eabswhich is
usually low for very short wavelengths(high energy)that passthrough the win-
dow and the counter without being absorbed,and high for long wavelengths
becauseof the increasedabsorptionby the window. Thus, the bestcountingeffi-
ciency is attainedwhen fw is minimum and fe is maximum. For example,the
counting efficiency of a scintillation counter for wavelengths0.02 to 0.20 nm
usually exceeds90%. For longer wavelengths,however, and with a beryllium
window ofO.127-nmthicknessabout 25% of the Ca (0.336 nm)and nearly 95%
of the Si (0.713 nm) radiation is lost. On the other hand, the counting efficiency
of a gas-flow proportionalcounterusing a P-lO (90% Ar-lO% methane)mixture
and a Mylar window 0.00625mm thick, is nearly 100% for CaKa and KKa radi-
ations, but drops to 60% for Fe (0.194 nm) and 30% for AI (0.834 nm).
INSTRUMENTATION
There are two main types of XRFS's. The wavelength-dispersivespec-

trometers(Fig. 7-1a), which utilize the diffracting propertiesof a single crystal
to dispersethe wavelengthsof the x-ray spectrumemitted from the sample,and
the energy-dispersivespectrometers (Fig. 7-1b) that use electronicsorting of the
pulsesof different amplitudeproducedby different energyx-ray photons.In the
latter, a multichannelpulse height analyzermeasuresthe pulse height distribu-
tion and subsequentlydisplaysthe spectrum.
x-ray tube
spectrometer
circle
\J
_- secondary
x-rays
v \ AI and A2 ' \ ::i( sample
collimator
detector
,
, A2
......---~~\
\_ ) detector
(a) Wavelength-dispersivespectrometer
EJ
A, counts- - - -
A2coWlts-- - - U
MeA ____ nf-- x-__
Si (Li)
detector
secondary
ray-s--...y,'Y
Aland A2
(b) Energy-dispersivespectrometer
Fig. 7-1. X-ray fluorescencespectrometers-inthis example,elements1 and 2 in the sampleemit

characteristicwavelengthsA.I and A.2' Thesewavelengthsare separatelymeasuredby crystal dif-
fraction in (a) or by pulse-heightanalysisin (b) (whereMeA =multichannelanalyzer).
Wavelength-Dispersive
Spectrometers
Thesespectrometersmay be built either as single-channelsequentialor

multichannel simultaneousspectrometers.Single-channelunits have only one
counterwhich is mechanicallycoupledwith the analyzingcrystal. Thus, when
the crystal is set at a particularBragg angle8, the counteris automaticallyset at
the correspondingangle 28. The various spectrallines of different elementsare
measuredsequentiallyby moving the counter (goniometer)from line to line
either manuallyor by a mechanicaldrive. In automaticsingle-channelspectrom-
eters,the angularpositions28" 282, ..., at which lines A." A.2, ... for the analyzed
elementswill be reflectedare presetor programmed.The counter movesrapidly
from position to position, and accumulatesenoughcounts for each elementto
makean accurateintensity measurement. Thesemeasurements are recordedon a
printer or sent to a computerfor conversionto concentrationsof the elements
involved. Multichannel simultaneousspectrometersare most efficient where a
predeterminedsuite of elementsis to be measuredrepetitively over a long peri-
od of time. They usually haveas many channels(crystalsand counters)as there
are spectrallines (elements)to be measured(as many as 30). For eachchannela

crystal and counterare fixed at the correct angularposition to measurea select-
ed spectralline. All the analyzingchannelsreceive the samefluorescentradia-
tion from the sample,while one nondispersivecontrol channelreceivesfluores-
cent radiation directly from a standardand servesto monitor the output of the x-
ray tube. Since the data are acquiredsimultaneouslya large numberof samples
can be analyzedin a short time.
The wavelengthdispersivespectrometers(WDS) could utilize either a flat
crystal or a curved (focusing) crystal for x-ray dispersion.The flat crystal is
mountedon the central arm of a rotating goniometerso that the Bragg angle 8
can be varied by simple rotation of the crystal mount (Fig. 7-2a). Beam diver-
genceis limited by primary and secondarycollimators consistingof a seriesof
parallel metal plateswhich are positionedin front of the x-ray sourcebetween
the sampleand/or the crystal detector.X-rays parallel to the platespassthrough
the collimator while those diverging significantly are absorbedby the plates.
This reduceddivergenceimprovesthe resolutionof the spectrometerbut source
x-ray intensity lossesin incident and dispersedradiation are unavoidable.The
closer the spacingbetweenthe collimator platesthe better the resolutionbut at
the expenseof greaterintensity losses.Therefore,a variety of interchangeable
coarse-and fine-spacedcollimators are employedto achievethe best analytical
conditiol).s. Fluorescentx-rays incident on the flat crystal are diffracted into the
counterby lattice planesparallel tothe crystal surface.Sinceno focusingoccurs,
the beamdispersedby the crystal is relatively broad, which necessitatesa rela-
tively wide counterreceiving slit.
In the curved crystal spectrometer(Fig. 7-2b) fluorescentx-rays incident
on the crystal are focusedand diffracted on a slit (in place of the collimator) in
front of the detector. The analyzing crystal has its reflecting planesbent to a
radius 2R equal to the diameterof the focusing (Rowland) circle and its surface
curved to a radius R equal to the radius of the Rowland circle. The essential
requirementsof this geometricconfigurationare for the x-ray source,the surface
of the dispersingcrystal, and the entranceslit to the detectorto all lie on the cir-
cumferenceof the circle so that
D = 2R sin 8 [11]
where8 = angle of incident tothe crystal x-ray beamand D = distancefrom the

emitting slit (ES) or the receiving slit (RS) to the crystal. This distanceis kept
equalby rotating both crystal and counterin sucha way that the counterrotation
angle is always twice as large as the crystal rotation angle. A variety of inter-
changeableslits is usedto accommodatethe proper resolution-intensityrequire-
ments.
In both flat and curve crystal configurationsthe angle 28 at which a par-
ticular wavelengthis reflected dependsonly on the d-spacingof the analyzing
crystal. Since the longest wavelengthsthat can be diffracted are equal to 2d,
small d-spacingcrystals are usedfor short wavelengths(elementswith high Z)
and large d-spacingcrystals are used for long wavelengths(low Z elements).
Someof the most commonlyusedanalyzingcrystalsare listed in Table 7-2.
sample
(a) Flat-crystalgeometry
o
sample
,
protective
R \ cover
I
I o
I
I
I I
\ I
\ .
focusmg I
I
,,
\ circle I
\ , I
"
... _--- ..... -"'" "
~
detector
(b) Curvedcrystal geometry
Fig. 7-2. WavelengthdispersiveXRFs with flat-crystal (a) and curved-crystal(b) geometry(ES =

emitting slit and RS =receivingslit).
Energy DispersiveSpectrometers
Thesespectrometershave becomewidely used since the late 1960s fol-

lowing advancesin electronicand digital processingtechnologythat have made
them convenientin operationand maintenanceand practicalfor routine spectro-
scopicdeterminations.The essentialpartsof thesespectrometersare: (i) a solid-
statedetector,usually a silicon(lithium) counterfitted with a field effect transis-
Table 7-2. Characteristicsof commonly useddispersioncrystals(Williams, 1987).

LowestZ detectable
(max e= 70°)
Crystal Abbreviation (hkl) 2d(A) K L Efficiency
Topaz (303) 2.712 23(V) 59(Pr) Average
Lithium fluoride LiF (220) 2.848 23(V) 57(La) High
Lithium fluoride LiF (200) 4.028 19(K) 49(In) Intense
Germanium Ge (111) 6.532 16(S) 40(Zr) Average
Quartz (1011) 6.686 15(P) 40(Zr) High
Quartz (1010) 8.50 14(Si) 37(Rb) Average
Pentaerythritol PET (002) 8.742 14(Si) 37(Rb) High
Ethylenediaine d-tartrate EDDT (020) 8.804 14(Si) 37(Rb) Average
Ammonium hydrogen ADP (101) 10.648 12(Mg) 33(As) Low
phosphate
Mica (muscovite) (002) 19.8 9(F) 26(Fe) Low
Potassiumacid phthalate KAP (1010) 26.6 8(0) 23(V) Average
Rubidium acid phthalate RAP, RbAP (1010) 26.1 8(0) 23(V) Good
Thallium acid phthalate TAP, TIAP (1010) 25.75 8(0) 23(V) High
Lead steratedecanoate PbSD 100 5(B) 20(Ca) Average
Lead melissate 160 4(Be) 20(Ca) Average
tor (FET) preamplifierboth cooled by liquid Nz, and (ii) a multichannelanalyz-

er (MeA). They are mechanicallysimple systemsbecausethey do not needthe
analyzing crystal required by the wavelengthspectrometers,but electronically
complexbecauseof the presenceof the MeA.
The silicon(lithium) detectorcollects the chargesproducedfrom the inter-
action of the emittedx-ray photonswith the drifted Si and the preamplifierinte-
gratesthem and converts them to low voltage pulseswith amplitudespropor-
tional to the energyof the emittedx -ray photons.Thesepulsesare amplified, fil-
tered for noise and convertedto numerical values (digitized) using a seriesof
amplifiers, single channelanalyzersand analog-digitalconvertersbefore being
directedto the MeA.
The MeA measuresand sorts the digitized pulsesinto a seriesof comput-
er memory registersor channels.Each channel representsa particular energy
interval of the x-ray spectrumin which correspondingpulsesare collected and
counted.Typical MeA's have1024channelsof lO-eV incrementsspanningx-ray
spectrain the rangeof 0 to 10.24keV (0.12 nm), which encompassalmostall of
the characteristicx-ray lines routinely used in XRF analysis.A timer, which is
usually a "live time" clock, controlsthe time intervalsover which countsare col-
lected and providesautomaticdead-timecorrectionby stoppingthe clock when
the system is temporarily insensitive to processfurther pulses. The processed
data can be output in a visual display or a printer as countsper channelor as a
graphicimageof the completespectrum.
The utility of thesespectrometersis basedon the excellentenergyresolu-
tion of the Si(Li) detector, which is much better than any other proportional
detectorand the ability of the MeA to perform rapid pulse-heightanalysismuch
fasterthan a single channelwavelength spectrometer. The MeA can measurethe
intensitiesof all spectrallines emitted from the samplesimultaneouslyand in a
very short time (usually 1-2 min), unlesselementsof very low concentrationsare
to be determined.
Advantages/Disadvantages
of Wavelength-Dispersive
Spectrometers
and Energy-DispersiveSpectrometers
One of the major advantagesof energy-dispersivespectrometers(EDS)

spectrometersis their speedand conveniencewith which they can be used to
completecomplexmultielementanalysisevenby the most inexperiencedopera-
tors. The absenceof a dispersingcrystal eliminatesproblems associatedwith
optical geometricrequirementsand crystal contaminationas well as errors aris-
ing from mechanicalwear and tear of the resettingcomponentsof the wave-
length-dispersivespectrometers (WDS), which ultimately lead to poor analytical
precision. In addition, since countsfor all elementsare accumulatedduring the
samemeasurement cycle, they can be obtainedfrom the samespot or areaof the
sample.The sequentialdeterminationof successiveelementsby WDS requires
extensivebombardmentof the target sampleareawhich may damageor create
diffusion gradientsin thermally sensitivesamples,thus causinga serioussource
of errorsin the intensity measurement.Alternatively, changingthe targetsample
areamay causesimilar errorsin nonhomogeneous samples, and using low-beam
currentsmay lead to unacceptablylow x-ray intensities.
The intensity of the fluorescentx-rays emittedfrom the sampleis general-
ly reducedin WDS before it reachesthe counterdue to the presenceof the col-
limator(s)and the inefficient diffraction ofthe analy'zingcrystal.Theselossesare
eliminated in EDS so that the radiation entering the counter is quite intense.
Alternatively, low-power x-ray tubesor low-intensity radioactivesourcescan be
usedto excite the sample.This low-power equipmenthas provided the impetus
behindthe developmentof miniature tubesand various types of simple portable
spectrometers of limited capability but useful in ore researchand environmental
investigations.
The resolution of EDS is better for short wavelengthsand high-energy
radiationwhile that of WDS is more efficient for long wavelengths(Ka lines of
elementslighter than Kr with Z = 36) and low-energyx-rays. This often yields
much betterimagesand more detailedresolutionat low beamcurrentswith EDS
and completelyinadequateresolutionwith WDS.
On the other hand, there are somepractical disadvantages associatedwith
EDS. Although the energy resolution of the silicon(lithium) detector is about
three times betterthan the gas-filled proportionalcounteralone, the presenceof
the dispersingcrystal and the focusing of the dispersedx-ray beamprovidesan
overall better resolution in WDS. This means that overlapping interferences
betweenadjacentspectralemissionsare more pronouncedand more difficult to
correctwith EDS.
Furthermore,WDS generallyhave betterpeakto backgroundratios, espe-
cially for low concentrationsand for light elementsbeyondthe effective work-
ing rangeof the EDS detector.Becauseincreasedelectronicnoisecontributesto
higher backgroundsin EDS, their ability to detect and measurelow concentra-
tions especially of light elements with low-energy spectra is significantly
reduced.
Problems also exist in the electronic processingof the silicon(lithium)
detectorsignals.The proportionalityrequirementbetweenx-ray photonenergies
and pulse amplitudesis disturbedfor count rates exceeding15 000 counts per
second(cps), thus causingpoor performanceat evenmoderatelyhigt total count
rates.This disadvantagebecomesmore seriousconsideringthat wb ile the total
countof a WDS reflects x-raysof a single wavelength,the total count of an EDS
correspondsto pulsesof all sizescollectedat the sametime. For example,if the
radiation emitted from a sampleconsistingof two elementsA and B produces
count rates of 150 000 and 1 000 cps, respectively,their intensity is measured
incorrectly becausethe total count rate exceedsthe linear rangeof the counter-
electronicsystem.In order to correct the problem, a reductionof power in the
excitationsourceby a factor of 10 will reducethe Ka count rate of the B element
to only 100 cps, which in retrospectmay not be adequateto resolve it from the
background.Furthermore,becauseof the precisionrequiredin pulseshaping,fil-
tering, and discrimination,the processis relatively slow, requiring deadtimes in
the rangeof 10 to 20 f.1S which are about 10 times greaterthan thoseof WDS. At
high-count rates even the "live clock" device is inadequateto. compensatefor
pulse pile-up effects, which may result in the production of spuriouspulsesor
enhancement of backgroundintensities.
The needfor constant coolingof the detector-preamplifiersystemwith liq-
uid N2 is somewhatinconvenientand the detectormay be seriously damagedif
the dewar is accidentallyallowed to run out of liquid N2 while the EDS unit is
on.
EquipmentSelectionand Safety
In spite of the readily available descriptionsof XRF spectrometersand

their componentsin the literature, often it is difficult for a prospectivebuyer to
decideon the relative merits of commerciallyavailableequipmentand the types
that are best suited to the analyst'sneeds.Obviously the determinantfactor in
this caseis the type of applicationand the objectiveof the laboratory.For exam-
ple, a process controltype, which specializesin the laboratoryanalysisof only a
few elementson a relatively similar matrix, would require a relatively simple
spectrometertunedto the elementsof interest.In contrast,a generalpurposeana-
lytical laboratory which analyzesmany elementsin dissimilar matrices may
require a more sophisticatedspectrometer.Another important factor in equip-
ment selectionis the sampleload, which may rangefrom a few samplesper week
to a few hundredsamplesper day. Generally, the higher sample load makesa
high-speedXRFS desirable.This requirement,however,dependson the sample
type and the required preparation.For ready-to-runsamplesor those requiring
minimum preparation,a high-speed,simple-to-operateXRFS may be ideal. For
samplesrequiring elaboratepreparationsand computations,the advantageof the
high speed has essentially no effect on the total analytical time involved.
Variations in the type of XRF equipmentand sample preparationalso require
operatorswith different training. While a processor quality control spectrometer
can havean operatorwith generalknowledgein lab proceduresand limited x-ray
training, a versatilespectrometerin a generalpurposelaboratoryrequiresa tech-
nician highly trained in practical x-ray spectroscopy.
The choiceof an XRFS alsowill dependon spaceandotherfacilities avail-

able (samplepreparation,storage,electric power, cooling capacity,ventilation,
computerinterfacing capabilities, etc.). Most XRFS's use either a radioactive
sourceor an x-ray tube for excitation. A radioactivesourcedoes not require a
power supply, and thereforeit is smallerin size and more versatile.However, it
requiresadditional safety measuresbecauseit cannot be turned off. Also, effi-
cient excitation of only a small range of spectrallines representinga few ele-
mentscan be accomplishedwith the properchoiceof radioisotopes.X-ray tubes
also are availablein a wide rangeof voltage ratings,with the high voltage rating
tubesbeing usedwith WDS and the low voltagetubeswith EDS. The high-volt-
age tubesrequire a longer and generally more complex power supply and cool-
ing systemthat increasesthe size and the cost of the spectrometer.
Sincethe x-ray sourceis most efficient in exciting elementswith Z slight-
ly less than that of the target, severalmethodsare usedto modify the radiation
emittedby the sourceincluding different voltage settings,using other targets,or
utilizing variousfilters. Provisionsfor suchmodificationsshouldbe built into the
instrument. Improvised modifications often are not effective and may create
safetyhazards.
As discussedearlier, WDS provide better resolutionfor low atomic num-
ber elements.To overcomethis limitation EDS are frequently equippedwith
minicomputersthat are able to provide deconvolutionand peak-strippingcl!-pa-
bilities for resolvingoverlappingpeaks.The degreeof effectivenessof the meth-
ods dependson the concentrationof the elementsmeasuredand the particular
algorithm used. Since radiation of only a particular wavelength(only one ele-
ment) is measuredby anyonesettingof the crystal in WDS, while the entire x-
ray spectrumis countedat onceby EDS, processingthe pulses(deadtime) takes
much longer in the latter system.Therefore,for accuratemeasurements of small
concentrationsEDS will require very long times and since most of the counts
will be coming from the matrix they will provide a very low countingefficiency.
However, for large amountsof severalelementspresentin a sampleEDS can
measureall of themsimultaneouslyin a relatively short time.MultichannelWDS
also could measureseveralelementsat a time, but they are generally presetso
that it is difficult to changethe elementsmeasured.
The maintenancere.]uirementswill vary dependingon the type of spec-
trometerusedbecausethe WDS are mostly basedon mechanicalsystemswhile
the EDS on electronicsystems.Someof the relatively simple maintenancecan
be doneby the laboratorypersonnel,but other maintenancetasksare more com-
plex and require trained specialistsprovidedby the vendor.
Several systemsof recording data are available from the simplest rate-
meter-stripchart recordercombinationto the microcomputercontrolled signal
output servicesusing teleprintersor screendisplays. The microcomputersalso
are usedfor datamanipulation(storage,corrections)as well as calculationof the
final concentrations.Since there is no generalconsensuson the best procedure
for correcting interelementinterferences,severalsoftware programsare avail-
able for this purpose,eachone with ratherspecificapplications.Therefore,com-
patibility of the programwith the spectrometerand computertype and the range
of the elementsof interestis necessary.Becausemost of the spectrometerman-
ufacturersdo not make the computer hardware and softwareusedwith the spec-
trometer, compatibility problemsfor certain applicationsmay arise even using
computerperipheralsprovidedby the manufacturer.
Besidesdata manipulations,anotheradvantageof the computer-controlled
systemis that it can operateunattended,thus allowing that time to be usedfor
other tasks,such as samplepreparationand record keeping.The availability of
the automaticsamplechangerthat is able to handlemany samplesat a time con-
tributes to big time savingsespeciallyin laboratorieswhere large numbersof
similar samplesare processed.
Although it is well known that radiationcould causebumsand physiolog-
ical changesin the humanbody, it is humannatureto be at times carelessin the
presenceof danger.Available statisticsshow that thereis at leastone accidentper
200 x-ray installations.Unfortunately,many of theseaccidentsoccur to people
with considerableexperience.Since the effects of radiation exposureare not
observeduntil considerabletime haspassed,it is imperativethat peopleusing x-
ray equipmenthavethoroughinstructionson x-ray safety.The operatorof x-ray
apparatusis generally exposedto two dangers,electric shock and radiation
injury. Both of thesehazardscan be reducedto negligible proportionsby proper
equipmentdesign and reasonablecare on the part of the operator.All modem
instrumentshave factory installed safety interlocks, which prevent accidental
exposureto x-rays. Therefore,it is importantthat theseinterlocksare maintained
functional by the x-ray user.
X-ray fluorescencespectrometersrequire severalhundredvolts for their
detector power supplies and several thousandvolts for the x-ray generator.
Therefore,specialprecautionsmust be usedto preventelectricalaccidents.Most
of the currently availablex-ray spectrometersare shockproofedand havesever-
al electrical interlocks to prevent radiation leakage.However, they should be
inspectedregularly to be surethat they are in goodworking order. Portablecoun-
ters (radiation survey meters)are available for surveyingvarious areasaround
the equipment forpossibleradiationleaks. Film badgesshouldbe worn regular-
ly by people who spend significant amounts of time near x-ray equipment.
Governmentregulationsregardingradiation safety are very strict. More infor-
mation aboutx-ray safetyregulationscan be found in specialgovernmentpubli-
cationswhile a useful summaryabout radiation units, survey meters,tolerance
levels and reportedaccidentshas beenpublishedby Jenkinsand Haas(1973).
ANALYTICAL CAPABILITIES
QualitativeAnalysis
Wavelength-and energydispersivespectrometersare both well suitedfor

qualitativeanalysisof elementswith Z ~ 9 within their detectableranges.A vac-
uum path is necessaryfor detectionof elementswith Z :<;; 20. The detectionof F,
O2, N, and C is attainablewith the useof very thin counterwindows, while win-
dowlesscountersshouldbe usedfor detectionof B and Be (Henke,1965).
The procedureinvolves the collection and recordingof samplespectraat

the desired wavelengthor energy region under specified operatingconditions
and the identification of the elementspresentfrom the position of emitted spec-
tral lines and relative intensities.The emitted spectral linesare checkedagainst
availablecomprehensivetablesof characteristicspectra.This processis usually
fast, especiallyif the spectralsearchis computercontrolled or an EDS is used
(capableof recordingthe entire spectrumwithin a few minutes).Generallyx-ray
spectra are relatively simple to interpret, but positive element identification
should be basedon the presenceand appropria'eintensity of more than a single
characteristicpeak(at least Ka and K~ lines).
Detectionlimits are dependenton samplecompositionand analyzingcon-
ditions. Norrish and Chappell(1977) quote the following approximatedetection
limits for silicate minerals, rocks, and soils with counting times not exceeding
200 s: C and 0 - 1%, F - 0.2%, Na and Mg - 100 mg!kg, Al - 30 mg!kg, S - 8
mg!kg, Sc-Mo - 1 to 3 mg!kg, and I through U - 2 to 6 mg!kg. Optimum sensi-
tivity conditions for low concentrationelementscan be achievedby carefully
optimizing excitation and detection conditions and effective use of the pulse
amplitudediscriminatorso that the backgroundradiation is kept as low as possi-
ble. In most casesthe latter is accomplishedby automaticcompensationof the
ampiifier gain or by the use of the appropriateattenuatoror specific analyzing
crystal. Characteristiclines from the x-ray tube anodeandpossiblecontaminants
also could appearon the emittedspectrum.Furthermore,for variousreasons,the
angularposition of a spectralline may not coincide preciselywith that listed in
the tables. This could be due to slight misalignmentof the goniometer,slight
crystal imperfections,temperaturechanges[particularly sensitive is the penta-
erythritol (PET) crystal], detectordrifting, and elementalcoordinationand oxi-
dation stateeffects.
Although EDS are particularly well suitedto qualitativeanalysis,their res-
olution for wavelengths>0.08 nm are inferior to thoseof WDS. Therefore,weak
spectrallines tend to be obscuredby relatively high backgroundsor overlapfrom
adjacentstrongerlines. This reducesthe overall sensitivity of thesespectrome-
ters and requirescomputer-based stripping techniquesto improve the resolution
of obscuredor overlappingspectra.An additionalconvenienceoffered by many
EDS spectrometers is the automaticidentificationof spectrallines by a data-pro-
cessingsystemdisplayedon a video monitor. This utility further enhancesthe
speedof qualitative EDS analysis.
QuantitativeAnalysis
The basic quantitativepremiseof XRF analysisis that the intensity (f) of
radiation emitted from an infinitely thick sampleis proportionalto the concen-
tration (C) of the element(s)presentin that sample.For a particularelementA
then this relationshipcan be written as follows
[12]
where, CA = concentrationof elementA, fA = the intensity of the characteristic

spectralline for the element,and KA = proportionalitycontent.Theoretically,the
KA could be calculatedfrom calibration standardsand the relationshipcould be

used to calculateelementalconcentrationsof unknown samplesby measuring
emitted x-ray characteristicintensities.Unfortunately,the proportionality factor
KA dependson the massabsorptioncoefficient, the physical and chemicalstate
of the sampleanalyzed,and enhancementeffects arising from the interactionof
the x-rays with the massof the sample.All thesefactors, but especiallythe mass
absorption coefficient and the physical state of the sample, produce matrix
effects, which causedeparturesfrom linearity that affect the accuracyof com-
parativeanalysesbetweenunknownsand standards.
Assumingan infinitely thick sample,the intensity of the characteristicflu-
orescentradiation for a specific elementemergingat the samplesurfacecan be
more realistically approximatedby the equation
K· C· p. 10
h=----=---::-- [13]
(Ill seccp+ Ilz sec.v)
whereK = proportionalityconstant,C = weight fraction of the elementanalyzed

p = density of the sample,10 = intensity of the primary beam,III and Ilz = linear
absorptioncoefficients for the primary fluorescent radiation, respectively,and
seccp and seclJI = complementsof the incident and reflecting angles.Substituting
Al = 1l1/p and A z Il z/p for the primary and t1uorescentmassabsorptioncoeffi-
cients, respectivelyin Eq. [13] we have
[14]
Assumingthere are no major absorptionedgesbetweenthe effective excitation

energy,the tube spectrum,and the energyof the absorptionedge,Al = kAz, and
a spectrometergeometrywith <1> = 30° and'JI = 60° (as is usually the case),then
seccp= 1 and sec.v = 2 and Eq. [14] is simplified to
[15]
due to predominanceof A z over A l . Consideringnow both, sampleand calibra-

tion standard,Eq. [15] can be written as
[16]
where x and s representvalues for sampleand standard,respectively.Equation

[16] can be modified to
[17]
or to
[18]
if samplesand standardshave similar composition{A 2(x) '" A 2(s)}' This is a sim-
ple linear relationshipwhich is commonly used to determinethe first approxi-
matecompositionof the sample.
Mass absorptioncoefficientsfor all the constituentsof the samplecan be
calculated from published tables or determined experimentally (Haukka &
Thomas,1977; Norrish & Hutton, 1977;West et aI., 1974). For more than a few
componentsin a samplethe use of a computerprogramis necessaryto perform
thesecalculations(Gunn, 1976; Oertel, 1971a,b).
The effects due to the physical state of the sample or microabsorption
effects arise from sample heterogeneity.This is more of a problem in coarse-
grained samplesand can be minimized by very fine grinding. Dilution of the
samplewith a diluent of a similar absorptioncoefficientto that of the samplealso
can be helpful if the elementsof interestare not presentin very low concentra-
tions. Microabsorptioneffects in soil sampleswhich are primarily composedof
light elements(Si, AI) can be overcomesuccessfullyby dissolvingthe samplein
a borateflux, which when fused eliminatesa significant portion qf particle size
problemsand matrix variations. Other diluents include cellulose,ion exchange
resins,and quartzor heavyelementabsorberssuchas La or Bi. Powdersamples
rather than fused samplesare preferredfor measuringtrace elementsbecause
dilution may reduceinstrumentsensitivity below the detectionlimits. Plant sam-
plesdo not usually requiredilution becausetheir organicmatrix actsas a diluent.
Extensivegrinding of plant samplesis not recommendedeither, becauseit may
causemarked changesin the intensity of the radiation emitted from some ele-
ments.Evenwith the abovecompensations, however,matrix effectsare not com-
pletely eliminatedand require proper correctionsbefore the data are considered
quantitativelyreliable.
If matrix correctionsare not employed,the resultingerrors may be severe
but most often they are in the orderof a few p~rcentage points or less. For many
practical applications, especially in semiquantitative approaches,analytical
errors of this magnitudeare much smallerthan samplingand preparationerrors
and, therefore, they can be tolerated. In such approachesthe measuredx-ray
intensitiesof the elementof interestare comparedwith standardsof similar com-
position. The standardscan be referencenatural mineral, rock, or soil samples
with known composition,or artificial mixturesof AI 20 3, Si02, Fe203and MgO.
Standardsfor analysisof plant samplesusually employ a cellulosematrix.
Standardcalibrationcurvesmay be developedby using externalor internal
standardsor by spiking the unknown sample with the element of interest.
Reagentchemicalsof high purity and in dehydratedforms should be used for
standardpreparation.Elementsusedas internal standardsshouldhave a spectral
line and absorptionedge close to that of the analyte element(s).External stan-
dardsshouldbe homogeneousand should have a compositionas similar as pos-
sible to the sample being analyzed. Suitable standardsfor quantitative XRF
analysisof soil and plant materials can be obtainedfrom the U.S. Geological
Survey (Flanagan,1986; Parker, 1969; Stem, 1976), CanadianGeologicalSur-
vey (Abbey, 1980). National Institute of Standardsand Technology,and various
spectrochemicalcompanies.
For samplesand standardsof similar c( mposition, the backgroundcontri-

bution remainsrelatively constantand therefore,approximatecompositionsof
the elementspresentcan be determinedbasedon the linear calibration of the
intensity of the emitted radiation vs. the elementalconcentration(Eq. [18]).
Theseapproximateconcentrationscan then be correctedfor matrix effects and
convertedto actualconcentrationsusing appropriatemassabsorptioncoefficient
functions. For most soil samples,XRFS can be usedto analyzenearly all major
and minor elementswith the sum of the oxidesand volatiles being almost 100%.
Although most of the samplesanalyzedby XRFS are consideredinfinitely
thick (thicker than the penetrationdepthof most energeticradiations),it is pos-
sible to use infinitely thin samplesin which matrix effects are minimal and the
intensity of the fluorescentradiationis usually proportionalto the elementalcon-
centration.It has beencalculatedthat absorptioneffects in such samplesessen-
tially vanish when samplesare uniform with thickness.;0.35/IlP (Chung et aI.,
1974), while enhancementeffects are normally negligible. Infinitely thin sam-
ples can be preparedfrom aqueousor polymethacrylatesuspensions(Gunn,
1976), air dusts(Hammerleet aI., 1973), precipitates(Stone,1963; Watanabeet
aI., 1972) and in mixtures with chelatingion resins(Blount et aI., 1973; Carlton
& Russ,1976). The detectionlimits in such samplesalso are improved because
of the minimum backgroundcontribution of the very thin substrates.
Sourcesof Error in QuantitativeAnalysis

The most significant errors in quantitativeXRFS are associatedwith: (i)
the operator,(ii) the samplingand preparation,(iii) the instrument,(iv) counting
statistics,and (v) interelementmatrix effects.
OperatorErrors_Operatorerrorsare potentially the most seriousin x-ray
spectrometricanalysisbecausethey encompassa variety of sourcesincluding
selectionof instrumentalconditions,preparationof standardsand samples,oper-
ation of the instrument,and processingand interpretationof the analytical data.
Errors madein any of thesephasesof analysiscan havedisasterouseffectson the
final results.In spite of the automationand standardizationof the XRF analysis
in most modernlaboratories,a good understandingof eachphaseof the analyti-
cal process,function of each instrumentcomponent,and the necessityfor good
maintenanceand periodic performancechecksis essential.
Some preliminary steps that can help minimize operator errors include
evaluationsof compositionalrangesof the samples, identification of elementsto
be determinedwith expectedlevels of accuracyand precision,selectionof suit-
able samplepreparationtechniques,and appropriatecalibrationstandards.Based
on .this preliminary experimental work the proper analytical conditions and
instrumentsettingswill be selected(choice of characteristicspectralline to be
measuredfor eachelement,type of analyzingcrystal, excitation source,type of
detectorand pulse amplitudediscriminatorsetting).
Although as a generalrule of thumb, the Ka line is normally usedfor light
elements(Z .; 33), the La for elementswith Z between34 and 80 and the Ma for
heavy elements(Z > 80), possiblespectralinterferencesand overlapsshould be
considered,especially in trace-elementanalysis. For example, the strongest
BaLa line (0.2776nm) is normally avoidedin traceelementanalysisbecauseof

proximity to the TIKa line (0.2750nm), which is much strongerin soil and geo-
logical samples.Instead the Ba42 (0.2404 nm) is used, which is efficiently
excitedby a chromium x-raytube.
In WDS the selectionof the proper analyzingcrystal, primary collimator
spacing,and Bragganglesettingdependson the elementsto be analyzed.A pre-
cise spectrometercalibration using reliable standardsis accomplishedby slow
scanningacrossthe peakof a particularelementto locate its maximum intensi-
ty, or by scanningacrossthe peakat a uniform rate and taking.readingsat 80%
of the maximumintensityat eithersideof the peak.The peakmaximumcentroid
can then be takenas the position halfway betweenthe two readings.In comput-
er-controlledWDS spectrometersthe position of the peakmaxima is computed
automaticallyfrom a seriesof intensity measurementsacrossthe approximate
position of a characteristicspectral line, which may last only a few seconds.
Crystal selection becomessomewhatcomplicatedonly in overlapping ranges
(Table 7-2), where the better resolutionof the high angle rangesmay be com-
promisedby lower dispersingefficiency or longeranalyticaltime. Coarsercolli-
matorsare normally usedfor major elementsbecauseof higher peakintensities
but for traceelementanalysiswhere,betterresolutionis critical fme collimators
( narrow slits) are preferred.
If there is a choice betweendetectors,a gas flow proportional counter
shouldbe usedfor wavelengths>0.14 nm, while a scintillation counteris more
suitablefor wavelengths<0.30 nm. For the overlappingrange (0.14-0.30nm)
the gas flow proportional counter may provide somewhatbetter resolution or
both detectorsin tandemcan be usedto enhancethe intensity of weak spectral
lines.
In selectingthe x-ray generatorand tube voltageand currentthe goal is to
achieveefficient excitationof the desiredelementsand optimumemissioninten-
sities (high peakto backgroundratios). Excessivelyhigh count ratesshould be
avoidedbecauseof increaseddeadtime, amplitudeshift problems,and unneces-
sary overloadof the x-ray tubes.Becausex-ray tube interchangeis not practical
in most cases,the selectionof a single anodefor the determinationof a certain
group of elementsis usually madebeforethe purchaseof the XRFS. For exam-
ple, for major elementanalysisin soil or geologicalsamples(usually elements
with Z ~ 26) a chromium anode is sufficient. For trace element analysis,
however,a rhodium, molybdenum,or gold tube may be necessary.Tube inter-
changes,wherenecessary,shouldbe kept to a minimumbecausethey deteriorate
the life of the tube and enhancechancesfor contamination.Alternatively, multi-
ple-anodetubes which are commercially available for some XRFS's may be
selected.
The operatoralso have to establishthe appropriatesettingsfor detector
voltage,amplifier gain, and discriminatorbaseline and window width for each
spectrum.Most of thesesettingsare optimized experimentally,with the most
commonprocedurebeing to selectand thereaftermaintainconstantthe detector
voltage,which providesan optimumrangeof amplituderesolutionandminimum
count-ratedependentpeak shifts. Fine adjustmentsof the amplifier gain or the
window baseline/widthcan then be made for each characteristicspectrumto
achievethe desiredamplitudedistribution and resolution.In most EDS systems,

a numberof optionsfor automaticsettingsare availabledependingon the spec-
tra being analyzed.
Sample RepresentationAnd PreparationErrors_ As is the case for

every analysisevery effort shouldbe madeso that the small subsampleon which
XRF analysisis performedis selectedand preparedto be truly representativeof
the whole analyticalsample.This is especiallycritical for XRF analysisbecause
evenin relatively largesamples(a few gramsin size) the depthpenetrationof the
primary x-ray beamis only a few micrometers,thus making the total sampleana-
lyzed usually less than 0.1 cm3. Before preparingthe samplefor XRF analysis
the analystmust considerand carefully selectthe form of sampleand the prepa-
ration techniquemost appropriateto each project. Since physical or chemical
heterogeneityof standardsor samplesare potentialsourcesof error, the samples
haveto be preparedin an identical fashion. The most difficulties are encountered
with powderedsoil or geologicalsamplesbecauseof their variable particle size
range and chemical composition. Compressingvery finely ground specimens
into durablepellets may reducethe problems,but for the determinationof light
elements(Al, Si, Mg) fractionationto a seriesof size fractionsis recommended.
Liquid and thin-film samplesalso can be analyzedsatisfactorily,provided they
are stableunderthe radiationof the primary beamand a He path is availablefor
determinationof lighter elements.Since fine-grinding cannotcompletelyelimi-
nate particle-sizeeffects for major elementdeterminationin soil and geological
samples,homogenizingthe sampleby fusion with an appropriateflux and cast-
ing the melt into a glassdisc has becomea commonpractice. Becauseof dilu-
tion effectsfrom the flux, however,pressedpowder pellets are preferredfor the
determinationof traceelementswith characteristicspectrallines <0.30 nm.
InstrumentalErrors.Theseare errorsassociatedwith the function of the

various componentsof the spectrometerand their short or long-term perfor-
mance stability. The detector/counterdead time system may require frequent
compensationor correction for count rate optimization of certain elements.
Furthermore,becausecomparativemeasurementsmade by XRF analysis are
normally performedat different times, they cannotalways be made under iden-
tical analytical conditions. Electronic as well as mechanicalsystemdrifts as a
function of time are commonand tend to increasewith the ageof the instrument,
regardlessof how well it is manufacturedor maintained.Instrumentalerrorsusu-
ally do not contributemore than 0.2% to the total analytical error, but constant
checksof the systemfrom time to time using the recommendedmanufacturers
standardsare recommendedto assureproperperformanceand reproducibility of
the instrument.
MeasurementAnd Statistical Errors. Generally errors in quantitative

XRFS are random and systematic.Systematicerrors are usually causedby dif-
ferencesin the physicalstate,and chemicalcompositionbetweenor within stan-
dardsand samples.They collectively causethe meanvalue of a seriesof mea-
surementsto be systematicallydifferent from the true analytical value. Methods
for treating theseerrors are discussedin the next section. Randomerrors arise
from sourcessuchas operatorinconsistency,electronicor mechanicalinstability

of the instrument,x-ray generationand emission,etc. The first two cannot be
physically controlled but x-ray generation,emissionand detectionerrorscan be
assessed so that their contribution to the final analytical result is kept at a mini-
mum.
Sincex-ray generationand detectionare randomratherthan uniform time-
dependentprocesses,the result of any intensity measurement will dependon the
time interval over which it was made.Therefore,the time intensity of an x-ray
line emitted from a samplecan only be precisely establishedby measurements
madeover an infinite period of time. Becausethis is not practical, the intensity
is estimatedfrom finite measurements and the measuringerror is evaluatedby
statisticaltechniques.Alternatively, if the operatorhas a preliminary idea on the
expectedcount rate, statisticalcalculationscan be madeto determinein advance
the countingconditionsrequiredto producea selectedprecisionlevel. Although
increasingcounting times may improve the statisticalreliability of the data, this
benefit may be outweighedby variation in analytical conditionsover long peri-
ods of time and increasedoperationand maintenancecosts.It is possibleto cal-
culate the optimum count rates and counting times that would improve analyti-
cal efficiency and determinefrom standardsof known compositionthe statistical
lower limits of detectionfor a given element.
CountingEfficiency-DetectionLimits
A few experimentalmeasurementsare adequatein order to calculatethe
counting time required to yield a desired level of random error or precision.
Alternatively, the analysiscan be performedin a fixed-count mode which pro-
videsthe countingtime necessaryto collecta presetnumberof counts,and there-
fore, a calculatedcount rate. Assuminga Gaussianx-ray distribution of N mea-
surementsabout a true meanvalue No, the relative standarddeviation E (count-
ing error) is given by
E (%) = 100/v'lV [19]
For example,if the count rate for a particularelementis 1000cps and the desired
precision(randomcountingerror) is less than 1%, the counting time requiredto
yield sucha precisionon the count rate could be calculatedfrom Eq. [19] as fol-
lows
N =(100/1)2= 10 000 counts. [20]
Therefore,the count time requiredto yield a 1% countingerror underthesecon-

ditions would be 10 000/1 000 or 10 s. Generally,an N-fold improvementin pre-
cision requiresa countingtime equal to n2, which makesit impract:calto indef-
initely improve precisionby extendingthe counting time. Also, since the count-
ing error dependson the numberof countscollected(Eq. [19]), it is more effi-
cient to operateon a fixed-count ratherthan a fixed-time mode althoughthe lat-
ter may be more convenientin certaincases.
In caseswherethe net intensity of a characteristicx-ray line requiresmore

thanone measurement (peak-background), the relativestandarddeviationt(net) of
the estimatednet count rate (Cp - Cb) is given by
[21]
whereCp andCb are count ratesfor peakand backgroundmeasurements, respec-

tively, and tp and tb are respectivecountingtimes. For a fixed time, tp =tb = Ttot/2
and Eq. [21] becomes
[22]
SubstitutingCp = Nplt and Cb =NJt whereNp andNb are countson the peakand
background,respectively,Eq. [21] becomes
[23]
From this, it is evidentthat precisionincreasesas the backgrounddecreases or as

the peakto backgroundratio becomesgreater.
For a fixed-count-mode,Np = Nb, Np = Cp • tp and Nb = Cb • tb, Ttot = tp +
tb, and tpltb = CJCp' Therefore,Eq. [21] becomes
[24]
A third methodof counting,called optimizedtime, is usedto achieveopti-

mal precisionby apportioningTtot betweenbackgroundand peak measurements
J
so that tpltb = (Cp - Cb)· For this mode,
1
E -
(net) - {i( rc; - ~) . [25]
At very low concentrationsCp approachesCb and countingtimes shouldbe

apportionedequally betweenpeak and back~ound measurements. Also, as the
Cp - Cb approacheszero the countingerror increases.This condition canbe used
to calculatethe statisticallower limit of detection,which is defined as the low-
est concentrationgeneratingCp significantly higher than Cb at Cb ± 20 or Cb ±
30 confidencelevels defined in termsof standarddeviation0 of the background
count rate. Therefore,at the 30 confidencelevel, the lowest limit of detection
(LLD) would occur when
Np - Nb =30 (Nb), [26]
or Cp - Cb = 3VCb/tb' [27]
Since Ttot =2tb, Eq. [27] becomes

Cp - Cb =3 • 2jCb/tb =6jCb/tb, [28]
and in concentrationterms, the lowest limit of detection
LLD = 6/m jCb/tb, [29]
wherem is the numberof countsper secondper unit of concentration(sensitivi-

ty) for the elementanalyzed.Equation[29] providesthe statistical lowerlimit of
detection,but not necessarilythe practical lower limit of determinationbecause
essentiallyat the statistical LLD the counting error is infinity (Cp - Cb = 0).
Therefore,for practical purposesthe practical lower limit of determinationis
usually taken to be about twice the statisticalLLD, which representsconcentra-
tions equivalentto about 60 of the backgroundcount.
The sensitivity (m) factor varies asa function of excitationefficiency, ana-
lyzing crystal, detectorresponse,matrix composition,etc., and it is usually bet-
ter in the intermediatewavelengthregion (0.1-0.3 nm). At shorterwavelengths
excitationis relatively inefficient due to higherbackgroundsand at longerwave-
lengthsabsorptioneffectsreducemeasurableintensities.It also shouldbe kept in
mind that limits of detectioncalculatedby this techniqueare basedonly on ran-
dom statisticalerrorsand do not take systematicerrors into account.Systematic
errors associatedwith nonlinear backgroundprofiles, which require multiple
backgroundmeasurements,and with interferencesfrom overlapping spectral
lines may have considerablygreatereffects on practical LLD than randomsta-
tistical errors.
Interferenceand Matrix Effects

Interferenceerrors in quantitativeXRF analysiscan be associatedeither
with the backgroundor other characteristicemissionline(s). Backgroundinter-
ferencecan be reducedconsiderablyby carefully adjustingthe pulse amplitude
discriminationsystem.This adjustmentusually increasessufficiently the peakto
backgroundratio in major element analysisso that the backgroundinterference
is minimal. However,in traceelement analysis the backgroundintensity must be
carefully establishedand subtractedfrom the peak intensity. The most common
procedure is to measurethe background intensity at two off-peak positions
equidistantfrom the peak, averagethe countsand subtractthem from the peak
intensity. The backgroundcount positions should be selectedso that they are
close to the peak, and are free of spectral interferencefrom other elements.
Assumingthat the backgroundunderthe analyticalpeak is linear or has a negli-
gible slope, this backgwund measurementtechnique is adequate.Nonlinear
backgroundsshouldbe correctedfor profile curvatureby graphicalextrapolation
of the backgroundprofiles on eitherside of the peakor by using backgroundcal-

ibration standardsfree of the analytical elementfrom which a correctionfactor
can be established.
Spectralinterferencesare usually not a problemin major elementanalysis,
but in certaincasesare sufficiently severeto require correctionsor adjustments.
Potential interferencein major elementanalysiscan be avoidedby selectingan
alternativebut lessintenseanalyticalline, howeverat low concentrationssuchan
alternativemay reducethe emissionintensitiesto undetectablelevels. Although
interfering (overlapping)lines do not have exactly the samewavelength,a por-
tion of one peak (tail) may coincidewith the centroid of the other. Under these
conditions,the measuredintensity of the analytical peak includesin addition to
the normal backgrounda componentof the interfering peak.Therefore,determi-
nation of the true peakintensity requiredthe estimationof both contributionsand
subtractionfrom the total peakintensity. This estimationis basedon proportion-
al contributions(peak+ background)of the interferingelement,establishedfrom
suitable standardsamplescontaining neither the analytical nor the interfering
elementand others containing known quantitiesof the interfering element,but
none of the analytical element. Examplesof such correctionsfor interference
betweenSr, Rb and Zr can be found in Norrish and Chappell(1977).
Matrix or interelementeffectsare a function of the compositionof the sam-
ple and usually result in an inexactly proportionalrelationshipbetweenthe emit-
ted x-rays and the concentrationof the emitting elements.Unlessinfinitely thin
samplesare employed,the interactionsbetweenthe sampleand the exciting radi-
ation within the finite depthof penetrationpreventlinearity from beingachieved.
Differential absorption,excitation and enhancementeffects causedby variable
samplecompositionare very complex and therefore,difficult to predict.
Some theoreticalmodels employing a variety of mathematicalequations
have beenproposedto compensatefor compositiondependenterrors, but their
application is complex and requiressignificant computing power and time. A
major drawback of this correction approachis that the correction parameters
used in the equation dependon prior knowledge of the sample composition,
which is the object of the analysis.Thus, an approximatecompositionis being
establishedfrom which the equationsand correctionparametersare set for cal-
culation of a corrected composition following several cycles of iterations.
Becausethe originally assumedsamplecompositionis questionableand absorp-
tion coefficientsfor light elementsor long wavelengthradiation include a con-
siderabledegreeof uncertainty,thesecorrectionproceduresmay at times involve
errors which exceedthe magnitudeof the matrix effects. Although theoretically
this may be the best approachto the problem, its complexity has directedXRF
analyststo other more pragmaticsolutions.Suchsolutionsinclude:
1. Toleratingmodesterrorsin caseswherecomparativechangesin com-

position are more importantthan excessiveanalytical accuracy.
2. Using standardsof the sameor similar compositionwith the samples
for repetitive work with similar samples.
3. Containing matrix errors within acceptablelimits by diluting with a
liquid or a solid that reducescompositionalinterferences.
4. Measuring the absorption coefficients directly or indirectly on the

samplesand standardsin an additionalseriesof measurements.
5. Utilizing infinitely thin samples(thin films of a few hundrednm in
thickness),which make matrix effectsnegligible.
6. Using variouscompensatingtechniques.
A more extensivedescriptionof someof theseapproachesis given in the
following section.
Linear CalibrationMethods.For many years themost popularcorrection

methods in quantitative XRF analysis have been those employing calibration
standardsfor developmentof calibration curvesfrom which the compositionof
unknown samplesis calculatedbasedon the emitted characteristicline intensi-
ties. These"in-type" modelsare still widely usedtoday in spite of their limita-
tions becauseof the ease of their application and the minimum computer
requirementfacilities. These methodsassumethat a linear relationship exists
between the intensity of the characteristicline and elemental concentration
becausethere is no significant changein total absorptionor enhancementover
the range of the calibration line, due to matrix matchingbetweensamplesand
calibration standards.In practice, however, there are three potential problems
with this approachthat needto be considered.
First, it must be realized,that in trying to minimize absorptioneffects by
limiting the total absorptionrangewe in fact put constraintson the rangeof the
calibration curve for certain elementswith high absorptioncoefficients. A sec-
ond potential problem lies in the selectionof the calibration standards,which
should reflect not only the analyterange but they shouldcompensatefor poten-
tial absorption/enhancement interactions. A third problem is the calibration
curve itself. Most often a good straight line fit of x-ray intensity data plotted
againstelementalconcentrationswith an acceptablelevel of deviation would be
sufficient to declarethe methodsatisfactory.In many cases,however,the curve
is not straightand the deviationsnot acceptable,due to either insufficient preci-
sion of the collected counts, inaccuratestandardcomposition,sample hetero-
geneity or excessiveabsorption/enhancement effects.All thesepotentialsources
of error shouldbe carefully evaluatedbefore anothermethodis adopted.
Dilution Methods. These methodsattempt to reduce matrix effects by

diluting the samplewith an absorberof low absorptioncoefficient and at the
sametime obviatepossibleheterogeneityproblems.Commonsolid diluentsused
in this methodinclude sodiumcarbonate,sodiumtetraborate,lithium tetraborate,
and boric acid, which are mixed with the sampleat certainratios and the mixture
is fusedto provide a homogeneous specimenin a glassdisk shape.Alternatively,
if the sampleis soluble,it can be diluted with Hel or othersolventsbeforeanaly-
sis. Use of liquid dilutents,however,precludes analysis in vacuo,and thus,great-
ly reduces sensitivityfor light elements.It should be kept in mind that this
method, while reducing absorptioneffects, also decreasesthe intensity of the
emittedcharacteristiclines by a factor approximatelyequalto the squareroot of
the dilution ratio. Therefore,it is unsuitablefor traceelementdeterminations.A
double dilution method proposedby Tertian (1972) is a powerful techniquefor

the analysisof mixtures of single elements,employingtwo dilutions of the stan·
dard and sample.The expressionutilized in this applicationtechniqueis
(q) (C;)/(C; - Ci)

= (CD (G)/(G - CD
[30]
wherePx and Ps are the weight fractions of a certain elementin the sampleand
C;
standard,respectively,Ci and are the count ratesobtainedfrom the sampleat
dilutions 1 and 2, and q and G are the respectivetwo count ratesobtainedfrom
the standard.To keepthe calculationssimple the sameweightsof sample/diluent
and standard/diluentare used. The dilution ratios must be selectedcarefully
becauseif the dilution is small, the intensity difference will be small and the
countingerror large, while if the dilution differenceis too large, the intensity for
the most diluted samplewill be so weak that the count error may be significant.
Furthermore,if there is no automaticdead-timecompensation,dead-timecor-
rectionsare essentialbecauseof significant count lossesin the leastdiluted sam-
ples.
Compensation Methods. Thesetechniquesutilize informationfrom the x-
ray spectrumof the sampleanalyzed;which is not associatedwith the character-
istic line intensities, in order to compensatefor matrix effects. Two common
methodsrepresentingthis approachare thoseemployingscatteredradiationmea-
surementsand thoseusing internal standards.The first techniqueis basedon the
fact that the primary x-ray radiation undergoesa certain degreeof scattering,
which is matrix dependent,and therefore,examinationof the scatteringcharac-
teristics of the samplecould provide a quick first assessment of possiblematrix
effects. This approachis particularly useful becauseit requiresvery few stan-
dards, but no special sample treatment,and once a correction model is estab-
lished, only one or two additional counting measurements can provide the nec-
essaryscatteringdata.
The primary tube radiationis scatteredat the surfaceof the sampleand also
at depth within it. The intensity of the scatteredradiation can be approximated
by the equation
K- s
h(scattered)'" A [31 ]
(scattered)
whereK = constantand s = compositiondependentmassscatteringcoefficient.

Becauseof wavelengthproximity betweencoherentlyand incoherentlyscattered
radiations,they could beconsideredas having the sameA value, which is essen-
tially a linear function of the correspondingmassabsorptioncoefficients over
certainenergyregionsand compositionalranges.Therefore,
A(scattered)'" Kl - A(sample), [32]

and combiningEq. [31] and [32] we have the relationship
IJI,..{scattered) '" K z • C, [33]
which relateslinearly the intensity ratio of the analyte emissionline and the scat-
teredtube line to the concentration(C) of a particularelementin the sampleinde-
pendentof matrix effects.A linear calibrationcurve compensated for absorption
effects can then be establishedby selectinga scatteredtube line near that of the
analyte line (without interferencefrom absorptionedges)and by plotting their
ratio against known concentrationsof calibration standards.Andermann and
Kemp (1958) useda scatteredradiation line of high intensity at 0.06 nm, while
Kalman and Heller (1962) usedbackgroundscatteredradiation near the charac-
teristic analytical line with very good precision.
The scatteredradiation methodfor compensationof matrix effects is used
primarily. for trace elementdeterminationin samplesand standardsof similar
composition.Its major drawbackis that it requiresadditional measurements for
the scatteredline, which essentiallydoublesthe analysistime. The methodhas
limited applicability to sampleswith grosscompositionalvariationsor analysis
of light elementsbecauseof increasedbackgroundcontributions,which areinde-
pendentof absorptioneffects.
The internal standardmethod involves the addition to the samp~e of a
known amountof anotherelement,which has a suitable emissionline and ab-
sorptionedgenearto thoseof the analyteelement.This guaranteessimilar matrix
effects for the emissionsof the analyte and the internal standard,and therefore,
a linear relationship betweentheir measuredintensitiesand concentrationsas
depictedin Eq. [34]
h (standard) =K • C(sample)
_---"_=_<..:.:.L.
__
[34]
h (internal standard) C (internal standard)
Obviously, the elementaddedas the internal standardshould not be pre-

sent,at least in measurablequantities,in both samplesand standardsand should
be addedin concentrationsthat will yield comparableintensitiesto thoseof the
analyte.A limitation of this techniqueis the preparationof homogeneoussam-
ples, especiallywhen the elementis addedin solid form, which makesuniform
dispersiondifficult and createssignificant microabsorptioneffects. For this rea-
son, this techniqueis not recommendedfor analyte wavelengths>0.3 nm. The
methodis well suitedto the determinationof one or two heavyelementspresent
at low to moderate concentrations in relatively uniform matrices,but not for a
numberof different elementsin the samesample,where multiple internal stan-
dardsmake the procedurerather complex and impractical.
Another internal standardapproachis the spiking of the samplewith cer-
tain quantitiesof the elementto be analyzed,thus maintainingidentical matrix
effects.In this case,If 11 and I z are the measuredanalytefluorescenceintensities
before and after addition of a known concentrationof the elementin question,
the raw concentration(C) of the sampleis given by the equation
X·RAY FLUORESCENCESPECI'ROSCOPY 195
c= 11· ac [35]
h-/l
a
where c = the addedelementalincrement.To maintain an adequateprecision
in the measurement of 12, ac shouldbe sufficient but not to the point that it will
alter significantly the massabsorptioncoefficient of the samplefor the analyte
emission.Although the methodis subjectto the samesamplehomogeneitylim-
itations as the internal standardtechnique,it is simpler and free of undesirable
interferencesfrom absorptioneffectsor edges,and thus,bestsuitedfor traceele-
ment concentrationdeterminations.
MeasurementOf Absorption Coefficients. For samples with wave·

lengths<0.3 nm (elementsheavierthan Ca) direct and reliable measurements of
absorptioncoefficients,especiallythosefor the fluorescentradiation(A2) which
are dominant, can be made by using pressedpelletized specimensof known
thickness (I), weight, and cross-sectionalarea, and by calculating the mass
absorptioncoefficient from the equation
In 10 -In I
A 2 =-....::......-- [36]
pi
where pi =weight of pellet/crosssectionalarea.

Mass absorption coefficients measuredon duplicate samplesgenerally
agreewithin a few percentagepoints, but severalsamplesof different thickness
may be necessaryto coverthe wavelengthrangebetween0.05 and0.3 nm. In the
absenceof major absorptionedgesin the sample,absorptioncoefficientsfrom
one wavelengthcan be usedto measureseveralelements.Thus, in soil and geo·
logical samplesthe absorptioncoefficient for ZnKa radiation could be usedto
measureNi and Cu but not Mn becauseof the FeKa absorptionedgebetween
ZnKa and MnKa radiations. Counting errors in measuringabsorptioncoeffi-
cientscan be kept small (-1 %) by maintaininga high 10/1 ratio (104), but when
I is small backgroundcorrectionsare necessary.The methodbecomesimpracti-
cal for high valuesof pi becauseof the excessivelylong countingtime required.
Other disadvantages of the techniqueinclude the specialexperimentalconfigu·
ration requirementto perform the measurements, and the unsuitability for sam·
pIes that cannotbe pressedinto pellets, for light elementswith low energy,and
for very heavyelementsfor which severaldilutions shouldbe employed.
MathematicalCorrectionModels. The developmentof numericalalgo-
rithms betweencharacteristicx-ray wavelengthemissionsand chemicalcompo-
sitions, basedon calculatedabsorptioncoefficientsin the early 1950s(Beattie&
Brissey, 1954; Norrish, 1959, p. 59; Lucas-Tooth& Pyne, 1964), were greeted
with little enthusiasmbecauseof the inadequacyof the then-availablecomputer
systems,but have found wide application with the advent of recent computer
technologies.This approachinvolves the use of regressionanalysis to solve
196 KARATHAl\ASIS & HAJEK
equations,the parametersof which are calculatedfrom samplesof known com-

position.
For a binary systemAB the concentrationof A can be calculatedfrom a
calibrationcurve developedfrom a seriesof calibrationstandards.According to
Eq. [15], this is only possiblewhen thereis a linear relationshipbetweenCA and
lA, which requiresthat A2 is constant,or the massabsorptioncoefficientsof A
and B for IA are identical.In mostcases,this relationshipis not linear, but hyper-
bolic, bowing upwardsor downwardsdependingon the relativemagnitudeof the
absorptioncoefficientsof A and B for lA, and possiblesecondaryenhancement
effects.The degreeof line curvatureis, therefore,a measureof the effect of B on
the emissionintensity of IA and can be usedto makematrix effectscorrections.
Concentratingprimarily on adsorptioneffects Eq. [15] can be written for
the binary AB systemas
[37]
whereaA andaB are coefficientsrepresentingtotal absorptioneffectsdue to ele-

mentsA and B respectively.SinceCA and CB =1 Eq. [37] becomes
[38]
[39]
and
[40]
This is a linear expression,which can be usedto correctthe measuredIA

provided that CB is known or approximatedand aA (B)' which representsthe
effect of elementB on the characteristicintensity of element'A undergiven ex-
perimentalconditions,is known or can be established.In reality, the CA and CB
valuesusedoriginally in this equationare the uncorrectedfirst approximations
of concentrationsof A and B, which are eventuallyrefined by a seriesof itera-
tion cycles.The coefficientsusedto expressthe natureandmagnitudeof oneele-
ment on the emissionintensitiesof anotherare termedalphafactorsor influence
coefficientsand can be derived by graphicaltechniques,regressionanalysis,or
calculationand refinementfrom tabulatedmassabsorptioncoefficients.
Multicomponent systemsprovide further complicationsand require the
assumptionthat binary coefficienteffectsare linearly additive,i.e., for a ternary
systemABC h =(KACA)/[aAC A + aBCB + acCc]. Although this assumptionis
not alwaysvalid, the use of this methodhas provided satisfactorysolutionsfor
many analyticalproblemsin XRFS, especiallyif combined~th othercompen-
sation techniques.Some of the most popular mathematicalmodels used for
matrix effects corrections include the Lucas-ToothlPynemodel (1964), the
Lachance-Traillmodel (1966), the Rasberry-Heinrichmodel (1974), the funda-

mental parametersmodel (Criss & Birks, 1968), the Claisse-Quintin model
(1967), the Norrish-Huttonmodel (1969), and variousothercombinationswhich
have beenreviewedby Tertian and Claisse(1982) and Rousseau(1987).
SAMPLE PREPARATIONTECHNIQUES
Soil, geological,and plant samplescan be analyzedby XRFS in a variety

of forms including loose powders,pressedpellets, fused disks, thin films, and
solution extracts.When trace elementconstituentsare the focus of the analysis,
various preconcentrationtechniquessuch as ion exchange,coprecipitation,or
chelation also may be necessary.The selectionof the appropriatemedium is
dependentupon the natureof the sample,the analyticalgoal, amountof sample
available,element(s)to be determined,detectionlimit requirements,time avail-
able and the reliability of the dataobtained(Bower & Valentine, 1986).
LoosePowders
Finely groundloose powdersamplesof soil or plant materialsare conve-
nient to prepareand quick to analyzebut their analysisis subjectto many errors.
Someof the mostseriousflaws are due to segregation/particle size effectsand/or
density gradientswithin the matrix developedduring the filling of the sample
cell. This type of problemscan be minimized by pulverizing the samplesand
standardsto homogeneousmatricesusing appropriatemills and grinding equip-
ment, and packingthe sampleto a reproducibledensity. In most cases,grinding
the samplesto lessthan 45 Jlm in size and tappingthe invertedcell with the sam-
ple a few times would sufficiently reduceparticle size effectsand provide a uni-
form packingdensity, unlessvery light elementssuch as Mg, Si or AI are to be
determined.The use of loose powder in XRF analysis is generally limited to
applicationsthat requirequick resultswith relatively low analyticalaccuracyand
precisionand has been especiallyuseful for the determinationof heavy metals
(usually heavier than Fe) presentin moderateto large quantities(Levinson &
dePablo,1976; Berry et aI., 1968).The lack of an in vaccuoanalysisoption with
loose powdersamplesis anotherdisadvantageof this technique.
PressedPellets
Pressedpellets are preparedlike the loose powdersand subsequentlyare
pressedinto a pellet or briquette using a press and a die setting that yields a
densersample,with a flatter surface.Although subjectto particlesize limitations
similar to the loose powder samples,the pressedpellet is a much more repro-
ducible preparationtechniquethan loosepowderswith lessrisk of contamination
of the instrumentwith dust,which enablesreadydeterminationsof elementssuch
as Ti, Cr and Mn in soil and geological samples.One disadvantageis that fre-
quentand/orprolongedexposureof the pelletsto the x-ray beamtendsto cause
gradual deteriorationof their surfaceand matrix instability, thereforerequiring
frequentpreparationof new standards.
The pelletscan be preparedwith or without backingor binder. Aluminum

or plastic cups are frequently usedfor backing, designedto producea briquette
with a plasticlike casingaroundthe samplemixture (Fabbi, 1972).Another pop-
ular procedurethat improvespellet stability involves the uniform mixing of the
pulverizedsamplewith about 15% by weight of a bindersuchas polyvinyl alco-
hol, cellulosenitrate, boric acid or other commercialmixtures, and pressingthe
mixture into a pellet (Goodman,1973; Norrish & Hutton, 1964).For more robust
briquettes a thermosetting binder such as methylmethacrylateor phenol
formaldehydecan be used,which after heating in a drying oven will causethe
binder to set or melt, thus making the pellet less susceptibleto crumbling or
breaking (Leake et aI., 1970). It is essentialthat a standardizedprocedurefor
preparationof both samplesand standardsis followed assuringidentical grind-
ing, mixing, sampleand binder weight, pressureand pressingtime conditions.
The pressurerequired to form a coherentsamplevaries with the nature of the
sample.Too low pressurewill provide easily breakablepellets,while a too high
pressurewill causeshearing.If necessary,a plasticfilm (Mylar, scotchtape)can
be used on one of the sides before pressingto prevent samplecontamination
(Kanaris-Sotiriou& Brown, 1969).
FusedDiscs
The mostcommonmethodfor preparationof soil or geologicalsamplesfor

XRF analysisis the fusion of the pulverizedsamplewith a flux (most often a
boratesalt), followed by castingof the melt in a mold to form a glassdisc that
would fit in the spectrometersampleholder.This technique eliminates many ana-
lytical problemsarising from particle size inhomogeneityand samplecomposi-
tion. Otheradvantagesinclude the increaseddurability of the glassdisks over the
pressedpellets,easystorageand reusability.The major disadvantages include the
dilution factor which affectsthe analysisof trace elements,the reductionof flu-
orescenceintensity of low atomic number elementsand the destructionof the
samplebefore analysis.
The fusion is accomplishedby mixing the analytical flux with the sample
in an appropriateratio (usually in the rangeof 1:1-10:1), and heatingthe mix-
ture to a high temperature(-1200°C),in an inert,heat-resistantcrucible. A sub-
sampleheatedto the temperatureusedfor the fusion is employedto estimatethe
content of volatiles. There are numerousvariations of the originally proposed
technique(Claisse,1956), in terms of selectingthe type of flux, flux-to-sample
ratio, samplesize, fusion time and temperature,disc size, castingtechnique,etc.
Although lithium tetraborate(Li 2B40 7) is almostuniversallyapplicableas a flux,
certain mixtures have proved to be more satisfactory for specific samples.
Generally, acidic fluxes (rich in Li 2B40 7) are best suited for dissolving Al 20 3,
alkali or alkaline earthoxidesor carbonates,while basicfluxes (lithium metabo-
rate, LiB0 2) are more suitablefor dissolvingacidic oxidessuchas Si02 or Ti0 2•
The melting point of the mixture is anotherconsideration.The optimum flux is
the one that reactscompletelyand quickly with the sampleand has the lowest
melting point. For this reason,mixtures of lithium tetraboratewith metaborate,
carbonateor other reagentsare often selected.Lower fusion temperaturesreduce
preparationtime and inhibit lossesof volatile elementsfrom the sample.Some

samples,however,especiallythosehigh in Zr02, require higher fusion tempera-
tures. Many procedureshave been published for fusing samplesinto a borate
glass disk (Howeknecht, 1983; Hutton & Elliot, 1980). Norrish and Hutton
(1969) useda flux consistingof a mixture of 47.03%anhydrouslithium tetrabo-
rate, 36.63% lithium carbonate,and 16.34% lanthanum oxide. Bennett and
Oliver (1976) useda mixture of Na2B407and NaN03 to increasethe solubility
of high metal concentrationsamples;Johnsonand Maxwell (1981) suggesteda
mixture of 50% Li 2B40 7, 30% LiF, and 20% NH4N03 for the determinationof
major, minor, and trace elementsin the samedisc, while Haukka and Thomas
(1977), Eustell and Willis (1990), and Lee and McConchie(1982) recommend-
ed low dilution methodsfor rock and mineral samples.
Thin Films
The use of thin films is often a convenienttechniquefor analyzinga small

amount of sample or measuringtrace constituentspreconcentratedby solvent
extraction,coprecipitation,ion exchangeor other procedures.The advantageof
the thin film is that it maintainsradiation absorptionto negligible levels, so that
the intensity of the secondaryfluorescentradiationis linearly proportionalto the
concentrationof the elementin question.The samplethicknessrequiredto meet
the negligible radiation absorptioncondition rangesbetween0.03 and 6 11m,
dependingon the analytewavelength(the longer the wavelengththe thinner the
film).
Thereare variousways of thin film preparationfor XRF analysis.Someof
the simplest include spreadingthe pulverized sampleas a thin layer on scotch
tape or betweensheetsof Mylar film stretchedover a samplecell and held in
placewith O-rings, or dissolving the sampleand evaporatingit on a Mylar film
or a piece of filter paper.Ion-exchange-impregnated papersor membraneshave
successfullybeen used for the determinationof sulfates (Schreiber & Pella,
1979), and other trace constituentsin geological samples(Blount et aI., 1973).
Coprecipitationgel techniquesfor trace elementanalysisin thin films have been
describedby Luke (1968), while the application of chelation techniqueshave
beenreportedby Knapp et al. (1975).
Liquid Samples
Liquid samplesfrom various extractionsalso can be usedfor XRF analy-

sis, especially for heavy metals present at moderate to large concentrations
(Bernardo et aI., 1985). This techniqueemploys special polyethylenesample
cells, which are filled with the liquid sampleand coveredwith a piece of film
(Mylar) spreadover the top and pusheddown with the snapring to form a drum-
like window. The cells are inverted to check for leakagesbefore being inserted
in the spectrometer.The main disadvantageof this techniqueis that it does not
allow the use of a vacuum,and the increasedprobability of spillage. However,it
has been used quite extensively for Fe determination in citrate-bicarbonate-
dithionite extractsof soil samples.For lighter elements,He gas can be usedin

the analyzingchamberto improve sensitivity.
METHODS FOR ANALYSIS OF MAJOR ELEMENTS
Preparationof GlassDiscs
Equipment
1. Clean, mechanicallydriven grinding equipmentfor grinding 2 to 5 g
of soil material to <250-Jlm particle size without contaminatingthe
samplewith the elementsbeing determined.Usually an agatemortar
and pestleare adequatebut ring andpuck mills or mixer mills also are
suitable.
2. A mixer/mill with various mixing vials and balls for homogenizing,
mixing, and blendingsamplesand standards.
3. Platinum cruciblesof 20- to 25-mL capacity, with platinum covers,
platinum-tippedtongsfor handlingthe crucible, and platinum-stirring
wire about0.75 mm in diameter.Cruciblesmadewith Pt-Rh-Au alloy
are not wettedby the fusion mixture and therefore,are easierto clean.
4. A blast (Meker) burner able to maintain a temperatureof about
1000°C,and nickel-chromiumtrianglesto hold the crucible,or a muf-
fle furnacewith temperaturecontrolledat 1200°C.
5. 1\vo electric hot plateswith smoothaluminum surfaceusedfor mak-
ing and annealingthe glassdiscs, respectively.Both shouldbe equip-
pedwith adjustabletemperaturecontrolsin the rangeof 200 to 400°C.
Asbestosmatscan be usedfor the annealinghot plate.
6. Aluminum plunger,brassrings and aluminumor carbideplatenswith
shapesand dimensionsshown in Fig. 7-3 [other devices,such as the
one designedby Kerrigan (1971) can also be used]. Although alu-
minum-platensare cheaperthan tho~e made from graphite,someAI
contaminationof the melt is possible.
7. Protectiveclothing, high temperatureresistantgloves, face mask or
safety spectacles.
8. Teflon beakers(50-mL capacity) and stirring bars used for cleaning
the crucibles.
9. A third electric hot plate-stirrerwith adjustabletemperaturecontrols
usedfor cleaning thecrucibles.
10. Envelopesand desiccatorsfor storing the glassdiscsunder vacuum.
Materials
1. Fusionflux LiB0 2, anhydro~, reagentgradeas suggestedby Vanden
Heuvel, 1965, or a mixture of Li 2B40 7, 47.03%, lithium carbonate,
Li 2C03, 36.63%,and lanthanumoxide, LaO, 16.34%,ready-madeor
preparedas describedby Norrish & Hutton, 1969). Lithium metabo-
I
1'-- 42.0 ImI diam.~
I
I I I I
I I I I e
I J+"31.Smm di"·4 I E
co
I I I I :;;
I II r~'~1mI
Brass Ring
Aluminum Plunger
'0
:-
'=~-='
I i!
I,
d,olll'-dT
,.- , /
Wood.,
handl.
I' on
N
I
. ...i..
BRASS RING
I
IE
E
....
'"
1-~;;:"1 I
I
_~ ,Li2 1'mm
002 1 7m", -.! i
1f-2~.lam
i1
I. ~
I
331T11ft
38mm _c 1 t lL 0
t 1_315mm_1 ~ 313""" _ I r
ALUMINUM (OR - ALUMINUM (OR STAINLESS
GRAPHITE) PLATEN STEEl) PLUNGER
Fig. 7-3. Assessoriesfor preparing lithium metaborate(a) and lithium tetraborate(b) glass discs
[with permissionfrom K. Norrish and J.T. Hutton, 1969; an accuratex-ray spectrographicmethod
for the analysisof a wide rangeof geologicalsamples(Geochim.Cosmochim.Acta 33:431-453.»)
rate fluxes of high purity are availablefrom a variety of chemicalcom-

panies including ICN Pharmaceuticals,Inc., Plainview, New York,
USA; Baker Chemical Co., Phillipsburg, New Jersey08865, USA;
Aldrich Chemical Company, Inc., Milwaukee, Wisconsin 53233,
USA, and others.The Li 2B40 7-LiC03-LaO ready-mixedflux is avail-
able from Ultra Carbon Corp., P.O. Box 747, Bay City, Michigan
48706,USA; the United Mineral & ChemicalCorp., 129 HudsonSt.,
New York, lOO13, USA; or Johnson-MattheyChemicals, Ltd.,
OrchardRoad, Royston,Herts, SG8 5HE, England.To ensureoxidiz-
ing conditionsNaN03 or LiN0 3 is addedto the flux at a rateof 1.33%.
All chemicalsshouldbe of the highestpurity available.
2. Nitric Acid 3% for dissolving flux residue left inside the crucibles
after the fusion. Alternatively, the cruciblescan be dippedin an anhy-
drous Na2C03 flux fused at lOOO°C in a muffle furnace and subse-
quently washedwith 6 M hydrochloric acid. The first procedureis
somewhatslower but lesstroublesome.
3. Standard samples prepared from certified reference materials of
known composition, which can be obtained from various sources
including: the National Institute of Standards and Technology,
Gaithersburg,Maryland 20899, USA; British Chemical Standards,
Newby, Middlesborough,Teeside TS8 9EA, England; Canadian
Certified ReferenceMaterials,CanadaCentrefor Mineral and Energy
Technology,Ottawa,Canada,KIA OG1; U.S. GeologicalSurvey,U.S.
Dep. of the Interior, Reston,Virginia 22092,USA, and others.
4. Pure Si02, A1Z03' FeZ03' and other elementaloxide samplesof high
purity for preparingblank disks, which will serve to measureback-
groundcountsfor other elements,or to be usedas standardsfor high
concentrationsof theseelements.
Procedure
Lithium Metaborate Flux
1. Obtain a representative2 to 5 g of pulverizedto <150 Jlm and homog-
enizedsample.Dry the sampleand flux at 1lOoC and maintain in a
desiccator.
2. Weigh between0.75 and 1.25 g of the oven-dry sampleor standardon
oneweighingpaperand on anotheran amountof anhydrousflux mate-
rial equalexactly five times the weight of the sampleor standard.
3. Transfera portion of the weighedLiB0 2 to a clean,labeled20- to 25-
mL platinum crucible to cover the bottom, add the weighedsample,
and then cover the samplewith the remainderof the weighedLiB0 2.
4. If sampleto be analyzedcontainsmore than a few tenthsof a percent-
ageof ferrous Fe (reducedsoil samplesdo), add about0.1 g of Na202
or NaN03 or LiN0 3 to the crucibleon top of the soil samplebeforethe
final portion of the weighedLiB0 2 is added.
5. Stir the mixture in the crucible with the platinum wire carefully to cre-
ate a somewhathomogenizedsample.
6. Tum on the electric hot plates and adjust the temperatureat 350 to
400°C. Place the aluminum-plungeron the hot plate and allow it to
reachthe hot plate temperature.
7. Placethe crucible containingthe sampleon the triangle over the blast
burnerwith the tongs,tilt the crucible towardsyou and cover with the
Pt lid allowing about 1/10 openingat the highest rim of the crucible.
The latter is doneto maintainoxidizing conditionswithin the crucible.
Apply oxidized flame to the far side of the crucible using a low flame
for the first few minutes to prevent the powder from blowing out of
the crucible, and then turning the flame up for maximum heating.
When most of the sampleis fused (-10 min), graspthe crucible with
the Pt-tippedtongs and swirl the melt to removethe bubblesfrom the
bottom and to hastenfusion of the remainingsolids. When the fusion
is complete(-20 min), useplatinum wire or a motor stirrer with a plat-
inum wire to stir the melt two to three times at I-min intervalsso that
a uniform melt is formed. Alternatively, swirl the crucible holding it
with the tongsseveraltimes during the fusion process.
8. When the melt is ready to pour, place the cold brassring on the hot
plate and pour the melt into the ring by inverting the crucible over the
ring while holdingit with the tongs. Quickly centerthe ring aroundthe
melt and take the hot aluminumplunger,place it in the ring, and press
gently for about 10 s to spreadthe melt evenly so that it fills the ring
dimensions.Removethe plungerand placeit backon the hot plate sur-
face, and remove the brassring from around the disc. Allow the disc
to cool down to the hot plate temperaturefor about 30 min before
transferringwith heatedtongs to the other hot plate for the annealing
process.The latter shouldlast for at ieast another4 h before the discs
are allowed to cool down to room temperatureby turning the hot plate
off. If the disc cracksor is unsatisfactory,remelt in the samecrucible
and repeatthe procedure.Becausethe discs cannotbe labeledwhile
they are still hot, they are positionedon the annealinghot plate in a
predeterminedorder. Once they are cooled to room temperature,they
can be labeled using a labeling tape glued to the surfaceof the disc
facing the hot plate.
9. If the disks have rough edgesor surfaces,they can be smoothedwith
a file. Use cleanpiecesof tissueto removethe filings and avoid touch-
ing the surface with the hands to prevent contamination.Store the
discs in labeledenvelopesin a desiccator.
10. To clean the used platinum crucibles, place a small magneticstirrer
inside, and immerse them completely in Teflon beakershalf-filled
with 4% HN03 and placedon a hotplate-stirrercontrolledat 80°C. An
overnighttreatmentusually producescleancruciblesready to use the
next morning. If a fastercleaningprocessis requiredthe cruciblesare
filled with Na2C03which when fused over the blast burner dissolves
the LiB0 2 glass residue. The melt is poured onto a containerwith
quartzsand,the crucible is cooleddown, and the Na2N03 residueleft
is dissolvedby immersing the crucible in 6 M HCl. Platinum wires,
stirring rods, and platinum-tippedtongsare cleanedsimilarly.
Lithium Tetraborate-Lithium Carbonate-Lanthanum Oxide Flux
1. Obtain a representative5- to lO-g sampleand grind to <150 ~m. Dry
the sampleat 110°C and the flux mixture at 400°C. Keep both in a
desiccator.
2. Weigh into a labeledplasticcontainerexactly 0.34 g of sample,1.80g
of flux, and 0.02 g of NaN03. Seal the containerand shakethe mix-
ture to producea homogeneousmixture.
3. Switch on the furnaceto lOOO°C, if required;a hot plateat 220°Cwith
the plunger,brassring, and platenson it; and a hot plate at 200°C on
which two asbestosmatsare placed.
4. Tip the fusion mixture into a platinum/goldcrucible, and melt either
over the burneror in the furnace.
5. When the mixture has melted,stir with a platinumwire stirrer, or mix
by swirling the crucible. This ensuresa thoroughmixing of the melt
and removesbubblesfrom the bottom.
6. When the melt is ready,placethe brassring aroundthe platen,pour the
melt into the centerof the platen, and immediatelybring the plunger
down onto the melt. You can then withdraw the plunger, removethe
brassring, and transfer the disc to the other hot plate betweentwo
asbestosmats.
7. Re-fuse any material remaining in the crucible into one piece, and
allow the crucible to cool. This material will then tip out of the cru-
cible and should be kept in caseit is necessaryto re-fusethe disc. If
necessary,clean the crucible and repeatthe procedurewith the next
sample.
8 When all the sampleshavebeenmadeinto discs,slowly tum down the
hot plate, allowing the discs to annealand cool betweenthe asbestos
mats. When cool, the discs may be filed to remove rough edgesand
shouldbe storedin separate,labeledenvelopes.
9. Cleanthe platinum/goldcrucible by melting someNa2C03in the cru-
cible andswirling the melt around.When the crucible is cool, placeit
in a containerof dilute HCI until the melt hasdissolved.Washthe cru-
cible with distilled water,dry it, andstoreit in its case, whichwill help
keepits shape.Cleanthe platinum wire in the sameway.
lO. Preparea blank from very pure Si02 in the sameway as the samples.
Use it to measurebackgroundcountsfor all oxides other than Si02.
For analysisof Si02, preparea blank from very pure alumina (AIZ0 3).
Comments
1. Samplesvery low in Si and AI (<10%) fused in LiB0 2 will partially
crystallize insteadof forming a suitableglassdisk. Such samplescan
X-RAY FLUORESCENCESPECTROSCOPY 20S
be spikedwith pure Si or AI referencematerialsso that they produce

a viable glassmixture.
2. Oxidizing conditions,obtainedby the addition of Na202, NaN03, or
LiN0 3 are necessary.Undersatisfactoryoxidizing conditions,Fe-con-
taining samplesusually produce yellow glass discs, while greenish
discs are formed undera reducingflame.
3. The weighing of flux and sample must be accurateto maintain the
ratio of flux to samplethat is usedduring the correctioncalculations.
If sampleamountsare limited, it is possible to producesatisfactory
discs from 0.25 g, but a similar reduction in flux must be made to
maintainthe ratio, or pure Si02 may be usedto make up the weight of
the sample.
4. The temperatureof the annealinghot plate should be adjustedto suit
the discs,becausetoo fast a rate of cooling can cause cracking.
5. for accurateresults,it is advisableto produceduplicatediscsfor each
sample.
6. Glass disks for taking blank readingsto correct for impurities in the
flux are preparedin the samemanneras above exceptthat pure rea-
gentsare addedto the flux in amountsto give the blank aboutthe same
absorptioncharacteristicsas the soil samples.The addedreagentfor a
particularblank must be totally free of the elementbeing dete~ined
and must form a glasswith the flux. A suitableblank for the analysis
of most soils for all elementsexceptSi canbe preparedby fusing pure
silica with the flux, while for Si analysis,a similar disc can be pre-
paredby fusing pure alumina with the flux. When standarddisks are
preparedusing more than one standardmaterial,particularly when one
material is usedin a small amount,the materialsare weigheddirectly
into the -platinum crucibles containing the first incrementof LiB0 2 •
This procedureeliminatessampletransfererrors.
7. During the fusion, the sampleand flux lose weight, and an accurate
measureof the total weight loss is neededfor the correctioncalcula-
tions. By drying the flux at 400°C and keepingit in a desiccatorprior
to the weighing procedure,one can assumethat weight loss due to the
flux is zero. The weight loss of the samplecan be determinedsepa-
rately by heating known weights to 1000°C and calculating weight
loss. If there is only a limited quantity of sample,it can be ignited to
find the weight loss and then usedas the sample.For a more accurate
but time-consumingassessmentof weight loss, the flux is weighed
into the crucible, fused at lOOO°C, allowed to cool, and reweighedto
find the weight loss due to the flux. The sampleand NaN03 are added,
and the whole weighed, re-fused,cooled, and weighed again to find
the weight loss of the sample.
Analytical Measurements
Special Equipment
1. X-ray emission spectrograph,with a vacuum or helium path and
coarsecollimator.
2. X-ray generator(high-voltage).
3. X-ray tubes with chromium, tungsten,molybdenum,or gold anodes
for WDS or Rh for the EDS.
4. Analyzing crystals: LiF(200), PET, thallium hydrogen phthalate
(TIAP), and (optional) Ge(III) for the WDS.
5. Detectors, scintillation counter and gas-flow proportional counter
usingAr -CH4 (90:10) gas,or silicon-dioxide detectorimmersedin liq-
uid N2 for the EDS.
6. Electronics, high voltage (EHT) supply, amplifier and preamplifier
circuits, pulse-heightselector,rate meter, recorder,scaler, and timer
(printer, video monitor, and multichannelanalyzerfor EDS).
7. One-micrometer(1 !lm) thick, aluminum-flashedMylar (polypropy-
lene) windows for the gas-flow counterwindows of the WDS.
8 Sampleholders.
9. Standardsand a blank madeup with a matrix(flux) similar to the sam-
ples being analyzed.
10. Fine and coarsereceivingcollimatorsfor flat crystal geometryunits.
The aboveitems are usually suppliedas completex-ray spectrographs,
but
basic setscan be supplemented.Commercialmanufacturersand suppliersof x-
ray spectrographsinclude Philips, General Electric, Rigalu (Danvers, MA),
Siemens(Cherry Hill, NJ), Jeol (Peabody,MA), Telsec(Southfield, MI), Kevex
X-Ray, Inc. (Scotts Valley, CA), Shimadzu Scientific Instruments, Inc.
(Columbia, MD), and others.Thesemanufacturersalso will supply most, if not
all, of the accessoriesrequired.
Procedurefor WavelengthDispersiveSpectrometers
1. Set up and align the instrument by following the manufacturer's
instructionsor the equipmentmanualsand fit the correct crystal, col-
limator, x-ray tube, and counter for the samples and the
elements/oxidesto be analyzedin accordancewith the generalcondi-
tions as set out in Table 7-3.
2. Switch on the instrumentand allow it to warm up for at least 1 h so
that the generator,electronics,and gas flow counterstabilize. During
this period, set the generatorat or close to the requiredkilovolts and
miIIamperes.
3. Determinethat the positionof the peakbeing usedfor analysisis locat-
ed exactly on the standardsample by scanningover the peak and
bisecting the angle betweenthe positions of half-peak height (Fig.
7-4). Evacuatethe sampleand crystal chamber,and adjust the gener-
ator, detectorvoltage, and pulse-heightselectorto give the maximum
peakto backgroundintensity (Fig. 7-4). Note all the settingsused.It
is useful to checkwith a standardsamplethat the intensity for given
conditions remainsconstantfrom day-to-day,becausethis is a check
on the overall condition of the spectrograph.
4. Carefully place the preparedsamplesand standardsperfectly flat in
the correct sample holders, and fit them into the spectrograph.
Evacuatethe sampleand crystal chamberif necessary.
Table 7-3. Instrumentalconditions and sample count rates for major silicate analysis using the
methoddescribedby Norrish and Chappell,1977 (Jones,1982).t
Oxide X-ray tube kV rnA Crystal Counts Sensitivity Counting time
S-l • %-1 % s
Na2°:j: Cr 40 20 TIAP 12 0.08 100
MgO Cr 50 25 TIAP 22 0.05 40
A1 20 3 Cr 45 25 PET 28 0.09 40
Si02 Cr 50 30 PET 32 0.07 40
P20 S Cr 50 30 PET 45 0.10 40
P2OS:j: Cr 44 20 Ge 200 0.008 50
K20 Cr 45 25 PET 1000 0.073 10
CaO Cr 40 20 LiF 1700 0.090 10
Ti0 2 Cr 40 15 LiF 3000 0.006 10
MnO W 40 20 LiF 2200 0.037 10
Fe203 Cr 40 15 LiF 420 0.067 10
Fe203 W 30 10 LiF 46000 0.050 10
Fe203 Au 50 20 LiF 2500 0.006 10
t Analyseswere madeusing Philips PW 1540spectrographwith 480-/lm primary collimator,K spec-

tra, vacuum, l-/lm polypropylenewindow, 5- to lO-V channelon a pulse-heightselector.
:j: As reportedby Norrish and Chappell(1977).
5. Recordthe peakcountsfor eachstandardand sampleand also for the

blank if one is used.Where the blank is not used(e.g., for the analy-
sis of Si02), since the blank is a pure Si02 sample,backgroundread-
ings are required,usually at +1 0 and _10. Figure 7-4 showsthe rela-
tionship of peak and backgroundintensities.For statisticalprecision,
Peak
position
Np Nt
1 Nb· ' •
-~)'7> 29
Fig. 7-4. Diagramshowingpeak position and peakcount rate {Np =Nt - Nb, where Nt =total count
and Nb = (N b+1" + Ntr-l)2, (Jones,1982)}.
aim for as high a count as possible,and adjust the counting times in

accordance withEq. [25]. The countingrate, however,shouldbe such
that thereis no pulse-heightshift, and you shouldlimit countingtimes
to a period within which instrumentdrift is negligible.
Comments
1. If only one tube is to be purchasedfor major element/oxideanalyses
of soils, the Cr tube is most suitable.The chromium tube can be used
for the analysisof Fe and Zr, although intensitieswill be lower than
with a tungstentube. The chromiumtube is not suitablefor the analy-
sis ofMn.
2. Severaltableswill convert elementalwavelengthsto 28 for the vari-
ous analyzing crystals. Thesetables should be consultednot only to
locate peak angles but also to check that peaks due to other ele-
ments/oxidesare not so close as to interferewith the peakbeing ana-
lyzed.
3. The principlesand usesof pulse-heightselectionare well describedin
Jenkinsand De Vries(1970),and the correctuseof pulse-heightselec-
tion can eliminate, or greatly reduce,problemsof overlappingpeaks
and interferencefrom other elements/oxidesthat can greatly enhance
peakintensities.
4. It is a matterof personalchoicewhetherto determinethe countsfor a
single wavelengthover all the samplesto be analyzedor to count for
a seriesof wavelengthsfor eachof the group of sampleswhich can be
loadedinto the samplechamberof the spectrograph.Whateveris pre-
ferred, checksshouldbe madeto ensurethat the position of a spectral
peakhas not drifted. Nor should there be instrumentdrift over a long
period of counting. It is usually best to include regular checkson a
standardsamplein any countingroutine.
5. The sample or crystal chamber (or both) must be evacuatedlong
enoughafter eachchangeof sample(or crystal) so that count ratesare
stable.
6. The conditionsin Table 7-3 should in many respectsbe a guide only.
The exacttube excitationwill dependon the natureof an analysisand
the conditionof the spectrograph,it is unwiseto exceedthe wattageof
an x-ray tube for a long time. The stability of the instrumentalso will
determinehow long it is possibleto count without encounteringinter-
ferenceor drift. It is often necessaryto compromisebetweengetting
as many counts as possible (by increasing the kilovolts and mil-
liampereson the tube, the gain on the detector,and time of the count-
ing) and avoidinglong-termdrift andothersporadicinterferences.The
exactnumberof countsper second(cps) per percentagepoint of each
oxide will vary greatly from one laboratory to anotherbut the sensi-
tivity, or lower limit of detection, indicates the theoretical level of
detectionfor that oxide (Table 7-3).
7. Referenceshould be made to the section which deals with counting

statisticsand associated errors. In a seriesof repeatedcounts,any that
vary from the meanby more than threestandarddeviationsshouldbe
rejected,becausea deviationof this magnitudeindicatesunacceptable
instability in the instrument.
Procedure for Energy Dispersive Spectrometers

1. Selectthe voltage (kV) and the tube current(!-LA) beforeanalysis.For
single elementanalysisa rule of thumb requiresthe kilovolt settingat
2.5 to 3 times the absorption edge energy for efficient excitation.
Therefore,for Fe (absorptionedge = 7.1) 20 kV should be efficient.
For multielementanalysis,
kV = 3/4 • E, + 3/2 Eh [41 ]
whereE[ = absorptionedgeof the lowestenergyline, andEh = absorp-

tion edgeof the highestenergyline to be analyzed.For a rangeof AI
(E, = 1.5 keV) to Fe (Eh =7.1 keV) Eq. [41] estimatesa kilovoltage of
11.8. If Fe is the major elementand AI the minor elementa kilovolt-
age of 10 would be sufficient. If both are major elementsor AI is the
major element,kilovolts of 15 to 20 would be the optimum voltage.
An optimum tube current is one that provides 10000 to 15000 cps.
Much higher countswill result in excessivedeadtime and loss of res-
olution, while much lower countsmay causecollection of inadequate
data.
2. Selectthe operationalkiloelectronvolt rangeand calibrate the system
using a spectrumof x-rays with two strong lines at known energies.
The line energiesshouldbe widely separatedwith a ratio 24:1. A typ-
ical calibration standardis an AI-Cu alloy disk. Calibration can be
done manually or automatically.The peak to backgroundratio of the
peakshould be at least5:1 and the counting rate in the rangeof 1000
to 5000 cps.The automaticcalibration mode performsa cycleof iter-
ations until the difference betweenthe location of the peak centroids
comparedto the theoreticalposition is less than or equal to 1 eV
3. Presetthe live fixed time or fixed count to the desiredlevel and pro-
ceed with the data collection processwhile the presetconditions are
satisfied.
4. Carefully place the sample and standardsin the sample holder and
insert them in the spectrograph.Evacuatethe sample chamberand
when the vacuumlevel reaches theappropriatelevel initiate the count-
ing process.Identify each samplewith a file name under which the
collecteddata are stored.
5. Plot the generatedx-ray spectrumor print grossor net intensitiesafter
correctingfor backgroundusing a linear or polynomial function pro-
vided by the manufacturer'sor other softwarepackage.
6. Obtain the quantitativecompositionof the sampleusing direct com-

parisonswith standardsand/orafter performingmatrix correctionsuti-
lizing the most suitableof the modelssuppliedby the manufacturer.
Calculationof Results
1. Calculatefor the standardand samplethe averagemeasuredcps over
a presetfixed time for the peak (Np ) and backgroundor blank (Nb ),
and the peak minus blank (Np - Nb ) or peak minus backgroundNp -
(Nb+ 1° _ Nb- 1o )!2.
2. Determinethe nominal concentrationof eachoxide (C%) in the sam-
ple from the equation
(Np - Nb ) standard
C%(sample)= (Np - Nb ) sample• [42]
C %(standard)
From the nominal concentrationscalculatedfrom Eq. [42] after cor-

recting for weight lossesduring the fusion process,the total composi-
tion of the samplecan be estimated,which ideally should total 100%.
If the sum of all oxide componentsis significantly lower or higher
than 100% (a good rule of thumb is more than 100 ± 5%), either there
are some missing componentsthat have not been measuredor there
are significant matrix effectswhich needto be compensatedfor.
3. Before attempting any matrix corrections determine whether dead
time (t) correctionscan improve the total recoveryof the sample.This
is accomplishedby converting the measuredcount (Nm ) to the cor-
rected count Nc with Nc - N m/(l - Nml) and using the corrected
countsin Eq. [42] to estimatethe oxide concentrations.
4. If the recoveryis still not satisfactory,selectthe appropriatemodel to
correct for matrix and interelementeffects. This is a tediousprocess
which requiresa significant effort, but it has beensimplified substan-
tially with the adventof microprocessors.Severalof thesemodelsare
being supplied now by the manufacturers.Using as an example the
Norrish and Hutton (1969) fusion procedurefor silicate analysisand
expressingthe nominal weight fraction (P) of eachoxide in the sam-
ple as a function of nominal concentration experimentallydetermined
by the linear approximation
P= C%/100, [43]
the correctedoxide concentrations(CA) for elementA in the sample

can be calculatedas follows
whereP =nominal weight fraction of the oxide in question,X =mass

absorptioncoefficient of the flux used for the characteristicwave-
length of the tube target;P A, PB, P e, ... = nominal weight fractions of

the oxidesin the sampleexperimentallyestimatedfrom linear approx-
imations; M A , M B , Me ... = correctedfactors for each oxide (essen-
tially the differencebetweenthe absorptioncoefficient of pure borate
glassand the massabsorptioncoefficient of elementsA, B, C ... for
the characteristicradiation used; P L = weight fraction for sample
weight lossesduring the fusion; and ML = correction factor for the
weight loss during the fusion for the characteristicradiation.
Norrish and Chappell(1977) presenteda table of experimental-
ly determinedand normalizedcorrectioncoefficientssuitablefor rou-
tine major elementalanalysisof silicate samples(Table 7-4). These
values are not true absorptioncoefficientsbut representthe effect of
borate glass on the massabsorptioncoefficient of the elementsana-
lyzed. Therefore,they are specificto the preparationprocedureand the
samples/standards/flux setsused by the authorsand should be recal-
culatedfor otherconditions.Using as an examplea sampleof compo-
sition Fe203= 5.32%, Ti0 2 = 3.98%, CaO = 2.50%, Si02 = 51.55%,
andAl 20 3 = 30.65%and a fusion loss of 5.00%determinedfrom a lin-
ear approximationanalysis,and following the procedureof Norrish
and Chappell(1977), the correctedpercentageof Fe203can be calcu-
lated accordingto Eq. [44] and Table 7-4 as follows
% Fe203= 5.32 {1.046 + [0.0532 (-0.027)]
+ [0.0398 (+0.146)] + [0.0250 (+0.134)] + [0.5155 (-0.065)]
+ [0.3065 (-0.074)] + [0.05 (-0.163)]} = 5.26% [45]
Similarly, calculatethe correctedpercentagesfor all elemental

oxides.This constitutesthe first iteration cycle.
5. Repeatthesecalculationsin a seconditeration cycle in which the orig-
inal approximated(nominal) weight fractions are substitutedby the
correctedvalues obtainedfrom the first iteration cycle. Continue the
corrective iterationprocessuntil it producesnegligible changes.In
practice,this is usually accomplishedafter two or three iterations.
6. The sum of the corrected percentagesof the elemental oxides and
volatiles (assumingall the elementspresentwere counted)should ide-
ally be within ±0.1% of 100%.
ANALYSIS OF MINOR AND TRACE ELEMENTS IN SOIL

AND PLANT SAMPLES
Preparationof PressedPowderPellets
Equipmentand Materials
1. Mechanicalgrinder, mixer mill or mortar and pestlemadeof materials
that will minimize contaminationby heavy metalsand able to pulver-
ize the sampleto a particle size of <50 )lm.
N
~
Table 7-4. Correctioncoefficientsfor major elementsin silicate analysist(Jones,1982).

y
LineKa X Fe203 MnO Ti0 2 CaO K 20 S03 P20S Si02 Al 20 3 MgO Na20 Loss
Fe 1.046 -0.027 -0.031 0.146 0.134 0.126 -0.060 -0.060 -0.065 -0.074 -0.090 -0.110 -0.163
Mn 1.045 -0.044 -0.044 0.146 0.135 0.130 -0.037 -0.063 -0.063 -0.074 -0.078 -0.100 -0.163
Ti 0.851 0.081 0.077 0.179 0.647 0.644 0.194 0.181 0.110 0.078 0.069 0.051 -0.132
Ca 0.865 0.090 0.092 0.065 0.110 0.723 0.201 0.182 0.128 0.105 0.068 0.051 -0.134
K 0.897 0.098 0.086 0.017 0.000 0.069 0.182 0.179 0.119 0.101 0.080 0.057 -0.139
S 0.894 0.086 0.074 0.002 -0.023 -0.037 -0.053 0.167 0.131 0.112 0.087 0.063 -0.139
P 0.896 0.108 0.094 -0.020 -0.037 -0.047 -0.059 -0.063 0.127 0.110 0.094 0.046 -0.139
Si 1.014 0.082 0.086 -0.034 -0.042 -0.055 -0.057 -0.061 -0.061 0.122 0.093 0.063 -0.158
AI 1.056 0.112 0.116 -0.032 -0.037 -0.048 -0.056 -0.060 -0.088 -0.072 0.116 0.058 -0.164
Mg 1.050 0.136 0.126 0.010 -0.021 -0.043 -0.046 -0.016 -0.070 -0.078 -0.084 0.080 -0.163
Na 1.032 0.158 0.142 0.041 0.005 -0.016 -0.034 -0.044 -0.053 -0.066 -0.081 -0.103 -0.161
t With permissionfrom J. Zussman1977. In this table the coefficientsare for concentration(weight fraction) of the oxide, not concentrationof the element.Thesecoef-
ficients apply only when the samplesare preparedaccordingto the Li 2B40,Li 2COr LaO proceduredescribedearlier. Similar, but numericallydifferent, coefficients
can be derived for different samplepreparations.With the exceptionof NaKa, the coefficientsare taken fromNorrish and Hutton (1969). The valuesfor NaKa have ~
beencalculated;Norrish and Hutton (private communication)have reducedthe coefficient of CaO on CaKa.
~
Ro
~
~
(a) Steel cylinder (b) Steel plunger

Snug fit in (a)
l
30.4 mm dlam. 30.4 mm dlam.
I .,
I· ·'
1
E
E
Ie
I
(C)~6mm
~J
bd
25.4mm
E
E
ill
10.28 mm
30.1 mm
r~.• mm·'l
(f) Perope. plunger
easy fit In (e)
E Polished both ends
E
&l
u....--......J
(0) Steel plunger as (f)
Note: Material for ali steel

(e) Aluminum ube partl II ComsteelKIO
Easy fit In (a) (tool steel)
Fig. 7-5. Accessoriesfor preparingpressedpowdersamples[with permissionfrom J. Zussman(ed.)

1977; Physical methodsin determinativemineralogy.(2nd ed.), Acad. Press,Inc. (London) Ltd.,
p. 263, copyright by Acad. PressInc. (London) Ltd.]
2. Hydraulic presscapableof reachingpressuresof up to 25 t.

3. Pellet-forming die set constructedof highly polished hardenedtool
steel with accessoriesas describedin Fig. 7-5 (Norrish & Chappell,
1977; Fabbi, 1972).
4. Boric acid, analytical gradereagentor chromatographiccellulosefor
backingpellets.
5. Plasticor paperenvelopesand desiccatorsfor storing pellets.
Most of the above equipment and materials are available
throughSpexIndustries,Inc., Edison,NJ 08820; Beckman-RIICLtd.,
London, SE26, England,and other manufacturers.
Procedure
1. Samplepreparation:(i) For soil samplesgrind a sampleof about 250
g in a 250-cm3 capacitymill for about5 min or if a mixer mill is used
for about 10 min. Generallya finely ground«50 !lm) 2-g samplewill
be adequatefor making an infinitely thick pellet suitablefor analyzing
most K spectrafor elementswith Z < 40 and all L spectra.(ii) Plant
materialsshouldbe oven dried and coarselychoppedor groundbefore
pulverizing a subsamplewith a small coffee mill. Oven dry again and
store in a desiccator.Usually a 2-g pulverizedand oven-driedsample
will producean infinitely thick pellet suitablefor the determinationof
elementswith Z :$; 30 (Jenkinset aI., 1966).
2. The following procedurehas been modified from that describedby
Norrish and Hutton (1964) and Baird (1970). Placea steelcylinder (a)
(Fig. 7-5) on the baseunit (c) of the die set. Insert a highly polished
steel disc (not shown in Fig. 7-5) and then fit in (a) the aluminum
inner tube (e). Pour in 2 g of sampleand spreadevenly by pressing
down gently with the Perspexplunger(t). Plant samplesmay require
considerablymore pressurethan soil samples.By pressingon the
plunger(t) or (g) and rotating the aluminum innertube,the sampleis
freed from themso that they can be withdrawn,while leaving the sam-
ple as a flat disc. Pour in about2 g of boric acid aroundthe edgeleft
by the aluminum tube and on top so that a sample is encased.The
amountof boric acid is not critical but it shouldbe enoughto form an
effective backing.
3. Insert steel plunger (b) in (a) and place the die system under the
hydraulicpress.Exert a pressurebetween5 and 10 t (dependingon the
type of the sample)for about1 to 2 min so that a cohesivepellet is pro-
duced.
4. After the pressureload hasbeenreleasedreplacethe baseunit (c) with
the hollow base(d) and pressthe plungeruntil the pellet is ejected.
5. Label the pellets on the backing with a felt-tip pen, place them in
envelopesand store themin desiccators.
6. Preparestandardswith compositionsimilar to the unknown ~amples
foHowing the sameprocedure.
Insteadof boric acid a few drips of polyvinyl alcohol solution (dissolved

in warm water) can be addedto the sampleand the mixture can be pressedin a
die systemwithout backing(Jones,1982). The procedurefor measuring fluores-
cent intensitiesfrom powderpresspellets and the calculationof elementalcon-
centrationsare similar to thosedescribedin the previoussection.
ElementalAnalysis of LoosePowderand Liquid Samples
Pulverizedsoil or plant samplesalso can be analyzedas loose powders

without pressingthem into a pellet. The powder is placedin specially designed
cells madefrom Teflon, polyethyleneor polypropylenematerialsand the cell is
coveredwith a Mylar film (usually 6 11m thick), which is held firmly in place
with a snap-ring.The film supportsthe samplewhen the cell is placedinverted
in the spectrographwithout interfering significantly with penetrationor absorp-
tion of the x-rays despitethe lack of a vacuumpath. Extensiveuseof this proce-
dure hasbeenmadein pedologicalstudiesinvestigatingdepthfunctionsof Ti and
Zr in order to evaluatesoil developmentand uniformity of parentmaterial.
Certain heavy metals also can be measuredby XRFS in liquid samples
derived from various extractionsutilizing the same sample cells and Mylar cov-
ers describedabove. One of the most commonapplicationsof this techniqueis
the determinationof Fe in citrate-bicarbonate-dithionite extracts.
Other applicationsinclude the determinationof total KzO contenton Mg-
saturatedsoil or clay samplespastedon glass slides used for x-ray diffraction
analysis(Karanthasis& Hajek, 1982), and cation exchange capacity determina-
tion on clay mineralsin powdersamples(Greenland,1974).
OTHER APPLICATIONS
Many other reports of analysisof soil geological and plant materialsby

XRFS can be found in the literature involving a variety of techniques,which are
suitablefor particularcases.Although it is not possibleto list all theseapplica-
tions, the following may prove helpful in particularanalytical problems.
X-ray fluorescencespectrometryhasbeenappliedto many aspectsof earth
sciencesincluding soil science,geology,mineralogy,and relevantenvironmental
problemswith remarkablesuccessand comparingvery favorably with the per-
formance of other analytical techniques (Ahmedali, 1989; Bartenfelder &
Karathanasis,1988). The versatility, easeand quicknessof the technique has
establishedit as a useful tool in multielementsurveysfor both major (Williams
& Rayner, 1977; Watts, 1977; Watatsuki et aI., 1977; Karathanasis& Hajek,
1983; Kirkland & Hajek, 1972; Low & Bristow, 1983; Subramanian,1979) and
trace elements(Drees & Wilding, 1973; Evans & Adams, 1975; Oertel, 1961;
Gilkes et aI., 1973; Childs, 1975; Bradleyet aI., 1978; Pottset aI., 1990; Erzinger
& Puchelt, 1982; Russ et aI., 1981; Kohno et aI., 1990). Table 7-5 summarizes
the analyticalconditionsfor XRFS, determinationof commonsoil elementswith
mean concentrations,samplepreparationand minimum detectionlimits (Jones,
1982).
Sulfur determinationby XRFS hasprovidedrapid and reliable data in acid
sulfate soils (Bloomfield et aI., 1968), various soil fractions (Brown & Kanaris-
Sotiriou, 1969; McLaren & Swift, 1977; Darmodyet aI., 1977; Randal& Sakai,
1983), and soil extracts (Roberts & Kochler, 1968). Several investigations
involving the analysisof Mn, Co, Zn, Cu, Fe, and Mo in soil samplesalso have
been conducted with satisfactory results (Williams, 1976; Wilkins, 1979;
Keramidas& Fanning,1976),while determinationsof Ti and Zr havebeenused
in pedogenicandsoil erosionstudies(Chapman& Horn, 1968; Bain, 1976; Kaup
& Carter, 1987; Marsan et aI., 1988; Murad, 1978; Olson & Beavers, 1987;
Sobecki & Karathanasis,1992). X-ray fluorescenceanalysisof Cr, Ni, Zn, As,
Table 7-5. Analytical conditionsfor XRF analysisof commonsoil elements,samplepreparationmethod,meanconcentrationsand detectionlimits by wavelengthdis- to.>
....
00-
persivespectroscopy(Jones,1991).t
Optimal radiation Analyzing Spectral Interelement Samplepreparation Mean Detection
Element & source crystal line 28 Detector* interferences method concentration limit
F Sc TIAP Ka 90.62 FPC Pressedpellet 200
Na Sc TIAP Ka 55.08 FPC Pressedpellet 0.6% 15
Mg Sc TIAP Ka 45.18 FPC Ca Glassdisk 0.5% 0.02%
AI Sc PET Ka 145.07 FPC Glassdisk 7.1% 0.01%
Si Sc PET Ka 109.21 FPC Glassdisk 33.0% 0.01%
P Sc GeIII Ka 140.90 FPC Glassdisk 0.06% 0.004%
S Sc GeIII Ka 110.68 FPC Pressedpellet 700
Cl Sc GeIII Ka 92.75 FPC Pressedpellet 100
K Cr PET Ka 50.69 FPC Glassdisk 1.4% 0.003%
Ca Cr LiF 200 Ka 113.06 FPC/SAC Glassdisk 1.4% 0.002%
Ti Cr LiF 200 Ka 86.12 FPC/SAC Glassdisk 0.5% 0.002%
Cr Au/W LiF 200 Ka 69.34 FPC/SAC V,Ba Pellet/glassdisk 100 <1
Mn Au/W LiF 200 Ka 62.96 FPC/SAC Cr Pellet/glassdisk 850 <1
Fe Au/W LiF 200 Ka 57.51 FPC/SAC Mn Pellet/glassdisk 3.8% <1
Co Au/W LiF200 Ka 52.79 FPC/SAC/SC Fe 8 <1
Ni Au LiF 200 Ka 48.66 FPC/SAC/SC Pressedpellet 40 <1
Cu Au LiF 200 Ka 45.02 FPC/SAC.SC Ni Pressedpellet 20 <1
Zn Au LiF 200 Ka 41.79 FPC/SAC/SC Au,Cu Pressedpellet 50 <1
As Mo/Au/W LiF 200 Ka 33.99 SC/SXC Pb Pressedpellet 6
Se Mo/Au/W LiF 200 Ka SC/SXC Preconcentration <1
Br Mo/Au/W LiF 200 Ka 29.96 SC/SXC Pressedpellet 5
Rb Mo/Au/W LiF 200 Ka 26.91 SC/SXC Br Pressedpellet 100 <1
Sr Mo/Au/W LiF 200 Ka 25.14 SC/SXC Pressedpellet 300 <1 ~
Zr Mo/Au/W LiF 200 Ka 22.55 SC Sr Pressedpellet 300 <1
Mo Au/W LiF 200 Ka 20.84 SC Preconcentration 2 ~
Cd Mo/Au/W LiF 200 KalLa 15.32/158.3SC/FPC Preconcentration 0.06 >
r:Il
Cs Cr LiF 200 La 91.72 FPC Pressedpellet ....
r:Il
Ba Au/W/Mo LiF 200 L~ 79.18 FPC Ti Pressedpellet 500 4 rI:o
La Au/W/Mo LiF220 La 138.58 FPC Preconcentration 30
Hg Au/Mo LiF 200 La 35.89 FPC Preconcentration 0.03--0.8 ~
Pb Au/Mo LiF 200 L~ 28.23 SC Sn Pressedpellet 10 5 t"j
~
t A Rh sourcealso can provide suitableexcitationover a wide rangeof elements.
* FPC =flow proportionalcounter,SAC =sealedAr counter,SC =scintillation counter,SXC =sealedXe counter.
~
Table 7-6. Analytical conditionsfor a WDS unit and detectableconcentrationrangesof commonelementspresentin ground plant materials(Kubota & Lazar, 1971).t
Element Plant material Analyzing crystal Wavelength Voltage Tube current Detector Path Concentrationrange
Ka BG kV rnA
i!
P Forageplant Quartz 6.155 6.108 50 40 FPC + PHA He 0.12--0.60%
S Forageplant NaCI 5.373 5.228 50 40 FPC He 0.05--0.60%
Cl Forageplant NaCI 4.729 5.228 25 20 FPC He 0.03--0.52%
K Forageplant Quartz 3.742 3.641 35 30 FPC He 0.10-4.00%
Ca Forageplant Quartz 3.359 3.641 35 30 FPC He 0.1~3.oo%
~
Cr Manure LiF 2.291 2.164 19 18 SC Air 0.01-1.17% ~
Mn Forageplant LiF 2.103 2.044 50 40 SC Air 15-120mglkg
Fe Forageplant LiF 1.937 2.044 50 40 SC Air 2~1oo mglkg
Co Woody plant LiF 1.791 1.860 50 40 SC Air 1.5--850mglkg
Ni Grain LiF 1.659 1.606 50 40 SC Air 0.3-33 mglkg
Cu Plant ash LiF 1.542 1.606 50 40 SC Air 63-522 mglkg
Zn Forageplant LiF 1.437 1.407 50 40 SC Air 12-50 mglkg
Sr Plant ash LiF 0.877 0.903 50 40 SC Air 2.5-400mgkg
Mo Forageplant LiF 0.710 0.731 50 40 SC Air 3-3OOmglkg
t WDS = wavelengthdispersivespectrometer,BG = background,FPC = flow proportionalcounter,PHA = pulse height analyzer,SC = scintillation counter.
N
...
-.I
Cu, and Pb, which are the main constituentsof many pollutantshas beenused
extensively in recent years and contributed reliable information in assessing
environmentalproblems(Wilkins, 1978; Piorek, 1990; Harding & Walsh, 1990;
Kendall et aI., 1984; Paveleyet al., 1988; Bowen, 1979). Although other impor-
tant potential pollutants such as Cd, Sn, and Hg are usually found below the
XRFS detectionlimits in environmentalsamples,preconcentrationtechniques
(precipitation, filtration, ion exchange,chelation) have greatly improved their
detectionand measurement(Murata et aI., 1984; Eidecker& Jackwerth,1987;
Van Grieken et aI., 1977). Most recently, the advent of synchrotronradiation
sourceshas allowed the sensitivity of XRFS to reachdetectionlimits as low as
0.02 mg kg-1 for most elementswith the utilization of much smaller samples,
thus essentially eliminating the need for preconcentrationbefore analysis
(Giauqueet aI., 1986).
Comprehensiveaccountsof x-ray emissionapplicationsfor the analysisof
plant materialshavebeenprovidedby Kubota andLazar(1971),and Norrish and
Hutton (1977). Someof thesereportsinclude detailedproceduresfor corrections
of matrix and interelementeffects,while otherssuggestthat thesecorrectionsare
importantonly whenthe massabsorptioncoefficientof the measuredelementfor
its own radiation is significantly greaterthan that of the matrix (Jenkinset aI.,
1966; Mudroch & Mudroch, 1977). Linear approximationshavebeenfound sat-
isfactory for a rangeof elementsin forage, grain, horticultural and woody sam-
ples (Ball & Perkins, 1962; Kubota & Lazar, 1971) and are in good agreement
with results obtained by atomic absorption spectroscopy.X-ray fluorescence
analysisof plant and other biological materialsfor S (Kubota & Lazar, 1967),
Cu, Cr, and As (Reistad& Petterson,1973), and a variety of heavy metalshas
beenreviewedby Brandt and Lazar(1958), Dixon and Wear (1964), Kubota and
Lazar (1971), Kornev et al. (1982), Rethfeld (1982), Manninen et al. (1980),
Valkovic (1980)andothers.Table7-6 summarizesanalyticalconditionsfor mea-
suring a variety of elementsin plant materials, their concentrationrange and
meancps ratios (Kubota & Lazar, 1971).
Standardsfor XRF analysis of plant materials are available from the
Natural Instituteof Standardsand Technoloy,but they also can be preparedfrom
cellulosematerialimpregnatedwith a solution containingknown concentrations
of the elementto be analyzed.The impregnatedcellulose material should be
dried at no°c and thoroughly mixed beforeanalysis.Suchstandardsare usual-
ly satisfactoryfor a rangeof samplesand elementswith Z :s; 30, but for certain
elementsseparatestandardsmay be required(Kubota & Lazar, 1967).
REFERENCES
Abbey, S. 1980. Studiesin "standardsamples"for use in the generalanalysisof silicate rocks and
minerals.Part6. 1979ed. Geol. Surv. Can. Pap.80-14. Geol. Surv. Can., Ottawa,Canada.
AhmedaJi,S.T. (ed.). 1989. X-ray fluorescenceanalysis in the geological sciences.Advancesin
methodology.Short CourseVol. 7. Geol. Assoc.Canada,Montreal, Canada.
Andermann, G.,andJ.w. Kemp. 1958. Scatteredx-rays as internalstandardsin x-ray emissionspec-
troscopy.Anal. Chern.30;1306-1309.
Bain, D.C. 1976.A titanium-rich soil clay. J. Soil Sci., 27:68-70.
Baird, A.K. 1970. Light elementanalysis.Adv. X-Ray Anal. 13:26-48.
X-RAY FLUORESCENCE SPECTROSCOPY 219
Ball, D.E, and D.E Perkins.1962. Plant analysisby x-ray fluorescencespectrograpy:Determination

of calcium and potassium.Nature (London) 194:1163-1165.
Bartenfelder,D.C., and A.D. Karathanasis.1988.A comparisonbetweenx-ray fluorescenceand dis-
solution methodsemploying lithium metaboratefusion for elementalanalysisof soil clays.
Commun.Soil Sci. Plant Anal. 19:471-492.
Beattie,HJ., and R.M. Brissey. 1954. Calibrationmethodfor x-ray fluorescencespectrometry.Anal.
Chern.26:980-983.
Bennett,H., and GJ. Oliver. 1976. Developmentof fluxes for the analysisof ceramicmaterialsby x-
ray fluorescencespectrometry.Analyst 101:803-807.
Bernardo,L.M., RB. Clark, D. Knudsen,and 1.W. Maranville. 1985. Analysis of cationic elements
in liquid samplesby x-ray. Commun.Soil Sci. Plant Anal. 16:823-835.
Berry, P.E, T. Furuta,and J.R Rhodes.1968. Particlesize effectsin radioisotopex-ray spectrometry.
Adv. X-Ray Anal. 12:612-632.
Bertin, E.P. 1975. Principles and practicesof x-ray spectrometricanalysis. 2nd ed. Plenum Press,
New York.
Birks, I.S. 1969. X-ray spectrochemicalanalysis.2nd ed. Intersci., New York.
Bloomfield, c., 1.K. Coulter, and R. Kanaris-Sotiriou. 1968. Oil palms on acid sulphatesoils in
Malaya. Trop. Agric. 45:289-300.
Blount, c.w., D.E Leyden,T.!. Thomas,and S.M. Gill. 1973.Application of chelatingion-exchange
resinsfor traceelementanalysisof geologicalsamplesusing x-ray fluorescence.Anal. Chern.
45:1045-1050.
Bowen, HJ.M. 1979. Environmentalchemistryof the elements.Acad. Press,London.
Bower, N.W., and G. Valentine. 1986. Critical comparisonof samplepreparationmethodsfor major
and trace elementdeterminationsusing x-ray fluorescence.X-Ray Spectrom.15:73-78.
Bradley,I., C.c. Rudefort,and C. Wilkins. 1978. Distribution of somechemicalelementsin the soils
of north west Pembrokeshire.J. Soil Sci. 29:258-270.
Brandt, C.S., and Y.A Lazar. 1958. Analysis of dried plant materialby x-ray emissionspectrograph.
J. Agric. Food Chern. 6:306--309.
Brondle, C.R., and AD. Baker. 1979. Electron spectroscopy:Theory, techniquesand applications.
Acad. Press,New York.
Brown, G., and R Kanaris-Sotirou.1969. Thedeterminationof sulphurin soils by x-ray fluorescence
analysis.Analyst (London) 94:782-786.
Carlson,T.A 1978. X-ray photoelectronspectroscopy.Acad. Press,New York.
Carlton, D.T., and J.C. Russ. 1976. Trace level water analysisby energy-dispersiveX-ray fluores·
cence.X-Ray Spectrom.5:172-174.
Castaing,R. 1961.The fundamentalsof quantitativeelectronprobe microanalysis.Adv. X-Ray Anal.
4:351-369.
Chapman,S.L., and M.E. Hom. 1968. Parentmaterial uniformity and origin of silty soils in north-
west Arkansasbasedon Zr and Ti contents.Soil Sci. Soc. Am. Proc. 32:265-271.
Childs, c.w. 1975. Composition of iron-manganeseconcretionsfrom some New Zealand soils.
Geoderma13:141-152.
Chung, EH., AJ. Lentz, and RW. Scott. 1974. A versatile thin film method for quantitativeX-ray
emissionanalysis.X-Ray Spectrom.3:172-175.
Claisse,E 1956. Accurate x-ray analysiswithout internal standard.Prelim. Rep. 327. QuebecDep.
Mines, Quebec,Canada.
Claisse,E, and M. Quintin. 1967. Generalizationof the Lachance-Traill methodfor the correctionof
matrix effects in x-ray fluorescenceanalysis.Can. Spectrosc.12:129-133.
Criss, 1.W., and L.S. Birks. 1968. Calculationmethodsfor fluorescentx-ray spectrometry-empiri-
cal coefficientsvs. fundamentalparameters.Anal. Chern. 40:1080-1086.
Darmody,R.G., D.S. Fanning,w.J. Drummond,and J.E. Foss.1977. Determinationof total sulfur in
tidal marshsoils by x-ray spectroscopy.Soil Sci. Soc. Am. Proc. 41:761-765.
Dixon, J.B., and 1.1. Wear. 1964. X-ray spectrographicanalysisof zinc, manganese,iron, and copper
in plant tissue.Soil Sci. Soc. Am. Proc. 28:744-746.
Drees,L.R, and L.P. Wilding. 1973. Elementalvariability within a samplingunit. Soil Sci. Soc. Am.
Proc.37:82-87.
Eastell,J., and J.P. Willis. 1990. A low dilution fusion techniquefor the analysisof geologicalsam-
ples: 1. Method and trace elementanalysis.X-Ray Spectrom.19:3-14.
Eidecker,R., and E. Jackwerth.1987. Multielementpreconcentrationfrom iron-containingsoils and
sediments.FreseniusZ. Anal. Chern. 328:469-474.
Erzinger, J., and H. Puchelt. 1982. Methods for the determinationof trace elementsin geological
materials.ErzemetaIl35:173-179.
Evans,LJ., and W.A Adams. 1975. Quantitativepedologicalstudieson soils derived from Silurian
mudstones.IV. Uniformity of the parentmaterial and evaluationof internal standards.J. Soil
Sci. 26:319-326.
Fabbi, B.P. 1972. A die for pelletizing samplesfor x-ray fluorescenceanalysis.X-Ray Spectrom.
1:39-40.
Flanagan,F.F. 1986. Referencesamplesin geology and geochemistry.u.S. Geol. Surv. Bull. 1582.
U.S. Gov. Print. Office, Washington,DC.
Giauque,R.D., J.M. Jaklevic, and A.C. Thompson.1986. Trace elementdeterminationusing syn-
chrotron radiation.Anal. Chern. 58:940-944.
Gilkes, R.I., G. Scholz,and G.M. Dimmock. 1973. Lateritic deepweatheringof granite.J. Soil Sci.
24:523-536.
Goodman,R.I. 1973. Rapid analysisof trace amountsof tin in streamsediments,soils and rocks by
x-ray fluorescenceanalysis.Econ. Geol. 68:275-278.
Greenland,DJ. 1974. Determinationof pH dependentchargesof clays using cesiumchloride and x-
ray fluorescencespectrography.p. 278--285.In S.N. Taichivov (ed.) Trans. Inl. Congr. Soil
Sci. Trans. 10,2. RISO BFAN, UFa, SSSR
Gunn, B.M. 1976. The use of computers in X-ray fluorescence analysis. X-Ray Spectrom.
5:175-177.
Hammerle,R.H., RH. Marsh, K. Rendan,RD. Giauque,and J.M. Jaklevic. 1973.Testsof x-ray flu-
oresCencespectrographyas a method for analysisof the elementalcompositionof atinos-
pheric aerosols.Anal. Chern.45:1939-1940.
Harding, AR., and J.P. Walsh. 1990. Application of field mobile EDXRF analysisto contaminated
soil characterization.Adv. X-Ray Anal. 33:647--654.
Haukka,M.L., and I.L. Thomas.1977.Total x-ray fluorescenceanalysisof geologicalsamplesusing
low dilution lithium metaboratefusion method.Matrix correctionsfor major elements.X-Ray
Spectrom.6:204-211.
Heinrich, K.F.J. 1961. Pulseheight selectionin x-ray fluorescence.Adv. X-Ray Anal. 4:370-381.
Henke, B.L. 1965. Some noteson ultrasoft x-ray fluorescenceanalysis-1O-100Aregion. Adv. X-
Ray Anal. 8:269-284.
Houseknecht,T.M. 1983. Fusion techniquesfor samplepreparationin x-ray fluorescenceanalysis.
Pit Quarry. 72:72-75.
Hutton, J.T., and S.M. Elliot. 1980.An accurateXRF methodfor the analysisof geochemicalexplo-
ration samplesfor major and traceelementsusing one glassdisc. Chern. Geol. 29:1-1l.
Jenkins,R 1974.An introduction to x-ray spectrometry.X-Ray Spectrom.3:28A
Jenkins,R 1976. An introduction to x-ray spectrometry.Heyden,London.
Jenkins,R, and J.L. DeVries. 1970. Practicalx-ray spectrometry.Philips Tech. Library, Macmillan
& Co., Ltd., London.
Jenkins, R., and DJ. Haas. 1973. Hazardsin the use of x-ray analytical instrumentation.X-Ray
Spectrom.2:135-14l.
Jenkins,R., P.W. Hurley, and Y.M. Shorrocks.1966. Plant mineral analysisby x-ray fluorescence
spectrometry.Analyst (London) 91:395-397.
Jenkins,R, RW. Gould, and Dale Gedcke.1981. Quantitativex-ray spectrometry.Marcel Dekker,
Inc., New York.
Johnson,W.M., and J.A Maxwell. 1981. Rock and mineral analysis.2nd ed. ChemicalAnalysisSer.
Monogr. Vol. 27. JohnWiley & Sons,New York.
Jones,AA 1982. X-ray fluorescencespectrometry.p. 85-121,In AL. Pageet al. (ed.) Methodsof
soil analysis.Part 2. 2nd ed. ASA and SSSA,Madison,WI.
Kalman, Z.H., and L. Heller. 1962. Theoreticalstudy of x-ray fluorscent determinationof tracesof
heavy elementsin a light matrix. Anal. Chern. 34:946-951.
Kanaris-Sotiriou,R., and G. Brown. 1969. Diminution of sulphur contaminationof powder speci-
mensin X-ray fluorescenceanalysis.Analyst (London) 94:780-782.
Karathanasis,AD., and B.F. Hajek. 1982. Revisedmethodsfor rapid quantitativedeterminationof
mineralsin soil clays. Soil Sci. Soc. Am. J. 46:419-425.
Karathanasis,AD., and B.F. Hajek. 1983. Transformationof smectiteto kaolinite in naturally acid
soil systems:Structuralandthermodynamicconsiderations.Soil Sci. Soc.Am. J. 47:158--163.
Kaup, B.S., and BJ. Carter. 1987. Determining Ti sourceand distribution within a paleustalfby
micromorphology,submicroscopyand elementalanalysis.Geoderma40:141-156.
Kendall, D.S., J.H. Lowry, E.L. Bour, and T.J. Meszaros.1984. A comparisonof trace metal deter-
minations in contaminatedsoils by XRF and ICAP spectroscopies.Adv. X-Ray Anal.
27:457-463.
Keramidas,V.Z., and D.S. Fanning.1976.Correctionof "tube contribution"interferencein the deter-

mination of heavy metalsby x-ray spectroscopyusing the "additions technique."Soil Sci.
Soc. Am. Proc. 40:857--860.
Kerrigan, G.C. 1971.A samplepreparationdevicefor x-ray fluorescence.J. Phys.E. 4:544-545.
Kimoto, S., S. Sato,H. Kamada,and T. Ui. 1966. On the primary x-ray analyzer.Adv. X-Ray Anal.
9:508-514.
Kirkland, D.L., and B.F. Hajek. 1972. Formuladerivationof the interlayeredvermiculite in selected
soil clays. Soil Sci. 114:317-322.
Knapp,G., B. Schreiber,and R.W. Frei. 1975. A simple concentrationprocedurefor tracemetalsfor
x-ray fluorescenceand atomic absorptionspectrometry.Anal. Chim. Acta 77:293-297.
Kohno, M., K. Tamura,and J. Azuma. 1990. Total analysisof major constituentsin soils by x-ray
fluorescencespectrometryusing calibration lines preparedwith soil referencematerials.
Pedologist34:37-46.
Kornev, E.A., AA Rimski-Korsakov, N.F. Bondarenko,v.v. Smirnov, AV. Malystenkov, V.I.
Kucheryuk,and N. Batygin. 1982. X-ray fluorescenceanalysisof traceelementsin plant and
soil samples.Sov. Agric. Sci. 11:5-6.
Kubota,J., and V.A Lazar. 1967. Routinex-ray emissionspectrographicanalysisof commonforage
plants. p. 93-107. In G.w. Hardy et al. (ed.) Soil testing and plant analysis,Part 2. SSSA,
Madison,WI.
Kubota, J., and V.A Lazar. 1971. X-ray emissionspectrograph:Techniquesand usesfor plant and
soil studies.p. 67--82.In I.M. Walsh(ed.) Instrumentalmethodsfor analysisof soils andplant
tissue.SSSA,Madison,WI.
Lachance,G.R., and R.J. Traill. 1%6. A practicalsolution to the matrix problem in x-ray analysis.
Cana.Spectrosc.11: 43-48, 63-71.
Leake, B.E., G.L. Hendry, A Kemp, AG. Plant, P.K. Harrey, J.R. Wilson, J.S. Coats,J.W. Aucott,
T. Liine1 and R.I. Howarth. 1970.The chemicalanalysisof rock powdersby automaticx-ray
fluorescence.Chern.Geol. 5:7--86.
Lee, R.F., and D.M. McConchie.1982. Comprehensivemajor and trace elementanalysisof geolog-
ical materialsby x-ray fluorescence,using low dilution fusions. X-Ray Spectrom.11:55-63.
Levinson, AA., L. dePablo.1975. A rapid x-ray fluorescenceprocedureapplicableto exploration
geochemistry.J. Geochem.Explor. 4:399-408.
Long,I.V.P. 1977. Electron probe microanalysis.p. 273-341.In 1. Zussman(ed.) Physicalmethods
in determinativemineralogy.2nd ed. Acad. Press,New York.
Low, AB., and 1.W. Bristow. 1983. X-ray fluorescencespectrometry:A useful tool in the chemical
characterizationof soils. S. Mr. J. Sci. 79:52-55.
Lucas-Tooth,J., and C. Pyne. 1964. The accuratedeterminationof major constituentsby x-ray fluo-
rescentanalysisin the presenceof large interelementeffects.Adv. X-Ray Anal. 7:523.
Luke, C.L. 1%8. Determinationof trace elementsin inorganicand organicmaterialsby x-ray fluo-
rescencespectroscopy.Anal. Chim. Acta 41:237-250.
Manninen,S., T. Paakkari,and A Ailta. 1980. An energydispersivex-ray fluorescencespectrome-
ter for traceelementanalysisof plant materials.Kemia-Kemi 7:107-109.
Marsan,F.A, D.C. Bain, and D.M.L. Duthie. 1988. Parentmaterialuniformity and degreeof weath-
ering in a soil chronosequence in northwesternItaly. Catena15:507-517.
McKinley, T.D.K, F.I. Heinrich, and D.B. Wittry. 1%6. The electron microprobe. John Wiley &
McLaren, R.G., and R.S. Swift. 1977. Changesin soil organicsulphur fractionsdue to the long term
cultivation of soils. J. Soil Sci. 28:445-453.
Mudroch,A, and O. Mudroch. 1977.Analysis of plant materialby x-ray fluorescencespectrometry.
X-Ray Spectrom.6:215-217.
Murad, E. 1978. Yttrium and Zirconium as geochemicalguide elementsin soil and stream sediment
sequences.J. Soil Sci. 29:219-223.
Murata, M., M. Omatsu,and S. Mushimoto. 1984. Solventextraction-microdropletx-ray fluores-
cenceanalysisof traceamountsof heavy metal ions. X-Ray Spectrom.13:83--86.
Norrish, K. 1959. The analysisof lead and zinc ores, and associatedmaterialsby x-ray fluorescent
spectroscopy.I. The determinationof Mn, Fe, Cu, Zn, As, Pb and Sb using borax fusions.
CSIRO Tech. Memo. Div. Soils no. 7. CSIRO, Melbourne,Australia.
Norrish, K., and B.W. Chappell.1977.X-ray fluorescencespectroscopy.p. 201-272.In J. Zussmann
(ed.) Physicalmethodsin determinativemineralogy,Acad. Press,London.
Norrish, K., and J.T. Hutton. 1964. Preparationof samplesfor analysisby x-ray fluorescentspec-
trography.I. Fusion in borateglass.II. Powdersamples.CSIRO Div. Soils Rep. 3. CSIRO,
Melbourne,Australia.
Norrish, K., and J.T. Hutton. 1969. An accuratex-ray spectrographicmethodfor the analysisof a
wide rangeof geologicalsamples.Geochim.Cosmochim.Acta 33:431-453.
Norrish, K., and J.T. Hutton. 1977.Plantanalysesby X-ray spectrometry.I. Low atomic numberele-
ments,sodiumto calcium. X-Ray Spectrom.6:6-11.
Oertel,AC. 1961. Relationbetweentrace-elementconcentrationsin soil and parentmaterial.J. Soil
Sci. 12:119-128.
Oertel, A.C. 1971a.Computerprogramsfor use in fluorescentx-ray analysesof silicated soils and
vegetablematter.CSIRO Div. Soils Tech. Pap.4. CSIRO, Melbourne,Australia.
Oertel, A.C. 1971b.Computerprogramsfor use in x-ray spectrography.Determinationsof traceele-
mentsof deadtime counters.CSIRO Div. Soils, Tech. Pap.5.
Olson,K.R, andAH. Beavers.1987.A methodto estimatesoil lossfrom erosion.Soil Sci. Soc.Am.
J.51:441-445.
Parker,A 1969. Some trace element determinations of the new U.S.G.S.silicate rock standards.
Chern.Geol. 4:445-449.
Paveley,C.F., B.E. Davies,and K. Jones.1988.Comparisonof resultsobtainedby x-ray fluorescence
of the total soil and the atomic absorptionspectrometry assayof an acid digestin the routine
determinationof lead and zinc in soils. Commun.Soil Sci. Plant Anal. 19:107-116.
Piorek, S. 1990. XRF techniqueas a methodof choice for on-siteanalysisof soil contaminantsand
wastematerial.Adv. X-Ray Anal. 33:639-645.
Poole,AR 1973. Determinationsof carbonin sedimentsby electronbeamx-ray excitation. X-Ray
Spectrom.2:165-168.
Potts, PJ., P.C. Webb, and J.S. Watson.1990. Exploiting energydispersivex-ray fluorescencespec-
trometry for the determinationof traceelementsin geologicalsamples.Anal. Proc.27:67-70.
Randall,PJ., and H. Sakai. 1983. Sulfur analytical methodsfor soils, plants,and animal samples.p.
255-269. In GJ. Blair and AR. Till (ed.) Sulfur in South-eastAsian and South Pacific agri-
culture. Aust. Dev. Assist. Bur., Armidale, New SouthWales,Australia.
Raspberry,S.D., and K.FJ. Heinrich. 1974.Calibrationfor interelementeffectsin x-ray fluorescence
analysis.Anal. Chern.46:81-89.
Reed,S.J.B. 1975. Electron microprobeanalysis.CambridgeUniv. Press,New York.
Reistad,T., and B. Pettersson.1973. Computerizedcorrectionfor elementinteractionin x-ray fluo-
rescenceanalysiswith specialreferenceto copper,chromiumandarsenic insolutionsandpre-
servedwood. X-Ray Spectrom.2:129-133.
Rethfeld,H. 1982. Determinationof tracesof heavy metalsin foods and feedingstuffs of vegetable
and animal origin, soil extracts,sewagesludges,and floor coatingsby meansof x-ray fluo-
rescenceanalysis.FreseniusZ. Anal. Chern.310:127-130.
Roberts,S., and F.E. Kochler. 1968.An x-ray spectrographicmethodof determiningtotal sulphurin
soil extracts.Soil Sci. 106:164-171.
Rousseau,RM. 1987. Painless XRFanalysisusing new generationcomputerprograms.p. 277-297.
In S.T. Ahmedali (ed.). X-ray fluorescenceanalysisin the geologicalsciences.advancesin
methodology.Geol. Assoc.Can.,Montreal, Quebec.
Russ,J.C., C.S. Barrett, P.K. Predecki,and D.E. Leyden (ed.). 1981. Energy dispersiveanalysisof
actinides,lanthanides,and other elementsin soil and sedimentsamples.Advancesin x-ray
analysis.Vol. 25. PlenumPress,New York.
Schrieber,B., and P.A Pella. 1979. Application of anion-exchangeresin-loadedfilters to the x-ray
fluorescencedeterminationof sulfate.Anal. Chern.51:783-784.
Sobecki,T.M., and AD. Karathanasis.1992. Sojls on hillslopes in acid gray and black shales.Soil
Sci. Soc. Am. J. 56:1218-1226.
Stem, W.B. 1976. On trace elementanalysisof geological samplesby x-ray fluorescence.X-Ray
Spectrom.5:56-60.
Stone,R.G. 1963. The determinationof Sr in tap water by x-ray fluorescencespectrometry.Analyst
(London) 88:56-58.
Strasheim,A, and M.P. Brandt. 1970. The use of direct electronexcitation for the analysisof geo-
logical samplesand comparisonof this techniquewith x-ray fluorescence.Spectrochim.Acta
25B:I-12.
Subramanian,V. 1979. Quantitativeanalysisof elementsin sedimentsand soils by x-ray fluores-
cence:a discussion.Clays Clay Miner. 27:305-306.
Tertian, R 1972. X-ray fluorescenceanalysisof liquid and solid solution specimens-atheoretical
account of the double concentration method and its performances.Spectrochim. Acta
27B:159-183.
Tertian, R, and F. Claisse. 1982. Principles of quantitative x-ray fluorescenceanalysis. Heyden,
London.
Valkovic, V. 1980. Analysis of biological material for traceelementsusing x-ray spectroscopy.CRC

Press,Inc., Boca Raton, FL.
Van Grieken,R.E., C.M. Bresse1eers, and B.M. Vanderborght.1977. Chelex-lOOion-exchangefilter
membranesfor preconcentrationin x-ray fluorescence analysis of water. Anal Chern.
49:1326-133l.
VandenHeuvel, R.C. 1965.Elementalanalysisby x-ray emissionspectrography.p. 771-819.In C.A.
Black et a!. (ed.) Methodsof soil analysis.Part 2. ASA and SSSA, Madison,WI.
Von Hevesy, G. 1932. Chemical analysisby x-rays and its applications.McGraw-Hili-Pub!., New
York.
Wakatsuki,T., H. Furukawa,and K. Kyuma. 1977. Geochemicalstudy of the redistributionof ele-
mentsin soil-I. Evaluationof degreeof weatheringof transportedsoil materialsby distrib-
ution of major elements among the particle size fractions and soil extract. Geochim.
Cosmochim.Acta 41:891-902.
Watanabe,H., S. Berman,and D.S. Russel!. 1972. Determinationof trace metalsin water using X-
ray fluorescencespectrometry.Talanta19:1363-1375.
Watts, S.H. 1977. Major element geochemistryof silcrete from a portion of inland Australia.
Geochim.Cosmochim.Acta 41:1164-1167.
West, N.G., G.I. Hendry, and N.T. Bailey. 1974. The analysisof slagsfrom primary and secondary
coppersmelting processesby x-ray fluorescence.X-Ray Spectrom.3:78-87.
White, E.W., and G.G. Johnson.1972. X-ray and absorptionwavelengthsand two-thetatables,2nd
ed. Am. Soc. Test. Mater., Philadelphia,PA.
Wilkins, C. 1978. The distribution of lead in the soils and herbateof West Pembrokeshire.Environ.
Poilu!. 15:23-30.
Wilkins, C. 1979. The distribution of Mn, Fe, Cu, and Zn in topsoils and herbageof North West
Pembrokeshire.J. Agric. Sci. 92:61"'{)8.
Williams, C. 1976. The rapid determinationof trace elementsin soils and plants by x-ray fluores-
cencespectrometry,1. Sci. Food Agric. 27:561-570.
Williams, C., and 1.H .Rayner.1977. Variability in three areasof the Denceworthsoil map unit. III.
Soil groupingbasedon chemicalcomposition.1. Soil Sci. 28:18(}"195.
Williams, K.L. 1987. Introduction to x-ray spectrometry.Allen & Unwin, London.
Zussman,J. (ed.). 1977.Physicalmethodsin determinativemineralogy.2nd ed. Acad. Press,London.
Published 1996
Chapter 8
Liquid Chromatography
M. A. TABATABAI, Iowa State University, Ames, Iowa
W. T. FRANKENBERGER, JR., University of California, Riverside,

California
The term chromatography has changedsignificantly since it was introducedby

Tswett about 100 yr ago. This term initially was used to describeseparationof
bandsof plant pigmentsextractedfrom greenleaveswith petroleumetheron cal-
cium carbonatepackedin a glass column. Tswett was interestedin a physical
methodof separationand testeda large numberof solventsfor extractingthe pig-
mentsand evaluatedmore than 100 solids substancescapableof selectiveretar-
dation of individual pigmentsthrough adsorption.While the term chromatogra-
phy was descriptiveof the colored band obtained,most of the chromatographic
methodsusedpresentlydo not involve separationof coloredcompounds,but the
principles are generally applicable. The original method of adsorption chro-
matographyand elution developmentof chromatogramsare still widely usedin
many laboratoriesfor both preparationand analyticalpurposes.Although the ini-
tial experimentsof Tswett have changed,and significant major advanceshave
beenmadein separationof gases,organicand inorganiccompounds,and ions, the
term chromatographywas retainedto describethe processof separatingcom-
poundsand ions by a variety of matriceson a large numberof columns.The his-
tory of chromatographyis coveredin severalsources.It hasbeenelegantlysum-
marizedby Ravindranath(1989). The history of liquid chromatography(LC) is
discussedat length by Ettre (1975, 1980). The phenomenonof chromatography
is probablyas old as natureitself becausemany of the geologicalprocesses,such
as soil and ore formation, sedimentationand diagenesis,migration and accumu-
lation of minerals and petroleum, canbe consideredto be accompaniedby, or
resultingfrom, chromatographicprocesses(Ritchie, 1966).
The term chromatographyis defined different ways, dependingon the
author.The InternationalUnion of Pureand Applied Chemistrydefinedthe term
as follows (IUPAC, 1974): "A method,usedprimarily for separationof the com-
poundsof a sample,in which the componentsare distributedbetweentwo phas-
es, one of which is stationarywhile the other moves.The stationaryphasemay
Book Seriesno. 5.
225
226 TABATABAI & FRANKENBERGER
be a solid, or a liquid supportedon asolid, or a gel. The stationaryphasemay be

packedin a column, spreadas a layer, or distributedas a film, etc.; in this defin-
ition chromatographic bed is usedas a generalterm to denoteany of the differ-
ent forms in which the stationaryphasemay be used.The mobile phasemay be
gaseousor liquid. Therefore,a chromatographicsystemconsistsof three com-
ponentsdescribedas solute, solvent, and sorbantor more appropriatelyas the
sample,mobile phase,and stationaryphase."
The processin liquid chromatography,in general,hasits roots in pioneer-
ing work in the areaof ion exchange,including the developmentof syntheticion-
exchangeresins.A review of the work publishedon thesetopics is beyondthe
scopeof this chapter,but information on the basic principles involved in the
operationof LC, including ion chromatography,is presented.
The recentadvancesin LC were possiblebecauseof the progressmadein
the developmentof stationary-phase technologyand instrumentation.While the
basicprinciplesof LC remainedessentiallythe sameover decades,high-perfor-
mance liquid chromatography(HPLC) is the outcomeof the developmentof
columnscontainingsmall (3- to 5-,um diam.) particlesof uniform size and shape
with narrow pore-sizedistribution, togetherwith the developmentof the tech-
nology of chemicalmodification of surfacefunctionality. This led to the devel-
opmentof solvent delivery systems,a wide rangeof analytical columns,a vari-
ety of on-line detectors,high-pressuresolvent pumping systems,sampleinjec-
tion systems,andsupportingtechnologythat resultedin modificationof an older
LC technology.In the past two decadesthis has beenknown as HPLC. It is a
highly versatile, rapid, and basically nondestructiveanalytical technique that
allows the separationand identification of a wide variety of organiccompounds
and inorganicions. The versatility of HPLC is due to a large numberof station-
ary phasesavailable in the analytical columns coupledwith an almost infmite
variety of mobile phasesand a wide range of general andspecific detectors.
When HPLC is coupled with a suitable fraction collection system, the com-
poundsof interestcan be recoveredfor further study.
Ion chromatography (IC)is a term that describesrecentmajor advancesin
the determinationof ions. Sinceits introductionin the mid-1970s bySmall et al.
(1975), researchin the areaof IC has made significant advancesin separation
and determinationof ionic species.Now, ion chromatographsare availablethat
feature high-speedseparationand continuousmonitoring by detector-analyzer
systems,which yield instantaneousreadoutof analyticaldata.
Many books have beenpublishedon HPLC and IC, including the devel-
opmentanduseof their components,and the potentialof thesetechniquesas ana-
lytical tools. The purposeof this chapteris to describethe instrumentsusedand
the methodsdevelopedfor analysisof soils andsoil solutions.Whenpossiblethe
applicationof theseanalyticaltechniquesto plant andwateranalysisalso will be
described.
PRINCIPLES
This sectionis not intendedto give a detailedaccountof chromatographic

or column theory. Accounts of generalchromatographicor column theory are
LIQUID CHROMATOGRAPHY 227
given by Karger et ai. (1973), Johnsonand Stevenson(1978), and Destefano

(1980). Much of the following discussionis revised from an earlier work by
Hassett(1982).
Chromatographicseparationis achievedwhen the solutesin the mobile sol-
vent phase demonstratedifferent affinities for the stationary solid phase, the
mobile phase,or both, resulting in different retentiontimes (t,) for the respective
compounds.The retentiontime for a given compound canbe explainedin terms
of the fundamental equation of chromatography(Karger et ai., 1973)
[1]
where to = elution time for unretainedcompoundsand k is the capacityfactor
k' = total amountof solute in the sationaryphase

total amountof solute in the mobile phase
The goal of chromatographyis to separatemixturesof compoundsinto sep-

aratebandsor peaksof individual compoundswithin a reasonableperiod of time.
The degreeof separationor resolution (R) of individual peaks is an important
chromatographicvariable. The resolutionbetweentwo peakscan be determined
from the following equation(Johnson& Stevenson,1978)
R = (t'2 - t,I)I[W2 + wI)l2]
R =211t,/(W2 + WI) [2]
where t,l and t,2 are the respectiveretention times of peaksone and two and WI
and W2 are peakwidths at the baselineobtainedby drawing tangentsto the peak
sides.Resolutionvaluesof 0.5 or greatershouldallow determinationof the num-
ber of peakspresent.Values of 0.4 or less may representunresolvedpeaks.
Column efficiency is relatedto retention time and peakwidths by the fol-
lowing equation(Johnson& Stevenson,1978)
N = 16(t,lw)2 = 5.5(t,lWI/2f [3]
whereN is the column efficiency in theoreticalplates,t, the compoundsretention

time, W the baselinepeak width, and wl/2 the peak width at l/2-peak height. In
practice,the useof wl/2 is recommendedbecauseof the problemsin estimatingW
for unresolvedor tailing peaks.
The value of N is independentof retention time, i.e., the value of N for a
peakeluting at 5 min would be approximatelyequal to the value of N for peaks
eluting at 10 or 20 min. This has the negativeimplication that longer retention
times result in greaterpeakedwidths (bandspreading).
The height equivalent to theoreticalplate (H, or HETP) is the preferred
measureof column efficiency (Johnson& Stevenson,1978) becauseit allows
comparisonbetweencolumnsof different lengths(L)
H =LIN =HETP. [4]
A smallervalueof H representsgreatercolumnefficiency,whereasa largervalue

of N representsgreaterefficiency of a particularcolumn.
Reducedplate height(h) takesinto accountthe particle size of the station-
ary phaseand is a pure measureof column quality independentof particle size
[5]
-'
where dp is the particle dianteter in millimeters. Column efficiency tends to
increasewith a deceasein particle size.
Peak asymmetrycan causemajor problemswith calculationsof column
parametersand with calculationof peak areas.Two types of tailing are recog-
nized. Thermodynamictailing results from the specific chemical propertiesof
the sample,solvent, and column packing. Kinetic tailing results from nonuni-
form column particles,poor column packingor both. An asymmetryfactor (b/a)
canbe calculatedby dividing the peakinto two portionsat maximumpeakheight
and taking the ratio of the partial peakwidths at 10% of peakheight maximum.
The 10% of peak maximum is an arbitrary point and has no theoreticalsignifi-
cance.A ratio of b/a equalto unity representsno peaktailing. Valuesgreaterthan
unity representtailing.
HIGH·PERFORMANCE LIQUID CHROMATOGRAPHY
Although liquid chromatographyhas been in use for many years, recent

advancesin technologyhaveled to rapid expansionin its applicationin analyti-
cal chemistry. Improvementsin solvent delivery systems(pulse free), packing
materials,betterunderstandingof the parametersaffecting chemicalseparation,
and the developmentof sensitiveand selectivedetectorshaveallowed HPLC to
becomean analytical techniquecomparablein speed,reproducibility and relia-
bility to gas chromatography(GC). The past developments,presentstatus,and
future trendsof HPLC are reportedby Brown (1990).
Some of the major advantagesof HPLC over GC are that, unlike GC,
which cannotbe usedfor determinationof somecompounds,HPLC can be used
for analysisof strongly polar compoundswithout conversionto derivatives,of
compoundswith low volatility, and of thermolabile materials. All chromato-
graphicmethods,however,are separativetechniquesand do not allow for posi-
tive identification. For identification, the sample must be analyzedeither by
anothertechnique(such as massspectrometry)or by the use of an appropriate
detectorin conjunctionwith the operationof an HPLC using different column
(containingdifferent packingmaterial)or solventsystems.
In all LC techniques,the samplecompoundsare separatedon separator
columns containing one of many possiblepacking materials. The eluent(s)is
subjectto high pressure,as in HPLC, or underlow pressure,as in IC. The detec-
tion is accomplishedby using one of severaldetectors;dependingon chemistry
of the compound(s)and the sensitivity desired.The peaksobtainedare recorded
SOLVENT
RESERVOIR
PUMP
SAMPLE
INJECTOR ANALYTICAL COLUMN
L . . - - - r - -...... - - - I RECORDER I
Fig. 8-1. Simple schematicdiagramof isocraticHPLC system.
on a strip chart recorderor integrator.An autosamplermay be employedfor high

volume applicationand unattendedoperation.
Instrumentation
Two basic types of HPLC instrumentsystemsare available: isocratic and

gradient.An isocraticsystemis capableof using only one solventconcentration
(Le., 85% methanol-15%water) for elution during a given chromatographicrun,
whereasa gradientsystemis capableof varying the solventconcentrationduring
the courseof a run.
A simple isocraticsystemconsistsof a single high-pressurepump, a man-
ual sampleinjector, an analytical column, a detector,and a recorder(Fig. 8--1).
In practice,the sampleis loadedwith a syringeinto the sampleloop (variable)of
the injector. The injector is then movedfrom load to inject, placing the samplein
line. The sample then moves with the solvent through the analytical column
where separationoccursand then to the detectorwhere the individual peaksare
sensed.The recorder provides a continuoustrace of the chromatogram.Peak
areasare usually usedfor determinationof sampleconcentrations.
A more highly versatile HPLC systemwith much greaterflexibility than
the simple isocratic systemalso is available.The systemmay be operatedas an
isocraticsystemor a gradientsystem(Fig. 8--2). One advantageof a gradientsys-
tem is that one eluentconcentrationcan be usedfor separationof highly mobile
compoundswith shortresidencetimes,then the eluentconcentrationcanbe mod-
ified to speedthe elution of the compoundswith longer residencetimes. Another
advantageof this systemis that samplescan be automaticallyinjected using the
microprocessorto restartany gradientprogramand the other componentsof the
system.This allows the operatorto load multiple samplesand then pursueother
SOLVENT SOLVENT
RESERVOIR RESERVOIR
1 2
CONTROLLER
PUMP1 PUMP2
ANALYTICAL COLUMN
Fig. 8-2. Simplified schematicdiagram of microprocessorcontrolled gradient HPLC system with

automaticsampleinjection, reportingintegrator,and fraction collector.
activities rather than having to constantly attend the instrumentand manually

inject samplesevery 20 to 30 min.
Columns
The column is the heart of the chromatographicsystem. It is usually a

cylindrical tube packedwith the stationaryphaseand marketedcompletewith
end fittings. The column material,dimensions,shape,packingmaterial, method
of packing, and the geometryof the end fittings are amongthe most important
factors that affect the performanceof the HPLC. A large variety of ready-to-use
columns of guaranteedperformanceare commercially available from a large
numberof sourcesand it is seldomnecessaryto preparethem in the laboratory.
Nevertheless,knowledgeof the factors affecting the column performancewill
help to derive the maximum benefit out of the column in terms of its efficiency
and life (Ravindraneth,1989).
The tube material most frequently used for the preparationof HPLC
columns is stainlesssteel, becauseof its strengthand inertnessunder normal
chromatographicconditions.The chromatographiccolumns available from the
supply housescome in different sizes,dependingon not just the convenienceof
packingand the size of packingparticles,but also on its use, preparativeor ana-
lytical. Chromatographicsilica gels are amorphous,porous solids that can be
preparedin a wide rangeof surfaceareafrom 2 to 8 x HP m2/kg. Averagepore
diametersrangefrom 50 to 250 A. Most modernsilicas usedfor bonded-phase
columnsare of the averageparticle diameterof 3 to 10 .urn. They can be either
irregularly shapedor spherical.Smaller particles lead to faster analysis times.
The preferredlength is 3 to 5 cm for columnswith 3-,um particles,and 5 to 10

cm for the 5-,um packing materials.Columns, 15 cm long with 12000theoreti-
cal platesare adequatefor most separations.The short columns(3-5 cm) offer
advantagesin analysistime, low solventconsumption,often better detectionlim-
its, and lower costs. For more demandingseparations,75 cm columns can be
used.
The packing material in the column is retainedby meansof microporous
frits. The pore-sizeor the frits shouldbe smallerthan the smallestparticle diam-
eter'spore size or abouthalf the meandiameterof the packingmaterialis usual-
ly adequate.The frit is held in placeby a nut of appropriategeometryand the col-
umn is connectedto the injection port and the detectorby meansof capillary
plumbing and Swage10k ferrule-nuts.The geometryof thesecouplingsis critical
becauseany deadvolume would adverselyaffect the column performance.Zero-
dead-volumeadaptorsfor all types of commercialLC systemsare available.
SeparationMethods
Four basic separationmethodsare usually employed in HPLC colums:

adsorption,partition, sizeexclusion,and ion exchange.Although it canbe argued
that all, exceptexclusion,representadsorptionmechanisms,the terminologywill
be retainedto be consistentwith the HPLC literature (Hassett,1982).
The adsorption mechanism achievesseparationthrough competition for
the solutebetweena polar stationaryphaseand a mobile phaseof variablepolar-
ity. This mechanismis most successfulin achievingseparationof polar solutes,
the more polar the solute, the longer will be its retentiontime. Ionic solutesare
too strongly retainedby the stationaryphase,whereasnonpolarcompoundsare
unretained by the column. There are many applications of adsorption chro-
matography, including separationof isomers and determination of polarity.
Column selectionis basedon different packings(pellicular vs. microporous)and
variousshapes(irregularvs. spherical).The major limitations to adsorptionchro-
matographyare: (i) long column-regeneration times, (ii) deactivationof silica by
water, and (iii) irreversibleretentionof somesolvents.
The partition mechanism achievesseparationthrough the competitionfor
the solute betweena nonpolar stationaryphase(silanols are chemical bonded)
and a mobile phaseof variable polarity. The lesspolar the solute,the longer will
be its retention time. This mechanismwas designedprimarily to achievechro-
matographicseparationof nonpolar solutes, but it has been used successfully
with polar and ionic compounds.This mechanismrepresentsthe most widely
usedand versatiletechniqueof HPLC. Horvath and Melander(1978) provide a
detaileddiscussionof this separationmechanism.
Size exclusion includesgel permeationand gel filtration chromatography.
There is no adsorptioninvolved becausemoleculesare separatedbasedon their
molecularsize. As the solutesmove through the column, the largestmolecules
will be unable to enter the pore volume, or may be entirely excluded,eluting
first. The smaller moleculeswill enter the pore volume and elute later. Size
exclusion chromatography(SEC) is more susceptibleto flow than any other
chromatography.Size exclusionchromatographyhasbeenconductedwith many
types of packing materials,including glass beads,porous silica, soft and rigid

gels, and modified silica columns.The primary applicationof SEC include sep-
arationof proteins,enzymes, and largepeptidesby}l-spherogeior TSK columns.
Ion exchange achievesseparationof ionic speciesthroughthe competition
of the ionic speciesin the solvent and the samplefor ion exchangesites on the
stationary phase(polystyreneor silica base). Ion exchangehas been used to
achieveseparationof ionic specieswith HPLC systems,but this mechanismis
primarily concernedwith the recentand rapidly growing field of ion chromatog-
raphy and is not discussedin detail in this chapter.
High-performance liquid chromatographyanalytical columns can be
groupedinto four classes:(i) normal phase,(ii) reversephase,(iii) polar-bonded
phase,and (iv) ion exchange.Normal-phasecolumns contain microbeadsof
polymerizedsilica with surfacehydroxyIs, andsilanol groupswith a slight acidic
character(Johnson& Stevenson,1978)
Si-OH.
Becauseof the solubility of microcrystallinesilica in water, water must be pre-

ventedfrom enteringa normal-phase column, or one will risk deactivatingthe
packing.The solventsfor normal-phasecolumnsinclude hexaneas the nonpolar
solventand isopropanol(IPA) as the polar solvent.By varying the ratio of hexa-
ne to IPA, one can obtain different degreesof solvent polarity and achievethe
desiredchromatographicseparation.Normal- phasecolumnsfunction primarily
by the adsorptionmechanism.
Reverse-phase columnscontain silica beadsthat have beencoatedwith a
CIS,CS, C6 or Cz [octadecylsilanederivatizedsilica column (ODS)] hydrocarbon
High surface coverageminimizes the problem with an aqueousmobile

phase,but vulnerability of the siloxane(Si-O-Si) bondlimits the usablepH range
to 2 to 7.5. The differencein selectivitybetweenalkyl groupsof different lengths
are not fully understood.With a given mobile phase, solvent retention and
hydrophobicselectivity are greateron the CIS packing. Separationson Cs and
other short-chaincolumnsare, however,more rapid and column efficiency may
be evenslightly great~r due to solvent masstransfer.For separationof polar mol-
eculesby reversephasetechniques,short-chainbondedphasesgenerally give
more symetrical peaks than long-chain phases.Typical solvents for reverse-
phasechromatographyinclude water-methanolor water-acetonitrile.Solvent
polarity canbe easilyvariedby changingthe watercontent.With high watercon-
tents,the more hydrophobiccompoundsare retainedto the greatestextent. Low
watercontentsresult in the more polar substances being retained.Reverse-phase
columnsare an exampleof the partition mechanism.Retentionof a compound
within the column is primarily relatedto its solubility in the solvent ratherthan
to its affinity for the surface.
Polar and ionic compoundscan often be successfully separatedby a
reverse-phase column. Organicacids can be separatedusing acetonitrileand an
aqueousbuffer (pH - 2.5). The low pH results in associationof the carboxyl

groupsof the acids, making them more hydrophobicand allowing separationby
reverse-phasechromatography.Ionic compoundscan be separatedby ion pair
chromatography.Ion pair chromatographyis a modification of reverse-phase
chromatography.A mixture of organic ions can be separatedusing a mobile
phase that contains an appropriate counter-ion (organic modifier such as
methanol, acetonitrile, or tetrahydrofuran)that has a hydrophobic side chain.
Different organic modifiers can be used to generatespecific functional groups
selectivity. The ionic portion of the counter-ionpairs with the organicions in the
sample,making the pair more hydrophobicand allowing separationby reverse-
phasechromatography(Eksborg& Schill, 1973; Kissinger, 1977).
Polar-bondedphasecolumnsare of two major types
Amino phase Si-O-Si-(CH2kNH2
Cyano phase Si-O-Si-(CH2kCH.
Due to the presenceof siloxanebonds,thesecolumnshave the samepH

restrictions as reverse-phasecolumns. Amino and cyano columns function by
both partition and adsorptionmechanisms.With the cyano columns,separation
dependson the solvent system,thus it is possibleto use a cyano column with a
wide rangeof polar compounds.A cyanocolumn is particularly useful in sepa-
rating membersof a homologousseries.The amino columns function as weak
anion exchangersor as selective basic polar adsorbents.With both of these
columns,normal- or reverse-phase solventscan be useddependingon the sepa-
ration desired.
Eluents(mobile phases)
Adsorption chromatographyis dominatedby two stationaryphases;silica

gel (representingthe normal-phaseliquid chromatography)and octadecylsilyl-
bondedsilica (for reverse-phase liquid chromatography).In normal-phasechro-
matography,a nonhydroxylic, less polar mobile phaseis generallypreferredfor
the separationof nonionic, medium polar small molecules;whereasreversed-
phaseLC is selectedfor compoundsbelongingto either extremesof the polarity
scale,the mobile phasesusedin the reversed-phase LC are generallywater-based
and many contain salts and medium polar, water-miscible organic solvents
(Ravindranath,1989). Becausethe retentionmechanismsof the solutesare lim-
ited by the choiceof either of the two most popularstationaryphases,the results
of the analysis is predominantlydeterminedby the selectivity of the mobile
phase;this is particularly true of the solute in the aqueousmobile phaserather
than its affinity to the adsorbent.The selectionof the compositionof the mobile
phaseis accomplishedby systematicstudiesof the effect of compositionchro-
matographicseparationof the analytes.In most cases,the mobile-phasecompo-
sition is recommendedby the columnsupply houses,and no experimentsneedto
be conductedto determinethe optimal compositionof the mobile phase.Only
HPLC solvents(ultrapurity grade)should be used.One should take into account
the ultraviolet (UV) cutoffs of solventswhen running agradient.Buffer solutions

shouldalwaysbe filtered (2,urn), because salts and bufferscancontainmicropar-
ticulates.When using a buffer systemor ion pairing reagents,do not allow the
mobile phaseto lie stagnantin the lines. Buffers and other reagentscan have a
corroding effect on stainlesssteel over time. For details about the mobile phas-
es, seeRavindranath(1989).
Detectors
A major developmentin HPLC hasbeenthe developmentof sensitive,on-

line detectorsystems.Unlike GC, in LC the physicalpropertiesofthe sampleand
mobile phasesare often very similar. This led to the developmentof two basic
types of detectorsfor use in LC systems(Hamilton & Sewell, 1977). These
involve: (i) the differential measurement of a propertycommonto both the sam-
ple and the mobile phase,and (ii) the measurementof a property that is specific
to the sample,either with or without the removal of the mobile phasebefore
detection. Examplesof the first type of detector, also known as bulk property
detectors,are the differential refractometer,conductivity, and dielectric constant
detectors.Soluteproperty detectorswhich do not require removal of the mobile
phasebefore detectioninclude the photometricdetectors(ultraviolet detectors,
visible photometers, infrared photometers, fluorescence photometers),and
refractiveindex detectors.The differential refractometeris the secondmost like-
ly usedLC detector,after ultraviolet detectors.This detectorcontinuouslymon-
itors the difference in refractive index betweenthe pure mobile phaseand the
mobile phaseplus the sampleas it elutesfrom the column. The main advantage
of refractometersis that, becausethey are bulk property detectors,they are of
universalapplication.The only requirementis that thereis a differencein refrac-
tive index betweenthe sampleand the mobile phase,and this can be as little as
10-7 refractive index units. This detector,however,is about two ordersof mag-
nitude lesssensitivethen the ultraviolet detectorsand cannotbe usedwith gradi-
ent elution. The sensitivity, definedas being equalto the noise level, is general-
lyon the order of 10-7 refractive index unit. Many other detectorsare available,
but their descriptionis beyondthe scopeof this chapter.
Examplesof the secondtype of detectorsare moving wire flame ioniza-
tion, electroncapture,and massspectrometrydetectors,which require removal
of the mobile phasebeforedetection(water is a difficult solventto removewith-
out losing the solute). These detectors,however, have several disadvantages
which have hinderedtheir acceptanceas LC detectors;theseinclude high cost,
bulkiness,relatively poor sensitivity, and operationalinconvenience(Hamilton
& Sewell, 1977).
The highly versatile nature of HPLC is primarily due to the number of
types of columns available for separationand the numberof types of detectors
for sensingthe individual compounds.Commercialdetectorscurrently available
include ultraviolet/visible(UVMS), fluorescence,electrochemical,radiometric,
refractive index, pulse amperometric, and conductivity.
Ultraviolet/visible detectors are basically UVNIS spectrometerswith
high-pressure(~4000 psi), low-volume standard,microbore or semiprepflow
UQUID CHROMATOGRAPHY 23S
cells. A variable wavelengthUVMS detectorhas the greatestutility of all the

detectorsavailable,althoughthe 254-nmsingle wavelengthdetectoris probably
in greatestuse.Improved sensitivitycan be accomplishedfor proteins,peptides,
phenols,and catecholaminesat 280 nm. In practice, a wavelengthis selected
(200-400nm) where the compoundsof interesthave reasonableabsorptionand
the solventsusedin elution have minimum absorption.The variablewavelength
UVMS detectordemonstrates greatadvantageover fixed wavelengthdetectors,
particularly when compoundshave similar residencetimes and poor resolution
results.When this occurs,the variablewavelengthdetectorcan often be set at a
wavelengthwhere only the compoundof interest absorbs.The variable wave-
length detectoralso can be usedto obtain ratios of absorbanceor peak areasat
two different wavelengths(dual channel).Comparisonof thesepeakarearatios
with ratios obtainedfor standard compounds is, in addition to residencetimes, a
meansof identification. Ultraviolet absorptioncan detectin the nanogramrange
of an injectedcompoundunder favorable conditions and is compatiblewith a
variety of solventwith gradientandstepelution. An UV photodiodedetectorcan
deliver low noise,low drift and high signal levelsfor sensitive,specificdetection
of UV chromophores.
Fluorescencedetectorsare basicallyfluorescencespectrophotometers with
appropriateflow cells. The main advantagesof fluorescenceare selectivity and
sensitivity. Fluorescencemeasuresthe amountof light emittedby a samplethat
hasbeenexcitedby light at a different wavelength.Fluorescenceis a widely used
method of detectionin HPLC, especiallyin biological areas.Few compounds
fluorescenaturally, it is usually necessaryto derivatizethe compoundwith a flu-
orescentcompound.Derivatizationor naturalfluorescencecan havegreatadvan-
tage, particularly when there is a strong backgroundof other nonfluorescent
compoundsthat would interfere with an UV NIS detector. High-performance
liquid chromatographyderivatization reagents (e.g., dansyl chloride, fluo-
rescamine,orthophthaldehyde)are available that will react with specific func-
tional groups: organic acids, thios, primary and secondaryamines,phenol, and
aldehydes.Fluorescencedetectorscan detectin the picogramrangeunderfavor-
able conditions.
Electrochemicaldetectorsmeasurecurrent voitammetrically or ampero-
metrically using solid electrodesor a dropping mercury electrode,respectively.
Use in reductivemode permitsdetectionof compoundssuchas estersand alde-
hydes,whereasuse in the oxidative mode permits detectionof compoundssuch
as catecholamines and aromatichydrocarbons.A thin-layer Kel-F-Graphiteelec-
trochemicaldetectorcapableof measuring2 to 7 p,g/L levels of certain carba-
mate pesticides has been developed by Anderson and Chesney (1980).
Electrochemicaldetectorsare compatibleonly with water and an aqueousmix-
ture of polar solvents.Refractiveindex (RI) detectorsalso havebeendeveloped.
Theseare usedprimarily by the food industry to measurefairly high concentra-
tions of sugars.Refractive index detectorsare restrictedto isocratic operation
and havedetectionlimits in the microgramrange.
The characteristicsof an ideal detectorfor HPLC include the following
(Hamilton & Sewell, 1977).The detectorshould:(i) havea high sensitivity (bet-
ter than 0.1 p,g of samplein 1 cm3 of mobile phase);(ii) responduniversally to
all solutes,or havea predictablespecificity; (iii) havea linear responseover sev-

eral orderof concentration;(iv) possesa low deadvolume; (v) be nondestructive;
(vi) be insensitive tochangesin temperatureand mobile phasevelocity changes;
(vii) operate continuously; and (viii) be reliable and convenient to use.
Unfortunately no single detectorcommerciallyavailable satisfiesall the above
criteria.
Recorders
Any strip chart recorderor integratorcan be usedfor recordingthe peaks.
The integratorshave the advantagein that they also report the areasunder the
peaks.It has beenthe experienceof the authors,however,that the peakheights
are bettercorrelatedwith concentrationthan are the areasunder the peaks.
Applications
The majority of the pesticidescurrently in use and their residues(after
degradation)are nonvolatile, nonpolarto medium polar compoundsthat can be
readily analyzedby HPLC. High-performanceliquid chromatographyretention
data for many pesticideshave been compiled (Lawrence, 1982). High-perfor-
manceliquid chromatographyis carried out on normal, but more frequently on
reversed-phase columns,using UV or fluorescencedetection,dependingon the
nature of the samples.Often postcolumnderivatization techniquesneed to be
used to form chromogenicor fluorogenic compounds,suitable for sensitive
detection.The continuing serial publication, edited by Zweig (1984) is a good
source of information in the field of pesticide extraction and anaylsis with
HPLC; the work describes detailed analyticalmethodologyfor the determination
of severalindividual pesticidesin a variety of samples.
Other applicationsof HPLC are in the areaof identification of phospho-
lipids in soils and sewagesludges(Stott & Tabatabai,1985), phytohormones
(Martens & Frankenberger,1991a), amino acids (Warman & Bishop, 1987;
Martens& Frankenberger,1991b),peptides(Warman& Isnor, 1989), aminosac-
charides,neutral saccharides,and glycuronic acids (Martens & Frankenberger,
1991c).
Comments
One of the most important requirementsof the HPLC is that the sample
injected for analysisshould be free of particulates.Loss of resolutioncan result
from a contaminatedprecolumnor analytical column. Reproducibility may be
affected by a contaminatedcolumn, an insufficiently conditioned column, or
microbial growth in the eluentswhen stored at room temperaturefor several
days.
ION CHROMATOGRAPHY
Ion chromatographyhas its roots in pioneeringwork in the area of ion
exchange,including the developmentof syntheticion-exchangeresins.A review
PRESSURE GAUGE
ELUENT
RESERVOIR
INJECTION
PORT
z 0::
::;;
:J
o
-.J ~
o
U
W
0::
<{
Cl.
W
,--1 RECORDER I
I
0::
Cl. (fJ I
I
EXCESS SAMPLE
WASTE
Ie system(Tabatabai& Basta,1991).
Fig. 8-3. Simplified schematicdiagramof suppressed-type
of the work publishedon thesetopics is beyond the scopeof this chapter,but

information on the basic principles involved in the operation of ion chro-
matographsis presented. Detailed information on Ie systemscan be found in a
book by Small (1989).
Instrumentation
Ion chromatographs can be divided into two major groups:thosethat oper-

ate on the principle of eluent suppression(dual-columnsystem)and thosewith
no suppressorcolumn (single-columnsystem).The electrical conductivity of a
solution varies with the concentrationof ions present,and this principle is the
basis of a universal Ie detectorsystem used for the determinatinoof all ionic
speciesin solution. Detailedcomparisonbetweeneluentsuppressedand nonsup-
pressedion chromatographyis presentedby Pohl and Johnson(1980) and Tarter
et al. (1987).
Eluent-Suppressed
System
The first suppressor-typeIe was marketed by Dionex Corporation

(Sunnyvale,CA). The basiccomponentsof a suppressed -type ion chromatograph
are shown in Fig. 8-3. For simplicity, the reservoirsof the eluentand water used
for regenerationof the suppressorcolumn and the valving systeminvolved in the
Ie are not shown. The instrumentemploysthe following components : (i) eluent
pump and reservoir;(ii) sampleinjection valve (the sampleloop can be adjusted
from about50,uL to severalhundred,uL); (iii) ion-exchangeseparationcolumn;
(iv) suppressorcolumn coupled to a conductivity detector, meter, and output
device; and (v) regeneratingpump with electronictimer and controls.
In the eluent-suppressedIe, the ion speciesare resolvedby conventional
elution chromatographyfollowed by passagethroughan eluentstripper,or "sup-
pressor," column, wherein the eluent coming from the separatingcolumn is
strippedor neutralized.Thus, only the ion speciesof interestleave the bottom of
the suppressorcolumn in a backgroundof HZC03, which exhibits a low conduc-
tivity, in the caseof anions,or water in the caseof cations,whereuponthey are
monitoredsubsequentlyin the conductivity celVmeter/recorder (integrator)com-

bination.The eluentflow rate can be varied by adjustingthe pump pressure,nor-
mally it is about 2 to 3 mLimin. An aliquot (2 mL) of a suspension-freesoil
extract is injectedby a plastic syringe into the IC. Alternatively, an autosampler
can be used to automatically inject samples.The Dionex IC models can use
either 0.5-or 5 mL samplevials. The sampleloop on the injection valve can be
adjusted,but normally a volume of 100ilL is used.The 2-mL volume is conve-
nient to ensureproperflushing of the injection valve loop and lines.
Several suppressordevices have been developed for suppressor-based
chromatography.These include the packed-columnsuppressor(Small et aI.,
1975), the hollow-fiber membrane suppressor(Stevens et aI., 1981), the
Micromembrane suppressor (Franklin, 1985; Stillian, 1985), and the suspension
(solid phasereagent)postcolumnreactionsuppressor(Gjerde & Benson,1990).
The first three types of suppressordevices were producedby Dionex Corp.,
Sunnyvale,CA, and the last device has beenusedwith the equipmentmarketed
by LACHAT Instruments(Milwaukee, WI). The original packed-bedsuppressor
has been improved and is now called the solid phase chemical suppressor
(SPCS).It is being marketedby Alltech Associate,Deerfield, IL. The problems
associatedwith the original packed-bedsuppressortechnique, such as band
z,
broadening,ion exclusion,and oxidation of NO are eliminated.The disposable
SPCSsimplifies the instrumentationrequired toperform suppressed-based IC by
eliminating the regenerationsystemsand the complex postcolumnreactionsys-
tems neededwith other suppressiontechniques(Saari-Nordhaus& Anderson,
1994). The lifetime of the SPCScartridgeis dependenton the ionic strengthand
flow ratesof the eluent,varying from 7 to 12 h.
In a variant of the suppressorcolumn system,the resin in the suppressor
column is replacedby an ion-exchangemembranein tubular form to condition
the eluentcontinuously(Stevenset aI., 1981). This membrane(sulfonatedpoly-
ethylene hollow fiber) acts exactly like the suppressorresin in that ions are
exchangedfrom the membranefor ions in the eluent system.The innovation is
that the membraneis regeneratedcontinuouslyby a gravity-fedflow of low-con-
centrationH2S04 (in analysisof anions)that continuouslyreplacesthe ions that
are exchangedonto the fiber with ions from the regenerant.Thus,separateregen-
eration stepsare eliminated.The replacementof the conventionalion-exchange
resin bed suppressorcolumn with the hollow fiber suppressorallows continuous
operationof an IC without varying interferencefrom baselinedips, ion-exclusion
effects, or chemicalreaction.Stevenset ai. (1981) concludedthat conventional
suppressorcolumn systemshave less band spreadingthan those using hollow-
fiber suppressors,and this results in slightly poorer resolution of early eluting
ions with the latter type of eluent suppressiontechnique.However, our experi-
enceis different. Furthermore,work by Weiss(1986)showedthat becauseof the
low deadvolume of a membranesuppressor,mixing and bandbroadeningeffects
are minimized and the sensitivity is generallyenhancedcomparedwith the more
traditional packedbed suppressor.Details of the theory of operationof the hol-
low-fiber suppressorare discussedby Weiss (1986), Stevens et ai. (1981),
Hanaokaet ai. (1982), and Small (1983). The stateof the art of ion chromatog-
raphy has beendescribedby Dasgupta(1992).
ELUENT
INJECTION RESERVOIR
PORT
SAMPLE LOOP
,---------1
I
RECORDER I
: CONDUCTIVITY
CELL
WASTE
Fig. 8-4. Simplified schematicdiagramof nonsuppressed-type

IC system(Tabatabai& Basta,1991).
Eluent-Nonsuppressed
System
Two alternativemethodsto that describedaboveare now availablefor ion

separationand determination.In both methodsno suppressorcolumn is needed
(single-columnsystem).Instead,moderatelyconductingeluentsare usedto elute
a variety of ions. One techniqueis a variation of conventionalHPLC, in which
silica-basedcolumn packings provide ion separations.In a second similar
approach,specially synthetizedmacroporouspolystyrene-divinylbenzene resins
with low capacitiesare coupledwith moderate-conductivitymono-or polyvalent
eluting ions (Smith & Chang, 1983). Dedicatedsystemsfor single-columnIC
have been introduced by Wescan Instruments, Inc. (Alltech Associates,
Deerfield, IL), Hewlett-PackardInstruments, Inc. (Palo Alto, CA), Waters
Associates(Milford, MA) and Brinkman Instrruments,Inc. (Westbury,NY). The
instrumentdistributedby Brinkman is manufacturedby Metrohm in Switzerland.
The basic componentsof the nonsuppressed-type (single-columnsystem)
ion chromatographare shown in Fig. 8-4. The techniqueemploysthe following
components:(i) eluent pump and eluent reservoir,(ii) sampleinjection valve (a
sampleloop of ~500,uL is normally used),(iii) ion-exchangeseparatorcolumn,
and (iv) conductivity detectorcoupledto an output device. In this system,a low-
capacityexchangecolumn and low-conductivityeluentare usedwithout the need
for a suppressorcolumn (Gjerde & Fritz, 1979, 1981; Gjerdeet aI., 1979, 1980).
Anions of interestelute in the hydrogenform (e.g., HCI, HN03, H2S04) against
a backgroundof ionized phthalateions. A numberof equilibria affect single-col-
umn chromatography(SCIC). Buffer ions (usually weak acid ions) equilibrate
with the free acid in solution. Both of thesespecies,in tum, equilibratewith their
bound forms at the surfaceof the stationaryphase(Jupille, 1987). Details of the
reactionsinvolved and factors affecting ion-exchangeseparationsin the nonsup-
pressedsystem,information on other typesof separations,and column technol-

ogy are presentedby Jupille (1987).
For the determinationof NH.t and alkali metals,the mobile phaseusedin
the nonsuppressed IC systemmust have a strong affmity for the ion-exchange
resin in order to displaceseparatedions from the analytical column. Maximum
sensitivity is achievedwhen the equivalent conductanceof the ionic species
gives a detectionsignal well abovethe eluentbackground(Gjerde et aI., 1979).
Dilute HN03 is usedfor the determinationof NH.t and the alkali metals(Fritz et
aI., 1980). Use of 10 roM HN03 (PH 2.1) has beenshown to give excellentres-
olution of monovalentcations with elution completein <6 min when a Vydac
401 TP cation-exchangecolumn (SeparationGroup, Hesperia, CA) is used
(Neito & Frankenberger,1985). Ethylene-diammoniumdinitrate (5 roM, pH 6.1)
competesmore strongly with divalent cationsin solution than doesHN03, thus
providing better resolutionand peaksymmetry for the divalent cations.Fritz et
aI. (1980)recommendeda solutionpH of 6.1 so that all carbonicacid specieswill
elute asbicarbonateandcauseno interferencewith the analysis.Methanesulfonic
acid (3 roM) also is an excellenteluent for analyzingLi, Na, NH4, K, Mg, and
Ca by the nonsuppressed-type Ie.
Backgroundconductanceand minimum detectionlimits of both alkali and
alkaline earth metals increasewith increasingconcentrationof the electrolyte
mobile phase(Iskandarani& Pietrzyk, 1982), whereasretentiontimes decrease
with increasingeluentconcentrationand decreasingresin capacity(k') (Gjerdeet
aI., 1979). The commercially available columns (e.g., Vydac 401 TP cation-
exchangecolumn) have relatively low k' [0.10 mole kg-I], although resins of
even lower k' have been synthesizedfor chromatographicseparationof ions
(Gjerde & Fritz, 1979; Gjerde et aI., 1979; Boyd et aI., 1954; Fritz & Story,
1974a,b).
Columns
Severaltypesof columnsare commerciallyavailablefor the ion-exchange

separationof the commoninorganicand organicanion via eluent-suppressed Ie.
Information on the type of resin usedand columnsavailablefrom Dionex Corp.
is summarizedby Johnson(1987).
In the nonsuppressed IC system,a low-capacity exchangecolumn and a
low-conductivity eluent are used. These include a macroporouspolystyrene
divinylbenzeneresin (Gjerde & Fritz, 1979b, 1981), or surface-quarterized sili-
ca (Girard & Glatz, 1981). A numberof equilibria affect the separationof ions
by the nonsuppressed IC system.Buffer ions (usually weakacid ions) equilibrate
with the free acid in solutions. Both of thesespecies,in tum, equilibrate with
their bound forms at the surfaceof the stationaryphase.Details of the reactions
involved and factors affecting ion-exchangeseparationsin the nonsuppressed IC
system,informationon othertypesof separation,and column technologyarepre-
sentedby Jupille (1987).
The columns used in eluent-suppressed-type IC were initially made of
glass.Presentcolumnsare madeof plastic. The performanceof thesecolumnsis
equivalent to or better than that of glass columns and there is no breakage.
Typical plasticcolumn internal diametersare from 4 to 9 mm, their lengthsvary

from 50 to 250 mm. At present,Dionex Corporationis the sole distributor of
thesecolumns.
Columns in single-column-type(nonsuppressed) IC can be glass,plastic,
or stainlesssteel. Most columns now in use are made of stainlesssteel. The
phthalateand benzoateeluentsused in nonsuppressed have pHvalues ranging
from three to seven;therefore,little corrosionis expected.The injection of sam-
ples with pH values greaterthan seven is not advisablebecausethe the silica
packing will degradeseverely.Sampleswith pH values greaterthan sevenare
normally treatedwith eluent until the proper pH balanceis achieved.
SeparationMethods
In the IC systems,the ion speciesare resolved by conventionalelution

chromatography.
Eluents
In the suppressed-based IC, the eluent used for determinationof anions

normally is a mixture of dilute NaHC03 and NazC03, althoughother dilute mix-
tures(e.g., NaHC03 + NaOH, NazC03 + NaOH) also are used(Johnson,1987).
The anionsare separatedand convertedto their strongacids in a backgroundof
HzC03, which has low conductivity. The presenceof strong acids in HzC03 is
measuredby a conductivity cell and reportedas a peakon a stripchartrecorder
or integrator.The eluentusedfor separationand determinationof alkali metalsis
dilute HCI, and in the caseof alkaline earthmetalsits a mixture of dilute HCI and
m-phenylenediamine dihydrochloride.The metalsare separated and convertedto
metal hydroxide in a backgroundof water.
In the nonsuppressed IC system,the eluentis a dilute organicacid (phthal-
ic, benzoic,or citric) are often used(Gjerde & Fritz, 1981; Jupille, 1987), with
phthalic acid being the most commonfor detectionof anionsbecauseof its wide
range of retention control (via pH adjustment) and equivalent conductance
(Jupille et aI., 1983). Anions of interest elute in the hydrogenfrom (e.g., HCI,
HN03 , HzS04) againsta backgroundof ionized phthalateions.
Detectors
The commonly used detectoris a high-sensitivity flow cell condcutivity

meter.The cell body is constructedof Kel-F plasticwith an internalvolume rang-
ing from 2 to 6.uL. The electrodesare madeof 316 stainlesssteel. The conduc-
tivity meter setting rangesfrom 0.1 to 10 000 .uS (Dionex Model 2002Ii). The
conductivity metersettingcommonlyused,however,is 3,10or 30.uS. The meter
signal is displayedon a digital readoutwith an advancedautozerocircuit off set-
ting the backgroundconductance.Conductivitymeasurements are quite sensitive
to temperaturefluctuations, and adequatetemperaturecontrol for stable base-
lines is desirable.Someof the early eluent-suppressed IC instrumentsmanufac-
turedby Dionex Corp. (e.g., Model 10) did not havethis temperaturecontrol, but
this has beenremediedin later models. To facilitate temperaturecompensation
(up to 80°) a thermistoris placedin the liquid line just after the electrodeof the
conductivity detectormodule (e.g., Dionex Model 2oo2i). The cell is driven by
a high-frequencyoscillatorfrom the main circuit board.The cell output drives an
amplifier, and changesin the ionic composition in the cell result in signal
changesto the amplifier. The signal causedby the presenceof conductiveions in
the cell, after temperaturecompensation, resultsin meter and recorder-pen
deflection.
The nonsuppressed instrumentshave many of the componentsof eluent-
suppressed-type instruments.The main difference betweenthese two types of
instrumentslies in the column packing, and the lack of regenerationpump and
timer in the nonsuppressed systems.
Many inorganic ions display strong absorbancein the lower rangeof UV.
In the past,thesewavelengthswere not readily accessibleto IC photometers.But
now with the availability of UV detectorsthat reachdown to 200 nm and less,
z,
inorganicanions(e.g., N03", NO Br-, 1-, Br03", 103", and S201-) can be deter-
z,
mined (Small, 1983). Although N03", NO and Br- in such diverse environ-
mentsas river and wastetreatmentwaters,rain, eutecticsalt mixtures, and sali-
va havebeendetermined,little information is availableon the use of UV detec-
tors for the determinationof theseor otherinorganicions in soils. Also, ions such
as SCN-, S203", and severalpolythionatespecieshave been measuredsuccess-
fully by using low-capacityresinsand NaCI04 as an eluent. Cortes(1982) used
silica-basedcolumnswith aminofunctional groupsfor the effectiveseparationof
both organicand inorganicanionsthat are UV absorbing.
Recorders
Seeprevioussectionon "Recorders."
Applications
Application of IC to soil analysiswas pioneeredby the senior author and

his associatesat Iowa State University in the late 1970s (Dick & Tabatabai,
1979). Since then severalpapershave appearedin the soil scienceliterature on
the use of suppressedand nonsuppressed IC systemsfor the determinationof
anions and cations in soil solutionsand exchangeablebasesin soils (Tabatabai
&Basta, 1991). Someof thesemethodshave beenapplied successfullyto plant
and water analysis (Busman et aI., 1983; Tabatabai& Dick, 1983; Basta &
Tabatabai,1985a,b,c;Goyal et aI., 1993; Kalbasi & Tabatabai,1985). The IC
systemshouldbe useful for a variety of methodsusedin soil and plant analysis,
provided that the reagentsusedin the proceduresare compatiblewith the basic
principlesof operationof the IC. As such,many of the currentmethodsusedin
soil and plant analysisproduceionic speciesin a backgroundof either highly
acidic mediaor high salt concentrations.Consequently,new approachesor mod-
ifications of current methodsare essentialbefore using the IC systemfor the
determinationof the ionic speciesproduced.A summaryof the various applica-
tions of the IC for analysis of soil, water, and plant samplesis reported by
Tabatabaiand Basta (1991). Both types of IC systemsare employed in many
applications.Eluent-suppressed IC provides lower detection limits for strong-
acid anions, gradient capability for complex samples, and compliance with
USEPA method 300.0. NonsuppressedIC provides lower detection limits for
cations(monvalentand divalent cationsor transition metals),weak acid anions,
and a wider choiceof eluentsand instruments.Application of IC for determina-
tion of total heavy metalsin soils and sewagesludgesare describedin detail by
Bastaand Tabatabai(1990, 1991) Numerousenvironmentalapplicationsof ion
chromatographyare summarizedby Frankenbergeret al. (1990).
REFERENCES
Anderson,J.L., and DJ. Chesney.1980. Liquid chromatographicdeterminationof selectedcarba-
mate pesticidesin water with electrochemicaldetection.Anal. Chern. 52:2156-2161.
Basta,N.T., and M.A. Tabatabai.1985a.Determinationof exchangeablebasesin soils by ion chro-
matography.Soil Sci. Soc. Am. J. 49:84--89.
Basta, N.T., and M.A. Tabatabai.1985b. Determinationof total potassium,sodium, calcium, and
magnesiumin plant materialsby ion chromatography.Soil Sci. Soc. Am. J. 49:76-81.
Basta,N.T., and M.A. Tabatabai.1985c. Determinationof potassium,sodium, calcium, and magne-
sium in na.tural watersby ion chromatography.1. Environ. Qual. 14:450-455.
Basta,N.T., and M.A. Tabatabai.1990. Determinationof total metalsin soils by ion chromatography.
Soil Sci. Soc. Am. 1. 54:1289-1297.
Basta,N.T., and M.A. Tabatabai.1991.Determinationof total metalsin sewagesludgesby ion chro-
matography.1. Environ. Qual. 20:79-88.
Boyd, G.E., B.A. Soldano,and O.D. Bonner. 1954. Ionic equilibria and self-diffusion rates in desul-
fonatedcation exchangers.J. Phys. Chern.58:456-460.
Brown, P.R. 1990. High-performanceliquid chromatography:Pastdevelopments,presentstatus,and
future trends.Anal. Chern. 62:995-1108A.
Busman,L.M., R.P. Dick, and M.A. Tabatabai.1983. Detenninationof total sulfur and chlorine in
plant materialsby ion chromatography.Soil Sci. Soc. Am. J. 47:1167-1170.
Cortes, H.J. 1982. High-performanceliquid chromatographyof inorganic and organic anions using
ultraviolet detectionand an amino column. J. Chromatogr.234:517-520.
Dasgupta,P.K. 1992. Ion chromatography:The stateof the art. Anal. Chern.64:775-783A.
Destefano,1.1. 1980. Performanceevaluation of liquid chromatographiccolumn. p. 1-14. In J.
Harvey, Jr., and G. Zweig. (ed.) Pesticideanalytical methodology.ACS Symp.Ser. 136. Am.
Chern. Soc., Washington,DC.
Dick, W.A., and M.A. Tabatabai.1979. Ion chromatographicdeterminationof sulfate and nitrate in
soils. Soil Sci. Soc. Am. J. 43:899-904.
Eksborg,S., and G. Schill. 1973. Ion pair chromatographyof organicammoniumcompounds.Anal.
Chern.45:2092-2100.
Ettre, L.S. 1975. The developmentof gaschromatography.1. Chromatogr.112:1-26.
Ettre, L.S. 1980. Evolution of liquid chromatography:a historical overview. p. 1-74. In C. Horvath
(ed.) High performanceliquid chromatography:Advancesand perspectrives.Vol. 1. Acad.
Press,New York.
Frankenberger,W.T., Jr., H.C. Mehra, and D.T. Gjerde. 1990. Environmental applicationsof ion
chromatography.J. Chromatogr.504:211-245.
Franklin, G.O. 1985. Developmentand applicationsof ion chromatography.Am. Lab. 17(6):65-80.
Fritz, 1.S., D.T. Gjerde, and R.M. Becker. 1980. Cation chromatographywith a conductivity detec-
tor. Anal. Chern.52:1519-1522.
Fritz, J.S., and J.N. Story. 1974a.Selectivity behaviorof low-capacity,partially sulfonated,macrop-
orous resin beads.1. Chromatogr.90:267-274.
Fritz, 1.S., and J.N. Story. 1974b. Chromatographicseparationof metal ions on low capacity,
macroreticularresins.Anal. Chern. 46:825-829.
Girard, J.E., and J.A. Glatz. 1981. Ion chromatographywith conventionalHPLC instrumentation.
Am. Lab. 13(1):26-35.
Gjerde, D.T., and J.V. Benson.1990. Suspensionpostcolumnreaction detection method for liquid
chromatography.Anal. Chern.62:612-615.
Gjerde, D.T., and J.S. Fritz. 1979. Effect of capacity on the behaviorof anion-exchangeresins. J.
Chromatogr.176:199-206.
Gjerde, D.T., and J.S. Fritz. 1981. Sodium and potassiumbenzoateand benzoicacid as eluentsfor
ion chromatography.Anal. Chern.53:2324-2327.
Gjerde,D.T., J.S. Fritz, and G. Schmuclder.1979.Anion chromatographywith low-conductivity elu-
ents.J. Chromatogr.186:509-519.
Gjerde,D.T., G. Schmuckler,and J.S.Fritz. 1980.Anion chromatographywith low-conductivity elu-
ents.II. J. Chromatogr.187:35-45.
Goyal, S.S., AAR. Hafez, and D.W. Rains. 1993. Simultaneousdeterminationof total sodium,
potassium,magnesium,and calcium in plant tissuesusing acid digestionand ion chromatog-
raphy. Agron. J. 85:1192-1197.
Hamilton, R.I., and PA Sewell. 1977.Introductionto high performanceliquid chromatography.John
Wiley & Sons,New York.
Hanaoka,Y., T. Murayama,S. Muramoto, T. Matsuura, andA Nanba. 1982. Ion chromatography
with ion-exchangemembranesuppressor.J. Chromatogr.239:537-548.
Hassett,lJ. 1982. High-pressureliquid chromatography.p. 123-131. In A L. Page et al. (ed.)
Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr 9. ASA and SSSA,Madison,WI.
Horvath, c., and W. Melander. 1978. Reverse-phasechromatographyand the hydrophobiceffect.
Am. Lab. 10:17-36.
InternationalUnion of Pure and Applied Chemistry. 1974. Recommendations on nomenclaturefor
chromatography.PureAppl. Chern. 37:447-462.
Iskandarani,Z., and DJ. Pietrzyk. 1982. Ion interaction chromatographyof inorganic anionson a
poly (styrene-divinylbenzene) adsorbentin the presenceof tetraakyl-ammoniumsalts.Anal.
Chern. 54:2427-2431.
Johnson,E.L. 1987. Eluent suppressedion chromatography.p. 1-22. In J.G. Tarter (ed.) Ion chro-
matography,Marcel Dekker, New York.
Johnson,E.L., and R. Stevenson.1978. Basic liquid chromatography.Varian Assoc.,Inc., Palo Alto,
CA.
Jupille, T. 1987. Single-columnion chromatography.p. 23-82. In J.G. Tarter (ed.) Ion chromatogra-
phy. Marcel Dekker, New York.
Jupille, T.H., D.W. Togami, and D.E. Burge. 1983. A single-columnion chromatographyaids rapid
analysis.WescanInstruments,Inc., SantaClara, CA
Kalbasi,M., andM.A. Tabatabai.1985.Simultaneousdeterminationof nitrate,sulfate,and phosphate
in plant materialsby ion chromatography.Commun.Soil Sci. Plant Anal. 16:787-800.
Kargar, B.L.,L.R. Snyder,and C. Horvath. 1973. An introduction of separationscience.John Wiley
& Sons,New York.
Kissinger, P.T. 1977. Commentson reverse-phaseion-pair partition chromatography.Anal. Chern.
49:883.
Lawrence,J.F. 1982. High-performanceliquid chromatographyof pesticides.Acad. Press,Inc., New
York.
Martens,D.A, and W.T. Frankenberger,Jr. 1991a.On-line solid-phaseextractionof soil auxins pro-
duced from exogenously-appliedtryptophan with ion-suppressionreverse-phaseHPLC
analysis.Chromatographia32:417-'422
Martens,D.A, and W.T. Frankenberger,Jr. 1991b.Pulseamperometricdetectionof amino acidssep-
aratedby anion exchangechromatography.J. Liq. Chromatogr.15:423-439.
Martens,D.A, and W.T. Frankenberger,Jr. 1991c.Determinationof saccharidesin biological mate-
rials by anion-exchangechromatographywith pulsedamperometricdetection.J. Chromatogr.
546:297-309.
Neito, K.F., and W.T. Frankenberger,Jr. 1985. Single column ion chromatography:II. Analysis of
ammonium, alkali metals, and alkaline earth cationsin soils. Soil Sci. Soc. Am. I.
49:592-596.
Pohl, C.A, and E.L. Johnson.1980. Ion chromatography:The state-of-the-art.J. Chromatogr.Sci.
18:442-452.
Ravindranath,R. 1989. Principlesand practiceof chromatography.JohnWiley & Sons,New York.
Ritchie, A.S. 1966. Chromatographyas a natural processin geology.Adv. Chromatogr.3:119-134.
Saari-Nordhaus,R., and J.M. Anderson,Jr. 1994. Ion chromatographicanalysisof anions using a
solid-phasechemicalsuppressor.Am. Lab. 26:28C-281.
Small, H. 1983. Modern inorganicchromatography.Anal. Chern.55:235A-242A.

Small, H. 1989.Ion chromatography.PlenumPress,New York.
Small, H., T.S. Stevens,and W.C. Bauman. 1975. Novel ion exchangechromatographicmethod
using conductimetricdetection.Anal. Chern.47:1801-1809.
Smith, F.e.,Jr., and Re.Chang.1983.The practiceof ion chromatography,JohnWiley & Sons,New
York.
Stevens,T.S., J.e. Davis, and H. Small. 1981. Hollow fiber ion-exchangesuppressorion chro-
matography.Anal. Chern.53:1488-1492.
Stillian, J. 1985. An improvedsuppressorfor ion chromatography.LC Magazine3:802-805.
Stott, D.E., andM.A. Tabatabai.1985.Identificationof phospholipidsin soils andsewagesludgesby
high-performanceliquid chromatography.1. Environ. Qual. 14:107-110.
Tabatabai,M.A., and N.T. Basta. 1991. Ion chromatogrpahy.p. 229-259.In K.A. Smith (ed.) Soil
analysis.Marcel Dekker, New York.
Tabatabai,M.A., and w.A. Dick. 1983. Simultaneousdeterminationof nitrate, chloride,sulfate,and
phsophatein naturalwatersby ion chromatography.J. Environ. Qual. 12:209-213.
Tarter,J.G.,E.L. Johnson,andT. JupiIIe. 1987.Comparisonof eluent-suppressed and single-column
ion chromatography.p. 83-86. 1.G. Tarter (ed.) Ion chromatography.Marcel Dekker, New
York.
Warman,P.R., and C. Bishop. 1987. Freeand HF-HCI ex;tractableamino acidsdeterminedby high-
performanceliquid chromatographyin a loamy sand.BioI. Fert. Soils. 5:215-218.
Warman,P.R, and RA. Isnor. 1989.Evidenceof peptidesin low-molecularweight fractionsof soil
organicmatter.BioI. Fert. Soils 8:25-28.
Weiss,J. 1986. Handbookof ion chromatography.Dionex Corp., Sunnyvale,CA.
Zweig, G. (ed.). 1984. Analytical methodsfor pesticidesand plant growth regulators.Vol. 1-13.
Published 1996
Chapter 9
Differential Pulse Voltammetry
LARRY M. SHUMAN, University of Georgia, Griffin, Georgia
Pulsevoltammetric techniquesare uniquely suited to analysisof soil solutions,

soil extracts,soil digests,andplant nutrientsolutions.They are useful to gain spe-
cific information that may not be attainableby any other method.Two particular
problemsaddressedby pulsevoltammetryare very low concentrationsof analyte
(<10-7 M) and determinationof the speciesof analyte.Thesetechniquesare read-
ily availablethrough commercialinstrumentsthat are not extraordinarilyexpen-
sive, but which require operatorexpertiseand are slow by comparisonto tech-
niquessuchas atomic absorptionspectrometry(AA), inductively coupledplasma
emission spectrometry(ICP), or automatedcolorimetric analysis. Thus, pulse
polarographictechniquesshould be consideredwheneverthey may be the only
way to obtain the necessaryinformation and wheneverrelatively small numbers
of samplesare being analyzed.
The generalelectroanalyticalterm is voltammetry, which encompasses all
current-voltagetechniquesinvolving the application of varying potential to the
indicator electrode in relation to a reference electrode, with the resultingcurrent
being measuredat the potential for oxidation or reduction of the analyte. The
more specific term polarography refers to voltammetry involving a dropping
mercury electrode(DME). Pulsetechniquesare designedto increasesensitivity
by samplingthe currentand increasingthe ratio of Faradiacto capacitivecurrent.
Anodic stripping voltammetry (ASV) is a techniquein which the analyte is first
preconcentrated at the electrodeby electrolysis.The analyteis then electrochem-
ically removedor "stripped" from the electrodeand the resulting current mea-
sured. Becauseof the preconcentrationstep, concentrationson the order of 10-9
M or lesscan be determined.(Refer to Table 9-1 for abbreviations.)
The DME offers several advantages over stationaryelectrodes.The elec-
trode doesnot accumulatesolid products,so the only importantvariablesare the
electrode potential, solution composition, and current. In addition, samples
requiring extremely strong reducing conditions can be analyzedbecauseof the
high overvoltage for the reduction of H+ or water on the mercury surface.
Becauseof this characteristic,analyzableions include alkali and alkaline earth
metals and difficult to reduceorganic matter (Meites, 1965). Theseadvantages
Book Seriesno. 5.
247
248 SHUMAN
Table 9-1. Abbreviations.

ASV Anodic StrippingVoltammetry
DCP Direct CurrentPolarography
DME Dropping Mercury Electrode
DPASV Differential PulseAnodic StrippingVoltammetry
DPCSV Differential PulseCathodicStripping Voltammetry
DPP Differential PulsePolarography
HMDE HangingMercury Drop Electrode
HPLC High-PerformanceLiquid Chromatography
IC Ion Chromatography
MTFE Mercury Thin Film Electrode
NPP Normal PulsePolarography
SMDE Static Mercury Drop Electrode
must be weighed againstthe disadvantages,however. Mercury is easily oxidiz-

able, so strong oxidizing potentials (>+0.5 V) are impossible to obtain. Also,
changesin electrodeareawith size of the mercurydrop give rise to varying cur-
rents. Pulsetechniqueshelp to eliminate thesecurrentvariationswith drop size,
however,by samplingcurrentfor eachdrop at the sametime in its life or for ASV
by using a single drop for the entire analysis.Many excellentreviewscan be con-
sulted for details of voltammetric techniques,including Osteryoung (1981),
Heineman(1981), and Wang (1985).
A generaldiscussionof ASV and differential pulse techniquesas well as
othertechniquesincluding direct current(DC) polarographywasgiven in the sec-
ond edition of this book (Street& Peterson,1982). Therefore,the current chap-
ter will include only a brief review to acquaintthe readerwith the techniquesand
then will stressmore recent methodsand their application to soil science.The
objectivesare to presentan overview of ASV and differential pulsepolarography
(DPP) and their applicationsin the areasof environmentalsoil analysis,and to
indicate ways soil scientists may use these techniquesto advantagein their
research.
DIRECT CURRENTAND NORMAL PULSE POLAROGRAPHY
Direct CurrentPolarography
The basic apparatusfor polarographyis the electrochemicalcell, depicted

diagrammaticallyin Fig. 9-1. The cell shown also is suitablefor DPP and ASV.
Specific characteristicsrequired for thesetechniqueswill be pointed out in the
discussionsbelow. The DME capillary is made of glass, usually coatedwith a
surfactantto decreaseentry of the solution into the capillary. It also is pointedto
help eliminate obstructionsto diffusion. The referenceelectrode (calomel or
Ag-AgCI) is isolatedfrom the solution by a salt bridge. The auxiliary electrode
is a simple platinumwire, and thereis an inlet and outlet for the purginggas,usu-
ally N2• The polarogramis recordedin terms of changesin current betweenthe
DME and the auxillary electrodeas the potential is varied betweenthesesame
electrodes.
DIFFERENTIAL PULSE VOLTAMMETRY 249
Dropping Hg or
HMO Electrode
@
Speed Control
~-Stirrer
Fig. 9-1. A typical polarographiccell.
The DC polarogramis characterizedby a broad band, the top of which

increasesin height at the characteristicpotential for the analyte and remainsat
this new level as the potentialcontinuesto increase(Fig. 9-2). The bandis caused
by increasesand decreasesin currentas the mercurydrop enlargesand eventual-
ly falls. The heightof the "wave" above the baselineis proportionalto the con-
centrationof analyte, correspondingto the eventualcondition where the metal
ions are reducedas fast as they reach the electrodeby diffusion. This current,
referredto as the diffusion current, is describedby the Ilkovic equation
[1]
whereid =averagediffusion current(f.1A), n =numberof electrons,D =diffusion

coefficient (cm2/s), m = massflow of the mercury (mgls), t = drop time (s), and
C =analyte concentration (mmol L- 1). If all cell parametersare held constant,the
diffusion current is proportional to the concentrationof the analyte. Other cur-
rents also flow in the cell in addition to this "Faradaic"current, the greatestof
which is the "Capacitance"current. The ratio of Faradaicto capacitancecurrent
limits the sensitivity of DC polarography.Below about 10-4 M the ratio becomes
progressivelysmaller, until the capacitancecurrent dominates.This limitation
was the impetus for the developmentof modem pulse methods,which improve
the ratio (Crow, 1988). For more detailedinformation on DC polarographycon-
sult the secondedition of this book (Street& Peterson,1982), or Crow (1988, p.
166-211).
250 SHUMAN
:...J... Zn 2+
,
1 ppm Zn' + and Pb ' +
i 5 lolA full scale
Pt>'+
Olff.r.ntlal Puis. /\~

~-------,,"
Normal Pulae
I-
Z
W
a:
a:
::l
u
-0.8 -0.4 0 .2
POTENTIAL
Fig. 9-2. Examplesof DC, normal pulse,and differential pulse polarograms.
Normal Pulse Polarography
As mentionedabove, pulse polarographyis that carried out at a DME.

Normal pulsepolarography(NPP) is the simplestof the pulse techniqueswhere,
at the start of eachpulse, no currentflows (Osteryoung& Schreiner,1988). The
waveform is shown in Fig. 9-3a,b, which indicatesthat the pulsed potential is
applied in stepsof increasingamplitudefrom the initial potential. Each pulse is
about 50 ms in duration, and is placedon the DME just as the drop reachesits
largestsize. At this stage,the areato volume is no longer changingrapidly. The
drop time is controlledby using an electromechanical "drop knocker" (Fig. 9-1).
The current is sampledduring the last 10 to 17 ms of the pulse,when the capac-
itative currenthasdecayedto nearzero, but the Faradiaccurrentis still high (Fig.
9-3d). Thus, the measuredsignal is no longer a band, but insteada "stepping"
line. It is nearly completely Faradaic,and increasesthe sensitivity by up to 10-
fold over DC polarography.
~
E
a.
~L, 20
NPP OPP
b.
iL,
E
40 msec
J )....&....;
11
t
D. E
~ I
)....&....;
12
c. POTENTIAL
PULSE
\ faradaic
\
CURRENT \
d. \
RESPONSE \
\
\ ~_-r- nonfaradaic
, '
'--' '-
" sample/
current
Fig. 9-3. Potentialwaveformsfor normal pulse (a, b) and differential pulse (a, c) polarography,and
current responseduring a pulse (0) (Osteryoung,1981; Heineman,1981).
The diffusion currentin NPP is the Cottrell current
[2]
whereF =the Farad(96 489 C/equivalent),A =electrodearea (cm2), tm =time at

which the current is measuredin secondsfrom pulse application, and where
remainingforms are as given for Eq. [1]. This equationagain indicatesthat the
currentis proportionalto the concentration,and that noneof the constantsshould
changeappreciablywith matrix. The currentdecreasesin proportion to tm-1/2, so
the measurementtime should be as small as possible to maximize the signal.
Times are in the rangeof 10 to 30 ms (Osteryoung,1981).
252 SHUMAN
DIFFERENTIAL PULSE POLAROGRAPHY
Principles
Differential pulse polarographyis similar to normal pulse polarography

except that a difference in current is measured,and gives a peak waveform
insteadof a wave. In DPP the samesquare-wavepulseis applied to the drop, but
the pulseshave a constantamplitude(commonly 50 mY) that are superimposed
on a relatively slow linear DC potential sweep(Fig. 9-3a,c). During the second
half of the pulse,the increasein currentis recordedas the differencebetweenthe
currentjust before the pulse and that in its final 10 to 17 ms (Meites, 1965). In
NPP, on the other hand,just the currentin the last 10 to 17 ms is recorded.Figure
9-2 providesa comparisonof the NPP and DPP waveforms.
In DPP the kinetics of the electrodereactionare assumedto be sufficiently
rapid that the reactionis diffusion-controlled.The currentin DPP is relatedto that
in NPP by
i DPP = i NPP [(0 - 1)/(0 + 1)] [3]
where0 =exp(-nFM/2) whereM is the pulseheight (mY) and nandF are as

for Eq. [1] and [2]. The ratio in Eq. [3] dependsonly on the numberof electrons
transferredand on the pulseheight M. It is always lessthan one. In practice,the
optimum pulseheight is from 50 to 100 m V for maximumsensitivity.
In DPP the magnitudeof the measuredcurrent is somewhatless than in
NPP, but the lower noise level and the resultantincreasein gain producea sig-
nificant increase in analytical sensitivity. For a two-electron reaction, DC
polarographyhas a sensitivity of a few micromolar. Normal pulse polarography
has detectionlimits in the rangeof 0.2 to 0.4 ).1M and DPP has a lower limit of
about 20 nM. One has to be cautiousabout kinetic complicationsin DPP, how-
ever,especiallyin naturalsystems.Organicmattercanchangereactionratesfrom
sampleto sample,and thus changethe peak height to concentrationratio. High
salt concentrationin the supportingelectrolyteproducesflat backgroundsin DPP.
However,in the analysisof environmentalsamplesit may not be desirableto add
salt becauseof the potential for contaminationor for the destructionof natural
equilibria and speciation.
Applications
Differential pulse polarographyis typically applied to the analysisof low

levelsof metalsin many different typesof solutions.In orderto usethe technique,
however,the necessaryanalytical parametersmust be known. The referencesin
Table 9-2 are intendedto provide someintroductionto the literaturefor specific
elements,while illustrating someapplicationsof DPP. Suchreferencesgive valu-
able information about important parameterssuch as the type and strengthof
backgroundelectrolyte,the electromotiveforce (EMF) or half-wave potentialof
the polarographicpeak,the initial scanpotentialand width of the scan,drop size
[where it can be controlledsuchas with the PrincetonApplied ResearchCorpor-
Table 9-2. Differential pulsepolarographicapplications.

Element Application Reference
AI AI-humics equilibrium Ritchie et aI., 1982
AI-chelateequilibrium Shumanet aI., 1991
Cu, Pb, Cd, Zn Analysis Polak & Janacek,1989
Ni Ni complexes Crow & Rose,1979
Pb Pb speciation Goncalveset aI., 1985
TI Analysis Hoeflich et aI., 1983
Ni Plant tissue,water Flora & Nieboer, 1980
ation's (Princeton, NJ) static-drop electrode], the expected current or peak

height, the analyte concentrationwithin the most accuraterange for DPP, and
possible interferences.If more than one metal can be analyzedusing a single
scan, the referencesalso are helpful in finding parametriccompromiseswhich
will give the bestresolutionandprecisionfor all analytes.Thus,soil chemistsare
provided with a multitude of information to serve as starting points for their
analysisof soil extracts,digests,or soil solutions.
The last 10 yr haveproducedmany reportsby electrochemistsand analysts
usingelectrochemicaltechniques.Thesedocumentthe trial-and-errorparametric
work necessaryto use DPP on specific samplesfor specific elements.The appli-
cation of DPP to analysis, speciation,and complexationresearchwill be dis-
cussedin more detail later.
ANODIC STRIPPINGVOLTAMMETRY
Principles
Anodic stripping voltammetry is probably the most widely used voltam-

metric techniquefor the analysisof metals in environmentalsamples.This is
becauseof its extremesensitivity and its ability to distinguishamongspeciesof
metals.The analyte,single or multiple, is depositedand concentrated electrolyt-
ically onto the electrode.It is then oxidized or reducedfrom the electrode,giv-
ing a larger limiting or peakcurrentthan possiblewith other techniques (Meites,
1965).
The metal ion in solution is reducedto zero oxidation state, forming an
amalgamwith the mercuryor, in someinstances,being depositedas an insoluble
salt. The time for the depositiondependson the concentrationof analyte, with
longertimes being necessaryfor lower concentrations.However,times over 2 or
3 min, shouldgenerallybe avoidedbecausethe metal diffusestoo deeplyinto the
mercury drop, thus broadeningthe stripping peak. Stirring the solution during
this step also must be reproducible,and slow enoughso as to not dislodge the
mercury drop. This step is followed by a 30-s rest period without stirring, to let
the amalgamcometo equilibrium. Then an anodicscan(more positive potential)
is applied and the current recorded.Different scanscan be usedincluding linear
sweep,differential pulse,and alternatingcurrent(AC), (amongothers).The scan
most often usedis differential pulse,becauseof the accompanyinggain in sensi-
254 SHUMAN
tivity. Sourcesof information on ASV include Heineman (1981) and Wang

(1985).
The electrodesmost often usedfor ASV are of mercury,either the hanging
mercury drop electrode (HMDE) or mercury thin film electrode (MTFE)
(describedin detail below). In someinstancessolid electrodessuch as platinum
or gold are employedinstead.The peak current at a HMDE using linear sweep
voltammetryis given by
[4]
where ip = peakcurrent OlA), CE is the analyteconcentrationin the drop (moles

L- 1), A is the electrodearea (cm2), n = number of electrons,D is the diffusion
coefficient in the Hg (cm2/s), and v is the potential scan rate (V/s) (Heineman,
1981). The correspondingequationfor the MTFE is
[5]
where L is the thicknessof the film (cm) and the others are asin Eq. [4]. The
MTFE gives sharperpeaksthan the HMDE becausethe mercury film is so thin
that the analyte cannotdiffuse far. In more concentratedsolutions,on the other
hand, the film can becomesaturatedwith the analyte, causinganothertype of
error. The aboveequationsshow that peakcurrent(Faradaic)can be increasedby
increasingthe scan rate. However, the capacitivecurrent also is proportional to
the scan rate with the problem being worse for the HMDE than for the MTFE.
The charging current ultimately limits the detection limit of the linear ramp
voltammetrytechnique.Even so, the techniqueis widely used(Heineman,1981).
As mentionedabove,the techniquesusedto increasethe sensitivity of ASV
include useof pulseor differential pulsemodes.The techniquefor pulsemethods
is similar to that for linear sweep,exceptthat pulse techniquesare describedfor
NPP and DPP are usedduring the stripping step.The peakcurrentfor differential
pulse anodic stripping voltammetry(DPASV) at a MTFE is
[6]
where tp is the pulse width (ms) and others as defined in Eq. [2] and [5]
(Heineman,1981). The peak current can be increasedby decreasingthe pulse
width. However, the pulse must be long enoughto allow decay of the charging
current before the current measurementis made.Cathodicstripping also is used
for a wide array of inorganic anions and organics.In this case,a film of salt is
depositedon the electrodesurface.The techniqueis useful for halides,selenide,
sulfide, and oxyanions such as molybdate, chromate,and vanadateas well as
many organics(Wang, 1985).
Somelimitations occur for ASV, but most can be overcome.One limitation
is that only about27 metalsor metalloidscan be determined(Ag, As, Au, Ba, Bi,
~~~~~~~~~~~~~~~~~~~~
Te, and Zn) (Wang, 1985). A secondlimitation involves analytesolubility in Hg.
If the solubility of the analytein mercury is exceeded,it will begin to plate on the
Table 9-3. Anodic strippingvoltammetryapplications.

Element Application Reference
As, Ni, AI, Se Analysis by ads.stripping Chianget aI., 1989
Cd,Cu,Zn Speciationin water Poltrowicz et aI., 1982
Co, Cu,Pb, Cd, Ni Analysis of natural waters Pihlar et aI., 1981
Cu, Pb,Cd Analysis Polak & Janacek,1989
Analysis of soil extracts Edmondset aI., 1980
Na,K Analysis Coetzee& Ecoff, 1991
Ni Analysis by adsorptionstripping Braun & Metzger,1984
Pb Speciation Goncalveset aI., 1985
Analysis of geologicalsamples Khasgiwaleet aI., 1972
Se (DPCSV) Analysis of soils, plants Forbeset aI., 1979
Sn Analysis of geologicalsamples Bond et aI., 1970
TI Analysis of geologicalsamples Calderoni& Ferri, 1982
Analysis Hoeflich et al.,1983
Analysis Liem et aI., 1984
Zn,Cd,Pb,Cu Analysis of waters Danieleet aI., 1989
electrode.This can producemultiple stripping peaks(Heineman,1981). A third

problem is that interferencesmay be observedin certain cases.Intermetallic
compoundscan be formed, stripping peakscan overlap, and there can be inter-
ferencesfrom organicmatter(Wang, 1985). Theseproblemswill be discussedin
further detail below.
Applications
Anodic stripping is particularly useful for total and speciationanalysisof

metalsand otherelements,and hasfound wide applicationin the analysisof nat-
ural watersas well as geological,biological, and soil samples.The listing of ref-
erencesin Table 9-3 is similar to those for DPP in Table 9-2. The entries are
intended to provide an introduction to the literature for specific elementsand
analyses.They are valuableresourcesfor parameterssuchas discussedprevious-
ly for DPP aswell as thosespecific to ASV including depositiontimes. Someref-
erencesin the table also report more recent innovationssuch as the paper by
Chianget al. (1989)which describesan adsorptivestripping method.Forbeset al.
(1979) also usedcathodicstripping to analyzesoils and plants for Se. Theseref-
erencescan be usedto initiate variousASV methodsfor plant, soil, and soil solu-
tion analyses.More detailswill be given in a later sectionon applicationof DPP
and ASV to soil analyses.
Modem VoltammetricTechniques
Modern techniquesare so diverse that a description of all is outside the

purview of this chapter.However,five methodsthat may be importantin the soil
and environmentalchemistrylaboratoryare listed in Table 9-4, along with refer-
encesin order to provide an introduction into the literatureand to illustrate some
actual applications.
Alternating current polarographyconsistsof superimposingan alternating
voltage of small amplitude (a few millivolts) on the DC voltage ramp (Zuman,
256 SHUMAN
Table ~. Modern voltarnmetrictechniques.

Method Elements Reference
AC Polarography Sn Bond et aI., 1970
Co,Ni Locatelli et aI., 1991
Pb, n, Sn Cammann,1978
ChemicallyModified Electrode(CME) Ni Baldwin et aI., 1986,
Ni Thomsenet aI., 1988
Cu Miwa et aI., 1984
PotentiometricStrippingAnalysis(PSA) Na,K Coetzee& Ecoff, 1991
Variousmetals Hansen,1991
Pb,Cu Madsenet aI., 1983
Cd,Pb,Tl Christensenet aI., 1982
Membrane-Covered
Film Electrode(MCMFE) Cd Stewart& Smart,1984
AdsorptionAnalysis Ni Kisser, 1988

Organicmatter Bersier& Bersier,1988
As, Ni, AI, Se Chianget aI., 1989
Co,Ni Locatelli et aI., 1991
Various metals Wang, 1989
1987). It is used in studiesof adsorptiveprocessesat the electrodesurfaceand

also can be usedanalytically for determiningmetals,since it helps to eliminate
baselineproblems(Cammann,1978).
Chemically Modified Electrode (CME) voltammetry makes use of the
reactivity of organicligandssuchas dimethylglyoxime(DMG) with metals.The
DMG is incorporatedinto a carbonpasteelectrode,with the analysisthen being
throughadsorptionstripping as describedbelow (Thomsenet aI., 1988).
PotentiometricStrippingAnalysis(PSA) eliminatesthe problemsassociat-
ed with currentmeasurements. The methodis similar to ASV, in that it involves
preconcentratingthe metal in the Hg by electrolysis.However,the metal is then
strippedoff chemically and the potential measuredas a function of time. This
providesboth quantitativeandqualitativeinformationaboutthe analyte(Hansen,
1991).
The Membrane-CoveredMercury Film Electrode(MCMFE) was devel-
oped for analysisof metals in natural waters, where organicsadsorbedat the
electrodesurfacecan interfere with the analysis.A dialysis tubing membrane
over the surfacehelpseliminateorganicswhile the voltammetryis carriedout by
conventionalDPASV (Stewart& Smart, 1984).
Adsorptionanalysisor adsorptionstrippingvoltammetryis similar to ASV,
except that the preconcentrationstep is nonelectrolytic.It is used for organic
matter and for organically complexedmetals (such as with DMG), where the
analyte cannot be electrodeposited.Wang (1989) has provided an excellent
review of adsorptionvoltammetry.
INSTRUMENTS AND ELECTRODES

Instruments
There are many commercialpolarographicinstrumentsavailable. Most
perform similar functions and differ primarily with respectto their degreeof
____o __ o ______________
_ _ • • _ _ _ _ _ _ _ __ _ _ _ _ __ _ _ • _ _ AM,
_________
Potential
Potentiostat
Programmer
r---Cur-re
nt -----'.
Follower
I '-f
Logic Cu rrent Knocker
and Output
Timing
x y
Fig. 9-4. Block diagramof a polarographicinstrument(Osteryoung,1981).
automationand sophisticationin data handling. Since the last edition (1986) of

this monograph,instrumentshavebecomemuch more versatile,combiningcom-
puterswith the polarographicequipmentto automatethe proceduresand provide
a variety of dataoutputs.The basicinstrumentremainsunchanged,however,and
follows the diagramgiven in Fig. 9-4. Thepotentialof the indicator or working
electrodeis controlledwith respectto a referenceelectrodeusing a potentiostat
and the resulting current is convertedto a voltage using a current follower. This
currentsignal then is processedin a currentoutput sectionby a logic and timing
section,which also controlsthe potentialprogrammer.This logic and timing sec-
tion also controls an electromechanical"drop knocker" when a DME is used
with pulse techniques.The noise in current measurements at low levels is mini-
mized by using current averagingover a measurementtime window. Modern
instrumentsoffer AC polarography,fast linear scanvoltammetry and dual-cell
operation in addition to DC polarography, NPP, DPP, ASV, and DPASV
(Osteryoung,1981).These instruments also are being increasinglyusedfor high-
performance liquid chromatography(HPCL) and ion chromatography(IC)
detectionwith specialelectrodesand other auxillary equipment.
The electrochemicalcell has becomefairly standardized,and is usually
madeas small as possibleto accommodatesmall samplesizes.A diagramof one
such cell is shown in Fig. 9-1. The removableglasscontainerhas a stationary
cap with holes for the indicator electrode,the referenceelectrode,the auxiliary
electrode,and the outgassingtube. For the most precisework, the cell is jacket-
258 SHUMAN
ed for temperaturecontrol. The cell is taperedtowards its bottom, with a small

flat areaat that point accommodatinga teflon-coatedstir bar for ASV. The stirrer
must havean ON-OFFswitch independentof the speedcontrol, in orderto repro-
duce stirring speedsaccurately.The referenceelectrodeis calomel or Ag-AgCI,
connectedto the cell via a salt bridge. The auxillary electrodeis a platinum wire,
usedto minimize errors due to cell resistance.
Indicator Electrodes
Mercury Electrodes
The traditional DME consistsof a mercuryreservoirattachedto a capillary
tube by flexible tubing allowing adjustmentof the drop size and frequency by
varying the capillarydiameterand the heightof the reservoir.Modern DME's use
a glasscapillary with a taperedtip and generally measureout the mercury drop
using a motorized plunger insteadof gravity. For example,the Static Mercury
Drop Electrode(SMDE) (Peterson,1979) actually dispenseseach drop for the
NPP and DPP techniques.This meansthat the drop size can be neara maximum
during most of the drop life, insteadof continually increasingas it does during
gravity flow (Peterson,1979).
The HMDE consistsof a similar glasscapillary, but it includesa device to
dispensereproducibledrop sizes.Early modelsuseda micrometerto producethe
drop and the literature often referred to the numberof micrometerdivisions to
indicate drop size. Modern automatedinstrumentsuse electromechanicalmeans
to producethe drop, as for the SMDE mentionedabove.The capillary must be
coatedwith a hydrophobicsubstanceto preventsolution from creepinginto it. A
commonprocedureinvolves washingthe capillary in 3 M HN03, rinsing, drying,
passingsiliconizing fluid through it (dichlorodimethylsilane in CCI4), and dry-
ing again.The main advantagesof the HMDE are the low residualcurrents,the
wide potential range,and the excellentreproductibility of its mercury drop size.
Disadvantagesin comparisonto the MTFE include limited stirring ratesbecause
of the possibility of dislodging the drop, low surface/volumeratios, broad strip-
ping peaks,and formation of intermetalliccompounds(Heineman,1981).
The other major type of mercury electrodeis the mercury thin film elec-
trode (MTFE), consistingof a thin film of mercury plated on carbon. The film
gives high sensitivity becauseof the larger surfaceareato volume ratio (3 orders
of magnitudelargerthan for the HMDE) and the fact that high-stirringspeedsare
possible.Sometimesthe electrodeis rotated.A carbondisk 2 to 4 mm in diame-
ter is epoxyedto a glassor Teflon tube (Fig. 9-5). Electronicconnectionis via a
copperor stainlesssteelwire passingthroughpowderedcarbonor mercury. Film
thicknessrangesfrom 10 to 1000 nm, with 100 nm being most common. The
mercuryfilm is platedat the sametime as the analyte,by adding1 to 5 X 10-5 M
mercuric nitrate to the solution. The substratefor the mercury film is usually
either "glassy" carbon, wax-impregnatedgraphite (WIG), or epoxy-bonded
graphite.Glassycarbonfinds the widest use.Even thoughthe MTFE is more sen-
sitive and precise than the HMDE, the HMDE is more versatile and reliable,
especiallyin the pulsemode (Wang, 1985).
TEFLON - - - T -
COPPER - -+----t+
WIRE
GRAPHITE
POWDER - - +-----4
GLASSY _ - - - j _
CARBON
Fig. 9-5. Diagram of a mercury thin film electrode(Wang, 1985).
Solid Electrodes
Solid electrodesare made of metals like platinum or gold, or of various
forms of carbon.They are usedwhen the applicationcalls for a potential range
outside that of mercury, usually for organic matter (Osteryoung,1981). Solid
electrodesalso are necessarywhen analyzingfor Au or Hg. Another type of solid
electrodeis similar to a MTFE exceptthat the metalis not mercury, but gold or
someothermetal. A gold film on carboncan be used,for example,to analyzefor
As (Wang, 1985).
GENERAL METHODS
The analytical samplefor opp or ASV must, of course,be in liquid form

and be free from organic or inorganic ions which causeinterferences.In some
casesthe interferencesare a part of the analytical procedurewhen working with
chemicalspecies.Samplesfor purely analytical procedures(as opposedto spe-
ciation) must have the organic matter removedby digestion, since the analytes
may be bound to them so strongly that they cannotbe reducedat the electrode.
Organicmatteralso can adsorbto the electrodecausingspuriouspeaks.It also is
desirableto dilute sampleswith analyteconcentrationsabove a few milligrams
per liter beforeanalysis.Eventualsamplesize would dependon the samplecell's
physicalcharacteristics.Most cells require samplesof from 10 to 30 mL.
Most electrochemicalanalysesshould be carried out in a supportingelec-
trolyte. This solution servesto adjust the pH to an optimum level, improve solu-
tion conductance,and minimize peak distortion during any stripping step.
Usually the salt content is brought to 0.01 M with KN0 3. The impurities intro-
ducedby addingelectrolytecan be minimized by pre-electrolyzingthe salt solu-
tion in a large vesselcontaininga mercurycathodeof large surfacearea.A neg-
ative potentialof about-1.5 V vs. a standardcalomelelectrodeis appliedand the
solution stirred for 24 h or longer. Metal impurities are removedby reduction
into the mercury cathode(Heineman,1981).
260 SHUMAN
For both DPP and ASV the solution must be deoxygenatedby outgassing.
Nitrogen gasis bubbledthrough the solution for a period of 5 to 15 min. The N2
is usually purified by first passingit through heatedcopperturningsor througha
vanadoussolution. To prevent evaporationof the sample, the N2 is passed
through a liquid of similar ionic strengthbefore enteringthe samplecell. After
outgassing,the N2 is continuouslyblanketedover the solution to preventair con-
tact during the analysis.
A DPP analysiscan be startedimmediatelyafter deoxygenating.For ASY,
however,the stirrer is startedand the depositionstep is carried out for an addi-
tional 1 to 30 min. Most depositiontimes are in the orderof a few minutes.Then
the stirring isstoppedand the solution allowed to equilibratefor 30 s beforestart-
ing the stripping step.
Two methodsare employedto convertthe peakcurrent(ip) to concentration
(activity). One is to preparea calibration curve, and the other is to use standard
additions.Standardadditionsare used for natural samplesto milJ.imize extrane-
ous matrix effects.After analysis,a small amountof a standardis addeddirectly
to the solution in the cell and the analysisrepeated.The formula for calculating
the original solution concentrationis
[7]
where i] = sample peak height (scale divisions), i2 = sample + standardpeak

height (scaledivisions),v = volume of standard addition(mL), V = original sam-
ple volume(mL), Cs = concentrationof standardsolution (mol L-I), and C = con-
centrationof the original sample(mol L- 1) (Heineman,1981).
APPLICATION TO SOIL SOLUTIONS AND NATURAL WATERS
Complexationand Speciation
Modem chemical analysis involves more than just determining the total
concentrationof an analyte.The "reactivity" of the element,its chemicalpoten-
tial (activity) and evenits oxidation state all are concernsof the analytical
chemist. Such values dependon the processesand propertiesof the elementin
solution, and especiallyon its interactionwith other soluble and colloidal com-
ponents.Soil solutions include a plethoraof inorganic ions, colloidal inorganic
and organicmatter,as well as solubleorganicand inorganicligands.It is certain-
ly not easyto determinethe activity of an inorganicanalytein this chemicalenvi-
ronment.The electroanalyticaltechniquesdiscussedabovecan be of greathelp in
sorting out the "free" uncomplexedion. They also may give information about
the lability (disociability) of the complexesformed, and aboutthe concentrations
of variousspeciesof the ion understudy.
In the early 1970s water chemistsstarted to apply DPP and ASV to the
determinationof complexedspeciesof metals. For example,Ernst et al. (1975)
usedDPP and DPASV to determinethe stability constantsof Cu, Pb, and Zn in
equilibrium with natural organic matter. Variations of peakcurrent with pH and
alkalinity gave information about the speciespresentfor comparisonwith equi-

librium models. They were able to determinethe stability constantsfor Cu and
Pb complexeswith carbonatefrom shifts in the peak potential. They were not
able to determineaccuratestability constantswith humic acid, however,because
the concentrationof humic acid in the diffusion layer was unknown. Such deter-
minations are basedon the assumptionthat the metal-ligand complex is not
labile. Thus, any metal beingdeterminedat the electrodesurfaceis assumedto be
already in solution, with none disassociatingfrom complexes.This assumption
may not hold in all cases,however.ShumanandWoodward(1973) usedASV and
complexometrictitration to determinethe CdEDTA (ethylenediaminetetraacetic
acid) formation constantsuccessfully.Shumanand Cromer(1979) usedthe same
techniqueto determineconditional stability constantsfor Cu-humic acid and
Cu-fulvic acid, after devising a methodto correct for kinetic dissociationof the
complexes.The dissociationof metal complexeswere rapid enoughthat some
dissociated,metal was actually being measuredat the electrode,causingan error
in the amountof "free" uncomplexedmetal determined.A more commonuse of
DPP and ASV is to determinegeneralspeciesor forms of metal designatedby
termssuch as "labile", "free" and "complexed"or adsorbedto colloids.
A techniquefor differentiatingforms of metal in naturalsolutionsis to ana-
lyze the solution using DPP or ASV without pretreatmentin order to determine
the labile or "free" uncomplexedforms, and then to treat the samplewith acid,
ultraviolet radiation, orpersulfateoxidation before analysisagain for the'total
metal content.A speciationschemerepresentativeof the usual approachfor nat-
ural watersis given by Florenceand Sately (1977). Free Cu, Pb, Cd, and Zn are
first determined,and then ultraviolet radiation is used to destroy organic matter
prior to determinationof total metalspresent.A further step is to passthe solu-
tion througha chelatingresin column to separateany metal containedon colloidal
matter from metal which can be chelatedby the resin. Another exampleof the
techniqueis illustratedin the work of Danieleet al. (1989), who usedDPASV for
naturalwatersto determinelabile and total Zn, Cd, Pb, and Cu. They determined
the metalsat natural pH and again at pH 1.5, in order to protonate any organic
ligandsand thus minimize metal complexation.
The metal complexingcapacityof soil solution also can be determinedby
titration, by incrementallyadding a standardmetal solution to the soil solution
and analyzingvia ASV after eachstep. The peak height plotted as a function of
addedmetal will show a suddenincreasewhen the metal-complexingcapacityof
the water is reached(Fig. 9-6). Plotrowicz et al. (1982) used this techniqueto
study organic-metalinteractionsin seawater.They concludedthat there is little
complexingof Cd, stronginteractionwith Zn, and varying degreesof interaction
with Cu. They also noted some differencesbetweenmarine and soil fulvic and
humic materials.Differential pulse polarographyhas been employedas well to
study the equilibria betweenhumic acids and AI (Ritchie et aI., 1982). Most of
the AI was found to be complexed.Shumanet ai. (1991) showedthat equilibri-
um models for AI-synthetic chelatescan be testedusing the DPP techniqueof
Ritchie et al. (1982).
Membranefiltration is often not a satisfactorymethod for distinguishing
betweendissolvedand particulateforms of metals,becauseof contaminationand
262 SHUMAN
COMPLEXIOMETRIC TITRATION
OF SOIL SOLUTION
.E
Ol
·iii
I
-'<
III
Q)
c..
Metal Concentration ________
Fig. ~. Typical graphof a complexiometrictitration to find the complexingcapacityof a soil solu-

tion sample.
adsorptionproblemsand the presenceof colloidal particlessmallerthan the filter

pores. Goncalveset a1. (1985) usedDPP and DPASV to determineeqUilibrium
contstantsfor the specific adsorptionof Pb on iron oxide, manganeseoxide and
silica oxide colloidal surface. No filtration was used and no pH buffers were
addedthat might alter the eqUilibria. Also, organicmaterialwas omittedfrom the
studiesbecauseof obviousinterferenceproblems.
Oxidation-Reduction
Polarographyis unique in that it also can be usedto determinethe oxida-

tion stateof a metal, becauseeach specieshas a different potential at which it
reactswith the mercuryelectrode.In working with anoxic or partially anoxicsoil
solutionsincluding thosesolutionswith increasedCO2 content,the solution must
be sampledin sucha mannerthat it doesnot comein contactwith the atmosphere.
The electrochemicalcell mustbe setup as a flow-through cell in orderto not mix
the solutionwith air. Shouldthe solution be exposedto air, solublegassessuchas
CO2 would be released,therebyincreasingthe pH and altering the speciespre-
sent.
Davisonet a1. (1988) described proceduresfor the determinationof reduced
forms of Fe(II), Mn(II), and S(-II) in anoxic waters.Theseprocedurescould be
appliedequally well to soil solutions.They usedNPP and DPP for the analyses,
with no electrolyteaddedin order that the chemicalequilibria in the solution not
be altered. Polarographictechniquesalso can be used to distinguish between
Fe(II) and Fe(III). The peaksfor Fe(II) by DPP have beenmeasuredat -1.35 V
for fresh water and -1.45 V for seawater.The peakpotential dependssomewhat
on other ions presentin the water as well. Fe(III) is usually so dilute as to be
undetectable,exceptfor somehydrolyzedspeciesor thoseheld in solution by nat-
ural organics.The oxidation stateof As also can be determinedby ASV. Even
thoughAs(III) may be the only form measuredby ASV, the As(V) in solutioncan
be convertedto As(III) chemically, the determinationrepeated,and the As(V)
found by difference(Heineman,1981).
Sampling f----------------------------------------------------------
!
0.45 }1m filtration Digestion
N 2 pressure, 1 bar Wet (HN0 3!H 2S0 4 + H P2)
!
UV-irradiation
mercury arc lamp
1-2 h
1
I Aliquotation for Determination ---------------------
r-
Cu, Pb, Cd, Zn by DPASV Cu, Hg by Ni, Co by DC or DPV
then Se(IV) by DPCSV DPASV at gold after accumulation
at HMDE, below 1jlg!1 at electrode at HMDE by DMG-chelate
MFE adsorption
Fig_ 9-7_ Flow diagramfor speciationof metalsin a naturalsolution (Pihlar, 1981)_
Oxygenis measuredat the mercuryelectrodeby the following reactionsin

neutralor alkaline solutions
O2 + 2H20 + 2e- = H20 2 + 20W [8]

H 20 2 + 2e- = 20W [9]
The first reactionoccursat a potential of -0.50 V vs. the standardcalomelelec-

trode and the secondat -0.90 V. The first wave is the one normally usedfor O2
analysis(Davisonet aI., 1988).
ChemicalAnalysis
Polarographictechniquescan be useful alternativesto AA, ICP, or colori-

metric methodsfor traceanalysisof metalsin soil extractsand soil solutions.For
example,the proceduregiven by Pihlar et ai. (1981)for analysisof naturalwaters
should be applicableto soil solutions or extracts(Fig. 9-7). The pretreatments
recommendedinclude filtration through a 0.45-J..lm membraneand treating to
removeorganicmatterby H20 2 and ultraviolet radiation.The sampleis then sub-
jected to DPASV for Cu, Pb, Cd, and Zn, using an HMDE. Another acidified
aliquot is usedfor DPASV at a gold electrodeto determineHg. Finally, this sec-
ond aliquot is neutralizedand then analyzedfor Ni and Co by DPP after accu-
mulation at an HMDE by DMG-chelateadsorption.The final techniquewas dis-
cussedaboveas adsorptionanalysis,and gives a detectionlimit of 1 ng/L for Ni.
264 SHUMAN
Sb
I 5.01lA
Pb
Cu
Cd
Fig. 9--8. Polarogramfor Cd, Pb, Cu, and Sb in a soil extract -0.1
(Edmondset aI., 1980). E(V vs Ag/ AgCI}
Copperand Pbcanbe determinedsimultaneously ata MTFE using DPASV

for soil extracts(Edmondset aI., 1980). Here, the metalsare extractedusing 0.05
M EDTA, which is subsequentlydestroyedby HN03 and H20 2 • The preconcen-
tration is carriedout at -1.0 V for 1 to 5 min, the stirring stoppedfor 30 s, and
the potential scannedfrom -1.0 V to 0.2 V (Fig. 9-8). The potential is then
steppedback to +1.0 V to strip off the mercuryfilm. The analyticalresultscom-
pare favorably to those from AA. Selenium also can be analyzed using
Differential Pulse Cathodic Stripping Voltammetry (DPCSY) at the HMDE
(Forbeset aI., 1979). The methodcan be appliedboth to soil and plant analyses,
following appropriate sample preparation. In the technique of Forbes et al.
(1979), the Se is volatilized and trappedin persulfate-sulfuricacid. The solution
is treatedwith HCI to destroyexcesspersulfateand convert Se(VI) to Se(IY). A
low level of Se is addedto the sample(0.253 J..lmol L-l or 0.02 ppm) before the
stripping analysisto placethe samplecurrentpeakheight on the linear portion of
the calibrationcurve.
Exotic trace metals like Tl also are good candidatesfor electrochemical
techniques.Although TI is not consideredan environmentalpollutant, it can
appearin cementdustsand is very toxic. Soils and rocks can be analyzedfor Tl
by digestingand then concentratinginto a small amountof acid. The sampleis
analyzedusing DPASV with a detectionlimit of 1 J..lglkg in a solid sampleusing
a depositiontime of 10 min (Liem et aI., 1984).Thus, trace metalsfound evenin
extremelylow levels in natural soils can be readily analyzedusing DPASV.
INTERFERENCESAND OTHER PROBLEMS
As for all analytical techniques,DPP and ASV have limitations that need
to be addressedby the analystso that the data obtainedare as accurateand pre-
cise as possible.The two problemsmost often encounteredare poor peakresolu-
tion, and adsorptionof organics.
In sampleswith overlappingpeaks,the largestpeakis often not the one of
analytical importance.The HMDE can exacerbatethis problem by producing
broad peaks as the analyte diffuses out of the mercury (Heineman, 1981).
Therefore,the MTFE gives better resolution.Other ways of coping with resolu-
tion are to shift potentialsby changingelectrolytes,or by using chelatingagents
to preferentially chelatethe interfering ion. Both of thesetechniqueswere sug-
gestedby Christensenet al. (1982) to separateTI and Pb peaks.Adding an excess
of EDTA chelatedthe Pb and Cd but left the TI in solution. A further methodto
help eliminate interferencesby other ions is to choosea depositiqnpotential for
ASV that is only as high as necessaryfor the analyte,thus eliminating interfer-
encefrom ions reducedat still higher potentials.
Minute amountsof surfactantsalso can producepeaksin DPP at the poten-
tials where they are adsorbedon the electrode surface (Osteryoung, 1981).
Although this phenomenoncan be exploited for analysisof organic matter and
forms the basisfor a methodtermed"tensammetry,"it cancausesignificant inter-
ferencesin metal determinationsfor natural watersor soil solutions.Adsorption
of surfactantscan shiftor obscurestripping peaksduring metal stripping analysis
(Bersier & Bersier, 1988). Organic matter also can producea broad background
peakwhich can makeevenstandardadditionsuselessin determiningthe concen-
tration of a metal.
Besidespeakoverlapand adsorptionof organics,therealso are other prob-
lems associatedwith polarographictechniques.Kinetic problems when deter-
mining "free" metalsin the presenceof organicmaterialhavebeendiscussedpre-
viously (Shuman& Woodward, 1972; Shuman& Cromer, 1979). Another prob-
lem involves the formation of intermetallic compoundsupon electrodeposition
(Heineman,1981; Baldwin et aI., 1986).The intermetalliccompoundsoften oxi-
dize at a differentpotentialfrom the individual metals,thus interfering with accu-
rate determinations.The problem can be avoidedif a metal is addedwhich pref-
erentially forms another intermetallic compound instead. For example, when
determiningZn, a Cu-Zn compoundcan causeproblems.One solution is to add
Ga3+ to form the more stable Ga-Cu intermetallic, thus leaving the Zn free
(Heineman,1981). Choosinga properelectrolysispotentialalso can be helpful in
avoiding intermetallicdeposition(Christensenet aI., 1982).
A possibleproblemwhen determiningion speciesis the pH at the electrode
surface.Reductionof Pb, Cd, and other metalsof interest to the corresponding
metal amalgamscan produce hydroxide at the electrodesurface.The resultant
problems are twofold. First, the results can be interpretedto suggestthat two
metal speciesare present,when in reality, there is only one. Second,the species
of interestcan appearto havea lower concentrationbecauseof the pH at the elec-
trode. Peak height is often extremely pH sensitive (Ritchie et aI., 1982), so a
small shift can lower the apparentconcentrationdrastically.
266 SHUMAN
CONCLUSIONS
Polarographictechniquesare becomingmore widely usedin various areas

of scientific investigation, especially in environmentalchemistry. Differential
pulse polarographyand ASV are excellenttools for identifying, characterizing,
and quantifying specific chemicalspeciesin untreatedsamples.As instruments
gain sophisticationin controlling analysisand in data manipulation,we should
seeincreasingapplicationof pulse electrochemicaltechniquesto soil chemistry.
Sophisticatedinstruments are already commercially available for reasonable
prices.
The techniquesdescribedare not difficult to implementfor soil extractsand
soil solutions, both with and without sample pretreatment.Soil scientistsare
encouragedto considerDPP and ASV when planning researchinvolving deter-
mination of metal speciation.Although such methodsare not "quick and easy,"
they are pow~rful techniquesfor examiningspeciesof metals in solution or fo.r
analyzingextremelylow metal concentrations.
ACKNOWLEDGMENTS
Contribution from the University of Georgia, Departmentof Crop & Soil

Sciences,Georgia ExperimentStation, Griffin, GA 30223-1797.This research
was supportedby Stateand HATCH funds allocatedto the GeorgiaAgricultural
ExperimentStations.
REFERENCES
Baldwin, R.P., J.K. Christensen,and L. Krugger. 1986. Voltammetricdeterminationof tracesof nick-
el(II) at a chemicallymodified electrodebasedon dimethylglyoxime-containingcarbonpaste.
Anal. Chern.58:1790-1798.
Bersier,P.M., and J. Bersier. 1988. Polarographicadsorptionanalysisand tensammetry:Toys or tools
for day-to-dayroutine analysis?Analyst 113:3--14.
Bond, A.M., T.A. O'Donnell, A.B. Waugh, and R.I.W. Mclaughlin. 1970. Use of polarographic
methodsfor the determinationof tin in geologicalsamples.Anal. Chern.42:1168-1172.
Braun, H., and M. Metzger. 1984. Analytical determinationof nickel in the environmentby adsorp-
tion voltammetryusing the mercury film electrode.Fres. Z. Anal. Chern.318:321-326.
Calderoni,G., and T. Ferri. 1982. Determinationof thallium at subtracelevel in rocks and minerals
by coupling'differentialpulse anodic-strippingvoltammetry with suitable enrichmentmeth-
ods. Talanta29:371-375.
Cammann,K. 1978.A critical copmarisonbetween flameless atomic absorptionspectroscopyand an
improved electrochemicalanodic stripping techniquein the caseof a rapid trace determina-
tion of lead in geologicalsamples.Fres. Z. Anal. Chern. 293:97-103.
Chiang, L., B.D. James,and R.I. Magee. 1989. Adsorptive stripping voltammetryof sometrace ele-
mentsin biological samples.II: Nickel, arsenic,and selenium.Mikrochim. Acta 2:149-156.
Christensen,J.K., L. Krugger, and N. Pind. 1982. The determinationof tracesof cadmium,lead, and
thallium in fly ash by potentiometricstripping analysis.Anal. Chim. Acta. 141:131-146.
Coetzee,J.F., and M.I. Ecoff. 1991. Potentiometricstripping analysisat microelectrodesin various
solvents and some comparisons with voltammetric stripping analysis. Anal. Chern.
63:957-963.
Crow, D.R. 1988.Principlesand applicationsof electrochemistry.3rd ed. Chapmanand Hall, London.
Crow, D.R., and M.E. Rose.1979.The electrochemicalbehaviourof the nickel(II) ion at the dropping
mercury electrode in the presenceof some complexing agents. Part 1. Electrochim. Acta
24:41-45.
Daniele,S., M. Baldo, P. Ugo, and G. Mazzocchin.1989. Determinationof heavy metalsin real sam-
ples by anodic stripping voltammetry with mercury microelectrodes.Part 2. Application to
rain and seawaters.Anal. Chim. Acta 219:19-26.
Davison, w., 1. Buffle, and R. DeVitre. 1988. Direct polarographicdeterminationof O2, Fe(II),
Mn(II), S(-II) and relatedspeciesin anoxic waters.Pure Appl.Chern. 60:1535-1548.
Edmonds,T.E., P. Guogang,and T.S. West. 1980. The differential pulse anodic stripping voltamme-
try of copperand lead and their determinationin EDTA extractsof soils with the mercuryfilm
glassycarbonelectrode.Anal. Chim. Acta. 120:41-53.
Ernst, R., H.E. Allen, and KH. Mancy. 1975. Characterizaitonof trace metal speciesand measure-
ment of trace metal stability constantsby electrochemicaltechniques.Water Res. 9:969-979.
Flora, C.J., and E. Nieboer. 1980. Determinationof nickel by differential pulse polarographyat a
dropping mercury electrode.Anal. Chern.52:1013-1020.
Florence,T.M., and G.E. Bately. 1977. Determinationof the chemicalforms of trace metalsin natur-
al waters,with specialreferenceto copper,lead, cadmiumand zinc. Talanta24:151-158.
Forbes,S., G.P. Bounds,and T.S. West. 1979. Determinationof seleniumin soils and plants by dif-
ferential pulsecathodic-stripping voltammetry. Talanta26:473-477.
Goncalves,M. de L.S., L. Sigg, and W. Stumm. 1985. Voltammetric methodsfor distinguishing
between dissolved and particulatemetal ion concentrations inthe presenceof hydrousoxides.
A casestudy on lead (II). Environ. Sci. Technol. 19:141-146.
Hansen, P. 1991. Heavy metal determination using potentiometric stripping analysis. American
LaboratoryApril, p. 52-56.
Heineman,W.R. 1981. Stripping voltammetry p. 125-150.In H.B. Mark, Jr., and 1.S. Mattson (ed.)
Water quality measurement.Marcel Dekker, New York.
Hoeflich, L.K, R.J. Gale, and M.J. Good. 1983.Differential pulsepolarographyand differential pulse
anodic stripping voltammetry for determinationof trace levels of thallium. Anal. Chern.
55:1591-1595.
Khasgiwale,K, R. Parthasarathy, and M. SankarDas. 1972. Determinationof lead in geologicalsam-
ples by anodicstripping analysis.Anal. Chim. Acta 59:485-489.
Kisser, R. 1988. Adsorptive stripping voltammetryof Ni(lI) using fast linear sweeps.Fres. Z. Anal.
Chern. 332:787-790.
Liem, 1., G. Kaiser, and M. Sager.1984. The determinationof thallium in rocks and biological mate-
rials at ng g-l levels by differential-pulseanodic stripping voltammetry and electrothermal
atomic absorptionspectrometry.Anal. Chim. Acta 158:179197.
Locatelli, c., F. Fagioli, and T. Gara. 1991. Peakresolution in the determinationof cobalt and nickel
by differential pulse and alternative current adsorption voltammetry. Anal. Chern.
63:1409-1413.
Madsen,P.P., I. Drabaek,and J. Sorensen.1983. The determinationof copperand lead in sediments
by potentiometricstripping analysis.Anal. Chim. Acta 151:479-482.
Meites, L. 1965. Polarographictechniques.2nd ed. Intersci. Publ., New York.
Miwa, T., L. Jin, and A. Mizuike. 1984. Differential-pulseanodic stripping voltammetry of copper
with a chemically-modifiedglassycarbonelectrode.Anal. Chim. Acta 160:135-140.
Osteryoung,J. 1981. Pulse polarographyp. 85-123. In H.B. Mark, Jr., and J.S. Mattson (ed.) Water
quality measurement.Marcel Dekker, New York.
Osteryoung,1.G., and M.M. Schreiner.1988. Recentadvancesin pulse polarography.Crit. Rev. Anal.
Chern. 19:51-S28.
Peterson,W.M. 1979. Static mercury drop electrode.American LaboratoryDecember,p. 69-78.
Pihlar, B., P. Valenta,and H.W. Numberg. 1981. New high performanceanalytical procedurefor the
voltammetric determinationof nickel in routine analysisof waters, biological materials and
food. Fres. Z. Anal. Chern. 307:337-346.
Plotrowicz, S.R., M.S. Young, J.A. Pulg, and M.1. Spencer.1982. Anodic stripping voltammetry for
evaluationof organic-metalinteractionsin seawater.Anal. Chern. 54:1367-1371.
Polak, J., and L. Janacek.1989. Application of polarographyin petrochemicalanalysis.TrendsAnal.
Chern. 8:145-150.
Ritchie, G.S.P., A.M. Posner,and 1.M. Ritchie. 1982. The polarographicstudy of the equilibrium
betweenhumic acid and aluminum in solution. J. Soil Sci. 33:671-677.
Shuman,L.M., D.O. Wilson, and E.L. Ramseur.1991. Testing aluminum-chelateequilibria models
using sorghumroot growth as a bioassayfor aluminum. Water Air Soil Pollut. 58:149-158.
Shuman, M.S., and J.L. Cromer. 1979. Copper associationwith aquatic fulvic and hjmic acids.
Estimationof conditional formation constantswith a titrimetric anodicstripping voltammetry
procedure.Environ. Sci. Technol. 13:543-545.
268 SHUMAN
Shuman,M.S., and G.P. Woodward,Jr. 1973. Chemicalconstantsof metal complexesfrom a com-

pleximetrictitration followed with anodicstrippingvoltammetry.Anal. Chern.45:2032-2035.
Stewart,E.E., and R.B. Smart.1984. Differential pulseanodicstrippingvoltammetryof cadmium(II)
with a rotatingmembrane-covered mercuryfilm electrode.Anal. Chern.56:1131-1135.
Street,J.J.,and W.M. Peterson.1982.Anodic strippingvoltammetryanddifferential pulsepolarogra-
phy. p. 133-148.In A.L. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron.
Monogr. 9. ASA and SSSA,Madison,WI.
Thomsen,K.N., L. Kryger, and R.P. Baldwin. 1988. Voltammetric determinationof tracesof nick-
el(11) with a mediumexchangeflow systemanda chemicallymodified carbonpasteelectrode
containingdimethylglyoxine.Anal. Chern.60:151-155.
Wang, J. 1985. Stripping analysis: Principles, instrumentation,and applications,VCH Publ., Inc.,
Deerfield Beach,FL.
Wang,J. 1989. Voltammetryfollowing nonelectrolyticpreconcentration.p. 1-88. In AJ. Baird (ed.)
Electro-analyticalchemistry:A seriesof advances.Vol. 16. Marcell Dekker,New York.
Zuman,P. 1987.Polarographicanalysis.p. 104-107.In S.P.Parker(ed.)Hill Encyclopediaof Science
andTechnology.Vol. 14, McGraw Hill, New York.
Published 1996
Chapter 10
Fourier Transform Infrared and Raman

Spectroscopy
C. T. JOHNSTON,Purdue University, West Lafayette, Indiana
Y. O. AOCHI, University of California, Riverside, California
Over the past 50 yr, vibrational spectroscopyhas provided a great deal of infor-
mation aboutthe structure,bonding,and reactivity of soil colloids. In early stud-
ies, primarily through the use of dispersiveinstruments,infrared spectroscopy
was usedextensivelyto identify soil constituentsand to elucidate structuralfea-
tures of both inorganic and organic components.The basisfor its utility in this
regardis that all soil componentsexhibit a vibrational spectrum.Unlike powder
x-ray diffraction methodsthat are sensitivemainly to crystallinecomponents,all
of the solid and liquid componentsfound in soil havea setof characteristicvibra-
tional bands.Thesebandscan be used as diagnostic indicators to confirm the
presenceof a particularconstituentor functional group. This approachhas been
the subject of numerousreviews including those of Morten~on et ai., 1965;
Farmer,1968, 1974; White, 1971, 1977; Ghosh, 1978; Brown and Clark, 1980;
MacCarthy and Rice, 1985; White and Roth, 1986; and Bloom and Leenheer,
1989.
Equally importantto the role it hasplayedin the identification and charac-
terizationof soil components,vibrationalspectroscopyhascontributedgreatly to
our understandingof chemicalprocessesinvolving soil and subsurfacematerials.
Vibrational motionsof the atomsin a moleculeor surfaceof interestare very sen-
sitive to small changesin their local environment.As interactionsoccur at the
solid-solutionor solid-vaporinterface,the molecularenvironmentsof the inter-
acting speciesare perturbed.These changes,in turn, can serve as diagnostic
propertieswhich may be used to explore surfacestructuresand to gain insight
into the interaction between surfaces and adsorbates.Diagnostic properties
includ:: the appearanceor loss of vibrational bands, changesin relative band
intensities,shifts in band frequency,or a changein the lineshapeof a particular
vibrational band. This information is most useful when correlatedwith data
obtained using other methods, particularly when those methods characterize
Copyright © 1996 Soil ScienceSociety of America and American Society of Agronomy, 677 s.
Book Seriesno. 5.
269
270 JOHNSTON& AOCHI
macroscopicpropertiesof the system,suchas thermodynamicstability constants

or reactionkinetics. Thesetypesof correlationsprovide a powerful approachto
understandingthe chemistry occurring at soil colloid surfaces(Motschi, 1984;
Sposito,1986; Johnston& Sposito,1987). The fate and transportof pollutants,
behavior of exchangeablecations near soil colloid surfaces,bonding mecha-
nismsof pesticides,specificadsorptionof anions,and the behaviorof waternear
mineral surfacesare all examplesof researchareasthat have benefitedfrom the
utilization of vibrational spectroscopy.Generaldiscussionsof theseapplications
can be found in Mortland, 1970; Farmer, 1971; Farmer and Russell, 1971;
Theng, 1974; Burchill et aI., 1981.
The applicationof vibrational spectroscopyto soil chemistryhasbeenhin-
deredin the pastby interferencesfrom water, impurities, and bulk soil materials.
A recentresurgenceof interestin this areais primarily attributableto the intro-
duction and availability of Fourier transform infrared (FfIR) spectrometers.
During the past 15 yr, FfIR spectrometershave replaceddispersiveinstrumen-
tation for virtually all applications.In comparisonto conventionaldispersiveIR
spectrometers, FfIR spectrometers are characterizedby intrinsically greatersen-
sitivity, increasedspectral precision and reproducibility, and faster spectral
acquisitiontimes. Consequently,a much broaderrangeof samplingaccessories
can be usedfor the analysisof solids,liquids, slurries,andgaseswith FfIR spec-
troscopy.Samplingaccessoriescurrently availablefor use with FfIR spectrom-
etersinclude diffuse reflectance(DR), cylindrical internal reflectance(CIR) and
related attenuatedtotal reflectance(ATR) methods,FfIR microscopy,photoa-
coustic spectroscopy(PAS), and hyphenatedFfIR-chromatographicmethods.
One areaof particularinterestis the use of in-situ samplepresentationmethods
that can provide nondestructiveinformation about the samplein aqueoussus-
pensionor in the presenceof water vapor.
The higher sensitivity of modem vibrational spectrometersand the
increasedversatility of their sampling capabilitieshave reducedor eliminated
many of the limitations encounteredpreviously in studiesof soil constituents.
Along with this enhancedpotential,however,hascomean increasedcomplexity
of both the instrumentationand the datait provides.The primary purposeof this
chapteris to provide a basicunderstandingof FfIR instrumentationand the fac-
tors that needto be consideredin its use to characterizethe structureand reac-
tivity of soil colloids. Since Ramanmethodsare highly complementaryto FfIR
spectroscopy,a discussionof Ramanspectroscopyalso is included in this chap-
ter. The emphasisfor both types of instrumentationis placed on those sample
presentationmethodsthat can be profitably appliedto the characterizationof soil
materialsand the chemistryoccurringat their surfaces.
THEORY
MolecularEnergy
The energy of a moleculeor crystal can be divided into four parts: elec-
tronic, vibrational, rotational,and translational.Vibrational transitions,the focus
FfIR & RAMAN SPECTROSCOPY 271
! Wd-mftared! Ne~-IR !
Wavenumber(cm-!) I I I I
10 100 1000 I()()()()
Wavelength
Wcronsijun) 1000 100 10
Frequency(Hz) I I
1O+!2 10+13 10-tl4
Fig. 10-1. Far-IR, mid-IR, and near-IR regionsof the electromagneticspectrum.
of this chapter, are defined as the periodic atomic displacementsof the con-
stituent atoms of a molecule or crystal about their equilibrium positions. The
complex, heterogeneousinorganic and organic constituentsfound in soil and
subsurfaceenvironmentsare characterizedby a large number of vibrational
bandsthat occur throughoutthe IR region. They occur as a result of the interac-
tion of the molecule or crystal with infrared (IR absorption)or visible (Raman
scattering)radiation.The IR spectralrangecovers10 to 10 000 cm-I and is sub-
divided into three subregionsshown in Fig. 10-1. Most applicationsof vibra-
tional spectroscopyare in the mid-IR spectralregion (400-4000cm-I ), howev-
er, soil-related applicationsin the far-IR (10-400 cm-I ) and near-IR (4000 to
10 000 cm-I ) regionsare being reportedmore frequently.
In the IR region, the wavenumber(V) is the practical unit of choice for
describing the electromagneticspectrum.The relationship of this unit to fre-
quencyand wavelengthis shown by
v I
'1- - - - - [1]
- (c/n) - A'
Wavenumber(v), expressedper centimeters,correspondsto the number of

waves in a l-cm section; frequency(v), expressedas cycles per second(S-I) or
Hertz (Hz), is the numberof vibrationsper unit time, wavelength(A) is the length
of one wave (units are usually ,urn); c is the velocity of light in a vacuum;and n
is the index of refraction.
Molecular Vibrations
Molecularvibrationsoccur at discreteenergiesin the IR region of the elec-

tromagneticspectrum and are referred to as vibrational modes or as normal
modesof vibration. The amount of energy requiredto excite a particular vibra-
tional mode dependson the type of periodic motion involved. Thesemodescan
be divided into two categories,internal vibrational modesand phonon modes.
Internal vibrational modescorrespondto the periodic motion of atomswithin a
crystal or molecule. They are described by three types of motion: (i) bond
stretching,(ii) bending, and (iii) torsional motions. Internal vibrational modes
are usually found in the 400 to 4000 cm-I range.Crystalline materialsalso have
272 JOHNSTON& AOCHI
A A A
,1\/ A tA
v v v
2 3 1
Fig. 10-2. The stretching(VI and V3) and bending(V2) vibrational modesof the water molecu!e.
vibrational transitions in the 10 to 400 cm-I region, termed phonon modes,

resultingfrom the movementof one unit-cell or functional unit relative to anoth-
er. In the past,this spectralregion «400cm-I ) wasnot highly accessibleand rel-
atively little is known about the low frequencyvibrational modesof most soil
constituents(Ishii et aI., 1967).Recently,however,FfIR spectrometers equipped
for far-IR measurements and low-frequencyRamanmeasurements havebecome
useful tools for probing this spectralregion (Michaelian, 1986; Herman et aI.,
1985; Fripiat, 1982; Prost& Laperche,1990; Laperche,1991).
The vibrational energies associatedwith the four types of molecular
motion (stretches,bends,torsions,and phononmodes) arevery different. A sim-
ple exampleof this fact is providedby the stretchingand bendingmodesof the
water molecule(Fig. 10-2).
The H-O-H stretchingvibrations (VI and V3) occur in the 3200 to 3700
cm-1 rangedependingon the physicalstateof water; i.e., vapor, liquid, or solid.
Less energy is requiredfor the H-O-H bendingmode (V2) which occurs in the
1600 to 1650 cm-1 region. A detailed description of molecular vibrationsis
beyondthe scopeof this chapter.The example(Fig. 10-2), however,servesto
illustrate the sensitivity to characteristicmolecular motions that makesvibra-
tional spectroscopysuch a powerful tool. The positionsand intensitiesof bands
observedin the infrared region of the spectrumare indicativeof the structureand
compositionof soil constituentsas well as the chemistryoccurringat interfaces.
Infrared Absorption
Vibrational transitionscan occuras a result of IR absorption,Ramanscat-

tering, thermalexcitations,and a numberof otherlesscommonprocesses.Direct
absorptionof IR radiationis the most commonmethodusedto study vibrational
transitions.In order for infrared absorptionto occur, two criteria must be met.
First, the frequencyof the IR radiation must matchthat of the vibrational mode.
Infrared radiationconsistsof an electricfield oscillatingat frequenciesof 1012 to
1014 Hz. If the frequencyof the incident IR radiationmatchesthe frequencyof
the internal movementsof the atoms correspondingto a particular vibrational
mode, then absorptionmay occur. The secondcriterion is that the vibrational
mode must producea changein the dipole moment.The interactionof an oscil-
FI'IR & RAMAN SPECTROSCOPY 273
Virtual State
hvo _
--r j- V=l - -+-'-- V= 1

---,-,--- v=Q _--l-_ v=Q
IR absorption Stokesscattering
Incident IR Beam TransmittedIR Beam IncidentVisible (laser)Source
rr1~5>
10
0
I
----~C> Sample
V
Sample
Detector
Inelastically
Scattered
Light
Detector
hvi hvo
Laser
Excitation
Line
Fig. 10-3. Schematicdiagramof IR absorptionand Ramanscatteringprocesses

.
lating electromagneticfield with a moleculewill push the positive chargesone

way and the negativechargesin the oppositedirection. If theseatomic displace-
mentscreateor induce a changein dipole momentin the samplethen the mole-
cule will absorb infrared energy and the correspondingvibrational mode is
termedIR-active. A schematicillustration of the IR absorptionprocessis shown
in Fig. 10--3.
Raman Scattering
Vibrational modes also can be studied using a fundamentally different

processknown as Ramanscattering.When photonsfrom a visible, monochro-
matic light source(e.g., an argon-ionlaser) interact with a sample,most of the
scatteredphotonsare not perturbed;i.e., they have the sameenergyas the inci-
dent photons.However, a small portion of the incident photons,roughly on the
274 JOHNSTON& AOCHI
order of 1 out of every 108, are inelastically scattered by the sample.

ConventionalRamanspectroscopyis basedon the processshown on the right
side of Fig. 10--3 which is termedStokesscattering.A small amountof energyis
transferredfrom the incident photon to the sampleresulting from a vibrational
transition from the ground stateto an excited level. The energyof this inelasti-
cally scatteredphotonwill be lessthan that of the incident photons,with the dif-
ferencein energiescorrespondingto the energyof the vibrational transition. A
small portion of the light scatteredby the samplewill be shifted to lower fre-
quenciesby discrete amounts correspondingto the vibrational energy levels
encountered.The position of a band in a Ramanspectrumis always givenin
wavenumbersand is measuredas the differencein energybetweenthe frequen-
cy of the laser (in cm-I ) and that of the inelastically scatteredband. Several
excellenttreatisescover both IR absorptionand Ramanspectroscopy,including
Wilson et aI., 1955; Long, 1977; Turrell, 1972; and Heffelfinger, 1971.
In order for a vibrational mode to be Raman-active(Le., observedin the
Ramanspectrum),the vibrational mode must producea changein the induced
molecular polarizability of the molecule.The conceptof molecularpolarizabili-
ty can be visualizedby picturing an electroncloud aroundthe vibrating atomsof
interest;changesin polarizability relate to how easily this electroncloud can be
deformedby the pulsating electromagneticradiation. Recall that the selection
rules for IR spectroscopyrequire a changein the induceddipole moment. One
practicalaspectof this differencein selectionrules betweenRamanand IR spec-
troscopy involves applicationsin aqueousmedia. Raman-activebandsof water
are comparativelyweakerthan their IR counterparts.Thus, the presenceof water
can be toleratedwith fewer problemsin Ramanspectroscopythan in IR spec-
troscopy.Unfortunately,the sensitivity of Ramanmethodsis generallylessthan
that of FIlR spectroscopy,so this advantagemay be of little benefit.
Vibrational Analysis
The typeof information that canbe obtainedfrom vibrationalspectroscopy

can be subdividedinto two broad categories.The first is directed towards the
identification of soil constituentsand the secondat characterizingthe chemical
and physicalpropertiesof soil materialsand their respectiveinterfacial process-
es. The simplestapproachis to examinethe vibrational spectrumof a soil mate-
rial for the presenceof diagnosticbandsthat indicatethe occurrenceof a partic-
ular component.This applicationis analogousto the use of powderx-ray pow-
der diffraction methodsto identify the presenceor absenceof a particularcrys-
talline solid phase.The observedband positionsfor a soil material can then be
comparedto the spectraof known referencematerialsor to tabulatedfrequencies
from literature references.Advantagesof this approachare that it does not
requireany detailedunderstandingof spectroscopy,is amenableto routine analy-
sis by a nonspectroscopist, and can be donequickly.
This modeof spectralinterpretationof vibrational spectracan be aided by
the useof groupfrequencies.The characteristicvibrational frequenciesassociat-
ed with certain submoleculargroups of atoms are termed group frequencies.
Thesediagnosticbandscan be usedto suggestthe presenceof a particularcom-
FI1R & RAMAN SPECTROSCOPY 275
ponent or functional group. Examplesof the submoleculargroups commonly

identified in soil-relatedapplicationsare listed in Table 10-1. Infrared analysis
of humic substances, for example,relies heavily on group frequenciesto indicate
the type and distribution of functional groups present.Invariably, a vibrational
band can be assignedto several possible functional groups. Thus, additional
experimentalcriteria must be used to confirm the assignment.In the case of
humic substances,correlationof the vibrational data with 13e nuclearmagnetic
resonance(NMR) data and elemental analysis can provide more definitive
assignmentsfor the vibrational bands(Johnstonet aI., 1994). One further diffi-
culty with relying upon group frequenciesaloneis that the bandsassociated with
a particularfunctional group can shiftas a function of pH, metal binding, or ionic
strength.
Vibrational spectraalso can be used to provide insight about molecular
structure or interfacial processes.This application requires a more detailed
understandingof vibrational spectroscopy.The most commontypesof diagnos-
tic propertiesused in these studies include shifts in position of a vibrational
mode,changesin the bandwidth or line shape,or the appearanceof new bands.
Sincethe spectralinformation of interestfor theseapplicationsis typically man-
ifested by a change in the vibrational spectrum,spectralanalysistools are fre-
quently required for theseapplications.Thesetechniquesinclude spectralsub-
traction, bas~line correction, band fitting, and deconvolutionmethods,and are
discussedin the section"SpectralAnalysis."
Table 10--1. Group frequenciesof soil constituents.

Components
Organic
O-H stretchingof carboxylic acids, phenols,alcohols 3500-3200
N-H stretchingof amines,amides 3400-3200
Aromatic C-H stretching 3150--3000
Aliphatic C-H stretching 2970--2820
C=O stretchingof carboxylic acids,amides,ketones 1750--1630
salts of carboxylic acids
asymmetricCOO- stretching 1650--1540
symmetricCOO- stretching 1450--1360
C-H bendingof -CHz- and -CH3 1465-1440
C-O stretching,O-H bendingof -COOH 1250--1200
C-O stretchingof polysaccharides 1170--950
Inorganic
Clay mineralsand oxides
O-H stretchingof structuralOH 3750--3300
O-H bendingof structuralOH 950---ll20
Si-O-Si stretching 1200-970
Sorbedwater
O-H stretching 3600-3300
O-H bending 1650--1620
Carbonates 1600-1300
900--670
1300-850
Phosphates 600-550
1200-1100
Sulfates 680-600
276 JOHNSTON& AOCHI
Fixed M irror
Fig. 10-4. Schematicdiagramof a FTIR showingthe source,beamsplitter,moving mirror, fixed mir-

ror, samplecompartment,and detector.
FOURIERTRANSFORM INFRARED SPECTROMETERS
Principlesof Operation
Fourier transform infrared spectrometersare superior to dispersive IR

spectrometersfor virtually all applications(Griffiths & de Haseth,1986). This
advantagecannotbe realized,however,if the key componentsare not appropri-
ately matched.The most importantcomponentsare briefly discussedbelow.
Interferometer
The interferometeris the central componentof a FTIR spectrometer.The
schematicof a Michelson interferometer-based FfIR spectrometeris shown in
Fig. 10-4. Light from the IR sourceis passedonto a beamsplitter,whereapprox-
imately half of the incident light is reflected onto a moving mirror, and the
remainderis transmittedthrough the beamsplitteronto a fixed mirror. The mov-
ing mirror introducesa phasedifference betweenthe reflected and transmitted
beamsby varying the pathlengthof one beamrelative to the other. Optical inter-
ferenceoccurswhen the two beamsrecombineat the beamsplitter.The resulting
modulatedIR beam is then passedthrough the samplecompartmentonto the
detector.Interferometersare designedso that the "moving mirror" travelswith as
little friction and tilt as possibleand at a constantvelocity. Two additional opti-
cal components are addedinside the interferometer.A low-power He-Ne laseris
usedfor frequencycalibration, and in somecasesdynamic alignment. In addi-
tion, a white-light sourceor laserquadrutureis requiredto determinethe point at
which the moving mirror and the fixed mirror are equidistant(called the zero
point of deflectionor ZPD). A numberof other interferometerdesignsare being
usedin addition to the Michelson interferometer.
Optical recombinationof the two beamsresultsin a complexoptical signal

that is calledan interferogram.The interferogramis the detectorresponsein volts
as a function of mirror displacementfrom the ZPD position. Representative
interferogramsare shownon left side of Fig. 10-5. When a Fourier transformis
applied to the interferogram,the data are convertedfrom the time domain into
the frequencydomain. Conversionof the interferograminto a single beam(SB)
spectrumproducesa spectrumin which the x-axis is in units of wavenumbers
(middle portion of Fig. 10-5; spectraC and D). Intensitiesin the SB spectracor-
respondto the quantitiesof IR energyreachingthe detector.
Detectors
Two typesof detectorsare commonlyusedin FfIR spectrometers for mid-
IR applications,quantumdetectorsand thermal detectors.The developmentof
quantum mid-IRdetectors,suchas the mercurycadmiumtelluride (MCT) detec-
tor, hassignificantly improvedthe sensitivity of FfIR spectrometers.The MCT
detector is a photoconductivedetectorthat measuresan increasein electrical
conductivity when illuminated. Detectorsensitivitiesare commonly represented
in termsof D* valuesthat expressthe level of responserelative to the noisepro-
duced in units of centimetersand Hertz squareroot per watt. Wide-bandMCT
detectorscover a wavenumberrangeextendingfrom 450 to 5000 cm-I , with an
optimum frequencyresponsein the 900 to 1200 cm-1 region, and have charac-
teristic D* valuesof 109 to 1011. Narrow-bandMCT detectors,on the otherhand,
cover a more limited range,about750 cm-1 to 4000 em-I, but havegreatersen-
sitivities (D* values of 1012_1013). In general,the higher the sensitivity of an
MCT detector,the more the spectralrangewill be restricted.In addition to high
sensitivity, MCT detectorsare characterizedby much fasterresponsetimes than
are thermal detectors.Thus, faster scanningmirror velocities can be usedwith
quantum detectors,resulting in faster scan rates and shorter data acquisition
times than can be obtainedwith thermaldetectors.
Pyrolectric devices, such as the deuteratedtriglycine sulfate (DTGS)
detector,measurechangesin temperature.They do not require cooling and pro-
vide a fairly uniform frequency responsefrom the far-IR through the mid-IR
region. In comparisonto MCf detectors,they are characterizedby lower overall
sensitivities(D* values of 106-1O~ and slower responsetimes. Despite these
limitations, they are usedfor many applications.They havethe addedbenefitthat
they do not requirecooling with liquid N2• Deuteratedtriglycine sulfatedetectors
fitted with polyethylenewindows also can be usedin the far-IR region from 80
to 500 cm-1• Bolometersare anothertype of IR detectorthat are more sensitive
than DTGS detectorsand also allow accessto a broaderrange of frequencies;
however,thesedetectorsare considerablymore expensivethan DTGS or MCT
detectors.The spectralrangesfor severaltypesof IR detectorsare listed in Table
10-2.
Beamsplitters
Modem FTIR spectrometersare often designedto cover more than one
spectral region. Emissivity of the source, optical transmissionand reflection
~
Interferograms Single Beam Spectra Absorbance Spectrum
0.7500
n--
LO
0).
Reference S8 0
.,....
<0
Spectrum C')
~
0.5000 LI 0)
C')
to
Reference IFGM II I 0
v
C')
C')
Sample IFGM
0.2500 LI r I 0
C\I
<0
.,....
Spectrum
I
I I I
o 2000 4000 6000 8000 500 1000 1500 2000 2500 3000 3500 4000 0.0000~o .~-- .!--- .!--- -'--- ,'. ....
# of Data Points Wavenumber Wavenumber
(Retardation)
Fig. 10-5. Illustration of stepsrequiredto producea ratioedabsorbancespectrum.A referenceinterferogramof the empty samplecompartmentis shownon the top left Ro
side; the interferogramof a montmorilloniteclay film is shownon bottomleft side; correspondingsingle beamspectraof referenceand samplein the middle. Ratioed
absorbancespectrumobtainedfrom files and.
Ig
e
Table 10-2. Spectralrangeof Ff sources,beamsplitters,and detectors(em-I).
10 100 500 1000 2000 5000 10000 15000 20000 25000 ~
1(0
I I I I I I I I I I
I
Sources
Globar - far-to mid-infrared
Tungsten-Halogen
i
~
Solid substrate
Beamsplitters
Mylar (3-25 11m thickness)
Ge coatedCsI (250-5000em-I)
Ge coatedKEr (400--6 000 em-I)
i
~
CaF2(1200-10000 em-I)
Quartz(3 800-40000em-I)
DTGS (75-600em-I) I
l Detectors
DTGS CsI (250-5 000 em-I) I
l
DTGS KEr (400-5 000 em-I) I
MCf-B (450--6 000 em-I)
I
MCf-A (700--6000 em-I) I
I InSb (2 000-10000 em-I) ]
PbSe(2000-11000 em-I)
I Silicon (8 600-25000 em-I) I

N
...;J
I,C
280 JOHNSTON& AOCHI
propertiesof the beamsplitter,and the spectralresponseof the detectormust all

be matchedfor the spectralregion of interest.Table 1(}"'2lists the spectralranges
for a suite of sources,beamsplitters,and detectors,extendingfrom the visible to
the far-IR region. The most commonly usedbeamsplitterfor the mid-IR spectral
region is a KBr beamsplitterwith a thin, uniform coatingof Ge or Si. Ideally, the
beamsplittershould reflect half of the incident light and transmit the remainder.
Coated KBr beamsplittersfail to transmit light above 8000 cm-I ; therefore,
applicationsextendinginto the near-IRand visible require the use of alternative
beamsplitter composition materials such as CaF2 and quartz (Table 1(}"'2).
Similarly, different sourcesmustbe usedfor different spectralregions.The point
to be made here is that careful attention should be paid to matching the appro-
priate source, beamsplitter, and detector combinations for different spectral
regions.
Data Collection
Unlike dispersive IR monochromatorsthat are designedto analyze the

sample and reference simultaneously, FTIR spectrometersare single-beam
instrumentsthat collect, process,and storesingle-beamspectra.The single-beam
spectrumrepresentsnot only the molecularpropertiesof the samplebut also the
optical efficienciesof all the componentsof the instrument.Consequently,bef9re
an absorbanceor transmittancespectrumcan be obtained,the single-beamspec-
trum of the samplemust be ratioed againstthat of a referencebackgroundspec-
trum. The referencespectrumis derived from the interferogramof the sample
compartmentor sampling accessorywithout the sample present.Typically, a
number between1 and 1000 scansare co-addedand stored as an accumulated
file. The numberof scansneededfor a particular application is dictatedby the
desiredsignal to noise ratio (SNR).
To illustrate this process,the stepsrequiredto obtain an absorbancespec-
trum are shown pictorially in Fig. 1(}"'5. Absorbancespectra [i.e., A(v)] are
obtainedby ratioing the intensity of the incident beam [Io(v)] againstthat of the
beamtramsittedthrough the sample [I(v)] accordingto Beer'slaw (Eq. [2]) as
shown below.
Sample
Io(v)
~D
I(v)
..
A(v) = 10glO ( -
Io(v)
-) [2]
I (v)
The referenceinterferogramof the empty samplecompartmentand the sample

interferogramof a montmorilloniteclay film are shownon the top and bottom of
the left side of Fig. 1(}"'5, respectively.All of the spectralinformation is encod-
ed within the interferogram, however, very little useful information can be

obtainedby visual inspectionof the data. Fourier transformof the interferogram
is requiredto convertthe data into a SB spectrum.The resultingSB spectrumof
the empty sample compartmentis shown in the upper spectrumof Fig. 10-5
(middle) and is referredto as the referenceSB spectrum.Spectraldata process-
ing routines of modern FTIR spectrometerscan automatically apply the fast
Fourier transform(FFI) transformto the interferogram,store the convertedSB
spectrumon the computer,and discardthe original interferogramwithout input
requiredby the user. In normal use,the referenceand sampleSB files are stored
as individual datafiles on the computer,however,the interferogramsare usually
not saved.In someapplications,such as kinetics, there is not sufficient time to
collect and processthe interferogramsduring an experiment.In this case, the
interferogramsare storedby the computerand processedat a later time.
After the referenceSB spectrumhasbeenobtained,the sampleis placedin
the samplecompartment,and a similar procedureis usedto collect the interfer-
ogram and correspondingsampleSB spectrum(in Fig. 10-5). The absorbance
spectrumshown in Fig. 10-5 (right) was obtainedby ratioing the sampleSB
spectrumagainstthe referenceSB spectrum according to Eq. [2]. The units along
the x-axis are in wavenumbersand are the sameas for the SB spectra.The units
along they-axis are in absorbance(or optical density)and indicatethe amountof
energy that is being absorbedby the sample. In this case, the structural-OH
stretchingband is observedat 3610 cm-1, the O-H stretchingband of sorbed
water occurs between2800 and 3600 cm -1, the O-H bending mode of sorbed
water occursat 1630 cm-I, the structuralSi-O and AI-O stretchingbands occur
in the 950- to 12oo-cm-t region, and the O-H bendingbandsof structural OH
groupsappearat 839 and 915 cm-t .
SpectralProcessingConsiderations
Choiceof SuitableReferenceSpectrum
The conditionsusedto obtain a reference-SBspectrumshouldmatchthose
usedto collect the sample-SBspectrumasclosely aspossible.Experimental con-
ditions that must not changebetweencollecting the sample-and reference-SB
spectrainclude the spectralregion being analyzed,optical resolution, apodiza-
tion function, electronicgain settings,size of the aperture,source,beamsplitter,
and purge status. In brief, all experimentalparametersthat would result in a
changein the SB spectrum,other than the changeresulting from absorptionor
reflection by the sampleitself, should remainfixed. A significant changein any
of theseparameterswill make the sample-SBspectrumincompatiblewith the
reference-SBspectrum.While the function of most of theseparametersis appar-
ent from either Fig. 10-4 or analogousparametersin other analytical instrumen-
tation, apodizationfunctions are uniqueto instrumentsthat require mathematical
transformationof the collecteddata. In the caseof FTIR, data for the interfero-
gram is acquiredas a function of mirror displacement.Since the moving mirror
travelsonly a finite distanceand then reversesits direction, the interferogramis
truncatedat the limits of travel. Bandsresultingfrom the Fourier transformation
282 JOHNSTON& AOCHI
of the datasetexhibit small side lobescalled"feet" or "pods."A wide variety of

apodizationfunctions can be applied to the interferogramdata set to minimize
theseundesirablespectralfeatures.In doing so, they also affect the shapeand
height Of the band itself.
In addition, many samplingaccessoriessuch as IR gas/vacuumcells, and
liquid cells absorb a considerableamount of IR radiation. To compensatefor
their IR absorptioncharacteristics,it is often desirable toinclude the empty sam-
ple accessoryin the samplecompartmentwhen the referenceSB spectrumis
obtained.In practice,SB spectracollectedunder identical conditionswill show
somevariation on aday-to-daybasisdue to changesin the exacttemperatureof
the IR source,purge status,etc. Thus, it is good practiceto collect a new refer-
ence spectrumduring each day of spectraacquisition, to compensatefor any
instrumentalchanges.
Trading Rules
The practiceof FI1R spectroscopyinvolves a trade-offbetweenresolution
and throughput(Griffiths & de Haseth,1986). Resolutionis defined in termsof
the minimum separationrequired between absorption bands that allows the
bandsto be distinguishedfrom one another.Optical throughputis a function of
the sizeanddegreeof divergenceof the beamat pointsof focus in the instrument.
Signal-to-noiseratio is a measureof the amountof signal producedat the detec-
tor relative to the noise level of the detector.Sufficient resolutionis neededto
observeall of the vibrational featurespresentin a particularsample.The use of
excessivelyhigh resolution,however,resultsin a reducedSNR and longer scan
times, without providing any additional information about the vibrational spec-
trum of the sample.Increasingthe optical resolutionby a factor of two resultsin
a 4- to 16-fold decreasein the SNR; thus, working at the lowest reasonableres-
olution value is preferable.The optical resolutionneededto representthe vibra-
tional spectrumof a sample shouldbe no greaterthan the width of the narrowest
band of interestin the IR spectrum.A practicalway to determinethe optimum
resolutionis to obtain severalsurveyFI1R spectraat different resolutionvalues
(e.g., at 1.0,2.0,and 4.0 em-t). The optimum spectrumcorrespondsto the spec-
trum obtainedat the lowest resolutionwhich doesnot degradethe quality of the
spectrum.The highestresolutionneededto characterizemostsoil constituentsis
2.0 cm-t ; however,a resolutionvalue of 4 or even8 cm-t is often sufficient.
Optical Saturationand ElectronicGain

The amountof energyreachingthe detectorin a FI1R spectrometershould
be matchedto the amountof electronicamplification appliedto the detectorsig-
nal. If too much light reachesthe detectorthen optical saturationoccursand the
analog-to-digitalconverteris overfilled. This problem is encounteredmost fre-
quently when nothing is presentin the samplecompartmentto attenuatethe IR
signal while referenceor backgroundspectraare being collectedor when ana-
lyzing sampleswhich absorblittle energy. When optical saturation occurs,the
detectorcannotrespondto a changein energyand the optical throughputmustbe
reduced.A reductionin optical throughputcanbe obtainedby decreasingthe size
of an aperturemountednear the sourceor by using optical filters or attenuators

(Fig. 10-4). Becausethe electronicgain and aperturesettingswill significantly
influencethe SB spectra,it is importantto keepthesesettingsthe samewhencol-
lecting referenceand sampleSB spectra.In obtaining FTIR spectraof soil and
colloidal materials,the oppositeproblemof too little light intensity to the detec-
tor is generallymore critical. In this case,electronicamplification (i.e., gain) of
the signal is requiredto fill the analog-to-digitalconverter(ADq. Enoughgain
shouldbe applied to fill the ADC but not so much as to result in saturation.The
objective is to add as much electronicgain as possiblewithout exceedingthe
working thresholdintensity of the detector.Although many of the gain settings
can be adjustedautomaticallyby the FTIR spectrometer,it is importantfor the
userto know what gain valueshavebeenusedduring spectralacquisitionso that
experimentscan be run undersimilar conditionsat a later time.
Water and CO2 in the vapor phaseabsorbstrongly in the IR region and
interferewith IR bandsof the sample.Watervapor is evidencedby hundredsof
rotational-vibrationalbandsin the OH stretching(3400-3900cm-I ) and H-O-H
bendingregion (1300-1900cm-I ). Similarly, CO2(g) has numerousabsorption
bandsin the c=o stretchregion (2300-2500em-I) region. For work in the mid-
IR region the instrumentand samplechambermust be purgedwith dry air or
evacuatedto avoid these interferences(Griffiths & de Haseth,1986). Typical
sourcesof dry air for purging FTIR spectrometersare air dryerswith or without
a CO2trap, liquid N2 boil-off, or compresseddry air or N2. The latter two sources
of dry air or N2 canbe costly for routine operation.Watervaporalsowill degrade
hydroscopicoptical components,such as KBr beamsplitters,so it is critical to
protect thesecomponentsfrom exposureto water vapor. Vapor phaseCO2 will
not damage optical componentsand can frequently be tolerated becauseit
absorbsin a regionwherefew otherIR bandsare present.Somewatervapor and
CO2 will typically remainafter purging,especiallyif samplesarebeingplacedin
and removedfrom the instrumentfrequently. One way to compensatefor these
interferencesis to store a representativevapor phasespectrumcontainingwater
and CO2• This spectracan then be subtractedfrom absorbanceor transmittance
files to "null out" the interfering bands.
SpectralAnalysis
Application of Beer'sLaw
For applicationsrequiring a quantitativeestimateof a particular compo-
nent, the familiar Bouguer-Beer-Lambertlaw (Beer's law) can be applied as
expressedbelow (Eq. [3])
(/Oe\;»)
A(v) = E(v)cd = 10g1O -_-
I(v)
[3]
whereA(v) is the absorbance valueof a bandassignedto the componentof inter-

v
est,E(V) is its molar absorptivityat wavenumber (em2/mmol), c is the concen-
284 JOHNSTON & AoeHI
tration of the component(mmoVem3), d is the thicknessof the sample(em),Io(v)

is the intensity of the incident beam,andI(v) is the intensity of the transmitted
beam.As definedin Eq. [3], A(v) correspondsto the peakheight of a particular
band.For complexmaterials,the heightof a particularbandfrequentlycannotbe
determineddue to spectraloverlap with other bands. In addition, peak height
measurements do not take into accountany changein lineshape(e.g., line broad-
ening) that may occur. This latter problem canbe avoided,in part, by measuring
the integratedintensity of a particular band or spectral region. The integrated
intensityA of a bandis definedby Eq. [4]
[4]
wherethe integrationlimits VI and V2 are chosento encompassthe bandof inter-

est. Beer'slaw can thenbe expressedas Eq. [5]
A =£cd [5]
where"£ is the integratedabsorptivity in units of centimetersper mole, c is the

concentrationof units of mole per cubic centimeter,d is the pathlengthin units
per centimeters.CommercialFfIR spectralprocessingsoftware generally pro-
vides a meansof integratingthe intensity of a particularband.
BaselineCorrection
Baselinecorrection techniquesare frequently applied to FfIR spectrain
order to determinepeak heights,compareone spectrato another,or simply for
plotting the spectrumwith a flat baseline.The simplestmethodusesa linear base-
line correctionacrossthe spectralregion of interest.For a single, well-resolved
band the baselineis drawn tangentto the spectrumso as to include the wings of
the selectedband.Oncethe baselinehasbeendrawn, the correctedabsorbanceof
the bandof interestcan be determinedby measuringthe height of a vertical line
from the maximumabsorbanceof the bandto the baseline.This methoddoesnot
work, however,when more than one bandcontributesto the areain the spectral
region of interest (Painteret aI., 1980). More sophisticatedbaselinecorrection
options include using a cubic spline or an exponentialfitting function. Accurate
determinationof peakheight is importantfor applicationsinvolving quantitative
analysis.A completediscussionof computerizedquantitativeinfrared analysisis
given by McClure (1987).
SpectralSubtractionand DifferenceMethods
The advent of computerizedIR spectrometers(FfIR and computer-con-
resultedin the birth of the digital subtraction
trolled dispersiveIR spectrometers)
techniquesapplied to IR spectroscopy.Thesespectralstripping techniquesare
FTIR & RAMAN SPECTROSCOPY 285
usedto remove the spectralcontribution of a particularcomponent,solvent, or

substratefrom a spectrumcontaining contributionsfrom severalcomponents.
Spectral subtraction methods also can be useful in the characterizationof
changesthat take placeas a result of a sorptionreaction,changein temperature,
pH, or someother variable. An illustration of this techniqueis provided by the
work of Jentyset al. (1989) who utilized spectralsubtractionmethodsto exam-
ine the behaviorof wateradsorbedon a silica-rich zeolite. In this case,the initial
spectrumof the dry zeolite was subtractedfrom spectrathat containedcontribu-
tions from sorbedwater. The differencespectraof the (zeolite + water) - (zeo-
lite) allowed the changesthat resultedfrom the sorptionof water to be examined
with minimal interferencefrom the spectrumofthe sorbent.Unfortunately,spec-
tral subtractionmethodsare prone to introducing spectral artifacts that result
from the subtraction process itself and have nothing to do with sample
(Hirschfeld, 1987). In practice, the criteria used to determinehow much one
spectrum can be removed from another are sUbjective. These criteria might.
include obtaining a "flat" baselineor "zeroing" out the absorbanceof a bandor
bandsof a particularcomponent.
Band Fitting
Whenseveralbandsoverlap,or when quantitativeinformation abouta par-
ticular lineshapeis desired,the spectracan be fitted to a synthetic mixture "Of
bandsusing a Gaussianor Lorentzianlineshape.A numberof softwarepackages
are availablecommerciallythat provide a wide variety of curve-fitting options.
The stepsinvolved in fitting a spectralregion to a syntheticspectrumare as fol-
lows:
1. Baselinecorrection(see"BaselineCorrection").
2. An initial guessof the number,position, width, and intensity for each
bandis typically required.A numberof softwarepackagesare available
that permit input of this information into the computerin an interactive
mode. The syntheticspectrumcan be comparedto the real spectrum,
and the goodness-of-fitcan be examinedby subtractingthe synthetic
spectrumfrom the "real" spectrum.
3. Optimization. Different computational algorithms can be used to
improve the fit of the syntheticspectrumto the "real" spectrum.These
algorithms include simplex optimization and nonlinear least squares
analysis(Hirschfeld, 1987).
The information generatedfrom band fitting generally includes the peak
height, bandwidth, integratedabsorbance,bandposition, and lineshapefor each
fitted bandin the spectrum.Recently,Bish andJohnston(1993) useda nonlinear
least-squaresanalysisband-fitting program to determinethe positions, intensi-
ties, bandwidths,and lineshapesof the OH-stretchingbandsin the FfIR spectra
of dickite obtainedas a function of temperature.Similarly to the spectralsub-
traction methods,caution must be used when fitting complex spectral regions
with syntheticbands.
286 JOHNSTON& AOCHI
Oi
~ ~ k
n
a P"B'[2Ju0-+"-!"
r fj m
p
bed e ~
1 g
q
~
Fig. 10--6. Schematicdiagramof Ramanspectrometershowing the laser(a), attenuator(b), calcite
wedge(c), polarizer(d), laser-focusingobjective (e), folding prism (f), sample(g), x-y transla-
tion stage(h), vertical adjustment(I), collection optics (j, m), mirror (k), television cameraand
imagingsystem( I), polarizationanalyzer(n), spectrometerentranceslit (0), spectrometer(p) and
detector(q).
Comparisonto RamanSpectroscopy
The Ramaneffect (Fig. 10-3) is basedupon light scatteringand is, thus,

fundamentallydifferent from IR absorption.The three principle componentsof
a Ramanspectrometerare the source(i.e., laser),collection optics, and detection
systemas shownin Fig. 10-6. Recentadvancesin lasertechnologyhaveconsid-
erably expandedthe choice of sources.In addition to the standardAr+ and Kr+
lasers,tunablediode lasers,Nd-YAG lasers,inexpensiveHeNelasers,solid-state
devices, Ti/sapphire laser, and dye-lasersare commercially available. Similar
improvementsin micropositioning devices have resulted in integratedsample
positioning devices that are coupled with optical microscopesand television
cameraimaging systems.One of the attractivefeaturesof Ramanspectrometers
is that the laserspot sizecan be focuseddown to about1.0-,umdiam. When cou-
pled with modem imaging systems,Raman spectrometerscan provide excep-
tional spatialdiscrimination.Recently,Ramanmicroprobe methods beenapplied
to clay and soil constituents.Thesespectrometersallow the sampleto be exam-
ined under a high-powermicroscopeshowing the actual position and focus on
the laseron the sample(similar to the operationof a single lens reflex camera).
Numeroustypes of detectionsystemsare available including scanningdouble
monochromatorswith photomultiplier tubes or polychromatorsystemsusing
diode-arraydetectors.For a more completegeneraldiscussionof Ramanspec-
troscopyseeWilson et aI., (1955), Turrell (1972), and Long (1977); for surface
chemistryapplicationsseeCampion (1987)and Johnston(1990).
SAMPLING METHODS
A wide diversity of samplepresentationmethodsare currently available

for analyzing solids, suspensions,liquids, and gases.The major accessories
availablefor FTIR spectrometerswith suggestedapplicationsare listed in Table
ITIR & RAMAN SPECTROSCOPY 287
Table 10--3. FTIR samplepresentationmethods.

Method Acronym Type of sample
Cylindrical internal reflectance CIR Aqueoussolutions
Mixed solvents
AqueousSuspensions of clay mineralsand oxides
Attenuatedtotal reflectance ATR Surfacesof strongly absorbingmaterials
Clay minerals
Polymers
Thin films
Diffuse reflectance DR Powders
Transmissionmethods TR
Self-supportingfilms Clay mineralsand hydrousmetal oxides
KBr pellets Quantitativeand routine analysisof soil constituents
Supporteddeposits Quantitativeand routine analysisof soil constituents
Liquid transmissioncells Aqueoussuspensionsof soil constituents
FTIR rnicrospectroscopy Il-FTIR Microcrystalline materials
Coatingson sandgrains
Fluid inclusions
Photoacoustic PAS Strongly absorbingsolids
Cryogenicsampling Clay minerals
Specularreflectance
Gascells and long path FfIR Analysis of vaporsfrom DTA
VOC determinationfrom landfills
10-3. Becausethe samplerequirementsof these accessories are diverse,few all

encompassingrecommendationscan be made regardingsamplepreparation.In
general,particles <2 pm in size will provide an optimum responseusing trans-
mission and DR FTIR methods.When mixturesof large (>25-.um particles)and
small «2.um) particlesare present,the spectralcontribution fromthe small par-
ticles may completely dominate the observed spectrum. Thus, particle-size
effects must be taken into consideration,especiallyfor whole soil materials.In
addition, strongly absorbingsamplesare problematicregardlessof the sample
presentationmethod. Photoacousticand attenuatedtotal reflectance methods
often provide the bestalternativeto examiningthesematerials.The main sample
presentationmethodsare describedin the following sectionwith emphasisgiven
to their applicationto studyingsoil and subsurfacematerials.
TransmissionMethods
TqlOsmissionsamplepresentationmethodsare the most commonly used

techniquesto examinethe IR spectraof soil constituents.Self-supportingfilms,
dispersionsin KBr pellets,and supporteddepositson IR windows are all suitable
for analysisby transmissiontechniques.In the discussionthat follows, each of
these methods and experimentsin which they have proved useful will be
described.Thesetechniquesare often the samplingmethodsof choicefor study-
ing soil constituentsprovided that the natureof the sampleand the type of infor-
mation desiredaboutthe sampleare compatiblewith the samplepreparationand
presentationstepsinvolved with the specific method.Extensivereviewsof trans-
mission techniquesand samplepreparationmethodologyalso can be found in
Griffiths and de Haseth(1986), White and Roth (1986) Colthup et al. (1990),
Hair (1967), Bell (1987), Farmer(1974), Van Der Maas(1972), Gadsden(1975).
288 JOHNSTON& AOCHI
Fourier transforminfrared microscopy,anothertechniquebasedon transmission

of IR radiation through the sample,is coveredseparatelyin the section"Fourier
TransformInfrared Microspectroscopy."
Self-SupportingFilms
Self-supporting films have been used extensively for sorption studies

involving soil colloids. Dependingon the shape, size,and size distribution of the
particlesit may be possibleto preparethin (5-2011m in thickness),self-support-
ing films that are ideal for FTIR analysis.Infrared spectraof representativesoil
and subsurfacematerialsthat havebeenstudiedasself-supportingfilms and their
sorption propertiesare listed in Table 10-4. Smectite-typeclays were used in
many of thesestudies,but the useof self-supportingfilms of goethite,boehmite,
lepidocrocite,palygorskite,silica, zeolites,and othersalso has beenreportedin
the literature(Table 10-4). The sheetlikeparticlesof smectitesare ideally suited
for preparingfilms becausethe lateral dimensions(i.e., along the a- and b-axes)
of the clay particlesgreatly exceedthat along the c-axis. Fibrous mineralssuch
aspalygorskite alsocanbe easily madeinto self-supportingfilms. In contrast,the
larger, blocklike particlesof kaolinite do not lend themselvesto this technique.
Self-supportingfilms provide major benefitsover other samplepresenta-
tion methodsfor studiesof sorptionmechanisms.Thesebenefitsinclude the fact
that preparationof the self-supportingfilm requireslittle or no grinding and no
compressionof the sample.The most important benefits are, however, that no
backgroundsubstrateis present,and that the active sorption sites of the sample
remain accessibleto sorbatechemicals.Therefore,in-situ sorption experiments
can be coupleddirectly with infrared analysis.This approachhas beenused to
study the vapor phasesorption of water, aromatic hydrocarbons,ammoniaand
otherorganicbases,pesticides,phenols,chlorinatedhydrocarbons,and N- andS-
containingunsaturatedcompoundson soil and clay minerals(Table 10-4).
Recently,a FTIR-gravimetriccell was usedto study the vapor phasesorp-
tion of organics and water onto self-supporting films of montmorillonite
(Johnstonet aI., 1991, 1992a,b;Tipton et aI., 1993). A schematicdiagramof the
cell is shownin Fig. 10-7. In theseinvestigations,a self-supportingfilm of mont-
morillonite was suspendedwithin the cell from a microbalance,and the atmos-
pherearoundthe samplewascontrolledby meansof a gas-vacuummanifold sys-
tem. Through the use of this approach,vapor-phaseadsorptionor desorption
isothermsand FTIR spectracould be collectedsimultaneously.The microbalance
providedgravimetricdataon the total massof the clay film and the massof the
sorbed species. Fourier transform infrared spectra taken during the sorption
process provided information aboutthe identity of the componentspresenton the
surface.Coupling the FTIR and gravimetric information thus allowed the indi-
vidual surfaceloadingsof eachcomponentto be determinedand the competitive
role of water in the sorption of organic vaporson montmorillonite to be estab-
lished (Johnstonet aI., 1991, 1992a).
The FTIR-gravimetric cell also provides a method to characterizethe
changesthat may occur in the sorbed speciesduring the adsorptionprocess.
Johnstonet ai. (1992b),for example,studiedthe sorptionof wateronto self-sup-
Table 10-4. Infrared studiesof soil and subsurfacematerialsusing self-supportingfilms.
Sorbent Solute Description Reference ~
RI>
Beidellite D 20 Selectivedeuterationand orientationof hydroxyl groups (Russellet aI., 1970)
Boehmite Catechol Sorption mechanism(s)of catecholon aluminum oxides (Fanner,1974)
Cations
Goethite Pyridine Surfaceacidity of goethite (Morterra et aI., 1984) ~
Goethite,gibbsite Chlorophenols Bonding of chlorophenolsto Fe and Al-oxides (Kung & McBride, 1991)
& boehmite
Hectorite Anisole Chemisorptionof anisoleon hectorite (Fenn et aI., 1973)
Hectorite Anisole Mechanismof chemisorptionon clays (Fenn et aI., 1973)
Hectorite Phenanthroline Orientationand interactionof adsorbedspecies (Berkheiser& Mortland, 1977) CIl
Layer silicates Orientationof structuralOH bond groups (Serratosaetaal., 1962)
~
n
o
Lepidocrocite D 20 Selectivedeuterationof surfacehydroxyl groups (Lewis & Fromer, 1986)
& Boehmite
~
Micas Orientationand absorptioncoefficient of structuralOH (MacCarthyet aI., 1987)
Montmorillonite Thiophene Interactionof unsaturatedheterocycliccompoundswith clay (Lorprayoon& Condrate,1982)
Montmorillonite Cyclic ammoniumcations Orientationof adsorbedspecies (Vansant& Uytterhoeven,1973)
Montmorillonite Alkylammonium Thermalstability of organoclays (Durand et aI., 1972)
Montmorillonite Amides Interactionof clay surfacewith cationic organicmolecules (Doner & Mortland, 1969)
Montmorillonite Amine cations Interactionof aliphatic and aromaticamineson montmorillonite (Chaussidon& Calvet, 1965)
Montmorillonite Aniline Chemisorptionof aniline on montmorillonite (Morealeet aI., 1985)
Montmorillonite Aniline Effect of acidity on clay-organicreactions (Yariv et aI., 1969)
Montmorillonite Aniline Sorptionand transformationof aniline and p-chloroaniline (Cloos et aI., 1979)
Montmorillonite Arene complexes Chemisorptionof methyl-substitutedbenzeneson montmorillonite (Pinnavaia& Mortland, 1971)
Montmorillonite Arene complexes Chemisorptionof unsaturatedorganicson clay surfaces (Mortland & Pinnavaia,1971)
Montmorillonite Benzene Polymerizationof benzeneon montmorillonite (Stoesselet al.,1977)
Montmorillonite Cu-ethylenediamine Orientationand interactionof adsorbedspecies (Burba & McAfee, 1977)
Montmorillonite Dimethoxybenzene Chemisorptionand formation of radical organiccationson clays (Johnstonet aI., 1991)
Montmorillonite Dimethylsulfoxide Orientationand interactionof adsorbedspecies (GarwoodJr. & Condrate,1978)
Montmorillonite H 20 Clay water interactions (Buswell et aI., 1973)
Montmorillonite H 20 Combinedsorptionand spectroscopicstudy of water-montmorillonite (Tipton et aI., 1993)
~
~

Sorbent Solute Description Reference
Montmorillonite Hydroxy-AI IR study of hydroxy-aluminuminterlayermaterial (Weismiller et ai., 1967)

Montmorillonite Li and Na IR study of chargereductionin montmorillonite (Spositoet ai., 1983)
Montmorillonite Li, Mg, Ca, and K IR study of chargereductionin montmorillonite (Calvet & Prost, 1971)
Montmorillonite NH3and H20 Surfaceacidity of smectitesas influenceby exchangeablecations (Mortland & Raman,1968)
Montmorillonite NH;t and crown ethers Intercalation,interaction,and surfacepropertiesof organo-clay (Casalet ai., 1984)
Montmorillonite Organiphosphorus pesticides Adsorption and interactionmechanismof pesticides (Camazano& Martin, 1983)
Montmorillonite p-xylene and H 20 Sorption of water andp-xylene by montmorillonite (Johnstonet ai., 1992a)
Montmorillonite Phenols Sorption and transformationof phenolson montmorillonite (Isaacson& Sawhney,1983
Montmorillonite s-Triazines Chemicaldegradationof s-triazinesby montmorillonite (Russellet ai., 1968)
Montmorillonite Sulfolane Influence of exchangeablecationson clay-organicreactions (Lorprayoon& Condrate,1981)
Montmorillonite Thiolane and Sulfur containing compoundinteractionswith montmorillonite (Lorprayoon& Condrate,1981)
tetramethylenesulfoxide
Palygorskite Pyridine Surfaceacidity and reversiblefolding of palygorskite (Blanco et ai., 1988)
Smectite 4-chloroanisole Abiotic transformationof 4-chloroanisoleon smectite (Govindaraj et ai.,1987)
Smectite Chloroethenes Polymerizationand dechlorinationof chloroethenes (Mortland & Boyd, 1989)
Smectite CO2 Adsorption and orientationof CO2 by smectite (McBride & Wesselink, 1988)
Smectite Dioxin Radicalformation and polymerizationof smectite (Boyd & Mortland, 1985)
Smectite H 20 Influence of exchangeablecationson hydration mechanisms (Prost, 1975)
Smectite Humin Interlayer formation of humin in smectites (Cloos et ai., 1981)
Smectite NHt Determinationof tetrahedralsubstitutionand surfaceheterogeneity (Chourabi& Fripiat, 1981) ....
Smectites Crown ethers Clay-organic andclay-waterinteractions (RuizHitzky & Casal,1986) g
Smectites Hydrazineand IR study of reductionand oxidation in smectites (Rozenson& Heller-Kallai, 1996) z
dithionite 8
z
1(0
g
==
-
Cahn Microbalilnce
Self SupportingCloy ri lm
Fig. 10-7. Schematicof the in-situ FtIRlgravimetric cell showing the electrobalanceand gas cell
coupledto the samplecompartmentof the FTIR spectrometer.In this illustration, a self-support-
ing clay film is suspendedfrom the balanceusing a hangdownwire. The pressurewithin the cell
is monitoredusing a capacitancemanometerpressuregauge.The pressurein the cell is controlled
by a vacuumpumpingstation and manifold.
porting films of montmorillonite exchangedwith different metal cations. The

vapor phasedesorptionisothermfor water from the surfaceof the clay is shown
on the left side of Fig. 10--8. Fourier transform infrared spectrain the H-O-H
bendingregion shown on the right side of the figure correspondto specific data
points along the desorptionisotherm. Johnstonet al. (1992b) showedthat the
position and molar absorptivity of this band are strongly influenced by the
amountof water sorbedon the clay film.
Fouriertransforminfrared studiesof self-supportingclay films can be cou-
pled with other spectroscopicmethodsas well. CombinedFTIR and ultraviolet
(UV) spectroscopies,for example, were used to investigate the strong color
changethat accompaniesthe adsorptionof certainorganiccompoundson miner-
ai surfaces.Sucha colorchangemay arisewhen unsaturated organic compounds
undergoclay-mediatedsingle electrontransferreactions.This reactionresultsin
the appearanceof a very strongelectronictransition in the UV -visible spectrum.
For thesecomplexes,it is desirableto observeboth the FTIR- and UV-visible
spectraof the clay-organiccomplex during the sorption process.A schematic
diagramof a modified gas cell designedfor this purposeis shown in Fig. 10--9.
The cell allows the use of both the IR or UV-visible optics without having to be
opened.This is accomplishedby mounting the film on a rotating stage that
allows orientationof the film in either the IR or UV-visible configuration.Thus,
tandemFTIR and UV -visible spectracan be collectedwhile the film is in a con-
trolled atmosphereor undervacuum(Johnstonet aI., 1991, 1992a).
Self-supportingfilms of soil colloids can be preparedby drying an aliquot
of an aqueoussuspensionof the colloidal materialonto a surfacethat is removed
prior to analysis.A dilute aqueoussuspensionof the soil colloid is preparedwith
a solids concentrationin the rangeof 0.5 to 5% (w/w). Care should be taken to
ensurethat the soil colloid is completely dispersedin the suspension.The vol-
ume of aqueoussuspensionused to preparethe film, usually ranging from 1 to
N
V2
, is
450
400 •
•
•
350
•
300
•
mg/g 250 •
•
2001 ••
••
•
150j •
•
100 •
I
•
c...
o
5"1'.~~ \~ =
:z
0'0 0.1 0.20.30.40.5 0.6 0.70.80.9 0.02:\ 1500 1600
a
:z
fI:o
P/Po
g
Fig. 10-8. Comparisonof the desorptionisotherm of water from a self-supportingclay film of Na-SAz-l obtainedat 24°C to the FTIR spectra(right side) of water ...
=
sorbedon the Na-SAz-l clay film. Each spectrumshownon the right side correspondsto one datapoint on the desorptionisotherm.
Top Vi ew
...
,,
,
,
,,
UV-v isible
UV-v isible Beam
Side View
Fig. 10-9. Schematicdiagram of controlled environmentFTIRJUV-visihle cell with rotating film

mount.
25 mL, will dependon the desiredareaof the self-supportingfilm. Ideally, the

self-supportingfilms should have a density of 1 to 3 mg cm-2 and a cross-sec-
tional area>1 cm2• Thicker films are sometimesneededfor experimentsthat
require handlingand mounting in a cell. Kung and McBride (1989a),for exam-
ple, reporteda densityof 13 mg cm-2 for their self- supportingfilms of goethite.
Materials on which self-supportingfilms can be preparedinclude Mylar,
polyethylenefilm (e.g., food wrap and storagebags), and aluminum foil. The
optimum materialwill depend,in part, on the particularsoil colloid. Typically, a
20- by 20-cm sectionof the substrateis attachedto a 25- by 2S-cmglassplate in
such a manner that it remains flat during the evaporationstep. This can be
accomplishedby taping it to the glass plate so that no wrinkles are present.
Polyethylenesheetscan either be tapedor held to the glassplate by a few drops
of water placedbetweenthe plate and the sheet(White & Roth, 1986). One- to
two-milliliter aliquotsof the aqueoussoil suspensionare then placedon the flat-
tened surfaceand allowed to dry. Each deposit will result in a dry deposit of
approximately 4 to 8 cm2 in size. After drying, the deposited film can be
removedby carefully running the sectionof sheetslowly over a knife edge.The
depositwill separatefrom the sheet.Self-supportingfilms formed in this manner
tend to be thicker at the edges.Thus, it is often desirableto use a smallerportion
cut from the middle of the depositin order to ensurea sampleof uniform thick-
294 JOHNSTON& AOCHI
ness.The self-supportingfilm can be mountedbetweentwo magneticplatesor

simply tapedto a metalor cardboardsurfacefor FTIR analysis.An exposedarea
1 cm in diametershouldbe maintainedfor sufficient throughputof infrared radi-
ation through the sample.
Another method to prepare self-supporting films for FfIR analysis
involves the use of a Millipore filter apparatus(Fernandezet aI., 1970).
Approximately25 mL of a dilute aqueoussuspension(solids concentration0.1%
w/w) is filtered through a 0.05-,urn polycarbonatemembranefilter (47-mm
diam.). The sampleis allowed to dry slowly on the polycarbonatefilter. After
drying, the film is removedby carefully running the filter-deposit over a knife
edge.This procedure canbe usedto prepareself-supportingfilms of soil colloids
and minerals that do not form uniform dry depositsusing the first method. A
third methodfor preparingself-supportingfilms of powderedsamplesis to press
the samplebetweenpolishedsteelplates.This methodis describedby White and
Roth (1986).
SupportedDepositson Infrared TransparentWindows

For soil materialsthat cannotbe preparedas self-supportingfilms, a sus-
pensionof the samplein aqueousor organic solvent can be evaporatedonto an
IR transparentwindows and analyzedas a supporteddeposit. Kaolinite and its
polymorphs,iron and aluminumoxides,and humic substances areoften analyzed
in this manner. It is important to work with particles <2 ,urn in size for FTIR
analysis.Particlesin the samesize range as the wavelengthof the IR radiation
(2.5-25 ,urn) tend to act as small mirrors, and as a result, transmissionof light
through the sampleis reduced.Methodsfor reducingthe particle size of sand-
and silt-size materialsare discussedin White (1977) and White and Roth (1986).
A 0.5- to l.S-mL aliquot of an aqueoussuspensioncontaining the <2-,um size
fraction is usedto form the deposit.Scatteringof light by the particlescan still
occur evenwith this fraction. The effect can be minimized by placing a drop of
oil such as Nujol on the sample;theseoils have an index of refraction near that
of the solid which minimizesoptical dispersion.
Highly orienteddepositsof kaolinite, goethite,and relatedmineralscan be
preparedas supporteddeposits (Serratosa et aI., 1962; Baes & Bloom, 1989;
Cambier,1986; Rouxhetet aI., 1977). This orientationcan be utilized to provide
information not obtainablefrom randomlyorientedsamples.lf the FTIR beamis
planepolarized,the supporteddepositcan be tilted at known anglesto determine
the orientationof structuralOH groups(the changein intensity of the IR spectra
with respectto the angle of incidenceis sometimesreferredto as pleochroism).
Thesemethodsalsocan be usedto determinethe orientationof adsorbed orinter-
calatedspecieson clay or oxide surfaces.
A wide rangeof IR transparentmaterialsare availablethat can be usedfor
supportingdepositsof soil colloids. The main factors to be consideredin select-
ing a particularIR window materialinclude the transmissionregion of the mate-
rial, cost, chemical reactivity, solubility in water, hygroscopicproperties,and
toxicity. A listing of some common IR window materials is presentedin Fig.
10-10.For depositsprepared fromaqueoussuspension,the most commonlyused
window materialsinclude AgCl, ZnSe,and ZnS. For spectralinvestigationslim-
0.1 05 10 50 10 50 IDOiL
10·125 A-OP
10.260 KDP n
04 eOROSILICA E GLASS rn
n:a:=Im GALLIUM ANTiMONIDE
1012 FlISEO III :A
CALCITE 5.5
I
104~ RU ILE
014 SAPPHIRE 65
N IU I ANA
~ INDIUM ARSENIDE
0
~ g LEAD SULFIDE (FILM) +
LEAD SElENIDE IFILMI+
I
AR 'U1rTi ANA rr
MAGNESIUM FLOR10£
50
m
r~
I
LEAD Tt.lLURIDEIFILM)'+
025
I MAGNESIUM OX IDE
In' 8 ( h TELLURIUM
1m
1 I AN-I
0.12 LITHIUM FLUORIDE 90
IC I~ AL IJJ.lFW:rR1ll£ I
lOG ARsENI RTSu~ W
I I/fOIUM HID 14
I IRTRAN- 2 IHI
~.GALLIUM ARSENIDE m
lU. --:.:..:--- ~lJkIll1- ,~~
\[[~:-~ ,.M!!UM FblI2..~~ ~
<019 SODIUM FLUORIDE 15
16..> INDIUM ANTIMONIDE t
A MIUM ~oE 1'-
025 LEAO FLUORIDE 16,
[l~Q~.:rllillli!QL I~
Q A SENI 01 I iSlTTNIUM GLASS 18>
1.0 AMORPHOUS SElENIUM 201
I E AlUM
1021 SODIUM CHLORIDE 26>
IL L I
021 POTASSIUM CHLORIDE ~O
0.21 KRS-6 ~5
0.42 THALLIUM CHLORIDE ~
.25 POTASSIUMBROMIDE 40
0.6 KRS-5 40'>
4 THALLIUM ROM I 40
~C 25 PO ASSIUM 10010 4
O. C IUM B; OMI
IUM 1010
0.25 IAMOND eo
0.1 0.5 1.0 5.0 10 50 100iL
Fig. 10-10. Transmissionregions of optical materials.The limiting wavelengthsfor both long and
short cutoff havebeenchosenas thosewavelengths(wavenumbers)at which a sample2 mm thick
has 10% transmission.
ited to the OH stretchingregion, glass coverslipscan be used. Other materials

may be usedif their IR absorptionbandsdo not interfere in the spectralregion
of interest.Evencertaintypesof plasticwrap, for example,havea minimal num-
ber of absorptionbandsin specific regionsof the infrared spectrum.
296 JOHNSTON& AOCHI
PotassiumBromide Pellets
For routine characterizationof soil colloids, the most commonly used
methodfor IR or FfIR analysisis the preparationof the sampleas a KBr pellet.
One of the major advantagesof the methodis that only a small amountof mate-
rial is needed;1 to 4 mg for routine analysis.Using micropelletmethods,detec-
tion limits as low as 1 to 20 ",g have beenreported(Luoma et aI., 1982). Also,
the spectraof KBr pellets are lesssusceptibleto anomalouslight scatteringdue
to a bettermatchof the indicesof refractionfor the KBr-mineral interfacecom-
paredto that of the air-mineral interface.This techniquealso is amongthe most
suitable samplepresentationmethodsfor quantitativeanalysisof mineral con-
stituentsprovided that the samplehas well-defined, discretebands. Potassium
bromide pellet methodshave been used extensively,for example,to determine
the amountof quartz and its polymorphsin multicomponentmixtures using the
threediagnosticbandsfor quartzat 695, 780, and 798 cm-1 (Tennenham& Lyon,
1960; Radulescu~D, 1976; Foster & Walker, 1984; Hlavay & Inczedy, 1985;
Flehmig & Kurze, 1973; Dodgson & Whittaker, 1973; Larsen et aI., 1972;
Mangia, 1975).
The samplepreparationstepsrequiredto makea KBr pellet include grind-
ing the sampleto reducethe particle size to <2 ",m, desiccationof the sampleto
removethe interferenceof sorbedwater, and finally compressionof the sample
under high pressure(9000-11000 kg of total force). Potassiumbromide pellets
are not well suited for sorption studieson soil colloids, and the presenceof the
solid KBr matrix obviously precludesin-situ sorptionstudies.Consequently,the
sorbent-sorbate complexmust be isolated,desiccated,ground,and pressedinto a
KBr pellet. Any of thesestepsmay significantly alter the bondingmechanismof
the sorbedspecieswith the sorbent.Thus, mechanisticdata suggestedby the
vibrational spectramay be largely due to artifactsresultingfrom the samplepre-
sentationmethod.Additional problemscan be anticipatedusing this methodfor
samplesthat are sensitive to dehydration, increasedpressure(e.g., pressure-
induced phasechange),or that may react with the KBr matrix itself. Despite
theselimitations, this methodhas beenusedextensivelyto characterizesoil and
subsurfacematerials.
Liquid TransmissionCells
Fourier transforminfrared spectraof solvents,solutes,and dispersedsus-
pensionsof soil colloids can be analyzedusing fixed-or variable-pathlength liq-
uid cells. Liquid transmissioncells havebeenusedto study the sorptionof organ-
ics on montmorillonite (Zhang et aI., 1990), characterizehumic substancesin
waterand D20 (MacCarthy& Mark, 1975),investigateionizationof solublecar-
boxylic acids(Drzymala& Kielkowska, 1985; Drzymala, 1985),and to examine
clay-waterinteractions(Salle de Chou et aI., 1980; Mulla & Low, 1983). These
cells consistof two IR transparentwindowsseparatedby a distancerangingfrom
2 ",m to 5 mm. Factorsto be consideredin the selectionof window materials
include their cost, useful spectralrange,chemical reactivity, toxicity, and solu-
bility in water. Desirablecell path lengthsare primarily determinedby the opti-
cal absorbanceof the solvent to be used.Water, for example,absorbsIR radia-
tion very strongly and limits the maximum path length of a liquid transmission
cell to about20 pm. Clogging and cleaningof thesecells can be problematicin
working with aqueoussuspensions of soil colloids becausethe size of the colloid
particlesis comparableto the path length of typical liquid cells. A discussionof
liquid transmissioncell designsand their applicationof soil materialsis given by
White and Roth, 1986. Attenuatedtotal reflectancemethods,such as the cylin-
drical internal reflectancecells, provide an attractive alternative for studying
solution-phaseFfIR spectra(seesection"AttenuatedTotal Reflectance").
SelectiveDeuterationMethods
A universalproblemencounteredin studiesof surfaceOH groupsis that of
distinguishingtheir vibrational modesfrom thoseof bulk structuralOH groups.
The relative proportion of bulk to surfaceOH groups typically is quite large;
thus, spectroscopictechniquesmust discriminate betweentwo very disparate
populationsof OH. A surface-sensitivevibrational technique,such as IR reflec-
tion-absorption spectroscopy(IRRAS), surface-enhancedRaman scattering
(SERS),or electronenergyloss spectroscopy(EELS), can be usedto overcome
this problem, but unfortunately restrictions apply to each method at present
which severelylimit its applicationto the study of soil colloids. A more useful
approachis to label surfaceOH groups selectively with deuterium(D) by iso-
topic exchangebetween IH in the solid and DzO (liquid or vapor). Although
otherisotopescan shift the vibrational frequencyof the OH group, the advantage
of using deuteriumis that similar surfacereactionsare expectedfor OD and OH
groups.If experimentalconditionsare arrangedsuitably, bulk OH groupscannot
exchangewith D and no changein the frequencyof their stretchingor bending
bandswill occur, allowing them to be distinguishedspectroscopicallyfrom sur-
face OH groups.A useful illustration of selectivedeuterationtechniquescomes
in the work of Lewis and Farmer(1986) who studied the isotopic exchangeof
OH groupson lepidocrociteand boehmite.
Diffuse Reflectance
Near-IR diffuse reflectanceinstrumentshave beenusedfor over 20 yr for
the quantitativedeterminationof the water contentin flour and starch(Hoffman,
1963).Thesedispersiveinstrumentsmeasuredthe diffusely reflectedradiation in
the near-IR region of the electromagneticspectrum.Practical utilization of the
diffuse reflectancetechniquein the more energy starvedmid-IR region had to
await the developmentof Fourier transforminstrumentation.
One of the primary advantagesof DR spectroscopyfor investigationsof
soil colloids is that the samples are commonly in the form of a powder.
Particularly when the focus of such investigationsis chemical activity at solid
surfaces,such samplesapproximatea natural state for clay minerals and other
soil materials. Furthermore,only minimal sample pretreatmentis required for
analysis.Finally, the potentialvalue of this techniquefor in-situ investigationsat
surfacesrelevantto soil colloids has been increasedsignificantly by the recent
commercial availability of controlled environmentchambersdesignedfor use
with DR accessoriesin FfIR instruments.In addition to the samplingflexibility
298 JOHNSTON& AOCHI
offered by DR methodsfor such studies,Griffiths and Fuller (1982) have pre-

sentedevidencethat sensitivity enhancementmay occur with this techniqueat
very low concentrationlevels and for speciesdepositedon the surfacesof solid
particlescomparedto transmissiontechniques.Comprehensivereviews of DR-
FTIR spectroscopyand the relevant factors involved are given by Fuller and
Griffiths (1980), Griffiths and Fuller (1982), Maulhardtand Kunath (1982), and
Chalmersand Mackenzie(1988).
Theory
Diffuse reflectance(DR or DRIFT) accessoriesfocus the beamleaving the
interferometerof an FfIR onto the surfaceof the powderedsample.If the radi-
ation is reflected and the anglesof the incident and reflected beamsare equal,
specularreflection has occurred.This type of reflected radiation containslittle
informationaboutthe sampleso DRIFf accessories are designedto minimize the
amountof specularcomponentreachingthe detector.The remainderof the inci-
dent radiationpenetratesthe sampleto a depththat dependson the reflective and
absorptivecharacteristicsof the sample.When this radiation is subsequentlyre-
emitted from the surface,it doesso in all directionsand is thus termed diffuse.
The diffusely reflected radiation may have undergoneabsorption,transmission,
reflection,and refractionwithin the samplematrix. Despitethe complexity of the
optical pathwayspossible,efficient collection of the diffuse radiationresultsin a
spectrumvery similar to a transmissionspectrum.
Intensitiesin diffuse reflectancespectraare usually presentedin terms of
the Kubelka-Munkfunction,fiR=), namedafter the developersof the theory most
commonly used to describethe phenomenon.In practice,R= is obtainedas the
ratio of the samplereflectanceto that of a referenceand this value is usedin the
relationship
[6]
Software is generally availableon FfIR instrumentsto perform the con-

version of intensitiesto Kubelka-Munkunits. If certain experimentalconditions
are met, this function can be linearly relatedto sampleconcentration.Thosecon-
ditions include: (i) a small specularcontribution to the collected radiation, (ii)
constantscatteringfrom the sampleover the concentrationrangeof interest,(iii)
the reflectanceof the sampleand the referencematerial are in infinite depth, i.e.,
do not changeas a function of sampledepth, and (iv) the referenceis nonab-
sorbing. The design of DR accessoriesand recommendedmethodsof sample
preparationare intendedto minimize deviations fromtheseconditions.
Accessories
CommerciallyavailableDR accessoriesare designedin one of two optical
geometries:(i) on-axis, and (ii) off-axis. Thesetwo configurationsare depicted
in Fig. 10-11. With the on-axisdesign,the sampleand the detectorforeopticsare
in the sameplane while in the off-axis type, they are in different planes. The
(al
MIRROR no IS CANTEO TO REFLECT

THE RAOIATION DOWNWARO TO THE
SAMPLE SURFACE IN THE MIDDLE.
INPUT OUTPUT
BEAM BEAM
Ibl
Fig. 10-11. Basic DRiFf designs:(a) on-axis(b) off-axis (reproducedwith permissionof Applied
Spectroscopy).
intent of the off-axis design is the deflection of the specularcomponentaway

from the mirror collecting the reflectedradiation with subsequentenhancement
of the diffusely reflected component.This samegoal is achievedin the other
design with the addition of a post that blocks out the specularcomponent.
Unfortunately,the postseverelyreducestotal energyreachingthe detector.In an
evaluationof the relative merits of thesetwo optical designsusing different par-
ticle-sizefractions and sampleconcentrationsof a commerciallyavailablesilica
sand,Yang et at. (1986)concludedthat specularrejectionandspectralquality are
essentiallythe samefor both types. Hembreeand Smyrl (1989), on the other
hand, found adherenceto Kubelka-Munk theory over significantly larger con-
centrationrangeswith the off-axis optics. Both typesof accessoriesare capable
of providing good-qualityspectraprovidedthat samplesare of small particlesize
and diluted in an alkali halide matrix.
300 JOHNSTON& AOCHI
Recently,environmentalcontrol chambersfor usein conjunctionwith the

diffuse reflectanceaccessorieshave become available. These chambersare
designedwith optical windows and gasportsthat allow spectraof samplesto be
taken in situ under vacuum,ambient,or high-pressureconditionsand various
temperatureregimes.They also allow the flow of inert gasor vaporphasechem-
icals throughthe samplechamber.Effective useof thesedevicesis difficult with-
out an accompanyingvacuum manifold systemfor purging or evacuatingthe
chamberandthe controlleddelivery of samplegas.Informationon the operation
and requirementsfor sucha systemcan be found in Smyrl et al. (1983).
Sample Preparation
Samplesare generallypreparedby grinding the analytewith an inert alka-
li halide at a concentrationof 1 to 10% in a Wig-L-Bug (SpectraTech Inc.,
Stamford,Cf) or mixer/mill for 1-3 min. This procedurereducesthe particle
size of both the analyteand the diluent and dispersesthe samplehomogeneous-
ly in the matrix material.Samplesanddiluentsalsocanbe groundseparatelyand
combinedafter grinding. The latter approachreducespossibleeffectsof sample
alterationby grinding with the diluent but also can result in a lesshomogeneous
sampleif the physical characterof the two componentsin the mixture is quite
different. This may particularlybe a problemwith humic substances that tend to
smearor agglomerateinsteadof dispersinginto uniform particlesupongrinding.
The ideal sampleis one that is intimately mixed, the particle sizesof both the
analyteand diluent are lessthan about30 !l1Jl in diameter,and similar particle-
sizedistributionsexist for all components.In-depthdiscussionsof sampleprepa-
ration for quantitativeanalysisby DRIFf spectroscopycanbe found in Hamadeh
et al. (1984) and Tevruchtand Griffiths (1991). The samplingcup of the acces-
sory deviceis overfilled with the mixture andthe top leveledwith a straightedge.
The SB spectrumof the sampleis then referencedagainstthat of the diluent
alone.Severalfactorscan affect the quality of the resultingspectraand the com-
mentsfollowing are intendedto addressthese.It shouldbe kept in mind that the
effectsof the parametersdependon the natureof eachabsorptionbandand may
not be the sameover the entire spectralrange.
Inorganic compoundscommon to soil, especiallyclay minerals, tend to
exhibit very strongabsorptionbands.If thesesamplesare not fmely groundand
diluted in an inert matrix, the appearanceof the spectrumin the region of the
strongbandswill be severelydistortedand intensity inversion may even occur
(reststrahlenbands).Similar spectraldistortion in the regionsof strongabsorp-
tion bandsmay be observedin the absenceof dilution for organicmaterialssuch
as humic substances.As the particle size of thesesubstancesis reduced,the
amountof specularcomponentis diminishedrelative to the radiation refracted
and absorbedby the sample.Absorptionconstantsalso increaseas particle-size
decreases.Both of thesefactors result in an increaseof the diffusely reflected
radiationas a function of decreasingparticlesize.
Matrix materialsmostcommonlyusedasdiluentsarethe alkali halidsKBr
and KCl. When cation exchangeis a problem,other diluentssuchas Si, Ge, or
diamondpowdercanbe used.Theselatter diluentstendto scatterstrongly,how-
ever,so the depthto which incidentradiationpenetratesinto the bulk of the sam-
pIe tends to be extremely small. Generally weaker diffuse reflectancespectra

thus result and usablespectralrangesare more limited. Whateverdiluent is used,
it is important that particle-sizedistributionsfor sampleand referencematerials
are as similar as possibleand the samplesare packedreproduciblyin the sample
cup.
Conditions under which the Kubelka-Munk function is linearly related to
concentrationare more closely approachedwhen the analyteis diluted to a high
degreein an alkali halide matrix. With the use of a diluent, the infinite depthcri-
terion for both sampleand referenceis usually met. Scatteringof radiation with-
in the bulk of the sampleis controlledby the propertiesof the matrix ratherthan
the sampleand is thus more likely to remainconstant.If the chemistryoccurring
at a solid surface is the subject of interest, however, the presenceof an alkali
halide may be undesirable.In thesecases,neat samplescan be usedbut quanti-
tative spectral interpretation must be approachedwith additional caution. In
regionsof strong absorptionof the substratematerial, distortion in the spectrum
or the appearanceof reststrahlenbandsis reducedby using the untreatedpowder
as the reference.Spectral features, however, arising from surface activity or
sorbedspeciesmay still be of limited quantitativeor even qualitative value in
these regions. With highly absorbingmaterials such as humic substancesthe
depth of penetrationinto the samplebulk can be very small becauseso much of
the incident radiationis absorbed.Spectralinformation on surfacespeciesis thus
severelylimited (Fraser& Griffiths, 1990).
Applications
Several recent publications demonstratethe potential usefulnessof DR
FTIR spectroscopyfor the characterizationof soils and soil constituents.Nguyen
et al. (1991) evaluatedthis techniquefor inorganicconstituentsas well as whole
soils. Spectrapresentedby theseauthorsreflect differencesin both organic and
inorganic compositionof severalsoils. As illustrated by Fig. 10-12, they also
demonstratethe effect of analyteconcentrationon the appearanceof reststrahlen
bands and spectral distortion. The very intense v(Si-O) absorption band at
approximately1100 cm-1 in the KEr pellet spectrumof a solonetzsoil appears
as an inverted band in the neat DRIFT spectrumof the samesample.Baesand
Bloom (1989) presentedDRIFT spectraof humic andfulvic acidsextractedfrom
a peat and comparedthem with KEr pellet spectra.Niemeyeret al. (1992) con-
cluded that while DRIFT spectrawere not useful for quantitationof humic sub-
stances,band intensitiescould be used to evaluatestructuralchangesoccurring
during humification processes.Information relevant to the characterizationof
humic materials also can be found in related studies of peat (Holmgren &
Norden, 1988) and coal (Fuller & Griffiths, 1980; Smyrl & Fuller, 1982).
An exciting potential applicationof DRIFT spectroscopyis in the investi-
gation of chemical activity at surfacesrelevant to soil science.Such investiga-
tions can be approached ineither of two ways. In the more conventionalof these,
solid surfacesare subjectedto somechemicaltreatmentand spectraof powdered
samplesbefore and after treatmentare compared.Using this technique,Nanzyo
and Watanabe(1982) confirmedearlier transmissionstudiesindicating the pres-
enceof a bridging binuclearsurfacecomplex for phosphatesorbedon goethite.
302 JOHNSTON& AOCHI
Q>
u
c:
'"<5
.0
til
.0
<
c
~ ~ ~ ~ ~ ~ 1~ lD ~ ~
Wavenumber
Fig. 10--12. Infrared spectraof a black earth:(A) neat DRIFf, (B) 3% in KBr DRIFf, (C) 0.3% in
KBR pellet (reproducedwith permissionfrom Australian Journal of Soil Research).
Kung and McBride(1989a,b)also utilized this techniquein their investigationof

interactionmechanismsof p-substitutedbenzoateson various iron oxides.
The secondapproachrequiresthe use of a controlled environmentcham-
ber that allows samplesto be treatedin situ and spectrato be collectedas surface
processesare occurring.Spectrapresentedin Fig. 10-13 demonstratethe results
obtainablethrough the use of this procedure.Thesespectrawere collectedafter
N2 saturatedwith 1,2-dibromoethane(EDB) had flowed through samplesof
illite, smectite,and kaolinite for 22 h (Aochi et aI., 1992). Desorptionof EDB
from the clay surfaceswas followed by collecting spectraas N2 alone flowed
throughthe clay samplesafter the sorptionperiod. Resultsof this study indicat-
ed that EDB accumulatedat the clay surfacesas a liquid phasecomprisedof two
rotational conformers.They further indicated that one of the conformers,the
gaucheform, was preferentiallyretainedby the clays. Thoseinterestedin using
this methodologyare strongly advisedto consult the literatureon DRIFT inves-
tigationsof the chemistryat surfacesof coal (Fuller et aI., 1985; Smyrl & Fuller,
1982, 1987) and catalytic agents(Van Every & Griffiths, 1991; Johnsonet aI.,
1988).
AttenuatedTotal Reflectance
In conventionaltransmissionspectroscopy,the samplethicknessmust be
very small so that sufficient light is able to reachthe detector.This requirement
ITIR & RAMAN SPECTROSCOPY 303
ILLI TE 1247
w
u
z
«
m
0::
o
U1
m
«
KAOLINITE 1247
w
u
z
«
m
0::
o
U1
m
«
w
u
z
«
m
0::
o
U1
m
«
Fig. 10--13. Desorption of EDB from clays after (a) 22-h sorption, (b) 1h, and (c) 2h desorption
(reproducedwith permissionfrom Environmental Science and Technology).
placessevereconstraintson the typesof liquid or solid samplesthat can be stud-

ied using thistechnique.For investigations insoil or environmentalsciencesthis
limitation is particularly troublesomesince samplesof interest tend to be opti-
cally opaque,anisotropic,and includewater as an importantcomponent.Because
water is a very strongabsorberof IR radiation,liquid transmissioncells suitable
for aqueoussolution studiesare limited to approximately15 to 20 flm in path-
length, difficult to clean, and not suitable for suspensions.Attenuated total
reflectancein many ways seemsto hold great promise for overcoming these
problems.With ATR, it is conceptuallyuseful to imaginelight rays within a crys-
tal reflecting from one surface to the other. With each reflection, the radiation
interactswith the samplein contactwith the crystal. Except for films of thick-
nesson the order of a few thousandangstroms,the extent of this interaction is
not dependenton the actual physicaldepthof the sample.This meansthat spec-
tra of bulk samplescan be obtainedsimply by bringing them in contactwith the
surfaceof the ATR crystal. Samplesfor spectroscopicinvestigationsby this tech-
nique could thus potentially include solids, solutions,or suspensions.
304 JOHNSTON& AOCHI
Theory
The following brief discussionis intendedas an aid to understandingthe
mannerin which differentparameterscan affect spectraobtainedusing ATR. A
thoroughtreatmentof the theorybehindthe techniquecan be found in the text by
Harrick (1979). The ATR experimentis basicallycomprisedof a crystal of high
refractive index, nl> in contact with a sampleof a lower refractive index, nz.
Radiation approachingfrom the densermedium is incident upon the interface
betweenthesetwo media.The fate of that radiationupon striking the interfaceis
determinedby the angleof incidenceas measuredfrom the normal to the surface.
If this angleexceedsthe critical angle,eo where (Eq. [7])
[7]
then all of the light is reflected back into the densermedium. At the point of
reflection, the incoming and reflected waves combine to establisha standing
wave normal to the surface.Within the crystal,the electricfield amplitudeof this
standingwave variesin a sinusoidalfashion.This electricfield continuesinto the
samplemedium,where it decaysexponentially.It is the interactionof this elec-
tric field with the samplepositionedthere that gives rise to the reflectanceatten-
uation or light absorptionwhich in tum resultsin an IR spectrum.
The most important factors in determiningthe strengthof this interaction
and, therefore,the intensity of the resulting spectrum,are the magnitudeof the
electric field at the interface and the depth to which it penetratesthe sample
medium. Both of thesefactors decreaseas the angle of incidenceincreases.The
depth of penetrationis, in addition, a function of other factors as shown by Eq.
[8]
[8]
where A is the wavelengthof the incident radiation,eis the angle of incidence,

nc isthe refractiveindex of the crystal, and nsc is the ratio of the refractive index
of the sample to that of the ATRcrystal. Typical values for dp are -0.1 A
(Griffiths & de Haseth,1986). From this equation,it is apparentthat the depthof
penetrationis less for ATR crystals of higher refractive index. In addition, the
depthwill be a function of the wavelengthof the incident radiation.The effect of
this wavelengthdependencyis a spectrumwhich differs from a transmission
spectrumin the relative intensity of absorptionbandsin different portionsof the
spectrum.Radiation of longer wavelengthwill penetrate moredeeply into the
sampleand as a result, bandsat thesewavelengthswill appearrelatively more
intense. Strong bands in ATR spectra also may appear somewhat distorted
becausethe refractive index of the sampleundergoessignificant changesin the
vicinity of an absorptionband. The effect of dispersionin the refractive index
nearabsorptionbandsis treatedextensivelyby Harrick (1979).
FTIR & RAMAN SPECfROSCOPY 305
Cylindrical Internal ReflectanceCell
T.r
7 .~
-:~~\~/~
~
Fig. 10-14a. Schematicdiagram of a cylindrical internal reflectanceATR cell.
Fig. 10-14b. Schematicdiagramof a trapezoidalATR accessory(reproducedwith permissionfrom

Applied Spectroscopy).
One interesting consequenceof the standing wave phenomenonis that

dipolesorientedin all directionsat the crystalsurfacewith respectto the incident
radiation are able to absorbenergy from the electric field generatedthere. The
amplitude of the electric field in different directions dependsnot only on the
angle of incidenceand refractive indices, but also upon the polarization of the
incident light. Absorption coefficients for all three directions may thus be
derived by comparingspectraobtainedusing nonpolarizedlight and light polar-
ized parallel and perpendicularto the plane of incidence.
Apparatus
The most commonATR crystal shapehas been the trapezoid.A conven-
tional design for an ATR accessoryusing this shapeis shown in Fig. 10-14
(Messerschmidt,1986). Mirrors are usedto focus the incoming beamon the end
face of the crystal and direct the exiting beam to the detector.As the incident
306 JOHNSTON& AOCHI
radiationpropagatesdown the length of the crystal, it interactswith the sample

in contactwith its surfacesat the points of reflection. In order to avoid spectral
distortion, the beamshouldenterthe crystal normal to the face. This meansthat
in the caseof this particularcrystaldesign,if a changein the angleof incidence
is desired,the crystal must be replacedwith one cut at a different angleand the
opticsadjustedaccordingly.
Numerousvariations on this basic crystal design have been offered by
manufacturerswith threebasicgoalsin mind: increasedsensitivity,accommoda-
tion to different typesof samples,and enhancedeaseof use.Crystal length can
bevariedto increaseor decreasethe numberof reflections.Hemicylindricalcrys-
tals allow the angleof incidenceto be varied over a wide rangebut allow only a
single reflection. In somedesigns,selectedfacesof the crystal are mirrored in
order to direct the exit beammore advantageouslyor to minimize the needfor
optical alignment. One recent design enjoying considerablepopularity for the
study of solutions and suspensionsis that of the CIRCLE cell, availablefrom
SpectraTech. This device incorporatesa rod-shapedcrystal with conical ends.
Enhancedsensitivity is claimedfor this apparatusbecausethe Cassegrainoptics
usedat both endsof the crystal make maximal use of the circular beamof an
FfIR instrument.It is designedto be very easyto usewith a wide variety ofsam-
ple typesand also can be usedas a flow cell.
As mentionedearlier,the depthof penetrationand, therefore,the intensity
of the spectrumalso is a function of the refractiveindex of the crystal. By using
a materialof lower refractiveindex, the depthof penetrationinto a given sample
can be increased.Common crystal materials include Ge, KRS-5 [Th(Br,I)],
ZnSe,ZnS, Si, and CdTe.It shouldbe kept in mind in selectingATR crystalsthat
thesematerialsdiffer not only in refractiveindex, they also differ in their useful
wavelengthrange,hardness,reflective losses,chemicalinertness,and resistance
to scratching.Theseall needto be consideredin selectingthe appropriateATR
crystal for the desiredexperiment.Propertiesof widely availableATR crystal
materialsaresummarizedin Table 10-5. Detaileddiscussionsof all of thesefac-
tors as well as optical designconsiderationscan be found in the text by Harrick
(1979). A large selection of crystal shapesand materials is available from
Harrick Scientific (Harrick Scientific Corp., Ossining,NY).
Table 10-5. Propertiesof ATR crystal materialscommonly usedfor mid-IR spectroscopy.

Useful
Refractive Hardness frequency
Material index (Knoop) range Properties
cm-1
Ge 4.0 550 5000-900 Hard, brittle, inert
Zinc selenide 2.42 150 20000-700 Soft, brittle, attackedby acid
Zinc sulfide 2.22 355 14000-1000 Hard, brittle, inert
Si 3.42 1150 9500-1500 Hard, inert
KRS-5 2.35 40 14000-400 Toxic, soft, attackedby acids,bases
Cadmiumtelluride 2.65 170 11 000-800 Brittle, inert
Amtir 2.5 170 11 000-800 Brittle, inert
Diamond 2.4 7000 45000-2500 Very hard, very inert
1600-FlR
Sample Preparation
The most importantfactor in an ATR experiment isthe degreeto which the
samplecontactsthe crystal element.With a typical element,the standingwave
interactswith the sampleto a depth of approximately1.5 pm (Rein & Wilks,
1982).While this interactionis replicatedat eachreflectionpoint, it is easyto see
that if the sampleis not in intimatecontactwith the crystalsurface,it is very pos-
sible to get no spectrumat all. Accessorydevicesdesignedfor use with solids,
therefore, include backing plates intendedto compressthe sample against the
crystal element.While this works well for malleablematerialssuch as synthetic
polymers,it is much more difficult to establishgood contactwith dry powders,
particularly if they are of coarseparticle size and hard (characteristicsthat con-
tribute also to the rapid degradationof the crystal surface).Some successhas
been achievedwith clay films formed by the depositionand subsequentdrying
of clay suspensionsonto glass plates or onto the surface of the crystal itself.
Thesefilms have an additional advantage in that the clay plateletsare oriented
with respect to the surface of the ATR element. Other investigators have
achievedgood contactwith aqueousclay suspensions (including deuteratedsus-
pensions)and pastes.
Surfacecontactis not as difficult to ensurewith aqueoussolutionsor col-
loidal suspensionsbut many other factors must be controlled in order to obtain
useful spectralresults.This is due to the needto usespectralsubtraction.While
spectralsubtractionis often useful in studiesof solids, it is almost universally
necessaryfor investigationsof liquid systemsbecauseof the presenceof bulk
solvent.The goal in sucha procedureis to subtracta referencespectrumof a sys-
tem componentthat most closely reflects its spectralactivity within the sample.
Water,of course,is generallythe solventof choiceand servesto illustrate the dif-
ficulties that may be encountered.The absorptionbandsof liquid water are rather
broad and intense and tend to mask valuable spectral information on other
speciesin the sample.The shapeof the water bandsis attributableprimarily to
H-bondingeffectswhich, in tum, are greatly affectedby temperature,pH, ionic
strength,solutecomposition,and the presenceof solid surfaces.All of thesefac-
tors must be consideredbeforeconclusionscan be drawn from spectraobtained
by subtraction.If spectralinformation is desiredin the immediatevicinity of the
water absorptionbands,water must be less than totally absorbing(equivalentto
0% transmittance)in thosespectralregions.This may be accomplished byreduc-
ing the depth of penetration with a crystal of a higher refractive index.
Unfortunately,it also resultsin lower sensitivity to the soluteof interest.
Applications
Applications of ATR to problemsin soil sciencehave beenrelatively lim-
ited. In one of the earliest reportedinvestigations,polarized light was used to
determinethe orientation of a lysine-vermiculitecomplex (Raupach& Janick,
1988). In this study, single crystals or clay films producedby depositionfrom
aqueoussuspensionwere pressedagainstthe flat surface of a hemicylindrical
ATR element.Spectrawere collectedfor both deuteratedand nondeuterated sam-
ples using parallel and perpendicularpolarizedradiation. The infrared informa-
308 JOHNSTON& AOCHI
tion was usedin conjunctionwith x-ray diffraction data to arrive at a configura-

tion for lysine moleculeson the silicate surface.This basic approachwas later
extendedto orientationstudiesof otherchemicalspeciesand otherclay minerals
(Raupachet aI., 1979, 1987; Raupach& Janick,1988). Studiessuchas theseare
only possiblewith uniaxially orientedclay samples.
In an unusualapplication of ATR spectroscopy,Mulla et ai. (1985) used
reflectancespectraof a clay suspensionin a dilute solution of D20 in H20 to
determinesurfaceareasof a numberof Na-saturatedmontmorillonites.Values
obtainedusing this methodologycorrelatedwell with thosefrom ethyleneglycol
monoethylether(EGME) determinations.Similar agreement,however,was not
found for surfaceareasof homoionic samplesof Upton montmorillonite when
the saturatingcation was not Na. In this study also, a hemicylindrical element
was selected.
By far the mostpopulardesignusedin recentATR investigations isthat of
the CIRCLE cell (Fig. 10-14). Tejedor-Tejedorand Anderson(1986) described
the application of this cell to investigationsof the goethite-aqueoussolution
interface.In this and in severalsubsequentpublications(consultTejedor-Tejedor
et aI., 1990; Tickanenet aI., 1991, for a discussionof methodologyand impor-
tant considerations),samplecomposition withinthe cell was varied with respect
to variablessuchas pH, ionic strength,and sorbatespeciesin order to character-
ize the surfaceof goethite, the state of water at the goethite-aqueous solution
interface, and interactionsof organic moleculeswith goethite at the molecular
level in the presenceof bulk water. The techniqueused is illustrated in Fig.
10-15. In this figure referenceaqueoussolution spectraof potassiumsalicylate
and a Fe/salicylatecomplex are comparedto the spectrum of salicylate ion
sorbedon the surfaceof goethite in aqueoussuspension.The interpretationof
spectraobtainedusing this methodologyis complicatedby severalfactors. The
most significantof theseis the fact that waterstrongly absorbsinfrared radiation
and its spectrumis thus difficult to subtractfrom samplespectra.Subtractionalso
is complicatedby variationsin the shapeof the absorptionbandsof water due to
changesin the variablesmentionedabove. Even when the interactionsof inter-
est are for componentsin true solution, problemscan arise becauserelatively
high concentrations(in general,>0.1%) are neededand interactionsoften occur
with the ATR elementsurfaceas well as betweenthe constituentsin solution.
Theseproblemsare compoundedin the caseof suspensionsby conformational
and surfaceeffects exertedby the suspendedsolid on both water and sorptive
molecules.Finally, spectral interpretationcan be hindered becausevibrational
assignmentsfor speciesin aqueoussolution are. notabundantin the literature.
Theseaspectsalso were discussedby Morra et al. (1989) with regardto charac-
terizationof humic acid in water.
FourierTransformInfrared Microspectroscopy
During the past 10 yr, IR microsamplinghas been revolutionizedby the

introduction of sensitive FTIR microscopes(Griffiths & de Haseth, 1986;
Chalmers& Mackenzie,1988; Messerschmidt,1988; Krishnan & Hill, 1990).
The application of this method to the characterizationof soil and subsurface
Fig. 10....15. Cylindrical internal reflectance(erR) FTIR spectraof a goethite-salicylatecomplexand

related aqueoussolution referencespectra(reproducedby permissionfrom American Chemical
Society).
materialsholds considerablepromisealthoughonly a limited numberof applica-

tions have been reportedin this area thus far (Johnstonet ai., 1990;Vry et ai.,
1988; Pironon et aI., 1990). The principal advantageof FTIR microscopyover
other methodsis the ability to obtain spatially resolvedFTIR spectraof small
particles or regions of complex, heterogeneous samples.An FTIR microscope
can be addedto most modernFTIR spectrometersas a samplingaccessory.The
microscopecan be coupledto the externalport of a FTIR spectrometer,or it can
be placedin the main sample compartment. In the latter configuration,the sam-
ple compartmentof the instrumentis then dedicatedto FTIR microsampling.
A schematicdiagramof a FTIR microscopeis shown in Fig. 10--16. The
modulatedIR beamfrom the interferometeris broughtinto the baseof the micro-
scope and focused onto the sample using an off-axis parabolic mirror. A
Cassegrainobjective (magnification is on the order of 30 to 40x for this objec-
tive) is usedto collect the IR beamthat is transmittedthrough the sample.After
passingthrough an apertureand polarizer (optional), the transmittedIR beamis
focusedon a small-areaMCT detector.
Fouriertransforminfrared microscopesare operatedby switchingbackand
forth betweenthe "visual inspection"and "IR analysis"mode. The visual mode
is used to visually position the sampleso the region of interestcan be selected.
Once this is accomplished,the microscopecan be switched to the IR mode to
obtain the IR spectrum.The accessoryis placedin the visual modeby sliding the
viewing eyepieceand folding mirror into the optical beam thus diverting the
310 JOHNSTON& AOCHI
I side view I I lronl view I
LN, Cooled MCT detector

'--r-I.~---' MCT detector
wire-grid polarizer
iris aperature
sample stage
mirror IR beam
Fig. 10-16. Schematicdiagramof a FTIR microscopesamplingaccessory.
image to the viewing eyepiece(consistingof a glassobjectivewith the magnifi-

cation of 4x to lOx). An illuminator source(e.g., quartz halogenlamp) optically
coupled to the microscope provides visible illumination of the sample.
Positioningof the sampleis accomplishedthrough the use of a standardmicro-
scopex-y translationstage,or optionally, a motorizedmicropositioningstagecan
be used. When the sampleis appropriatelypositioned,the region of interest is
isolatedby meansof a rectangularor circular aperturethat servesto maskout the
unwantedportionsof the sample.By this means,the IR beamis only allowed to
passonly through the spatialregion of interest.In practice,the minimum dimen-
sion for the apertureis on the orderof 5 to 10fml. Severeproblemsrelatedto the
limited amountof light passingthrough the sample(optical throughput)and dif-
fraction effects are encounteredfor smallerapertures.Diffraction effects can be
expectedfor aperturevaluesof lessthan 30{1m, however,they can be minimized
by collecting a referenceSB spectrumusing the same aperturesettings. For a
more complete discussion of diffraction effects and optical nonlinearities in
FTIR microscopysee Chalmersand Mackenzie,(1988) and Krishnan and Hill
(1990). Once the sampleis positionedand the aperturefixed, the visible illumi-
nation optics are then switchedout so that the transmittedIR beamis passedto
the MCT-detectorand the spectrumcan then be taken.
Many FTIR microscopesalso can operatein an external reflection mode
which is useful when the sampleis too thick or opaquefor a microtransmission
study. In this configuration,the IR beam is focusedonto the surfaceof interest
and the reflectedcomponentis collected into the microscope.The band shapes
observedin reflectancespectracan often be distorted becausethey are much
more subject to anomalousdispersioneffects than transmissionspectra.These
effects are causedby the rapid and significant variation in the samplerefractive
index that occurs in the vicinity of an absorptionband. The Kramers-Kronig
FrIR & RAMAN SPECTROSCOPY 311
transformationcan be used to correct for these distortions (Krishnan & Hill,

1990; Chalmers& Mackenzie,1988; Gerson& Chess,1988).
Fouriertransforminfrared microscopyhasbeenusedrecentlyto determine
the orientationof structuralOH groupsin single crystal particlesof kaolinite and
dickite (Johnstonet aI., 1990). PolarizedFfIR spectrawere obtainedfor single
crystal specimensof dickite and kaolinite. The polarizationbehaviorof the OH
stretching bands of dickite is illustrated in Fig. 10-17. Previous attempts to
locate the positionsof the H atoms in kaolinite and dickite using various struc-
tural methodsdid not result in an unambiguousdescriptionof the OH groups.
The spectral data obtained from the microscope,on the other hand, provided
direct information about their crystallographicorientation.The size rangeof the
kaolinite particlesusedwas 5 to 15.um acrossthe 001 face and 10 to 50.um for
the dickite particles. In the caseof kaolinite, the bandsshowedless of a polar-
ization effect than expectedon the basisof the crystal structureand indicatedthe
Keokuk kaolinite crystalsexaminedwere twinned (JohnstonetaI., 1990).
Low TemperatureStudies
Recentstudieshaveshownthat cooling certainclay mineralsto nearliquid

He (12 K) temperaturesresultsin significantly improved resolution(Brindley et
aI., 1986; Prost, 1984; Prost et aI., 1987; Prost et aI., 1989; Bish & Johnston,
1993). The OH stretchingbandsof kaolinite, for example,narrow in line width
and shift in position. The room temperatureFfIR transmissionspectrumof the
poorly crystalline sample is characterizedby four bands in the OH-stretching
region in Fig. 10-18. Upon cooling this sampleto 12 K, however, thebandsnar-
row in line width and reveal the presenceof sevendiscretebands.The addition-
al bandsin the low temperatureFTIR spectrumof the poorly crystalline materi-
al are attributedto a dickite impurity, as well as stackingdisorders.The presence
of the impurities was only evidentin the low temperaturespectrum.
Cooling samplesfor IR or Ramananalysiscan be accomplishedusing sev-
eral different methods.The simplestand leastexpensivemethodusesa liquid N2
cryostat in which the sample is cooled by a thermal connectionto a liquid N2
reservoir.The samplestagemust be evacuatedto a moderatevacuum(<102 Pa)
to avoid unwantedcondensationon the sample.Potassiumbromide pellets and
supporteddepositson IR windows can easily be accommodatedby low temper-
ature cryostatsand displex cold-fingers.
Closed-cyclehelium refrigeratorsalso can be used to cool samplesfor
spectroscopicstudies.They are more expensivethan liquid N2 cryostats,but they
can be used to cool samplesto lower temperature(approximately12 K). After
the sampleis mountedon the tip of the cold-finger, the cell is evacuatedand the
compressoris turned on. The temperatureof the sampleis monitoredby a ther-
mocouplemountednearthe sampleand the temperatureis maintainedby a small
heatingelementlocatednearthe tip of the cold-finger. Becausethesedevicesare
closed-cycle,no liquid N2 is requiredfor cooling. Temperaturesof 12 K can gen-
erally be obtainedafter a 1- to 2-h period. Liquid He cryostatsprovide another
meansof cooling the sample.A more completediscussionof low temperature
FfIR methodsis given by Griffiths and de Haseth (1986).
~
...w
A B
3623 3623
0.7000
~ "/~'
.'
,
: '
:... :
............ .;';
0.6000
<D
90
"c:
«l
.0 3656
'-
5l 0.5000
.0 60
«
30
0.4000
o
0.3000 3550
3550 3750 ....o
Wavenumber (cm- 1) Wavenumber (cm- 1) =
z
Fig. 10-17. Polarizedsingle-crystalFfIR spectraof Ouray dickite in the 3550 to 3750 cm-1 region. Stackplot showingspectrawith no y-offset with absorbancescale ~z
shownon left. (b) Samespectrain which listed polarizationanglescorrespondto angle betweenelectric vector of the transmittedinfrared beamand crystallographic ~
a-axis of the dickite crystal.
g
....
=
3750 3800
'v\tlvenumber
Fig. 10-18. TransmissionFTIR spectraof a disorder,poorly crystalline Georgiakaolinite specimen

as a function of temperature(330, 252, 168, 88, and 8 K).
RamanMicroprobe
A complementarytechnique to FfIR microspectroscopyis the Raman

microprobe.As illustrated in Fig. 10-6, this method combinesa Ramanspec-
trometerwith a high-poweredoptical microscope.ConventionalRamanscatter-
ing is basedupon the excitation and inelastic scatteringphotonsin the visible
portion of the electromagneticspectrum.Thus, IR mirrors (i.e., Cassegrainian
optics) requiredfor FfIR microspectroscopycan be replacedby superioroptical
transmissionmicroscopes.Consequently,the imagequality that can be obtained
using a Ramanmicroprobeinstrumentis greatly superiorto that obtainedusing
a FfIR microscope.Second,Ramanspectroscopyis basedupon light scattering
as opposedto optical transmission.Thus,samplesthat are too thick or opaquefor
FfIR microtransmissionspectroscopycan readily be obtained using a Raman
microprobe.A third benefit is that the lasercan be focusedto an areaof approx-
imately 0.5 pm. This value comparesto the minimum aperturesize of 10 p.m
requiredfor FfIR microspectroscopy.Thus, considerablygreaterspatial resolu-
tion can be obtainedusing Ramanmicroprobemethods.
Despite these advantages,Raman spectra of natural materials can be
severely restrictedby backgroundfluorescencefrom the sample or impurities
that canmaskthe Ramansignal. Another commonproblemis relatedto low sen-
sitivity. A seriouslimitation of Ramanspectroscopyas a surfacetechniqueis the
intrinsic inefficiency of the scatteringprocessitself. For a typical sample,on the
orderof only 1 out of every 108 photonsincident upon the samplewill be Raman
scattered.Thus,a greatexcessof photonsare requiredto produceenoughRaman
scatteredphotons to be detected. With a sensitive detection system such as
chargecoupleddevice (CCD) or an image intensified array detector,only a few
1M
....
"'"
339
270
<6,
~G
i\ 74'
, ~~
".IUIII/ I IV 7'7
.
~\ .\
jl'
I I. "
I II ' I 10"
33.
\" ".,...! 750
\".~,-., " '~""v·,},,·_.• .,,
c
525 1\\ 7'7 /:
270 . '
/III \I
"
"5 ~
2
I~
"
t \ "1
68
V
" "
J I "<. 8'6
-i'-), J' \ "
• I , ,
~j\ I \....
, j J ~,." . . .,'. . .~., . . , ... . . ....,"_~~ B
' 33. "7 \ 7 50
~ '21
~
248 1\ j I 525 ! \ 7" 8.' 7 t\
271 "5 VI I i
f2 ,,_ I ~ d-." ," \~.J '.,",\ , ••
,fA 10 '" '" " ""'~.,.~, A
Lr__-,__-.__-.__-,,-__, -__. -__. -__,-_I D ....

200 400 600 800 1000 1 1
35S0 3600 36120 3~O 36160 36~O 3700 3/20 37i40 g
Z
Wavenumber
Fig. 10-19. Single crystal Ramanspectraof Ouray dickite in the 200 to 1200 cm-1 and 3575 to 3750 cm-1 region, for comparison,the ¥fIR absorbancespectrumof ~
Ouray dickite is shown at the bottom right side of hte OH stretchingregion. PI>
~
tensof milliWatts of laserpower,or less,are requiredto observea Ramanspec-

trum. The actualflux of photonsfocusedonto the small (0.5-,urn)laserspotsize
is quite large. Thus, the high photonflux canprovide enoughenergyto degrade
or alter the sorbedspecies.
Ramanmicroprobemethodshave beenusedto study fluid inclusionsin
rocks (Burruss et al., 1989), identify carbonate,aluminum oxide, and alumi-
nosilicateminerals(Alvarez et al., 1976; Hermanet al., 1987, 1987; Rodgers,
1993), examine interlayer water in muscovite and vermiculite (Haley et al.,
1982), and, more recently, to examinethe single crystal Ramanspectraof clay
minerals.As a prototypical applicationof this technique,single crystal Raman
spectrawere obtainedusing a Ramanmicroprobeinstrumentare shownbelow.
Individual crystalsof an Ouraydickite specimen(10--50,urnacrossthe 001 face)
were mountedon a very thin glassfiber drawn from a glassrod. The fiber was
then mountedon a combinationrotation-translationstagethat permitsthe single
particleto be manipulated(rotatedand translated)while viewing underneaththe
microscope.
The correspondingRamanspectrain the OH stretchingregion are shown
in Fig. 10--19. The intensity of the OH stretchingbandsobservedin the bottom
two spectraare comparableto the spectra obtained using the FfIR microscope,
the top spectrumcorrespondsto a single dickite crystal on edge. The drastic
changein intensityof the OH stretchingbandsresultsfrom the orientationof the
dickite crystal. Of greatersignificancefor soil and subsurfacematerials,this
methodprovidesa meansof identifying individual crystallinephases,amorphous
oxide or organiccoatingson sand-and silt-sizedparticleswith a spatialresolu-
tion of about1 pm.
ACKNOWLEDGMENTS
The authorswould like to thank Walt Farmer,Brian McNeal and Willie

Harris for their critical review of the manuscriptand helpful suggestions.
REFERENCES
Alvarez, R., R.E. Cramer,and I.A. Silva. 1976. Laserramanspectroscopy:A techniquefor studying
adsorptionon aluminumsesquixide,gibbsite.Soil Sci. Soc. Am. J. 40:317-319.
Aochi, Y., W.I. Farmer,and B.L. Sawhney.1992. In-situ investigationof EDB sorption/desorption
processeson clay mineral surfacesby diffuse reflectanceinfraredspectroscopy.Environ. Sci.
Techno!.26:329-335.
Baes,A.U., and P.R. Bloom. 1989. Diffuse reflectanceand transmissionFourier transform infrared
(DRlFl) spectroscopyof humic and fulvic acids. Soil Sci. Soc. Am. J. 53:695-700.
Bell, A.T. 1987. Infrared spectroscopyof high-areacatalytic surfaces.p. 105-134.In J.T. Yatesand
T.E. Madey (ed.) Vibrational spectroscopyof moleculeson surfaces.New York, Plenum
Press.
Berkheiser,V.E., and M.M. Mortland. 1977. Hectorite complexeswith Cu(ll) and Fe(II) - 1,10-
phenanthrolinechelates:Clays Clay Min. 25:105-112.
Bish, D.L., and C.T. Johnston.1993. Rietveld refinementand Fourier transform infrared spectro-
scopicstudy of the dickite structureat low temperature.Clay Clay Min. 41:297-304.
Blanco, c., J. Herrero,S. Mendioroz, and J.A. Paraes.1988. Infrared studiesof surface-acidityand
reversiblefolding in palygorskite.Clays Clay Min. 36:364-368.
316 JOHNSTON & AOCHI
Bloom, P.R., and JA l.eenheer.1989. Vibrational, electronic,and high-energyspectroscopicmeth-

odsfor characterizinghumic substances. p. 411-423.In M.H.B. Hayeset al. (ed.) Humic sub-
stances.Vol. 2. JohnWiley & Sons,New York.
Boyd, SA, andM.M. Mortland. 1985.Dioxin radicalformation and polymerizationon Cu(II)-smec-
tite. Nature(London) 316:532-535.
Brindley, G.W., C. Kao, J.L. Harrison,M. Lipsicas,and R. Raythatha.1986. Relationbetweenstruc-
tural disorderandothercharacteristicsofkaolintesanddickites. ClaysClay Min. 34:239-249.
Brown, A, and E.R. Oark. 1980. Infrared analysisof geologicmaterials.J. Geol. Educ. 29:92-95.
Burba, J.L., and J.L. McAtee. 1977. The orientationand interactionof ethylenediaminecopper(II)
with montmorillonite: Clays Clay Min. 25:113-118.
Burchill, S., M.H.B. Hayes,and OJ. Greenland.1981.Adsorption.p. 284-299.In OJ. Grenlandand
M.H.B. Hayes(ed.) The chemistryof soil processes.JohnWiley & Sons,New York.
Burruss,R.C., T. Ging, and D. Carlson.1989.Ramanmicroprobeobservationsof carbonandoxygen
stableisotopesin geologicmaterials.MicrobeamAnal. 24:173-175.
Buswell, AM., K. Krebs, and W.H. Rodebush.1937. Infrared studies: III. Absorption bands of
hydrogelsbetween2.5 and 3.5 micrometers:1. Am. Chern.Soc. 59:2603-2605.
Calvet, R., and R. Prost.1971.Cation migration into empty octahedralsitesand surfacepropertiesof
clays. OaysOay Min. 19:175-186.
Camazano,M.S., and M.J.S. Martin. 1983. Factorsinfluencing interactionsof organophosphorus
pesticideswith montmorillonite.Geoderma29:107-118.
Cambier, P. 1986. Infrared study of goethites of varying crystallinity and particle size: I.
Interpretationof OH and lattice vibration frequencies.Clay Min. 21:191-200.
Campion,A 1987. Raman spectroscopy. p. 345-417.In J.T. Yatesand T.E. Madey (ed.) Vibrational
spectroscopyof moleculeson surfaces.PlenumPress,New York.
Casal,B., E. Ruiz-Hitzky, 1.M. Serratosa,and J.J. Fripiat. 1984. Vibrational spectraof ammonium
ions in crown-ether-NH4+-montmorillonitecomplexes.J. Chern. Soc. FaradayTrans. One.
80:2225-2232.
Chalmers,J.M., and M.W. Mackenzie. 1988. Solid sampling techniques.p. 105-188. In M.W.
Mackenzie(ed.) Advancesin appliedFourier transforminfrared spectroscopy.JohnWiley &
Sons,Chichester.
Chaussidon,J., and R. Calvet. 1965. Evolution of amine cationsadsorbedon montmorillonite with
dehydrationof the mineral.1 Phys. Chern.69:2265-2268.
Chourabi,B., and J.J.Fripiat. 1981.Determinationof tetrahedral substitutions and interlayersurface
heterogeneity from vibrational spectra of ammonium in smectites. Clays Clay Min.
29:260-268.
Cloos, P., C. Badot, and A Herbillon. 1981. Interlayer formation of humin in smectites.Nature
(London) 289:391-393.
Clothup,N.B., L.H. Daly, andS.E.Wiberley. 1990.Introductionot infraredandRamanspectroscopy.
3rd ed. Acad. Press,Boston. 1990.
Dodgson,J., and W. Whittaker. 1973. The determinationof quartz in respirabledust samplesby
infrared spectrophotometry- I. The potassiumbromide disc method. Ann. Occup. Hyg.
16:373-387.
Doner, H.E., and M.M. Mortland. 1969. Intermolecularinteractionin montmorillonite: NH-CO sys-
tems.Clay Clay Min. 17:265-270.
Drzymala,J. 1985.Chemicalequilibria in the oleic acid-water-NaCIsystem.J. Colloid InterfaceSci.
108:257-263.
Dryzmala,J., and M.M. Kielkowska. 1985. Infrared spectrophotometricanalysisof acidified aque-
ous sodiumoleatesolutions:Spectrochim.Acta, 41(7):949-950.
Durand,B., R. Pelet,and J.J.Fripiat. 1972.A1kylammoniumdecompositionon montmorillonitesur-
facesin an inert atmosphere.Clay Clay Min. 20:21-35.
Farmer,V.C. 1968. Infrared spectroscopyin clay mineral studies.Clay Min. 7:373-387.
Farmer,V.C. 1971.The characterizationof absorptionbandsin clays by infrared spectroscopy.Soil
Sci. 112:62-67.
Farmer,V.C. 1974.The layer silicates.p. 331-359.In v.c. Farmer(ed.)The infrared spectraof min-
erals.Min. Soc.,London.
Farmer,V.C., andJ.D. Russell.1971. Interlayercomplexesin layer sillicates:The structureof water
in lamellerionic solutions.Trans.FaradaySoc. 67:2737-2749.
Fenn, D.B., M.M. Mortland, and TJ. Pinnavaia.1973. The chemisorptionof anisoleon Cu(lI)-hec-
torite. Clay Clay Min. 21:314-322. .
FrIR & RAMAN SPECTROSCOPY 317
Fernandez,M., lM. Serratosa,and W.D. Johns.1970. Perturbationof the stretchingvibration of OH

groupsin phyullosillicatesby the interlayercations.p. 163-167.In lM. Serratosa(ed.) Proc.
Conf. Reunion Hispano-Belgade Minerales de la Arcilla, Madrid, Spain. 1-3 June. Ana!.
Proc. Compt. Rend.
Flehmig, w., and R. Kurze. 1973. Quantitativephaseanalysisby infrared spectroscopyof mineral
mixtures. NeuesJahrb.Min. Abh. 119:101-112.
Foster,R.D., and R.F. Walker. 1984. Quantitativedeterminationof crystallinesilica in respirable-size
dust samples byinfrared spectrophotometry:Analyst (London) 109:1117-1127.
Fraser,DJJ.,and P.R Griffiths. 1990. Effect of scatteringcoefficient on diffuse reflectanceinfrared
spectra.Appl. Spectrosc.44:193-199.
Fripiat,1.1. 1982. Application of far infrared spectroscopyto the study of clay mineralsand zeolites.
p. 191-210.In 1.1. Fripiat (ed.) Developmentsin sedimentology.Vol. 34. Elsevier, New York.
Fuller, E.L., N.R Smyrl, and R.L. Howell. 1985. Chemistry and structureof fuels: Reactionmoni-
toring with diffuse reflectance infrared spectroscopy.p. 225-252. In J. R Durig (ed.)
Chemical,biological and industrial applicationsof infrared spectroscopy.JohnWiley & Sons,
New York.
Fuller, M.P., and P.R Griffiths. 1980. Infrared microsamplingby diffuse reflectanceFourier trans-
form spectrometry.App!. Spectrosc.34:533-539.
Fuller, M.P., I.M. Hamadeh,P.R. Griffiths, and D.E. Lowenhaupt.1982. Diffuse reflectl\nceinfrared
spectrometryof powderedcoals. Fuel 61:529-536.
Gadsden,lA 1975. Infrared spectraof minerals and related inorganic compounds,Butterworths,
England.
GarwoodJr., G.A, and R.A CondrateSr. 1978. The IR spectraof dimethyl sulfoxide adsorbedon
severalcation-substitutedmontmorillonites:Clay Clay Min. 26:273-278.
Gerson, DJ., and C.A Chess. 1988. Polymer composite failure analysis by infrared reflectance
microspectroscopy.p. 73-84. In RG. Messerschmidtand M.A Harthcock (ed.) Practical
spectroscopyseries.Vol. 6. Marcel Dekker, Inc., New York.
Ghosh, S.N. 1978. Infra-red spectraof some selectedminerals, rocks and products.J. Mater. Sci.
13:1877-1886.
Govindaraj,N., M.M. Mortland, and S.A. Boyd. 1987. Single-electron-transfer mechanismof oxida-
tive dechlorination of 4-chloroanisole on copper(II)-smectite. Environ. Sci. Techno!.
21:1119-1123.
Griffiths, P.R., and M.P. Fuller. 1982. Mid infrared spectrometryof powderedsamples.p. 63-129.In
RJ.H. Clark and R.E. Hester (ed.) Advancesin infrared and Raman spectroscopy.Vol. 9.
Heyden,London.
Griffiths, P.R, and J.A de Haseth. 1986. Fourier transform infrared spectrometry.John Wiley &
Sons,New York.
Hair, M.L. 1967. Infrared spectroscopyin surfacechemistry.Dekker, New York.
Haley, L.v., I.W. Wylie, and J.A. Koningstein. 1982. An investigationof the lattice and interlayer
water vibrational spectralregionsof muscoviteand vermiculite using Ramanmicroscopy:A
Ramanmicroscopicstudy of layer silicates.J. RamanSpectrosc.13:203-205.
Hamadeh,I.M., S.A. Yeboah, K.A. Trumbull, and P.R. Griffiths. 1984. Preparationof calibration
standardsfor quantitativediffuse reflectanceinfrared spectrometry.Appl. Spectrosc.38:486.
Harrick, NJ. 1979. Internal reflection spectroscopy.Harrick Sci. Corp., Ossining,NY.
Heffelfinger, R.E. 1971. Determinationof chrysotile in airborne asbestosby an infra-red spectro-
graphictechnique.Atmos. Environ. 5:565.
Hembree,D.M., and H.R Smyrl. 1989. Anomalousdispersioneffects in diffuse reflectanceinfrared
Fourier transformspectroscopy:A study of optical geometries.Appl. Spectrosc.43:267-274.
Herman,R.G., C.E. Bogdan,and AJ. Sommer.1985. Laser Ramanmicrobe study of the identifica-
tion and thermaltransformationsof somecarbonateand aluminosilicateminerals.p. 113-130.
In R.L. Snyderet al. (ed.) Advancesin materialscharacterization.Vo!' 2. PlenumPress,New
York.
Herman,RG., C.E. Bogdan,AJ. Sommer,and D.R. Simpson.1987. Discriminationamongcarbon-
ate minerals by Raman spectroscopy using the laser microprobe. App!. Spectrosc.
41:437-440.
Hirschfeld,T. 1987. Computerizedinfrared: The needfor caution.p. 169-179.In G.L. McClure (ed.)
Computerizedquantitative infrared analysis.ASTM Spec. Publ. 934. ASTM, Philadelphia,
PA
Hlavay, 1., and J. Inczedy. 1985. Pellet preparationfor quantitativedeterminationof inorganic solid
substances by infrared spectroscopy.Spectrochim.Acta 41:783-787.
318 JOHNSTON & AOCHI
Hoffmann, K 1963. Moisture determinationin solids by meansof infrared reflection. Chern. Ingr.
Tech. 35:55--62.
Holmgren, A, and B. Norden. 1988. Characterizationof peat samplesby diffuse reflectanceFTIR
spectroscopy.Appl. Spectros.42:255-262.
Isaacson,PJ., and B.L. Sawhney.1983. Sorption and transformationof phenolson clay surfaces:
Effect of exchangeablecations.Clay Min. 18:253-265.
Ishii, M., T. Shimanouchi,and M. Nakahira.1967. Far-IR absorptionspectraof layer silicates.Inorg.
Chim. Acta 1:387-392.
Jentys,A, G. Warecka,M. Derewinski,and J.A Lercher. 1989. Adsorptionof water on ZSM5 zeo-
lite: J. Phys.Chern. 93:4837-4843.
Johnson,S.A, R-M. Rinkus, T.C. Diebold, and V.A. Maroni. 1988. The use of diffuse reflectance
infrared spectroscopy for studies of molecular sieve catalysis. Appl. Spectrosc.
42:1369-1375.
Johnston,C.T., S.F. Agnew, and D.L. Bish. 1990. Polarizedsingle-crystalFourier-transforminfrared
microscopyof Ouray dickite and Keokuk kaolinite: Clay Clay Min. 38:573-583.
Johnston,c.T., W.M. Davis, C. Erickson,and J. Delfino. 1994. FTIR spectroscopiccharacterization
of humic substances.p. 145-152.In N. Senesiand T.M. Miano (ed.) Humic substancesin the
global environmentand implicationson humanhealth. Elsevier,Amsterdam.
Johnston,C.T., T. Tipton, D.A Stone, C. Erickson, and S.L. Trabue. 1991. Chemisorptionof p-
dimethoxybenzene on Cu-montmorillonite.Langmuir 7:289-296.
Johnston,C.T., T. Tipton, S.L. Trabue,C. Erickson,and D.A Stone.1992a.Vapor phasesorptionof
p-xylene on Co and Cu exchanged SAz-1 montmorillonite. Environ. Sci. Technol.
26:382-390.
Johnston,C.T., G. Sposito,and C. Erickson. 1992b. Vibrational probe studiesof water interactions
with montmorillonite.Clay Clay Min. 40:722-730.
Johnston,C.T., and G. Sposito.1987. Disorderand early sorrow:Progressin the chemicalspeciation
of soil surfllces. p. 89-100. In L.L. Boersma (ed.) Future developmentsin soil science
research.SSSA, Madison,WI.
Johnston,C.T. 1990. Fourier transforminfrared and Ramanspectroscopy.p. 121-155.In D.L. Perry
(ed.) Instrumentalsurfaceanalysisof geologic materials.VCH, New York.
Krishnan, K, and S.L. Hill. 1990. FTIR microsamplingtechniques.p. 104--167.In J.R. Ferraroand
K. Krishnan (ed.) PracticalFourier transform infrared spectroscopy.Industrial and laborato-
ry chemicalanalysis.Acad. Press,Inc. San Diego.
Kung, KH.S., and M. McBride. 1989a.Coordinationcomplexesofp-hydroxybenzoate on Fe oxides.
Clay Clay Min. 37:333-340.
Kung, KH.S., and M. McBride. 1989b.Adsorptionof para-substituted benzoateson iron oxides.Soil
Sci. Soc. Am. J. 53:1673-1678.
Kung, KH.S., and M.B. McBride. 1991. Bonding of chlorophenolson iron and aluminum oxides.
Environ. Sci. Technol. 25:702-707.
Laperche,V. 1991. Etude de I'etat et de la localisationdescationscompensateurs dansles phyllosil-
icates.Ph.D. diss. Univ. Paris.
Larsen,DJ., LJ. Van Doenhoff, and J.V. Crable. 1972. Quantitativedeterminationof quartz in coal
dust by infrared spectroscopy.1. Am. Ind. Hyg. Assoc.33:367-372.
Lewis, D.G., and V.c. Farmer.1986.Infrared absorptionof surfacehydroxyl groupsand lattice vibra-
tions in lepidocrocite (gamma-FeOOH) and boehmite (gamma-AIOOH). Clay Min.
21:93-100.
Long, D.A 1977. Ramanspectroscopy.McGraw-Hili, New York.
Lorprayoon,v., and R.A CondrateSr. 1981. Infrared spectraof sulfolaneadsorbedon cation-substi-
tuted montmorillonites.Clay Clay Min. 29:71-72.
Lorprayoon,v., and R.A CondrateSr. 1982. Infrared spectraof thiopheneadsorbedon H-montmo-
rillonite. App. Spectrosc.36:696--698.
Lorprayoon,v., and RA CondrateSr. 1983. Infrared spectraof thiolane and tetramethylenesulfox-
ide adsorbedon montmorillonite. Clay Clay Min. 31:43-48.
MacCarthy,P., and H.B. Mark. 1975.Infrared studieson humic acid in deuteriumoxide: I. Evaluation
and potentialitiesof the technique.Soil Sci. Soc. Am. Proc. 39:663--668.
MacCarthy,P., and J.A. Rice. 1985. Spectroscopicmethods(other than NMR) for determiningfunc-
tionality in humic substances.p. 527-560.In G.R Aiken et al. (ed.) Humic substances in soil
sediment,and water: Geochemistry,isolation, and characterization.John Wiley & Sons,New
York.
MacCarthy,P., R.W. Klusman,and J.A Rice. 1987. Water analysis.Anal. Chern.59:308R-337R.
Luoma, G.A, L.K Yee, and R Rowland. 1982. Determinationof microgramamountsof asbestosin
mixturesby infrared spectrometry.Anal. Chern.54:2140-2142.
Mangia,A 1975. Determinationof .alpha.-quartzin atmosphericdust. Comparisonbetweeninfrared
spectrometryand x-ray diffraction techniques:Anal. Chern.47:927-929.
Maulhardt, H., and D. Kunath. 1982. Diffuse reflectancespectroscopyin the infrared. Talanta
29:237-241.
McBride, M.B., and L.G. Wesselink. 1988. Chemisorptionof catecholon gibbsite, boehmite,and
noncrystallinealuminasurfaces.Environ. Sci. Technol. 22:703-708.
McClure, G.L. 1987. Computerizedquantitativeinfrared analysis.p. 1-186. In G.L. McClure (ed.)
ASTM Spec.Tech. Publ. 934. ASTM, Philadelphia,PA
Messerschmidt,RG. 1986. A new internal reflection elementdesignfor high optical throughputin
FTIR Appl. Spectrosc.40:632-635.
Messerschmidt,R.G. 1988. Infrared microspectroscopy.p. iii-282. In RG. Messerschmidtand M.A
Harthcock(ed.) Practicalspectroscopyseries.Vol. 6. Marcel Dekker, Inc., New York.
Michaelian,KH. 1986.The Ramanspectrumof kaolinite #9 at 21 deg. C. Can.J. Chern.64:285-289.
Moreale,A, P. Cloos, and C. Badot. 1985. Differential behaviorof Fe(III)- and Cu(II)-montmoril-
lonite with aniline: I. Suspensionswith constantsolid:liquid ratio: Clay Min. 20:29-37.
Morra, MJ., D.B. Marshall, andC.M. Lee. 1989. FTIR analysisof Aldrich humic acid in water using
cylindrical internal reflectance.Commun.Soil Sci. Plant Anal. 20:851--867.
Mortenson,J.L., D.M. Anderson,and J.L. White. 1965. Infrared spectroscopy.p. 743-770.In CA
Black (ed.) Methodsof soil analysis.Part 1. Agron. Monogr. 9. ASA, Madison,WI.
Morterra, c., C. Mirra, and E. Borello. 1984. Ir spectroscopicstudy of pyridine adsorptiononto
alpha-FeOOH(goethite).Mat. Chern.Phys. 10:139-154.
Mortland, M.M. 1970. Clay-organiccomplexesand interactions.Adv. Agron. 22:75-117.
Mortland, M.M., and S.A Boyd. 1989. Polymerization and dechlorination of chloroetheneson
Cu(II)-smectitevia radical-cationintermediates.Environ. Sci. Techno!. 23:223-227.
Mortland, M.M., and TJ. Pinnavaia.1971. Formation of Copper(II) arenecomplexeson the inter-
lamellarsurfacesof montmorillonite. Nature Phys. Sci. 229(3):75-77.
Mortland, M.M., and KV. Raman. 1968. Surface acidities of smectitesin relation to hydration,
exchangeable-cation and structure:Clay Clay Min. 16:393-398.
Motschi, H. 1984. Correlation of EPR parameterswith thermodynamicstability constantsfor
Copper(II)complexes.Cu(II)-EPR as a probefor the surfacecomplexationat the water/oxide
interface.Colloids Surf. 9:333-347.
Mulla, D.J., and P.E Low. 1983. The molar-absorptivityof interparticlewater in clay-watersystems.
J. Colloid InterfaceSci. 95:51--60.
Mulla, D.J., P.E Low, and C.B. Roth. 1985. Measurementof the specific surfacearea of clays by
internal reflectancespectroscopy.Clay Clay Min. 33:391-396.
Nanzyo,M., and Y. Watanabe.1982. Diffuse reflectanceinfrared spectraand ion-adsorptionproper-
ties of the phosphatesurfacecomplexon goethite.Soil Sci. Plant Nutr. 28:359-368.
Nguyen, T.T., LJ. Janik, and M. Raupach.1991. Diffuse reflectanceinfrared Fourier transform
(DRIFT) spectroscopyin soil studies.Aust. J. Soil Res.29:49--67.
Niemeyer,J., Y. Chen,and J.-M. Bollag. 1992. Characterizationof humic acids,composts,and peat
by diffuse reflectance Fourier-transform infrared spectroscopy.Soil Sci. Soc. Am. J.
56:135-140.
Painter,P.C., R.W. Snyder,J. Youtcheff, P.H. Given, H. Gong, and N. Suhr. 1980. Analysis of kaoli-
nite in coal by infrared spectroscopy:Fuel 59:364--366.
Pinnavaia,TJ., and M.M. Mortland. 1971. Interlamellar metal complexeson layer silicates. I.
Copper(II) arenecomplexeson montmorillonite.J. Phys. Chern. 75:3957-3962.
Pironon,J., A Rochdi, O. Barres,A Burneau,P. Landais,M. Pagel,and B. Poty. 1990. Applications
of FTIR microspectroscopyin the earthsciences.p. 281-290.In E.E Vansant(ed.) Proc. Int.
Conf. Workshopin FTIR Spectroscopy,Antwerp, Holland. Univ. Antwerp, Holland.
Prost, R. 1975. Interactionsbetweenadsorbedwater moleculesand the structureof clay minerals:
Hydration mechanismof smectites.p. 351-359.In S.W. Barley (ed.) Proc. Int. Clay Conf.,
Mexico City. Appl. Publ. Ltd., Wilmette, IL.
Prost,R. 1984. Etude par spectroscopieinfrarougea bassetemperaturedesgroupesOH de structure
de la kaolinite, de la dickite et de la nacrite.Agronomie4:403-406.
Prost, R., A Dameme,E. Huard, and J. Driard. 1987. Low temperature(300-5K) IR study of struc-
tural OH groupsof kaolinite, dickite, and nacrite. p. 17-23. In L.G. Schultz(ed.) Proc. Int.
Clay Conf., Denver, CO. 1985. Clay Min. Soc., Boulder, CO.
320 JOHNSTON& AOCHI
Prost,R., A Dameme,E. Huard,J. Driard, andJ.P.Leydecker.1989.Infraredstudy of structuralOH

in kaolinite, dickite, nacrite,and poorly crystalline kaolinite at 5 to 600 K. Clay Clay Min.
37:464-468.
Prost, R., and V. Laperche. 1990. Far-infrared study of potassiumin micas. Clay Clay Min.
38:351-355.
Radulescu-D,N. 1976. Detectionand quantitativedeterminationof quarzin multi-componentmin-
erai mixtures by infrared spectroscpoy(quartz, kaolin, orthoclase).Staub-Reinhalt. Luft.
36:195-201.
Raupach,M., and L.I. Janik. 1988. Polarizedinfrared study of anilinium-vermiculiteintercalateI.
Spectraand models.J. Colloid InterfaceSci. 121:44~65.
Raupach,M., P.F. Barron,andJ.G.Thompson.1987.Nuclearmagneticresonance,infraredand x-ray
powder diffraction study of dimethlsulphoxideand dimethylselenoxideintercalateswith
kaolinite. Clay Clay Min. 35:208-219.
Raupach,M., P.G. Slade,L. Janik,and E.W. Radoslovich.1975.A polarizedinfrared and x-ray study
of lysine-vermiculite.Clay Clay Min. 23:181-186.
Raupach,M., w.w. Emerson,P.G. Slade.1979.The arrangementof paraquatboundby vermiculite
and montmorillonite.J. Colloid InterfaceSci. 69:398-408.
Rein, A, and P. Wilks. 1982. Cylindrical internal reflection cell. Am. Lab. 14:152-154.
Rodgers, K.A. 1993. Routine identification of aluminum hydroxide polymorphs with the laser
Ramanmicroprobe.Clay Min. 28:85-100.
Rouxhet,P.G., N. Samudacheata, H. Jacobs,and o. Anton. 1977. Attribution of the OH stretching
bandsof kaolinite. Clay Min. 12:171-178.
Rozenson,I., and L. Heller-Kallai. 1976. Reductionand oxidation of Fe+3 in dioctahedralsmectites
1: Reductionwith hydrazineand dithionite: Clay Clay Min. 24:271-282.
RuizHitzky, E., and B. Casal.1986. Intracrystallinecomplexationby crown ethersand cryptandsin
clay minerals.p. 179-189.In R. Setton(ed.)Chemicalreactionsin organicandinorganiccon-
strainedsystems.Reidel Publ., New York.
Russell,J.D., M. Cruz,J.L. White, G.W. Bailey, W.R. Payne,J.D. Pope,andl.l. Teasley.1968.Mode
of chemical degradationof s-triazines by montmorillonite: Science (Washington, DC)
160:1340-1342.
Russell,J.D.,v.c. Farmer,and B. Velde. 1970.Replacementof OH by 00 in layersilicatesandiden-
tification of the vibrationsof thesegroupsin infra-red spectra.Min. Mag. 37(292):869-879.
Salle de Chou,J., P.F. Low, and C.B. Roth. 1980.Absorptionof infrared radiationby 0 20 and HOO
mixed with montmorillonite.Clay ClayMin. 28:111-118.
Serratosa,J.M., A. Hidalgo, and J.M. Vinas. 1962. Orientation of OH bonds in kaolinite. Nature
(London) 195:486-487.
Smyrl, N.R., and E.L. Fuller. 1982. Chemistryand structureof coals:Diffuse reflectanceIR Fourier
transformspectroscopy(DRIFT) of air oxidation. p. 133-145.In E.L. Fuller (ed.) Coal and
coal products: Analytical characterizaitontechniques. ACS Symp. Ser. 205. ACS,
Washington,DC.
Smyrl, N.R., and E.L. Fuller. 1987.Diffuse reflectanceinfrared Fouriertransform(DRIFI) spectro-
scopicstudiesof coals: In situ studiesof acetylation.Appl. Spectrosc.41:1023-1028.
Smyrl, N.R., E.L. Fuller, and G.L. Powell. 1983. Monitoring the heterogeneous reactionof lithium
hydride and lithium hydroxidewith waterandcarbondioxide by diffuse reflectanceinfrared
Fouriertransformspectroscopy.Appl. Spectrosc.37:38-44.
Sposito,G. 1986.Distinguishingadsorptionfrom surfaceprecipitation.p. 217-228.In J.A Davis and
K.F. Hayes(ed.) Chemicalprocessesat mineral suifaces.ACS, Washington,DC.
Sposito, G., R. Prost, and J.P. Gaultier. 1983. Infrared spectroscopicstudy of adsorbedwater on
reduced-charge Na/Li montrmorillonites.Clay Clay Min. 31:9-16.
Stoessel,F., J.L. Guth, andR. Wey. 1977.Polymerisationde benzeneen polyparaphenylene dansune
montmorillonitecuivrique. Clay Min. 12:255-259.
Tejedor-Tejedor,M.I., and M.A Anderson.1986. "In situ" attenuatedtotal reflection Fouriertrans-
form infrared studiesof the goethite-aqueous solution interface.Langmuir2:203-210.
Tejedor-Tejedor,M.I., E.C. Yost, and M.A. Anderson.1990.Characterizationof benzoicand pheno-
lic complexesat the goethite/aqueous solution interfaceusing cylindrical internal reflection
Fouriertransforminfrared spectroscopy.Part I. Langmuir6:979-987.
Tennenham,W.M., and R.I.P. Lyon. 1960.Infraredtechniquesin the identificationand measurement
of minerals.Anal. Chern.32:1630-1634.
Tevrucht, M.L.E., and P.R. Griffiths. 1991. Quantitative investigation of matrices for diffuse
reflectanceinfrared Fouriertransformspectrometry.Talanta38:839-841.
Theng,B.K.G. 1974.The chemistryof clay-organicreactions.JohnWiley & Sons,New York.
Tickanen, L.D., M.1. Tejedor-Tejedor, and M.A. Anderson. 1991. Quantitativecharacterizationof

aqueous suspensionsusing attenuatedtotal reflection Fourier transform infrared spec-
troscopy:Influenceof internal reflection element-particle interactions on spectralabsorbance
values.Langmuir 7:451-456.
Tipton, T., e.T. Johnston,S.L. Trabue,e. Erickson, and D.A. Stone. 1993. GravimetricIFTIR appa-
ratus for the study of vapor sorption on clay films. Rev. Sci. Inst. 64:1091-1092.
Turrell, G. 1972. Infrared and Ramanspectraof crystals.Academic Press,London.
Van Der Maas,1.H. 1972. Basic infrared spectroscopy.2nd ed. Heyden& Son LTD, London.
Van Every, K.W., and P.R. Griffiths. 1991. Characterizationof diffuse reflectanceFTIR spectrome-
try for heterogeneous catalyststudies.Appl. Spectrosc.45:347-359.
Vansant, E.F., and J.B. Uytterhoeven.1973. The adsorptionof aromatic, heterocyclic and cyclic
ammoniumcationsby montmorillonite. Clay Min. 1O:61---{)9.
Vry, 1.K., P.E. Brown, and 1. Beauchaine.1988. Analysis of individual fluid inclusions by micro-
FTIR spectroscopy.MicrobeamAnal. 23:201-202.
Weismiller, R.A., J.L. Ahlrichs, and J.L. White. 1967. Infrared studiesof hydroxy-aluminuminter-
layer material. SoilSci. Soc. Am. Proc. 31:459-463.
White, J.L. 1971. Interpretationof infrared spectraof soil minerals: Soil Sci. 112:22-31.
White,1.L. 1977. Preparationof specimensfor infrared analysis.p. 847-864.In 1.B. Dixon and S.B.
Weed (ed.) Minerals in soil environments.SSSA, Madison,WI,
White, 1.L., and e.B. Roth. 1986. Infrared spectrometry.p. 291-326.In A. Klute (ed.) Methodsof
soil analysis.Part 1. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Wilson, E.B., J.e. Decius, and P.e. Cross. 1955. Molecular vibrations: The theory of infrared and
Ramanvibrational spectra.Dover PubI. Inc., New York.
Yang, P.w., H.H. Mantsch, and F. Baudais. 1986. A critical evaluation of three types of diffuse
reflectanceinfrared accessories.Appl. Spectrosc.40:974-978.
Yariv, S., L. Heller, and N. Kaufherr. 1969. Effect of acidity in montmorillonite interlayerson the
sorptionof aniline derivatives.Clay Clay Min. 17:301-308..
Zhang, Z.Z., D.L. Sparks,and N.e. Scrivner. 1990. Acetonitrile and acrylonitrile sorption on mot-
morillonite from binary and ternary aqueoussolutions.Soil Sci. Soc. Am. 1.54:1564-1571.
Published 1996
Chapter 11
Electron Spin (or Paramagnetic)

Resonance Spectroscopy
NICOLA SENESI, Universitti di Bari, Bari, Italy
Electron spin resonance(ESR) spectroscopy,also termedelectronparamagnetic

resonance(EPR) spectroscopy,is a nondestructive,noninvasive,highly sensitive
and accurateanalyticalmethodthat is applicableto the study of chemicalspecies
which have one or more unpairedelectrons.Thesespeciesinclude organic free
radicalsand radical ions, and transition metal ions with incompletelyfilled elec-
tronic d-shells.Suchspecieshave magneticmomentsarising from the spin of the
unpairedelectron(s),with sometimesadditional contributionsfrom orbital elec-
tronic motions, that render them paramagneticand, therefore, able to produce
ESR (or EPR) signalsin both the solid and solution states.
Organic free radicalsare indigenouscomponentsof humic substancesthat
are the most abundantand most chemicallyand biologically active fraction of the
nonliving organic matterin the soil and soil-relatedsystems.Paramagneticmetal
ions are widespreadin the soilsystemin a variety of solubleand insolubleorgan-
ic-associatedand mineral forms which are involved in practically all phenomena
that occur in the soil. Further,the use of ESR labels,traps and metal probesmay
provide information of great importancein many chemical,biological and pho-
tochemical reactions involving organic free radicals in soil and in processes
occurringat the solution-solidinterfaceson soil minerals.
THEORY AND PRINCIPLES
The Resonance Phenomenon
The spin angularmomentumassociatedwith an electronleadsto a circula-

tion of its chargedistribution and henceto the productionof a magneticfield, the
magnitudeof which can be symbolizedby an effective magneticdipole moment,
fie' which is given by
[1]
SegoeRd., Madison,WI 53711,USA. Methods o/Soil Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
323
324 SENESI
where g is a dimensionlessconstantcalled the Lande or spectroscopicsplitting

factor (or magnetogiricratio, i.e., the ratio of the magneticmomentto the angu-
lar momentumpossessedby the electron); Bis the electronic Bohr magneton
(given by eh/41tmc, wheree and m are the chargeand massof the electron),and
S = 1/2 (h/2 1t).
Electron spins are generally paired in atomic and molecular orbitals of
most substanceswhich, therefore, show a net spin and electronic magnetic
momentequal to zero and are nonmagnetic.In contrast,chemicalspeciescon-
taining one or more unpairedelectron(s),such as organicfree radicalsand most
transition metals,show magneticproperties,that is, they are paramagnetic.
In zerofield, the magneticmomentsof unpairedelectronsin a samplehave
randomizeddirections. In the presenceof a magnetic field, H, the electron
momentsare aligned parallel or antiparallelto the field, giving rise to two dis-
crete energy statesof the electron.The basic physical phenomenonunderlying
the ESR experimentis referred to as the Zeemaneffect, that is, the interaction
betweenthe electron magneticmoment,f-le' and an external magneticfield, H,
producingthe splitting of the energylevels of the unpairedelectrons.If the stat-
ic magneticfield of strengthH is applied in the z-direction,i.e., parallel to the z-
axis, the Zeeman interaction leadsto an energyfor an unpairedelectrongiven by
[2]
wheref1z is the electronmagneticmomentin the direction of the conventionalz-

axis, and Mz is the componentof the electron spin angular momentumin the
direction of the applied magneticfield, H (z-axis). The componentof the elec-
tron spin angularmomentummay assumetwo discretevalues,+ 1/2 and -1/2, in
dependence of the two possibleorientationsof the electronspinS in the magnetic
field, that are parallel (high energy)or antiparallel(low energy)to the H direc-
tion. Thus, the two spin statesare not energeticallyequivalent, the difference
(separation)increasinglinearly with the magnitudeof H (Fig. 11-1), according
to
[3]
In a samplecontainingunpairedelectronsin the thermodynamicequilibri-

um in a magneticfield of value Ho, a populationdifferenceexists betweenthe
two energylevels given by the Boltzmannlaw
NJN_ = exp (-g BHoiK T) [4]
whereT is the temperature,K is Boltzmann'sconstant,andN+ andN_ refer to the

populationin the upper(Mz = +1/2) and lower (Mz = -1/2) levels, respectively.
This leadsto an excesspopulationin the lower level. A transitionof the unpaired
electronfrom the stateof lower energyto that of higher energycan be induced
by the oscillating magneticfield componentof an electromagneticradiation of
proper frequency,Yo, applied perpendicularto the static magneticfield H, at a
certain field strengthHo where the energyequationfor resonanceis met
a b
! • +112 I,M
~
~
Z
11,- 9 _ H. - A! >-
~
c.:> 1I\=g /I H.
a:
...
>-
~ I ze:::::::=::::: i
c.:>
CII: ~
...Z ~
... l'"l
! - -1/2 I}M
Absorption
curve ~
~
o o
~
ESR
Signal
t
H.
He
Applied magnetic field ;H Applied magnetic field ;H
Fig. 11-1. Effect of an appliedmagneticfield, H, on the energylevels(E) of the two spin statesof an electron(S =1/2). ESR transition(s)at Vo =gfjHoIh, actualabsorp-
tion curve, and observedfirst-derivative ESR spectrumfor the casesof no nuclearinteraction(a) and interactionwith a nucleushaving I = 3/2 (b), e.g., Cu2+. The
nuclearhyperfine splitting is given by A/g~. ~
U1
326 SENESI
[5]
This is known as the "resonancecondition." Absorptionof radiationenergythus

occursand is detectedas an ESR signal that is recordedas the first derivativeof
the absorption(Fig. 11-1a).
However,a net absorptionof electromagneticradiationand, therefore,sig-
nal detectioncan occur only if the population of the lower level is maintained
greaterthan that of the upper level. Provided the magnitudeof the alternating
field is not too large, this population difference (Le., steady-stateconditions
expressedby Eq. [4D is maintainedby a mechanismof "spin-latticerelaxation."
This processenableselectronsthat have been promoted to the upper level to
return rapidly to the lower level by dissipatingenergyto their surroundings(the
lattice). If the relaxationtimes are long, the high radiation intensity will tend to
equal the numberof spins in both levels, that is, to "saturate"the spin system
(power saturationeffect). This will cause"homogeneous"broadeningof signal
line widths with loss of line resolution and reduction, or loss of net energy
absorption,that is, of signal intensity (seealso later in "Precaution,Sensitivity,
Resolution").
Spectral Parameters
The ESR signal is extremelysensitiveto the natureand conditionsof the

local environmentabout the absorbing electron. The effective magnetic field
experiencedby an electron(Heff) is thus the result of two terms: (i) the applied
magneticfield generatedby the spectrometermagnet(Happl); and (ii) the local
perturbationsof the field producedby the electron'senvironment(Hloc)
[6]
Heff determinesthe separationof the Zeemanenergylevels. This implies that the

resonantfield strength,Ho, that is the position of the ESR signal, and the shape
of the spectrumwill dependon the environmentalconditionsin the vicinity of
the electron. These effects often result in spectral patterns(Fig. 11-1b) more
complicatedthan the single-linespectrum(Fig. 11-1a),thus providing consider-
able information aboutthe molecularstructureof the systemstudied(seelater in
this chapter).
The most importanteffectsoccurringin the spin systemthat influence the
position and pattern of the ESR spectrumare the "electron Zeeman,""nuclear
hyperfine," "ligand superhyperfine,"and "nuclear quadrupole" interactions.
Theseeffects are relatedto a numberof spectralparameters,including the elec-
tronic g- factor (Eq. [1 D, and the nuclearhyperfineand superhyperfinecoupling
constants (see later in sections on "The Hyperfine Splitting and "The
SuperhyperfineSplitting"), that can be determinedin favorable circumstances
(seelater in this text).
A convenientmeansof representingan ESR spectrummathematicallyis
the quantummechanicaloperator"spin Hamiltonian,"which is a sum of a num-
ber of terms involving the previously cited spectralparametersand the electron
ELECTRON SPIN RESONANCESPECTROSCOPY 327
andnuclearspin operators.If a spin Hamiltoniancanbe fitted to the experimen-

tal spectrum,the parametersso obtainedcan be usedto makedeductionsabout
the chemicalandstructuralpropertiesof the sample.A detailedtreatmentof spin
Hamiltonianand a discussionof the physicochemicalmethodsusedto relatethe
obtainedESR parametersto molecularpropertiesof the sampleare outsidethe
scopeof this chapterand can be found elsewhere(amongothers,Carrington&
McLachlan,1967; Abragam& Bleaney,1970; Wertz & Bolton, 1972; Swartzet
aI., 1972; Atherton, 1973; Gordy, 1980).
The g-Factor
The g-factor gives the position of the transition at a given magneticfield

for a particularmicrowavefrequencyandcanbe easilyderivedfrom Eq. [5]. For
a free electrong =2.00232.According to Eq. [6], the resonanceposition of the
electronis shifted from this value by the "electron Zeeman"effect that arises
from the interactionof unpairedelectronswith the externalmagneticfield. The
observedg- factor is thereforedependenton the molecularpropertiesof the para-
magneticspecies.The main sourceof local magneticfields that causedeviation
of theg- factor from the free-electronvalue is the orbital magneticmomentorig-
inating from spin coupling to excitedelectronicstates(spin-orbit coupling).
For manyorganicfree radicalsthe g-valueis closeto 2.00anddoesnot dis-
tinguish well betweendifferent radicals.However,for transition metal ions g is
often characteristicof a particular ion or of a particularvalencestateof an ion.
For most paramagneticspeciesthe spin-orbit coupling is anisotropic,that is,
dependenton the orientationof the molecule relative to the externalmagnetic
field. This implies a g-factor typically anisotropic,i.e., orientationdependent,
and often showing rhombic or axial symmetry. The g-tensor is completely
describedby the threeprincipal components,go;, gyy, andg= alongthe threeaxes
x,y,z (x,y,z-system)that are generallycoincidentwith molecularsymmetryaxes.
An isotropicg -factor (gxx = gyy = gzz) is exhibitedby octahedral,tetrahedral,or
cubic symmetries.Specieswith "axial symmetry,"such as Cu2+ and V4+, have
oneprincipal or threefoldaxis of symmetry,conventionallythe z-axis,andequiv-
alentx- andy-axes.Thesespeciesexhibit two g-values,usually labelledgil (=g=
the g-valueparallel to the symmetry-,or z-axis),andgJ. (= gxx = gyy, the g-value
perpendicularto the z-axis in the x,y-plane).The threeprincipal g-valuesare all
different (gxx"# gyy"# gzz) for paramagneticspeciesthat haveno principal axis of
symmetry.
"Rigid-limit" spectrawith resolvedg~ and gJ. values are systematically
obtainedfor single crystals.However, most natural systems(e.g., soil minerals
and organic colloids) are investigatedin ESR as a "powder" matrix containing
the paramagneticspeciesin randomly orientedparticles.This implies broaden-
ing of the resonancelines and, possibly,loss of information. Nevertheless,with
sufficient anisotropy the principal componentsof the g-tensormay often be
determinedfrom the line shapeof the "powder" ESRspectrum.Anisotropy of
the g-tensoris often averagedfor paramagneticspeciesin solutionby rapid mol-
eculartumbling (faster than the ESR time scale).A single isotropic g-value is
thus exhibited, given by giso' or go = 1/3 (gxx + gyy + gzz). "Rigid-limit" spectra
328 SENESI
with resolvedg, - and g.1. valuescan be, however,obtainedeven in solution for

paramagneticspeciesof high molecularweight that cannotundergorotational
motion rapidly enoughto averagethe g- factor anisotropy(e.g., complexesof
humic substances with Cu2+ or V02+).
The g-values provide information on the nature of the paramagnetic
species,the symmetryof the metal ion in the samplematrix and on its ground
state,that dependon its location in the matrix lattice.
The HypertineSplitting
In additionto the positionof a resonance,anotherextremelyimportantfea-
ture of ESRspectrais the occurrenceof so-called"hyperfinestructure"(or split-
ting). This arisesthroughthe "nuclearhyperfme,"or dipole-dipoleinteractionof
the magneticmoments,fin, of magneticnuclei, which are nuclei with nonzero
spin (I:F- 0) suchas Cu (I =3/2), Mn (I =5/2), and V (I =7/2), with the magnet-
ic momentof the unpairedelectron,Pc. The interactionbetweenthe electronand
nuclearmagnetsis a mutualoneof magnitudedependingon Pc • fin andis, unlike
the positionof the centerof the resonance,independenton the appliedfield, Ho.
Permanentlocal fields arising from magneticnuclei thus alter the value of Heff
and split eachelectronspin level into 2I + 1 components.The interactionof an
unpairedelectronwith a single magneticnucleusthereforeresultsin a set of 2I
+ I equallyspacedhyperfine-structurelines replacingthe singleline resonancein
the ESR spectrum.For example,four lines will appearfor Cu (Fig. 11-lb), six
for Mn, and eight for V.
The splitting of the hyperfine componentsis generally approximatedby
A/gP, whereA is the magnitudeof the nuclearhyperfmeinteraction,the so-called
"hyperfine coupling constant,"arising from dipole-dipoleinteractionsbetween
the electronandits nucleus.The parameterA, like g, alsoexhibitsan orientation-
dependence(i.e., anisotropy).For transition metalsin solution, isotropic hyper-
fine splitting appearsdue to averagingof the anisotropicinteractionby the rapid
thermalrotation of paramagneticspecies.
The valuesof the hyperfmesplitting constantsare very useful to charac-
terize the natureof paramagneticspeciesand molecularsymmetryof transition
metal ions possessingmagneticnuclei.
The hyperfine splitting may be affectedby the presenceof different iso-
topesof the elementthat havedifferent nuclear spins,suchas63euand65Cu. The
effect is the productionof two partially superimposedquadrupletsin the region
gl of the Cu2+ ESR spectrum(Senesiet aI., 1985).
The SuperhypertineSplitting
A superhyperfinestructuremay be observedin an ESRspectrumwhen the
so-called "ligand superhyperfine" interaction arises between the magnetic
momentsof ligand nuclei and that of the unpairedelectronin transition metal-
ligand complexes.This may occur if the ligands have nuclear spin and the
unpaired d-electronsof the metal can be partially delocalizedon the ligand
atoms.
The most commonnucleusgiving rise to superhyperfinestructuresin the
ESRspectrais 14N, which hasa nuclearspin,I =1. Thus,each hyperfmeline is
split into threeapproximatelyequallyspacedcomponentsof equalintensity. Very

complexspectra,often difficult to be analyzed,are obtainedwhen more than one
N-ligand is involved in metal complexation,particularly if the ligands are not
equivalent.
This phenomenon,although resulting in similar apparenteffects on the
ESR spectrum,is differentiatedfrom hyperfine splitting, since in this casethe
electronspin interactswith ligand nuclei other than its own nucleus.
ForbiddenTransitionLines
"Forbidden" transitionscan occur, and related resonancelines appearin
the ESRspectrum,due to "mixing" of hyperfine levelsproducedby the so-called
"nuclear quadrupole" interaction. This arises from the nuclear quadrupole
moment(for example,of the Mn nucleus,I = 5/2) and the electric field gradient.
The interaction,besidesshifting the relative positions of the nuclear hyperfine
levelsand thus alteringthe observedhyperfinesplittings,also"mixes" the hyper-
fine levels, so that "forbidden" transitionscan occur.
Generally,quadrupoleeffects are very small and rarely observedin pow-
der and frozen solution spectra.
THE ESR EXPERIMENT
Instrumentation
A block diagram of a typical ESR spectrometeris shown in Fig. 11-2.

Commonspectrometersoperatein the X-band or Q-band(microwave)frequen-
cy regions of the electromagneticspectrum,correspondingto -9 GHz (wave-
length -3.0 cm) and to 35 GHz (wavelength1.1 cm), respectively.The basic
componentsof the most commonly used X-band ESR spectrometerare: (i) an
electromagnetsupplying a static, or direct current (DC), continuousmagnetic
field that can be varied in strength,usually in the range0 to 1 tesla[1 tesla(T) =
104 Gauss]; (ii) the source of microwaves, usually a vacuum tube oscillator
called Klystron which provides a monochromaticcoherentsource of electro-
magneticradiation with a frequencytypically around9.5 GHz that is held con-
stantto within 1 part in 106; and (iii) a resonantor "microwave" cavity, which is
a hollow- conductingbox generallycuboid with dimensionsdesignedto support
a standingmicrowavepattern,where the sampleis placedand irradiated.
The sourcefrequencycan be swept either side of the centerfrequency by
about ±100 MHz for the purposeof tuning the microwavecircuit and cavity. The
Klystron is coupledby a waveguideto the resonantcavity and locked electroni-
cally to the resonancefrequency.A circulator, incorporatedbetweenthe Klystron
and the cavity, ensuresoptimum transfer of the microwave power from the
Klystron to the cavity and fromthe cavity to the detector.The cavity is interfaced
to the waveguideby a small iris diaphragmwhoseeffective diametercan be var-
ied by the movementof a suitabledevice until all the incident microwavesare
absorbedby the cavity. In this condition, the cavity is said to be "critically cou-
330 SENESI
~~s<:~·1
I~ ,
,U I
I '
L._ . _ . _. ~
I
I
. ---,
I
L._._._._._. _.~
Fig. 11-2. Block diagramof a typical X-band ESR spectrometer(adaptedfrom Thomson,1990).
pled." The spectrometeris usually operatedas closely as possibleto this condi-

tion. The cavity plays an important role in providing the ESR techniquea good
sensitivity. Since magnetic dipole transitions are weak, storageof microwave
energyby the cavityallows the intensity of the radiationfield at the sampleto be
maximized.Further,the cavity designensuresa standingwave patternwherethe
magneticfield componentof the radiation field is maximized and the electric
field componentis minimized at the sample.This avoids dielectric loss by the
samplevia electric dipole absorption,that would lead to loss of sensitivity and
the cavity failing to resonate.
In the practical ESR experiment,the frequencyof the radiation is usually
fixed and the external applied magneticfield is continuouslyswept in order to
bring the Zeeman separation,g~Ho, into resonancewith the energy of the
microwavefield, hvo. Most ESR spectrometersprovide for repetitive sweeping
of the magneticfield through the resonance,allowing the employmentof a com-
puter to averagetransients.Sensitivity is thus increased,since the signals add
proportionally to the numberof sweeps,while the noise addsas the squareroot.
When the resonancecondition (Eq. [5]) is met, the absorption of
microwaveenergyby the samplechangesthe effective coupling of the cavity to
the waveguide,and hencethe amountof microwavepower reflectedby the cav-
ity. This causesan imbalancein the microwavecirculatorarms,and the radiation
is reflected from the cavity to the crystal detector.Since the current variation
therebyinducedin the detectorgives a weak signal, this is greatly increasedby
modulating the intensity of the reflected microwave at low frequency (usually
100 KHz). This is obtainedby superimposingupon the externalmagneticfield

Ho a secondmagneticfield of a few tens of milliTesla which is modulatedplus
and minus at this frequency (100 KHz). Electronic detection of the reflected
microwavebeamis carried out by phase-sensitive detectionlocked to the mag-
netic field modulationfrequency.As a consequence, the output of the detector-
amplifier system,that is, the ESR spectrum,is displayedas the first derivativeof
the absorptionpeak and is usually recordedon a chart recorderas a function of
the applied external magnetic field, H (Fig. 11-1). Occasionally,the second
derivative is plotted to improve the resolutionof closely lying lines.
A detailed description of experimentaltechniquesof ESR spectroscopy
and of the factors affecting ESR spectralsensitivity and resolution,that will be
briefly discussedin the sectionon "Precautions,Sensitivity, Resolution"may be
found elsewhere(amongothersPoole, 1967; Alger, 1968; Ingram, 1969).
SamplePreparation
Samplepreparationfor the ESR measurementis relatively simple, since
the techniquecan analyze samplesin any form, that is, liquid, solid powder,
amorphouspolymer, frozen glass,or single crystal. Generally,solid powders(or,
rarely, single crystals)are packedinto a quartz ESR tube with an inside diame-
ter of a few millimeters. Aqueoussolution or suspensionsamplesare placedin
capillary tubesor specially designedflat cells to reducedielectric absorptionof
microwave radiation by the water molecules.For organic solvents(a dielectric
constantlessthan 10), standardcylindrical sampletubes(an internal diameterof
about3 mm) are used.In any case,tubesor cells madeof highly purequartzwith
no defectsare suitableas samplecontainers,havingthe advantageof no ESRsig-
nal and very low dielectric loss. Clay films also can be used by mounting the
films on flat quartz"tissuecells."
Degassing the sample before ESR analysis is sometimes required to
remove paramagneticO2 moleculeswhich can broadenESR spectraby dipole-
dipole interaction.Dipolar broadeningalso can occur if the concentrationof
paramagneticspeciesin a sampleis too great. In this case,the samplemust be
magneticallydiluted in a diamagneticmatrix. This allows the electronspinsto be
sufficiently isolatedfrom one anotherso as to minimize spin-spincoupling that
may broadenthe spectrum.To prevent line-broadening,paramagneticcenters
concentrationsof 10-2 to 10-3 M are suitable for solids, whereasin solution,
wherespectrallinewidthsare often very narrow, a concentrationnot greaterthan
10-4 M is generallyrecommended.
Precautions,Sensitivity, Resolution
After the samplehasbeenprepared,it must be correctly placedin the cav-
ity, which generallyhasa detectionregion about2 cm high, with the greatestsen-
sitivity at the centerof the cavity. Precautionsmust be taken to avoid signal dis-
tortion and loss of resolutionor sensitivity by improperinstrumentsettings.The
sensitivity of the techniquedependsprimarily upon the numberof spins present
in the sampleplaced in the cavity, but the needfor retaining magneticdilution
may be a limitation. Typically, a modemX-band spectrometer can detect1015 to
332 SENESI
1016 spins,whereasa Q-bandspectrometercan achievea factor of one order of

magnitudehigher. This implies a sensitivity of 10-7 to 10-8 molesat the X-band
and 10-9 molesat the Q-band.In practice,1 mL of solution which is 10-4 to 10-5
M in spins is ideal at the X-band. The sensitivity also dependsupon the cavity
size and thus upon the microwavefrequencyof the source.
Although the signal output of the instrumentis generally proportional to
the squareroot of the microwavepower level, the power cannotalwaysbe set at
maximum. The choice of power is critical as paramagneticspecieswith long
relaxationtimes become"saturated"at high power, with a reductionin intensity,
or loss of signal. Sensitivity is generally improved by setting the modulation
amplitude at higher values, but it should not exceeda fraction of the peak-to-
peak linewidth of the resonance,otherwisedistortion of the spectralline shape
can occur.
Accordingto Curie'slaw, maximumsensitivityfor paramagneticspeciesis
attainedat the lowest possiblesampletemperature.Thus, facilities attachedto
the cavity may be required for temperaturecontrol of the sample.All radical
speciesif sufficiently stablecan be observedreadily at room temperature(RT).
This is the casefor organic free radicalsin humic substances.Rapidly relaxing
paramagneticspecies,such as transition metal ions with d" configurationwhere
n =2 to 8, generallyexhibit an efficient spin-orbit coupling, that is, the spin sys-
temsare efficiently coupledto the lattice, thus producingshortspin-latticerelax-
ation times. The short lifetimeof a given spin stateproducesa broadeningof res-
onancelinewidths that accountsfor ESR spectrabeing too broad to be detected
at RT for sometransition metal ions, such as Fe2+ and Ni 2+. Cooling at liquid N
(bp = 77 K) or liquid He (bp = 4.2 K) temperatureand use of special cooling
devicesfor the sampletubes are often required for transition metal ions to be
observedby ESR. In many systems,however,the environmentof the paramag-
netic speciesmay be alteredby drastic temperaturechanges,therebyinducing a
modification of the ESR spectrum.
The extremelyhigh dielectric loss of water at microwavefrequenciesrep-
resentsa major disadvantageof working at RT in aqueoussystems.For this rea-
son also, most ESR measurements are made at reduced(liquid N or liquid He)
temperatureon frozen aqueoussolutions. Large metal-organicmolecules(e.g.,
mt:tal-humicsubstancecomplexes),however,cannottumble rapidly in solution,
thus the ESR spectrumat RT is similar to that observedfor powdersor frozen
solutions,with the previouslymentioneddisadvantages of lower sensitivity.
The major limitation of an ESR experimentis the resolution, which is
determinedby the width of the componentlines that may overlap to such an
extent that information is lost. The width of a resonanceline is affectedby two
mechanisms,namely, homogeneousand inhomogeneousbroadening.The most
important homogeneousbroadeningmechanismis "spin-lattice relaxation" by
which an electronthat has absorbed energy,and thereforeoccupiesa higherener-
gy level, returnsto the groundstateby losing energyto its environment(Le., the
"lattice"). This is a spontaneous processthat requiresa finite time to occurcalled
"spin-lattice relaxationtime," that is strongly temperaturedependent(seeabove
in this section),and affectsdirectly the width of the resonanceline. Large values
ELECfRON SPIN RESONANCESPECfROSCOPY 333
of spin-lattice relaxation time result in more narrowly defined Zeemanenergy

levels, and transitionsthereforeoccurover a narrower rangeof Happb resultingin
narrowerlinewidths. Obviously, short relaxationtimes will producethe opposite
effect, i.e., the linewidth broadening,as consideredearlier in this section.
Homogeneous broadening also may occur as a consequenceof
"microwave-power saturation" effects that may arise if a sufficiently high
microwavepower is applied. This tendsto equalizethe electronpopulationsof
the two levels, that is, the numberof electronsexcitedfrom the groundstatemay
equal the number that return. In this case,as the microwave power increases,
absorption of microwave energy and, thus, signal intensity decreasesand
linewidth increases.In the absenceof saturation,the ESRsignal intensity should
increaseas the squareroot of the microwavepower.
Inhomogeneousbroadeningarisesfrom nonuniformities in the magnetic
field throughout the sample resulting from other neighboring paramagnetic
speciesor the magneticmomentsof neighboringnuclei, or from dipolar interac-
tions betweenunlike spins. Theseeffects are random in direction and are often
referredto as "spin-spin interactions."The result is a merging of the individual
resonantlines or spin packetsinto a single overall line or envelope,with loss of
line resolution in the hyperfine and superhyperfinestructure,and related infor-
mation.
Two particularcasesof inhomogeneousbroadeningare worth mentioning
here.First, if the paramagneticspeciesare identical, an energyexchange between
the specieswill occur, which shortensthe lifetime of the individual speciesin a
given state.This effect may representa major factor in extremelybroadeningthe
resonancelines and, possibly,nonobservationof ESR spectrafor highly concen-
trated paramagneticmetal ions, such as Cu, rapidly tumbling in solution at RT.
Second,the effect of nearby nuclei with magneticmomentsmay be to in homo-
geneouslybroadenthe resonancelines, so that the hyperfine interaction is not
resolvedand a mergedsingle line is obtained.This effect is often observedfor
organic free radicals in humic substances(see later in the section on
"Interpretation,Examples,and Commentson Electron Spin ResonanceData").
A third form of inhomogeneousbroadening(g-strainbroadening)can arisefrom
local variations in the position of neighboringspecies,which producesmodifi-
cation in the g-factorsof the individual paramagneticspecies,thus affecting the
value of the magneticfield at which the resonanceoccurs.
More sophisticated magnetic resonancetechniques, namely, electron
nucleardouble magneticresonance(ENDOR) and electronspin echo(ESE), can
be used to overcomesome difficulties causedby inhomogeneousbroadening
(seelater in the sectionon "Electron Spin Resonance-Related Spectroscopies").
Physico-MathematicalDeterminationand TheoreticalInterpretation
of SpectralParameters
Once the ESR spectrumhas beenmeasured,valuesof spectralparameters

must be determinedthat can be relatedto chemicaland structuraldetails of the
sample,such as the nature of the paramagneticorganic or metal species,metal
oxidation state,site symmetryand type of metal binding.
334 SENESI
To determine accuratelyand rigorously the ESR (or spin Hamiltonian)

parameters(refer to the sectionon "SpectralParameters"for their physical sig-
nificance),the experimentalspectrumshould be comparedto a computer-simu-
lated spectrumfor the speciesbeing studied. The latter spectrumis calculated
with the use of trial parametersand the expressionsfor the magneticfields at
which transitionsoccur. A new set of trial parametersis thus chosen,the spec-
trum is simulatedagain, and it is then comparedto the experimentalone. This
procedureis repeateduntil satisfactoryagreementis found betweenthe two spec-
tra.
In practice,the elaboratecomputersimulation proceduremay be avoided
by the relativley simple, eventhough not rigorouscomputationof g-factorsand
hyperfine and superhyperfineconstants,directly from experimentaldata. These
data can be accuratelyderived from the experimental ESRspectrumand from
spectrometersettingsusedin the measurement,accordingto standardequations.
These equationswill be describedin details later in this text, separatelyfor
organic free radicals (see section on "Electron Spin ResonanceData Calcula-
tions") and paramagneticmetal ions (see section on "Calculation of Electron
Spin ResonanceParameters").
Oncethe ESR parametersare obtained,they do not give any direct insight
into the structuraland bondingfeaturesof the responsibleparamagneticspecies.
To obtain this, it is necessaryto relateg andA valuest<:> the electronicstructure
and symmetryof the site of the speciesstudied.This implies the useof more or
less elaboratephysicochemicalapproachesbasedon the "crystal field" model
approximation,or "ligand field" and "molecularorbital" calculations.A discus-
sion of thesemethodsis beyondthe scopeof this chapterand can be found else-
where(amongothers,Abragam& Bleaney,1970).
The considerablycomplextaskof performingelaboratedmathematicalcal-
culationsbasedon a rigorous physicochemicalapproachmay be completelyor
partially avoided in some instances.Empirical comparisonand correlation of
experimentalESR spectralparameterswith ESR data referredto in the scientif-
ic literatureon similar model and synthetic systemsor natural compounds(e.g.,
metal proteins,coals,organicpolymers,metalsadsorbedon pureclay, etc.), often
provide the information of chemicaland structuralinterestto the soil scientist.
APPLICATIONS TO SOIL SYSTEMS
Whole Soil
The ESR spectraof soils commonly show a very broad and intenseferro-
magneticresonancepeakcentredat approximatelyg = 2.00 (Vonsovskii, 1966).
Ferromagneticresonancerefers to the phenomenonof microwave radiation
absorptionby strongly magneticmaterialsthat are commonly abundantin soils.
Iron oxides and hydroxides,and mineralsof Mn, Ti and other transition metals
are amongsoil mineralscapableof "ferromagneticresonance"and can produce
ESR spectra.StrongESR signal intensity is exhibited(g - 2) at RT by ferrimag-
netic maghemiteand magnetite(Angel & Vincent, 1978).
A collection of randomly oriented crystals, as well as polycrystalline

solids, producevery broad linewidths due to severalcontributing mechanisms.
The usual mechanismsthat accountfor the observedESR linewidths of para-
magneticspecies,i.e., spin-latticerelaxationand spin-spininteraction(seepre-
viously in sectionon "Precautions,Sensitivity, Resolution"),are only insignifi-
cantly responsiblefor the line broadeningin these systems.In a collection of
powderedparticleswith different shapesand randomorientationrelative to the
appliedmagneticfield, demagnetizingfields andmagneticanisotropyof the indi-
vidual crystallites are the main mechanismsthat contribute to shifting in the
effective magneticfields and the resonancefield position, thereby creating a
range of resonancefield values that broadensignificantly the signal linewidth
(Craik, 1971). High porosity and the presenceof defectsand/orimpurities in the
crystalsfurther increasesignallinewidth.
Electronspin resonancespectraof whole soil can, therefore,provide very
limited chemicaland structuralinformation on soil componentsand properties.
OrganicParamagneticSpeciesin Soil
Organicparamagneticspecies,commonlytermedorganicfree radicals,are
mostly presentin soil as constituentunits indigenousto the molecularstructure
of humic substances(HS). Both humic acid (HA) and fulvic acid (FA), that are
the most importantHS fractionsof soil organicmatter,are relatively rich in free
radicals, representingone of their most peculiar reactive properties(Senesi&
Steelink,1989; Senesi,1990a,b).
ElectronSpin ResonanceSpectra
In the usual experiment,the ESR spectrumof organic free radicals is
obtainedat RT on solid samplesof unfractionatedHS, or more commonly, HA
or FA fractions, or their subfractions,placed in the resonantcavity packedin
columns in suitable ESR quartz tubes. For dissolvedsamples,usually in water
solvent, a specialESR flat cell is used.The magneticfield is sweptover a rela-
tively narrowscanrange(generally5-10 mT), throughthe field at which the free
electronresonates(g =2.00232).Most organicfree radicalsresonatein fact, at a
field (about340 mT) correspondingto g-valuescloseto this number,that is evi-
denceof a "spin-only" magneticmomentobservedfor electronsin s-orbitals(L
= 0). Typical spectrometersettingsand operating functionsusedin the measure-
ment of this spectrumare: microwavefrequencyclose to 9.5 GHz; microwave
attenuation,13dB, correspondingto a microwave power ot about 10 mW, at
which the signal is generallyfound to be least saturated;and modulationampli-
tude, 0.63 mT.
The typical ESR spectrumof humic and fulvic free radicalsis featuredby
a single-line resonance,devoid of any structure(Fig. 11-3a), while a partially
resolvedstructureis rarely observed(Fig. 11-3b).
ElectronSpin ResonanceData Calculations

The analysisof the ESR spectrumprovidesat most three typesof spectral
parametersof importancefor the characterizationof the nature,origin, and prop-
336 SENESI
O.5mT
Fig. 11-3. 1Ypical single-line ESR s~ctrum of soil humic acids and fulvic acids (a) and four-line
spectrumobservedfor someacid-boiledhumic acids(Athertonet aI., 1967).
ertiesof the free radical: the g-value,the width of the absorptionline and, rarely,
the hyperfmestructure.QuantitativeESR also canbe usedto obtain the concen-
tration of unpairedelectrons,that is, of paramagneticcenterspresentin the sam-
ple.
The g-value can be accuratelyapproximatedfrom the magnitudesof the
magneticfield at which resonanceoccursfor the sampleand for a standardof
known g-value, usually N,N-diphenylpycrylhydrazyl(DPPH) diluted in pow-
deredKCI (gDPPH=2.0036).A small amountof standardcontainedin a capillary
tube can be tapedto the sampletube and the signal of the standardis usedas a
field "marker." SinceVo is identicalfor both, the sampleand standard,g is read-
ily calculatedfrom the ratio of field positionsusingthe relationshipderivedfrom
Eq. [5]
[7]
whereu (unknown)andk (known) refer to the sampleand standard,respective-

ly, and Hk can be calculatedfrom Eq. [5] usingVo andgk.
The width of the resonanceline, or line width, MI, is generallymeasured
in teslaor in gauss(1 gauss=10-41),as the peak-to-peakseparationof the first-
derivativeESR signal.
The hyperfmesplitting is measuredas the separation(in gaussor in tesla)
betweenthe hyperfinelines of the spectrumon calibratedchartpaper.A standard
with widely separatedhyperfinelines and accuratelyknown hyperfinesplittings
can be usedto checkthe accuracyof the magneticfield calibration.
Absolutemeasurements of spin concentrationrequireknowledgeof a large
numberof factors,all of which may be considerablyin error or may vary during
the experiment.Spin concentrationis, therefore,usuallyestimatedby comparing
the ESR signal of the samplewith that of a standardchar containinga known
contentof paramegneticcenters,e.g., the "strong pitch" suppliedby the manu-
facturer,or DPPH.Sincethe areaunderthe ESRabsorptioncurveis directly pro-
portional to the numberof paramagneticcenterscontributing to the resonance,
the spin concentrationof the sample(spinsper gram)canbe determinedby com-
ELECfRON SPIN RESONANCE'SPECfROSCOPY 337
paring its signal areawith that of the standard.Becausethe spectrometeroutput

is in the form of a first derivative curve, a double integration should be per-
formed to obtain the area. Since the line shapesof the sampleand standardare
identical (Lorentzian shape), the comparison can be made more simply by
assumingthat the areais proportionalto the productof the height(h) and square
of the width (w) of the first derivative signal. For a valid comparisonof signal
areas,the linewidths and spin concentrationsof sampleand standardshould be
comparable.If hyperfinesplitting of the sampleoccurs,the areaunderthe hyper-
fine lines must be summedto obtain the totalareato be considered.The follow-
ing equationcan then be used to calculatethe spin concentration,in spins per
gram, in the sample
(spin per gram)u = [(spin/pergram)k (hwZ)u (gain)k]/[(hwZ)k (gain)u qu] [8]
where u and k are as above (Eq. [7]), gain is the relative signal amplifier gain
usedand qu is the weight (in grams,moistureand ash free) of the sample.
Becausethe measurement of the spin contentin the sampledependson the
comparisonwith a standard,the procedurerequiresthat the sampleand standard
be measuredin an identical environment,namely, samematrix and samegeom-
etry, equal volumes, same temperature,and same spectrometersettings (e.g.,
samemodulation amplitude) should be used. In the ideal case,the experiment
should be conductedby measuringsimultaneouslyequal volumesof solid sam-
ple and standardplacedtogetherin the cavity. However, in most casesthis is not
possible. Thus, a double cavity instrumentor concentricsampletubes can be
used. Most ESR experiments,however, are performedby using separatetubes
for the sampleand standard,placing them alternativelyin exactly the sameposi-
tion in the resonantcavity. In the caseof solution samples,the choiceof solvent,
pH, and time affect the free radical contentmeasured(Senesi& Steelink, 1989).
Otherfactorsthat affect accuracyin the spin contentmeasurement are powersat-
uration, that must be avoided,and the shapeof the curves,that shouldbe similar
for the sampleand standard(Lorentzianshape).For example,shoulderregions,
that canbe pronouncedwhen associatedwith broadsignals,are difficult to quan-
tify in obtainingthe signal area.
Despitetheselimitations, a reasonablereliability and comparabilitycan be
expectedin the relative changesin spin concentrationsand other ESR parame-
ters measuredin a certain samplesubjectedto a sequenceof variationsin phys-
ical or chemicalfactors, suchas time, pH, temperature,irradiation, redox condi-
tions, acid-hydrolysis,methylation, and interaction with other chemicals.To a
lesser extent,the measurementof g-valuesand linewidths suffersfrom the unre-
liability inherentin its comparisonwith standards.
Interpretation,Examples,and Commentson ElectronSpin

ResonanceData
The g-valuecan provide preliminary information concerningthe chemical

natureof organicfree radicals.The g-valuescommonlymeasuredfor HA andFA
originatedfrom soils and related materialsdo not differ significantly one from
338 SENESI
another,and generally range between2.0030 and 2.0050, thus suggestingthat

free electronsare associatedwith similar structural componentsin HA and FA
moleculesof any source(Senesi& Steelink,1989; Senesi,1990a,b).The g- val-
ues obtainedare consistentwith indigenoussemiquinoneradical units possibly
conjugatedto aromatic rings (g = 2.0041 for 9,1O-anthraquinone),although a
contributionfrom methoxybenzene radicals(g-valuesrange,2.0035-2.0040)and
N- or S-associatedradicals(g = 2.0031 - 2.0037)cannotbe excluded(Blois et
aI., 1961). Lower g-valueswould result from partial delocalizationof the free
electronfrom the 0 atom of the semiquinoneto aromaticC atomsof the conju-
gated aromaticnetwork. A g-value as low as 2.0025 is reportedfor anthracene
(Blois et aI., 1961).
The linewidths of the ESR signal of organicfree radicalsalso do not show
any particular trend in relation to the nature and origin of the HA or FA. They
generallyrange between3.5 and 7.5 x 10-4 T for solid samples,whereaswater
solutionsof HS alwaysexhibit narrowerlinewidths(2.0-3.0x 10-4 T) (Senesi&
Steelink, 1989; Senesi,1990a,b).This effect may be ascribedto rapid tumbling
of moleculesin liquids, great freedomof rotation and low degreeof interaction
with neighboring molecules. Fulvic acids usually show linewidths slightly
greaterthan HA of the samesource.Small differencesin linewidths of ESR sig-
nals may result from a variablenumberof unresolved,superimposednarrow res-
onancesat slightly differing field values,which merge into a single, relatively
broad ESR line. Line broadeningmay thus reflect either a partial delocalization
at variousextentsof the unpairedelectronfrom the semiquinoneonto the conju-
gated aromatic C atoms, and/or unresolvedhyperfine interactionsof the free
electronwith a numberof neighboringaromaticor aliphatic H nuclei. Otherfac-
tors that can influencethe ESR signallinewidthare the concentrationof free rad-
icals and stateof aggregationof the sample,solvent-soluteinteractions,interac-
tions with metal ions, power-saturationeffects,and temperature.
Unfortunately, in most casesthe ESR spectraof humic free radicals are
lacking hyperfine structure(Fig. 11-3a), which rendersimpossibleany further
descriptionof the chemicaland structuralenvironmentaroundthe radical itself.
Somedegreeof hyperfinestructurehasbeenfound, however,in the ESR spectra
of some soil HS (reviewed by Senesi,1990a). For example,a three-line spec-
trum, which was rationalizedin terms of two separateradical species-asemi-
quinone-typeradical and a quinhydrone-type-radical-isreported (Steelink&
Tollin, 1962).A four-line hyperfinestructure(Fig. 11-3b) attributedto the inter-
action of the unpairedelectronwith two nonequivalentH nuclei, was observed
for a numberof acid-boiledHA originatedfrom acid soils and peats(Atherton et
aI., 1967).The triplet spectrumobservedfor an oxidized soil FA was ascribedto
the interactionof the unpairedelectronon the 0 atom of a semiquinone unitwith
two adjacentequivalentH nuclei (Senesiet aI., 1977a).
The concentrationof unpairedelectrons(free radicals)is probablythe most
important piece of data that can be obtainedfrom the ESR spectrumof HS and
the most frequently cited ESRdata,but it is probablythe most difficult and least
accurateof ESR measurements(see previously in section on "Electron Spin
ResonanceData Calculations").The spin contentmeasuredfor soil HA and FA
of various nature and origin varies within more than two ordersof magnitude,
generallyranging between1016 and 1018 spinsper gram. Fulvic acid usually has
one-third to one-fifth the spin contentof HA from the samesource(Senesi&
Steelink,1989; Senesi,1990a,b).Spin concentrationsof FA measuredin solution
at neutral pH are always higher than thosemeasuredin the solid state. Caution
shouldbe exercised,however,in evaluatingand comparingthe spin contentsof
HS, becausetheir valuesare highly dependenton both the instrumentaland envi-
ronmentalconditions usedduring the ESR experimentand the intrinsic matrix
characteristicsof the sample.
FactorsAffecting Spin Contentin Humic Substances

The free radical concentrationin HS is affectedby severalenvironmental
factors and experimentalconditions,including pH, ionic strength,stateof aggre-
gation, hydrolysis, alkylation, redox potential, irradiation, and temperature.
Although relatively little information can be obtainedon the structureand prop-
ertiesof HS relying on absolutevaluesof ESRdata,the measurement of changes
in the ESR dataof HS subjectedto variationsof one or more of the above-men-
tioned experimentalfactors can furnish valuableinsightsinto the molecularfea-
tures and chemical reactivity of HA and FA. This matter has been recently
reviewedin details(Senesi& Steelink, 1989; Senesi,1990a,b).A brief summa-
ry of the resultsobtainedwill be presentedhere.
A pH increase,chemicalreduction,acid hydrolysis, or visible- and ultra-
violet (UV)-light irradiation in either the solid stateor water solution at various
pH values, enhancesthe total organic free radical concentrationof HS, while
leavingalmostunalteredboth the g-valueand the linewidth of the ESRsignal. In
all cases,exceptacid hydrolysis, the observedincreaseis not sustainedin time,
that is, the spin contentreturnsgradually,and almostreversibly,to a value simi-
lar to that beforethe treatment.Theseresults suggest that short-lived,"transient"
free radicalsof substantiallythe samenature of native, stable radicals, can be
producedin HS upon anyof the mentionedtreatments.
Exposureto increasesin temperatureup to 450°C or to increasesin high-
energy(gammarays) irradiationcausesa markedincreasein spin content,broad-
ening of linewidth, and a decreaseof g-value in solid HA. Thesechanges,that
are accompaniedby loss of 0 and increasein C atoms, probably result from
homoJyticbond-cleavageand delocalizationof the free electronfrom the semi-
quinonic 0 atomsto the aromaticC system.
Chemicalor electrochemicaloxidationgenerallyproducesa time- and pH-
dependentdecreasein the free radical content in HA and FA solutions, that is
apparentlyreversedby a reductiontreatment.A decreasein spin contentalso is
observedupon an increasein neutral electrolyte concentrationin FA and HA
solutions at pH close to neutrality. No changesin g-values or linewidths are
observedas a consequence of any of the describedtreatments.
Contradictoryresultshavebeenobtained forthe effect of alkylation on the
free radical contentof HS. The spin contentof podzolicsoil HA and FA decreas-
es dramatically upon methylationwhereasan increasein spin contenthas been
measuredfor a wide selectionof other soil HA and FA.
340 SENESI
StructuralImplicationsof ElectronSpin ResonanceResults

Freeradical concentrationof HS can be usefully correlatedto otherchem-
ical and structuraldata. In particular, itis correlatedpositively to the EJE6ratio
(ratio of absorbances at 465 and 665 nm), absorbanceat 465 nm, atomic C/H and
O/C ratios, 0 percentage,phenolicOH content,and IR absorptionsof aromatic
structures,and negativelyto H percentageand aliphatic IR absorption(reviewed
by Senesi,1990a).This providesevidencethat the free radical contentis direct-
ly relatedto the dark color and degreeof aromaticity,molecularcomplexity and
particle size of HS. A good correlationalso appearsto exist betweenfree radical
concentrationand degreeof humification in peats(reviewedby Senesi,1990a).
No specific correlationhas beenvalidated,however,betweenspin content and
type, natureor origin of HA or FA.
AccumulatedESR data strongly supporta quinone-hydroquinonesystem
indigenousto the HS moleculeasthe principal responsiblefor the generationand
maintenanceof semiquinoneradicalsin HS. All of the ESR propertiesof HS are
consistentwith this model that can be simply representedby the fundamental
reversiblemechanismbasedon an electron donor-acceptor(or charge-transfer)
systeminvolving the quinone(Q)-hydroquinone(H2Q) moieties(i.e., a quinhy-
drone system),coexistentwith semiquinoneradicals (HQ') and radical anions
(Q~) (Senesi,1990a)
2H+
Q + H2Q .... 2 HQ' .... 2Q'- [9]
20H-
A quinonecontentof 0.5 mmole per gram observedfor some HA would

theoretically yield up to 6 X 1020 spins per gram of semiquinoneanion in the
presenceof a baseand a suitable donor group. This will increasesubstantially,
from two to five ordersof magnitude,the relatively low amountof stablepara-
magneticcenters (10 16_1018 spins pergram) in HS, with obviousrelevantimpli-
cationson their chemicaland biochemicalreactivity.
Semiquinonesalso can be generatedby the partial reductionof quinonesor
photoirradiationof quinones in solution, in the presenceof proton (electron)
donors.The large decreasein spin contentobservedin podzolic HA and FA as a
consequence of selectiveblocking of phenolic OH groupsby methylation,con-
firms thesegroupsas the most importantelectrondonorsresponsiblefor the for-
mation and existenceof free radicalsin HS.
Interactionwith OrganicChemicalsand Metal Ions

Electron spin resonancespectroscopycan be applied to the study of the
interactionof HS with organic chemicals,such as pesticides,for evaluatingthe
role of organic free radicals in binding mechanismsinvolving free radical
species,such as charge-transfer(electron donor-acceptor)processesand cross-
coupling reactions.
The free radical contentof model electrondonor-acceptorsystemsis gen-
erally directly related to the tendencyof these systemsto form and exist. The
increasesin free radical concentrationmeasuredin the interaction products

betweenHA ands-triazineor substitutedureaherbicidesthus providedevidence
of the occurrenceof an electron donor-acceptormechanismin the interaction,
with the formation of a chargetransfercomplex.
In the caseof chlorophenoxyalkanoicherbicides,the productsof interac-
tion with HA showeda considerabledecreasein free radical concentration.This
suggestedthe occurrenceof homolytic cross-couplingreactionsbetweennative
humic free radicalsand free radical intermediatespossiblygeneratedin the pre-
liminary chemical,photochemical,and/orbiological degradationof the herbicide
molecule.This matterhasbeenrecently reviewedin detail by Senesi(1990a).
Electron spin resonancespectroscopyalso can be usedto study the effect
of interactionof metal ions with HS or. free radicals.A recentreview has been
provided on this matter (Senesi,1990a). Addition of several transition metal
ions, including Fe3+, Cu2+, and Mn2+ to solutionsof HA or FA had the effect of
decreasingthe free radical content. In studiesof podzolizationprocesses,ESR
data suggestedthat recently formed low-molecularweight HS cannot undergo
further polymerizationprocessesbecausetheir free radicalscombinewith Fe3+
and move to the Bh horizon. In anotherstudy, ESR spectroscopyrevealedthe
formation of semiquinoneradicalsby single electrontransfersoccurringat iron
oxide, e.g., goethite, and manganeseoxide surfacesduring the oxidation of
hydroquinone.Electronspin resonancespectroscopyhasbeenusedto character-
ize the natureof HS formed throughthe synthesis(oxidative polymerization)of
phenolicand other organiccompoundsin the presenceof naturalclays,soils, soil
oxides,and other metal ion catalysts.
ParamagneticTransitionMetal Ion Complexeswith Soil Organic

and Mineral Materials
Electronspin resonancespectroscopyis an importantmethodfor detecting

the presenceof paramagnetictransition metal ions in soil organic and mineral
colloids and for investigatingtheir structuresand properties.All metal ions with
an odd numberof unpairedelectronsshould give ESR spectra.Further, since a
transitionmetal ion can usually be broughtinto a variety of oxidation levels, it is
often possibleto arrangeconditions to elicit an ESR signal from a metal ion.
Electron spin resonanceanalysiscan provide useful and, in somecases,unique
informationon the presenceand identity of a given type of metal ion and its oxi-
dationstateandpossiblechanges,on the environmentof the metalbinding site(s)
(including ligand type, coordinationand symmetry),and degreeof stability of
metal complexes.
The ESR techniquecan be successfullyapplied to the study of structural
metal ions, such as Fe, in soil mineralsand, especially,of complexesformed by
soil organic matter fractions-includingHA, FA, and plant litter-with para-
magneticmetal ions of importanceto agricultureand to the environment,name-
ly, Fe, Cu, Mn, and V. This matter has been reviewed in detail by Senesi
(1990a,b).
It shouldbe noted,however,that the absenceof an ESRsignal in a sample
doesnot prove the absenceof paramagneticspecies.Thereare a numberof con-
342 SENESI
ditions underwhich it is not possibleto obtain an ESRsignal. Relaxationeffects

causedby power saturation or other causes(see section on "Precautions,
Sensitivity, Resolution") can preventthe observationof a spectrum.The ESR
signalis sometimesnot detectablein metalionswith an evennumberof unpaired
electronsbecausezero field-splitting arising from low symmetrycrystal fields
can move the resonancetransitionout of the frequencyrangeof most spectrom-
eters.If the metalions occurin pairs,thenthe ESRsignalsareoften undetectable
unlessthe pair of metal ions togetherpossessan odd numberof electrons.
Electron Spin ResonanceSpectra

The ESR spectraof metal complexeswith soil organicor mineral compo-
nentsare usually obtainedat either RT on solid powdersamples,or at liquid N
temperature(77 K) on either solid powdersor frozen (77 K) solution samples,
placed in suitable ESR quartz tubes. Usually, the magnetic field is initially
scannedover a wide range(from 0-1 1), then an enlargedspectrumis recorded
over a narrower field (0.2 or 0.1 1) comprising the region where resonances
appear,to allow for their detailedanalysisand interpretation.Measurementcon-
ditions and operatingfunctionscommonlyemployedare: microwavefrequency,
eitherapproximately9.2 GHz (spectraat 77 K) or approximately9.8 GHz(spec-
tra at R1); microwaveattenuation,13 dB (microwavepoweroscillatesbetween
9.0 and 9.2 mW); and modulationamplitude(0.63-4m1), accordingto the type
of metal ion being measured.
Four examplesof typical ESR spectraobtainedfor metal-organiccom-
plexes naturally occurring in isolatedsoil organic fractions are shown in Fig.
11-4 and 11-5.
Calculationof ElectronSpin ResonanceParameters

Threetypesof spectralparameterscan be obtainedfrom analysisof para-
magneticmetal spectra:(i) the g-value(s)of the metal(s)presentin the sample;
(ii) the hyperfinecoupling constant,A, if the metal nucleushas a nonzerospin
and a nuclearhyperfinestructureis apparent;and (iii) the ligand superhyperfine
splitting, if the unpairedelectronof the metalion is delocalizedby partial cova-
lence onto magnetic nuclei of surroundingligands and if the related ligand
nuclearsuperhyperfinestructureis observed.Rarely are resonancelines due to
"forbidden transitions"observed,e.g., for Mn2+.
The direct computationof g-valuesandhyperfmeand superhyperfinecon-
stants,A, canbe determinedfrom the experimentalspectrumdataandfrom spec-
trometersettingvaluesusedin the measurement, accordingto the standardequa-
tions (adaptedby Wertz & Bolton, 1972)
g =hvolPHo =0.714484vofHo [10]
whereVo (MHz) is the accuratelymeasuredvalueof the microwavefrequencyat

the resonancecondition,Ho is the value of the magneticfield at which the reso-
nanceis centered(on calibratedchart paper),h is the Planck'sconstantand Pis
the Bohr magneton;and
\
\
Free Radical
vo+/ Mn 2+
/
Free Radical
Fig. 11--4. Representativewide scanrange(800 m1) ESR spectraat 77 K of a Mollisol humic acid
from IHSS Referenceand Standardcollection (a; a', higher gain) (Senesiet aI., 1989a),a Paleosol
humic acid (b) (Senesi& Calderoni,1988), a loam soil humic acid (c) (Senesiet aI., 1989b),and
a decomposingleaf litter aqueousextractfrom a forest soil (d) (Spositoet aI., 1988).
A(cm-l) = A(MHz)/c = 2.80247(a g)/(c ge) = 0.46976610-

4 ag [11]
wherege (=2.00232)is the g-valuefor the spin of the free electron,g is calculat-
ed by Eq. [10], a is the hyperfinesplitting measuredas the peak-to-peaksepara-
tion (in 10-4 T) betweenthe hyperfinelines in the experimentalspectrumrecord-
ed on calibratedchart paper,and c is the speedof light in a vacuum.Accuracyof
magneticfield calibration of chart papershould be checkedby using a suitable
standard.
Examples,Interpretationand Comments
Ferric Iron. Electronspin resonancespectraof HA, FA, and decomposing
litter layersgenerallyexhibit an asymmetrical,isotropic resonanceline at about
g = 4.2 consistentwith high spin (five unpairedd-electrons)Fe3+ held in tetra-
hedral or octahedralsites of low-symmetry (rhombic) ligand field (Fig. 11-4)
(reviewedby Senesi,1990a,b). TheESR signaloftheseFe3+ complexesdoesnot
specify the chemical nature of the ligands, althoughg-valuesnear 4.2 are con-
sistentwith Fe3+ complexeswith 0 functional groups,possibly carboxylic acids
and/or polyphenols.This form of Fe exhibits considerableresistanceto proton
and metal exchange,and to chemical reduction, thus suggestingthat Fe34 is
strongly bound and protectedin inner-spherecomplexesin HS. A signal at g =
4.3, attributableto structural Fe3+, has been observedin layer silicatessuch as
344 SENESI
b
1+
Cu
IT~
13O mT
L-..J
IOmT
.,
: Iii! :', I',
Fig. 11-5. SameESR spectra asin Fig. 11-4b,c,d,recordedon an enlargedscanrange(200 mI).
kaolinites,vermiculites,micasand smectites(McBride et aI., 1975). This signal,

generallycomposedof an anisotropicand an isotropic g-factor, is suggestedto
arisefrom octahedralFe04(OH)zgroups.
A very broad signal centred near g = 2 is often exhibited by HS (Fig.
11-4a')(reviewedby Senesi,1990a,b).Very probably,this resonancearisesfrom
extendedspin-spin coupling of various neighboring paramagneticmetal ions
most likely Fe3+, absorbingin this region. Iron in suchsitesis easily reducedby
chemicalagentsand easily extractedby complexingagents,thus suggestingthat
it is weakly boundon externalsurfacesof HS. Soil clays,vermiculites(Le., nat-
Fig. 11-5. Continued.
urally weatheredmicas), and weatheredphlogopite often demonstratea strong

ferrimagneticresonanceat g =2 that tend to obscureother spectraldetails and
might be attributed to ferrimagneticclustersof structural Fe3+ and/or to ferric
oxides (McBride, 1986).
Two weak resonancesat low-field positions (near g = 9 and g = 6) are
sometimesobservedin ESR spectraof HS (reviewedby Senesi,1990a,b).The
former resonanceprobablyarisesfrom Fe3+ in siteswith nearorthorhombicsym-
metry, and the latter from high-spin Fe3+ in largely distorted,axially symmetric
crystal fields.
No direct ESR evidenceof Fe2+ specieshasbeenobtained forsoil compo-
nents.
Divalent Copper. In addition to the organic free radical and Fe3+ reso-
nancesdiscussedearlier, soil organic materials often exhibit an anisotropic,
rigid-limit spectrumof the "axial" type in the g = 2 region, with resolvedgil and
gl. componentsof the g-tensor(describedin the sectionon "The g-factor") (Fig.
11-4b,c,and ll-Sb,c). This patternis ascribedto the presenceof Cu2+ (reviewed
by Senesi,1990a,b).Since the nuclearspin of both Cu isotopes,63CU (69.2%)
and 65CU (30.8%), has a value of I = 3/2, the ESR spectrumshould be split into
four (Le., 21 + 1) featuresboth at g~ and gl.. However,only the componentat g~
is generallyresolvedpartially into a quadruplet,while the splitting of the gl. com-
ponentis too small to be resolved.
The ESR parametersof Cu2+-HS complexes,either naturally occurring or
obtainedsyntheticallyby Cu2+ ion-doping of HS samples,are in any casecon-
sistentwith a dx 2 _ y2 groundstatefor Cu2+ held in inner-spherecomplexesin the
HS matrix, with ligands arrangedin a squareplanar(distortedoctahedral)coor-
dination site (tetragonalsymmetry).The experimentalvaluesof theseparameters
346 SENESI
generallyindicatebinding sitesfor Cu2+ in HS involving eitheronly 0 function-

al groups (carboxyls, phenolic hydroxyls, carbonyls and, often, water mole-
cules),or both 0 and N ligands,or evenonly N atoms(i.e., a tetraporfyrinsite)
(reviewedby Senesi,1990a,b).The participationof N in the binding of Cu2+ by
HS also is supportedby the resolvedpatternobservedin somecasesat g.L' deriv-
ing from superhyperfmecouplingof the Cu unpairedelectronto N ligand nuclei
(I =1).
The measurt<dspectralparametersalso provideevidenceof a high covalent
bond contribution (i.e., delocalizationof the unpairedelectron toward the lig-
ands) for Cu2+ in HS. The combinedeffects of the crystal field splittings and
covalencylead to the following empirical orderof decreasinggl- and g.L-fac-
tors for different donor atoms:0 > N > S. Similarly, the valuesof the hyperfine
coupling constants,All andA.L' that are relateddirectly to the degreeof covalen-
cy and g- factors, increaseand, obviously, their absolutevalues, 1A111 and IA.LI,
decreasewith increasingelectronegativityof the ligand atoms (0 < N < S)
(Jameson,1981). However, small deviations from squareplanar coordination
will shift All to smallervalues,while largerdeviationsalso will shift gil to small-
er values(closerto g =2) (Peisach& Blumberg,1974).A goodcorrelationexists
betweenthe gll- andAll-values and eitherthe Cu2+ loadingor the coordination
environmentof model and naturalorganics(McBride, 1989). There is evidence
that covalent bonding, as indicat~d by low g-valuesand large hyperfme con-
stants,is favoredby low Cu2+ loading in HS, wherecomplexationto amine-type
N groupsis preferredto O-containingligands. In contrast,ESR parametersmea-
suredfor Cu2+ at high loadingsin HS indicatenot very covalent,althoughrigid,
binding of Cu2+, largely to O-containingligands,probablyas inner-spherecom-
plexeswith a higher degreeof mobility of Cu2+.
An accurateanalysisof the Cu2+ ESR patternat gil' possiblywith the aid of
a computersimulatedspectrum,may reveal that in somecasesit consistsof two
(or more) superimposedquadruplets,often very difficult to be differentiated,
eachcomponentarising from a different type of local environmentfor Cu2+. This
is evidenceof different classesof binding sites for Cu2+ in HS (reviewed by
Senesi,1990a,b).
Vanadyl Ion. More complicated, but relatively well-resolved spectra

exhibiting a richly structuredpatternat aboutg =2 are observedfor somenatur-
al HA or FA and/or their fractions (Fig. 11-4c) (reviewedby Senesi,1990a,b).
Analysis madeon an enlargedspectrumof the region (Fig. 11-5c) indicatesthe
presenceof two distinct, overlappingrigid-limit spectraof the "axial" type. One
comprises the typical anisotropic pattern of complexed Cu2+ previously
observed,the other consistsof the superimpositionof two hyperfine octuplets,
correspondingto the parallelandperpendicularcomponentsofV4+ (nuclearspin,
I = 7/2). The complexity of the spectrummay require computersimulation for
the accuratedeterminationof the ESR parameters,which are consistentwith a
V02+ rigidly boundin HS as an inner-spherecomplexin a squareplanarcoordi-
nation site.
Electronspin resonancedataprovide information on different ligand bind-
ing sitesavailableto V02+ in HS. Vanadyl complexesnaturally occurring insoil
HS are characterizedby relatively strongligand fields and high covalency,being

consistentwith phenolateor possibly N ligands.In contrast,complexesobtained
by V02+ -doping of HS involve weakerligand fields and lower covalencyindi-
cating that V02+ is primarily bound to surface carboxylate groups while it
remainspartially hydrated,thus resulting in relatively labile and exchangeable
forms.
Electron spin resonancedata also indicatedthat catecholgroupsare prob-
ably mainly responsiblefor the reductionof the metavanadateion, V03-, which
is the most stable form of V under typical soil conditions, to V02+ by HS in
acidic media. Further, ESR evidencehas beenobtainedfor the complexationof
the obtainedV02+ in forms that are protectedfrom oxidationat very high pH val-
ues, at which this speciesis thermodynamicallyunstable(reviewed by Senesi,
1990a,b).
Divalent Manganese.The ESR spectraobtainedfor decomposinglitter
samplesof different origins (Fig. 11-4d, and 11-5d) and someHA and fraction-
ated FA feature a well-resolvedisotropic patternconsistingof 6 almost equally
spacedprincipal lines and, possibly, 10 secondarylines (correspondingto for-
bidden transitions)of lesserintensity (reviewedby Senesi,1990a,b).The ESR
parametersof suchspectraare consistentwith high-spin hexahydratedMn 2+ (I =
5/2) in outer-spherecomplexes,bound by electrostaticforces to six 0 atomsof
negatively chargedcarboxylateand phenolategroups in a distorted octahedral
environment.
Spin Derivatizationin Soil Chemistry
Spin Labels
The spin-labelingtechnique,that is the attachmentof a simple and stable
paramagneticspeciessuch as the nitroxide radical to a substance,is a powerful
ESR-basedmethodfor evaluatingthe dynamicsof macromoleculessuch as HS
in solution (Swartz et aI., 1972; Berliner, 1976). Analysis of the line shapesof
the ESRspectrumof a spin-labeledhumic polymer may revealthe rotationalcor-
relation times of the macromolecule,and the types of molecularmotion in solu-
tion. These may provide, in tum, unique information on a number of related
propertiesof HS, including the state of aggregation,moleculemorphology and
conformation,relaxation times, and micellar character.Spin labels also can be
usedto investigateby ESR the anisotropicmotion on mineral surfaces(Berliner,
1976), thus providing information on the natureof surfaceadsorptionprocesses
not obtainableby other methods.The techniqueis sensitive,permitting adsorp-
tion of small quantitiesof the spin label to be studied,and allows fully wet sys-
tems to be analyzed.
An interestingillustration of this techniqueis the use of two organicanion-
ic nitroxide spin labelsprovidedwith eithera carboxylateor an organophosphate
functional group, to study the nature of bonding to noncrystalline alumina,
boehmiteand gibbsitein aqueoussuspensions (McBride, 1982a).Analysis of the
ESR spectraof theseradical speciesrevealedthat both are adsorbedrapidly onto
the high-surfaceareaalumina and boehmite,whereasonly the organophosphate
348 SENESI
is adsorbedon gibbsite.The rotationalcorrelationtimescalculatedfrom the ESR

spectraindicated that a loss in rotational motion of the radicals accompanied
adsorption,with the greatermotional restriction observedfor the carboxylate.
The degreeof motional restrictionof the organic radical anions on the fully
hydratedsurfacessuggestedthatboehmiteadsorbsthe carboxylatelargely by lig-
and exchangeof a single surfaceOH, whereasthe organophosphate probably
forms a bidentatebond. The moderateloss in rotational motion and easeof
exchangeabilityof the carboxylateboundon non-crystallinealumina suggested
weak bondingby nonspecificelectrostaticforces in addition to ligand exchange
of surfaceOH.
Spin Traps
The techniqueof spin trapping involves reaction of a short-lived radical
with an exogenouscompoundto producea relatively stable radical(Thomson,
1990).A goodspin trap shouldreactrapidly with radicalsto form aproducthav-
ing an ESRspectrumthat canthen be usedto identify unambiguouslyand quan-
tify the initial short-livedradical trapped.Unfortunately,no spin trap availableat
presentcancompletely satisfythesecriteria. l\vo groupsof compoundsare most
useful, namely nitrosoarenesor nitroalkanes,such as nitrosobenzeneor 2-
methyl-2-nitrosopropene,and nitrones, such as 5,5-dimethylpyrroline1-oxide
(DMPO) and phenyl-N-t-butylnitrone(PBN).
The spin-trappingtechniqueis, therefore,suitedto providestructuralinfor-
mation on the transientfree radical speciesproducedin HS by light irradiation,
reduction,or raising the pH (seesectionon "FactorsAffecting Spin Contentin
Humic Substances").This information could be obtainedby examiningthe ESR
spectrumof the radical trapped.So far, no studiesare availableon the applica-
tion of this techniqueto HS.
Metal Spin Probes

Electron spin resonancespectroscopyis a powerful tool for obtaining
information about the molecular structure and dynamics of the paramagnetic
metal probestructureand dynamicsof the paramagneticmetal probeinteracting
with a surface.Lineshape,linewidth, and the g-value allow one to distinguish
betweena complex freely tumbling in solution and an immobilized adsorbed
species.
Humic SubstancesComplexationChemistry. Electron spin resonance
studieson synthetic metal complexespreparedby addition of a paramagnetic
metal "probe," e.g., Cu2+, Mn2+, and V02+, to natural soil organic components
can provide further information of molecular and quantitative value on the
"residual" complexingcapacityof thesesystemstoward metal ions and on the
stability of formed complexesunder physical and chemicaltreatmentssuch as
proton and metal ion exchangereactions(reviewed by Senesi,1990a,b).For
example,whenFA are dopedwith 5.5 to 50.1% Fe3+, the g =2 signalis enhanced
relative to the g = 4 signal, suggestingthat additional Fe3+ are preferentially
bound to surfacesites by FA (Senesiet aI., 1977b). Evidenceof inner-sphere
complexesof Mn 2+ coordinatedoctahedrally,possibly with carboxyl, phenolic

hydroxyl and/or carbonyl, has been obtained for some peat HA and soil HA
doped with Mn 2+ at high pH (>8) or high temperature(>50°C) (Lakatos et aI.,
1977; McBride, 1982b).
QuantitativeESR spectroscopyhas beenapplied to measurethe dynamics
of motion of FA-V02+ chelatesin solution and to obtain information on their
molecularconformationand aggregationproperties.Rotationalcorrelationtimes,
that are a measureof the rate of moleculartumbling in solution, were obtained
by the analysisof ESR spectrallinewidthsand combinedwith gel filtration chro-
matographydata to assessthe approximatemolecular weights of FA fractions
and stoichiometry of V02+ complexes formed by these FA (Templeton &
Chasteen,1980).
QuantitativeESR spectroscopyalso has beenusedto determineweighted
averageequilibrium constants(Kc) for watersolubleMn 2+-FA complexes,which
were in excellent agreementwith Kc values determinedby an ion-exchange
method(Gambleet aI., 1977). The ESR methodwas found more sensitive,con-
venient,and faster than the ion exchangeprocedure.
Ion Exchangeon Layer Silicates.Electron spin resonancespectroscopy
confirmed that divalent and trivalent transition metal ions such as Cu2+, Mn2+,
V02+, and cr3+ retain their inner hydration sphereas well as a high degreeof
rotational mobility on exchangesites of layer silicate clays, offering direct sup-
port for the involvementof nonspecificelectrostaticforces only and for typical
ion exchangebehaviorin the absenceof metal hydrolysis(reviewedby McBride,
1989). For example,ESR spectraof exchangeableCu2+ and Mn 2+ on kaolinite
indicate that planar Cu(H20)l+ are oriented parallel to the surfaces(McBride,
1976). At high relative humidity, solutionlike spectraare observed,suggestinga
high degreeof mobility of Cu2+ and Mn2+ exchangecationson kaolinite surfaces.
However, if the interlamellarspacingon the clay is limited to the equiva-
lent of one or two molecularlayers of water, a high degreeof motional restric-
tion of the hydratedmetal ions is observed.This hasbeenelegantlyillustratedby
an ESR study of Cu2+ adsorbedon hectorite, an expanding2:1 clay mineral
(McBride, 1986).When thisclay is air dried, an orientation-dependent rigid-limit
spectrumof Cu2+ is obtained,and the gil and go- componentsof the g-tensorare
resolved.The gil componentis split into four hyperfine lines, while the splitting
of the go-componentis too small to be resolvedin this spectrum,as a result of
the pronouncedanisotropyof the hyperfinesplitting. Evidently, the z-axis of the
planarCu(H20)g+ then tumblesrapidly enoughto completelyaveragethe g- ten-
sor and the hyperfine splitting anisotropy.This resultsin an ESR spectrumfea-
turing a single, broad,symmetricalline at RT.
Chemisorption on Mineral Surfaces. Transition metals are retained
largely in nonexchangeable forms on mineral surfaceswhen presentat tracelev-
els in soil, that is, metal oxides and hydroxides as well as amorphousalumi-
nosilicatesprovide surfacesites for chemisorptionof transition metals. Direct
evidencefor surfacecoordinationof metalscan be obtained fromESRspectraof
the adsorbedparamagneticions. It is clearly proven by ESR that oxide-sorbed
metalsare locatedat isolatedsurfacesites,exceptat high metal loadings,and are
350 SENESI
boundrigidly in comparisonto metal ions sorbedon exchangesitesof layer sil-

icate clays.
For example,the ESR spectrumof Cuz+ is altered on adsorptionof alu-
minum hydroxide and oxyhydroxides,with the g-value decreasingand the ~I
value increasingrelative to the free Cu(HzO)l+ (McBride, 1982c).This shift can
be interpretedas increasedcovalencyof ligand-metalbondingor as an increase
in crystal-field splitting.
Electron spin resonanceevidencehas beenobtainedthat chemisorptionof
Cuz+ occursat isolatedbondingsites on noncrystallineAl(OHh and microcrys-
talline AlOOH, probably by formation of one or two direct bondsbetweensur-
face Al-O groupsand Cuz+, that is favored at high pH. Edgesand defectson sur-
faces of noncrystallineoxides and, to a lesserextent, on crystalline oxides fea-
ture OH groupscoordinatedto single Al atoms,with the formal chargeof their
o atomsnot fully balancedby structuralcations.Thus, they tend to protonateor,
alternativ.ely,to be compensatedby a multivalent cation suchas Cuz+.
In contrast,the dominant(001) surfacesof gibbsiteand boehmiteare con-
sideredto be inactive in chemisorptionbecausethe OH groupsof thesesurfaces
are bridgedto at leasttwo Al3+ and, thus, are electrostaticallysatisfiedand have
no tendency to further coordinate with metal cations. However, ESR spec-
troscopy has proven that at low pH gibbsite chemisorbssmall amounts of
monomericCuz+ «0.5 mmole 100 g-l) which is orientedwith its z-axis p'erpen-
dicular to the (001) planesof the mineral (McBride et aI., 1984).
At pH > 5, the appearance of a broadfeaturelessresonanceand the dimin-
ishing of the rigid-limit spectrumrevealdifferent, possibly hydrolyzedand poly-
merized forms of Cuz+ on the surface. Exposure of the low-pH, gibbsite-
chemisorbedCuz+ to NH3 vapor does not produce desorptiondespite the dis-
placementof OH- and HzO ligandsby NH3. The large decreasesin g-valuesand
increasein ~Id-values indicatethat adsorbedCuz+ may form a complexwith NH3
while remainingrigidly bound to the oxide surface,that is at leastone Cu-O-Al
bond exists.
Thesefindings are confirmed in Cuz+ adsorptionexperimentsconducted
on microcrystallinegibbsiteand boehmitein the presenceof the chelatingligand
glycine (McBride, 1985a). Electron spin resonanceresults indicate that Cuz+
absorbsin the form of ternary complexesin which the metal coordinatessimul-
taneouslywith a surfacehydroxyl 0 and one (on gibbsite)or two (on boehmite)
glycine molecules,with the orientation of the Cu z-axis normal to the (001)
sheetsof the minerals.
Electron spin resonanceanalysisrevealedthat Cuz+ is adsorbedby allo-
phanesand imogolite as a monomeron two distinct typesof surfacecoordination
sites (Clark & McBride, 1984). The preferred sites are likely adjacentAlOH
binding Cuz+ by a binuclearmechanism,whereasweakertype of binding sites
occur at isolatedAlOH or SiOH groups.Electron spin resonancefurther reveals
that the bound ion is rigid, which is indicative of a direct bond (chemisorption)
betweennonexchangeable Cuz+ and 0 atomson the surface.Also in thesecases,
NH3 was able to readily displaceHzO and OH-ligandsfrom chemisorbedCuz+,
leading to the formation of ternary Cuz+-ammonia-surfacecomplexes.Further
evidencefor these mechanismswas obtainedfor Cuz+ adsorptionon titanium
dioxide (Bleam & McBride, 1986). The Cuz+ chemisorbsstrongly at low pH to

sites that retain it by a bidentatemechanism,whereasat higher pH a weaker
complex is formed with single Ti-OH groups.
The ESR techniquealso has been used to study Cuz+ adsorptionby alu-
minum oxidesand allophanesas affectedby phosphate (Clark & McBride, 1985;
McBride, 1985b).Electronspin resonanceresultssuggestthat phosphatecoordi-
nates with the axial position of a surface-boundCuz+, thereby producing a
ternary complex. High levels of sorbedphosphateon aluminumhydroxidessup-
pressesCuz+ adsorption,apparentlyby blocking the coordinationof Cuz+ to sur-
face AlOH groups.Evidenceby ESR hasbeenprovided thatVOZ+ is coadsorbed
with phosphateon boehmite and aluminosilicatesas an inner-spherecomplex
(McBride, 1987).
The ESR spectroscopyhas been used recently to probe the changeof the
ligand environmentof cr3+ upon complexationwith various inorganicanionsat
the solid/waterinterfaceof hectorite and montmorillonite (Charlet & Karthein,
1990).Adsorptionof cr3+ is strongly affectedby ligandspresentin solution.The
ESR spectraof the surfaceobtainedin the presenceof selenite,phosphate,and
P- provided evidenceof ligand coadsorptionwith formation of ternary surface
complexes.
ElectronSpin Resonance Parameters
andThermodynamicConstants.
Metal spin probescan be usedto estimatethermodynamicstability constantsof
metal-surfacecomplexesfrom the ESR parametergil' An examplewhich illus-
trates this possibility is representedby Cuz+ complexeswith hydrous-Alz0 3,
TiO z and somesilicasin the presenceof bidentateligands(Motschi, 1984).A lin-
ear relationshipbetweenthermodynamiccomplexformation constantsof square
planarCuz+ complexesin aqueoussolution and the correspondingESR parame-
ter gil was found. A decreaseof the gil value by 0.1 units correspondsto an
increasein thermodynamicstability by about eight orders of magnitude.The
potential of the model was demonstratedfor a seriesof ternary complexesof
Cuz+ with bidentateligandson o-Al Z0 3, for which speciationof Cuz+ and stabil-
ity constantswere obtained.On thesebases,a major revision of currentconcepts
of cation adsorptionon layer silicatesis possible.
ELECTRON SPIN RESONANCE·RELATED SPECTROSCOPIES
Analysis of the ESR spectrumcan provide information about the identity

of paramagneticspecies,their electronic and geometricstructureand the rota-
tional or translationalmotion. It also is particularly valuablefor the study of the
free radical intermediatesor productsof chemical,photochemical,and biochem-
ical reactionsand of valencestate and site symmetry changesof paramagnetic
metal ions: The principal limiting factor of information obtainedby ESR is the
resolutionof spectraltransitionsthat occursparticularlyfor paramagneticspecies
in powder samples.Two methodsare potentially very useful to overcomethis
limitation: (i) ENDOR spectroscopy,that is the major double resonancemethod
used, basedon a combinationof ESR and nuclearmagneticresonance(NMR)
352 SENESI
techniques(Kevan & Kispert, 1976); and (ii) ESE spectroscopy,that is a time-

domainelectronmagneticresonancemethod(Kevan & Schwartz,1979).
Either ENDOR or ESE representa useful tool in determiningthe compo-
nentsof the hyperfineand superhyperfinematrices,that is, they can extendthe
resolutionof the ESRexperimentby resolvinghyperfine,or superhyperfine,con-
tributionsto inhomogeneously broadened lines in circumstanceswherethe split-
ting betweenthesecomponentsis lessthanthe width of the individual spin pack-
ets. For example, the hyperfine splitting may lead to line broadeningif the
unpairedelectronis highly delocalizedand interactingwith many nuclei which
causesa loss of details.Further,ENDOR canbe appliedto resolvebroadsingle-
line ESR spectrain studiesof randomly orientedsystems,such as frozen solu-
tions, in which an ESR experimentis unableto provide the requiredresolution.
Although the ESEexperimentalapparatusandspectralanalysisis more complex
than for ENDOR, the former is a useful alternativeto the latter when the split-
tings are small and conventionalENDOR is difficult to perform.
ElectronNuclearDouble ResonanceSpectroscopy
Electron nuclear double resonancespectroscopymeasuresthe flip of a

nuclearspin by a radiofrequencyfield, detectedby a changein the intensity of
the ESR signal arising from the electronto which that nucleusis coupled.In the
ENDOR experiment,the magneticfield Ho is centeredon oneof the ESRtransi-
tions and irradiatedwith high intensity microwavepower until the signal is sat-
urated(Eq. [5]). A varying radiofrequency(VENDOR) is then introducedinto the
metal coils of the microwavecavity andthe intensity of the selectedESRtransi-
tion is measuredwhile the radiofrequency is swept over several tens of
megaHertzin orderto inducenucleartransitionsaccordingto the suitableselec-
tion rule. For an effective electronspin of 1/2 and a nucleusof nuclearspin I =
1/2, two ENDOR transitionsareobservedwhenthe resonanceenergyis satisfied
in the equation
MENDOR = h Vo ENDOR = 1/2 ~zzl :!: gn 13n Ho [12]
whereA zz is the z-componentof the hyperfinecoupling constantof the electron

with a neighboring nucleus (see previously in section on "The Hyperfme
Splitting") when the externalmagneticinductionfield Ho is appliedalongthe gzz
tensordirection, which is, in tum, parallel to the z-axis of the nuclearhyperfine
tensor;gn is a dimensionlessconstantcalled the nuclearg-factor; and 13n is the
nuclearmagneton.
For a nuclearspin I = 1 (e.g., 14N), four ENDOR transitionsshould be
observedand an ENDOR spectrumconsistingof two pairs of equally separated
lines should be obtained.The ENDOR spectrumcan, thus, "fingerprint" the
typesof interactingnuclei, describedby gn13n. Line intensitiesare generallyone-
tenth to one-hundredthlessintensethan ESR lines.
The ENDOR spectroscopyis a highly specializedand complextechnique.
However, the sensitivity and easeof carrying out experimentsis improving
rapidly, also due to the introductionof new designsof microwavecavity, called
loop-gap resonators.They require much smaller amounts of sample, can be

wound with an external radiofrequency,and can be placed inside a cryostat.
Further,ENDOR providesa meansof obtainingNMR signalsfrom paramagnet-
ic samples,e.g., the observationof 57Fe NMR in paramagneticsamples.
ElectronSpin Echo Spectroscopy
The advent of powerful computersin the field of NMR spectroscopy

enabledthe introductionof pulsedsourcesfollowed by Fourier transformof the
nuclearechoes[nuclearspin-echoFourierTransform(Ff)-NMR] with enormous
advantagesfor the NMR method.In the field of ESRspectroscopy,the spin-echo
techniqueshould allow, in principle, magneticrelaxation times to be measured
directly and, more importantly, very weak dipolar hyperfine interactionsto be
measuredeven in powder samples.This would permit the molecularsurround-
ings of the paramagneticspeciesto be proven.However,a fundamentaldifficul-
ty ariseswith electronspin echoes,namely, the speedwith which phasecoher-
enceof electronspinsis lost. Sincean electronis coupledmuch more strongly to
its chemicalenvironmentthan a nucleusis, the electronrelaxeson a timescalean
order of magnitudeshorterthan a nucleusdoes.This intrinsic difficulty can be
overcomeby the use of high-powermicrowave sourcesdesignedto pulse into
cavities with rapid decay times, so that detectionwithin a few tens of nanosec-
onds of a pulse is possible.This time is sufficient to enablethe detectionof an
echo from a set of electronspins refocusingafter a 90° pulse. Therefore,a free
inductivedecaycan be built up stepwiseusing varying timedelaysbetweenpuls-
es.
In practical pulsed ESR experiments,electron spin echoesare generated
most commonlyby a pulsesequenceconsistingof a 90° focusingpulsefollowed
by a precessiontime and then a 180° spin flip pulse which causesthe spin to
refocuswithin anotherprecessiontime period. The refocusingproducesa burst
of microwaveenergycalled an echo,whose intensity is measuredas a function
of the time betweenthe two pulsesto generatethe echo delay envelopewith a
time constantwhich gives a transversemagneticrelaxationtime. In many cases
the envelopeof the train of spin echoesis not a smoothexponentialdecay,but is
modulatedby weak dipolar hyperfine interactionsbetweenthe relaxing electron
and neighboring nuclei with nonzero spin to which the electron is coupled.
Fourier transform of the modulateddecay, called electron-spinecho envelope
modulation (ESEEM or ESEM) expressesthe results in the frequency domain
and the resultingpeaksgive directly the hyperfinecouplingfrequenciesof neigh-
boring nuclei. The best spectraare yielded by nuclei which are dipolar coupled
to the electronand which have nuclearquadrupolemoments.
Analysis of the ESEM spectracan thus provide significant structuralinfor-
mationwhich is generallynot availablefrom the ESRspectrum alone,due to res-
olution limitations. Typically, the ESEM pattern can be analyzedto yield the
numberof approximatelyequivalentinteractingmagneticnuclei as well as their
distanceof interaction,togetherwith any small isotropic hyperfinecoupling that
may be present.
354 SENESI
Both relaxation times that characterizethe two magneticrelaxationphe-

nomenaof paramagneticspecies canbe measuredby the ESE techniques.These
times are: (i) the spin-lattice relaxation time, Tb that characterizesthe rate of
radiationlesstransitions between the spin systemand the thermal motion of the
lattice or surroundings(see also in sectionon "The ResonancePhenomenon");
and (ii) the spin-relaxationtime, denotedby T2, that is due to energyexchange
betweenspinsin the system.The transverserelaxation directlymeasuredin pow-
dersby ESE is betterdescribedas a phasememory time sinceit is not generally
identical to T2• However,the true T2 in solids can be obtainedby a specialESE
method (Kevan & Schwartz, 1979). Although both types of relaxationshave
beenusedvery little for paramagneticspeciescharacterizationor to give struc-
tural or chemicalreaction information, this is an areawith significant potential
for developmentin soil chemistrystudies.
Comments
The ESE techniqueis in many ways complementaryto the ENDOR tech-

nique, in that the former allows detectionof more distant nuclei with hyperfine
couplingsin the low-frequencyrange,0 to 15 MHZ, where ENDOR transitions
are very weak. PulsedENDOR methodsalso have beenreported.The introduc-
tion of,new designsof microwavecavities transparentto radiofrequency.radia-
tion, will determinewith little doubt the areaof pulsedESR growth in the future.
Electron nucleardouble resonanceand ESE techniqueshave not yet been
applied to strictly soil chemistry studies, and it is in this area of ESR spec-
troscopywhere major scientific activity is expectedto occur.
REFERENCES
Abragam,A, and B. B1eaney.1970. Electron paramagneticresonanceof transition ions. Clarendon
Press,Oxford.
Alger, R.S. 1968. Electron paramagneticresonance:Techniquesand applications.Wiley-Intersci.,
New York.
Angel, B.R., andW.E.J. Vincent. 1978.Electronspin resonancestudiesof iron oxidesassociatedwith
the surfaceof kaolins. Clays Clay Miner. 26:263--272.
Atherton, N.M. 1973. Electron spinresonance.Halsted,London.
Atherton, N.M., PA Cranwell, AI. Floyd, and R.D. Haworth. 1967. Humic acid I. ESR spectraof
humic acids.Tetrahedron23:1653--1667.
Berliner, L.J. 1976. Spin labeling: Theory and applications.Acad. Press,New York.
Bleam,w.F., and M.B. McBride. 1986.The chemistryof adsorbedCu(II) and Mn(II) in aqueoustita-
nium dioxide suspensionsI. Colloid InterfaceSci. 110:335-346.
Blois, M.S., Ir., H.W. Brown, and I.E. Maling. 1961. Precisiong-value measurements of free radi-
cals of biological interest.p. 117-131.In M.S. Blois, Ir. et al. (ed.) Free radicalsin biologi-
cal systems.Acad. Press,New York.
Carrington,A, and AD. McLachlan. 1967. Introduction to magneticresonancewith applicationsto
chemistryand chemicalphysics.Harper& Row, New York.
Charlet, L., and R. Karthein. 1990. Study of inorganic Iigand-chromium(III)-surfaceternary com-
plexesby ESR spectroscopy.Aquat. Sci. 52:517-527.
Clark, C.I., and M.B. McBride. 1984. Chemisorptionof Cu(II) and Co(II) on allophanesand imogo-
lite. Clays Clay Miner. 32:300-310.
Clark, C.J., and M.B. McBride. 1985. Adsorption of Cu(II) by allophaneas affectedby phosphate.
Soil Sci. 139:412-421.
Craik, OJ. 1971. Structureand propertiesof magneticmaterials.Appl. Phys. Ser. no. 1. Pion Ltd.,
London.
Gamble, D.S., M. Schnitzer, and D.S. Skinner. 1977. Mn(II)-fulvic acid complexing equilibrium
measurements by electronspin resonancespectrometry.Can. 1. Soil Sci. 57:47-53.
Gordy, W. 1980. Theory and applicationof electronspin resonance. Wiley-Intersci., New York.
Ingram, DJ.E. 1969. Biological and biochemical applicationsof electron spin resonance.Hilger,
New York.
Jameson,R.F. 1981. Coordinationchemistryof copperwith regardto biological systems.p. 1-30. In
H. Siegel (ed.) Metal ions in biological systems.Vol. 12. Dekker, New York.
Kevan, L., and L. Kispert. 1976. Electron spin double resonancespectroscopy.Wiley-Intersci., New
York.
Kevan, L., and R.N. Schwartz. 1979. Time domain electron spin resonance.Wiley-Intersci., New
York.
Lakatos, B., T. Tibai, and J. Meisel. 1977. ESR spectraof humic acids and their metal complexes.
Geoderma19:319-338.
McBride, M.B. 1976. Origin and position of exchangesites in kaolinite: An ESR study. Clays Clay
Miner. 24:88-92.
McBride, M.B. 1982a.Organicanion adsorptionon aluminum hydroxides:Spin probestudies.Clays
Clay Miner. 30:438-444.
McBride, M.B. 1982b. Electron spin resonanceinvestigationof Mn 2+ complexationin natural and
syntheticmoleculesand soil organics.Soil Sci. Soc. Am. J. 46:1137-1143.
McBride, M.B. 1982c. Cu2+-adsorptioncharacteristicsof aluminum hydroxide and oxi-hydroxides.
Clays Clay Mineral. 30:21-28.
McBride, M.B. 1985a. Influence of glycine on Cu2+ adsorption by microcrystalline gibbsite and
boehmite.Clays Clay Miner. 33:397-402.
McBride, M.B. 1985b.Sorptionof copper(II) of aluminumhydroxidesas affectedby phosphate.Soil
Sci. Soc. Am. 1. 49:843-846.
McBride, M.B. 1986. Magnetic methods.p. 219-270. In A. Klute (ed.) Methodsof Soil Analysis.
SSSABook. Ser. 5., Part 1. 2nd ed. ASA, CSSA, and SSSA, Madison, WI.
McBride, M.B. 1987. Ternary V02+ ligand-surfacecomplexeson boehmiteand noncrystallinealu-
minosilicates.J. Colloid InterfaceSci. 120:419-429.
McBride, M.B. 1989. Reactionscontrolling heavy metal solubility in soils. p. 1-56. In B.A. Stewart
(ed.) Advancesin soil science.Vol. 10. Springer-Verlag,New York.
McBride, M.B., A.R. Fraser,and WJ. McHardy. 1984. Cu2+ interactionwith microcrystallinegibb-
site. Evidencefor orientedchemisorbedcopperions. Clays Clay Miner. 32:12-18.
McBride, M.B., T.J. Pinnavaia,and M.M. Mortland. 1975. Perturbationof structural Fe3+ in smec-
tites by exchangeions. Clays Clay Miner. 23:103-107.
Motschi, H. 1984. Correlation of EPR parameterswith thermodynamicstability constantsfor cop-
per(II) complexes.Cu(II)-EPR as a probe for the surface complexationat the water/oxide
interface.Colloid Surf. 9:333-347.
Peisach,J., and W.E. Blumberg. 1974. Structuralimplications derived from the analysisof the elec-
tron paramagneticresonancespectraof naturaland artificial copperproteins. Arch.Biochem.
Biophys. 165:691-708.
Poole, C.P. 1967. Electron spin resonance.A comprehensivetreatise on experimentaltechniques.
Wiley-Intersci., New York.
Senesi,N. 1990a. Application of electron spin resonance(ESR) spectroscopyin soil chemistry. p.
77-130. In BA Stewart(ed.) Advancesin soil science.Vol. 14. Springer-Verlag,New York.
Senesi,N. 1990b. Molecular and quantitativeaspectsof the chemistryof fulvic acid and its interac-
tions with metal ions and organic chemicals.Part I. The electron spin resonanceapproach.
Anal. Chim. Acta 232:51-75.
Senesi,N., D.F. Bocian, and G. Sposito. 1985. Electron spin resonanceinvestigationof copper(II)
complexationby fulvic acid extractedfrom sewagesludge.Soil Sci. Soc. Am. J. 49:119-126.
Senesi,N., Y. Chen, and M. Schnitzer.1977a.Hyperfine splitting in electronspin resonancespectra
of fulvic acid. Soil BioI. Biochem. 9:371-372.
Senesi,N., S.M. Griffith, M. Schnitzer,and M.G. Townsend.1977b.Binding of Fe3+ by humic mate-
rials. Geochim. Cosmochim.Acta 41:969-976.
Senesi,N., and C. Steelink.1989.Application of ESRspectroscopy tothe study of humic substances.
p. 373-407. In M.H.B. Hayes et al. (ed.) Humic substances.Vol. 2. John Wiley & Sons,
Chichester,England.
Steelink, c., and G. Tollin. 1962. Stable free radicals in soil humic acid. Biochem. Biophys. Acta
59:25-34.
356 SENESI
Swartz,H.M., I.R. Bolton, and D.C. Borg. 1972. Biological applicationsof electronspin resonance.
WiIey-Intersci., New York.
Templeton,G.D., and N.D. Chasteen.1980. Vanadium-fulvic acid chemistry: Conformationaland
binding studiesby electronspin probestechniques.Geochim.Cosmochim.Acta 44:741-752.
Thomson,A.I. 1990. Electron paramagneticresonanceand electronnucleardouble resonancespec-
troscopy.p. 295-320.In D.L. Andrews (ed.) Perspectivesin modemchemicalspectroscopy.
Springer-Verlag, Berlin.
Vonsovskii, S.B. 1966. Magnetic resonancein ferromagnetics.p. 1-11. In S.V. Vonsovskii (ed.)
Ferromagneticresonance.PergamonPress,London.
Wertz, I.E., and I.R. Bolton. 1972. Electron spin resonance:Elementary,theory and practical appli-
cations.McGraw-Hili, New York.
Published 1996
Chapter12
X-Ray Photoelectron Spectroscopy
R. K. VEMPATI, T. R. HESSAND D. L. COCKE, Lamar University,

Beaumont, Texas
Surfacesof mineralsplaya pivotal role in the adsorption,dissolutionand precip-

itation of ions in the soil environment.Prior to the developmentof advancedana-
lytical instrumentation,surfaceprocesseswere studiedusing solution chemistry.
In recentyears,the developmentof surfacesensitiveanalytical techniques,e.g.,
Auger electronspectroscopy(AES), ion scatteringspectroscopy(ISS), low-ener-
gy electrondiffraction (LEED), secondaryion massspectroscopy(SIMS), x-ray
photoelectronspectroscopy(XPS), scanningprobemicroscopy(SPM), etc., have
allowed direct probing of surfacephenomena.Of thesesurfacetechniques,XPS
is the most popular amongsoil- and geochemistsbecauseit provideselemental,
chemicalstateand semiquantitativecompositionalinformation.
The informationgleanedfrom XPS is extremelyuseful in the determination
of surfaceelementalcomposition(Bancroft et aI., 1977; Gonzalezet aI., 1988;
Seyama& Soma,1988); site occupancy(Seyama& Soma,1988); oxidation and
reductionchemistry(Stucki et aI., 1976); chemicalweathering(Hochella, 1988;
Inskeepet aI., 1991); Lewisand Br0nstedacid sites(Boradeet aI., 1990); adsorp-
tion of cationsand anions,chemicalbondingand surfacereactivity (Koppelman
et aI., 1980; Vempati et aI., 1990a, b) and differentiation of exchangeableand
nonexchangeable clay components(Seyama& Soma,1988). For more examples
and detail applications,the reader is advised to refer to reviews by Hochella
(1988), Perry (1990) and Cocke et al. (1994). These reviews discuss elegant
applicationsof XPS to geo- and soil chemistry research.For the most part, the
referencescited in this chapterand thosein the reviews concernXPS studieson
soil constituents,e.g., clays, Fe oxides, AI oxides, Mn oxides, carbonates,etc.
However, some surfaceanalysiswork on soil colloids and sedimentshas been
reported(Seyama& Soma, 1985) but the use of XPS for such studies is not a
commonpractice as it is extremely difficult to trace the origin of the elemental
signalsto a specific mineral. A particularelementof a certainoxidation statecan
display small shifts in its binding energy signals dependingon the mineral or
chemical situation.Thus for a complexmixture of materials,suchas soil and soil
colloids, there are often overlappingsignalsfrom the sameelementsthat are dif-
Book Seriesno. 5.
357
358 VEMPATI ET AL.
ficult to interpret. Furthermore,XPS spectraof multielement systemscan be

muddledby the variouselements'spectralpeaksoverlappingoneanother.Unlike
the problemwith the small chemicalshifts of a single element,this problemmay
be solvedby: (i) using a different spectralline for analysis,(ii) using a different
x-ray sourceif there are problemswith an Auger peakoverlappinga core level
peak,or (iii) decomposingthe overlappingpeaksinto individual componentsvia
a peak-fitting routine and then subtractingthe offending peak.For thesereasons,
studieson the individual soil componentsor simple mixturesare often favored.
For surfaceanalysis,XPS is popularbecause:(i) elementalidentification,
chemical state and semiquantitativecompositionalinformation for the all ele-
mentsin the periodictable exceptH and He becauseof their low photoionization
crosssection,can be determined;(ii) depthprofiling can be performedeither by
using a low take-off angle XPS method(nondestructive)or by Argon ion sput-
tering using an ion gun (destructivemethod); (iii) amorphousand crystalline
samplesmay be investigated;(iv) conductingand nonconductingmineralsmay
be analyzed,and (v) very small samplesize is needed(on the orderof 10 ng). In
doing surfaceanalysisthereare somedisadvantages and theseare relatedto how
the sample may be changedduring the analysis. Most surface analysis tech-
niques,XPS included,require a high vacuumenvironmentand utilize somesort
of probesuchasphotonsor electrons.This cancreateproblemsfor hydratedsur-
faces becausethey are susceptibleto dehydrationdue to high vacuum require-
ment, althoughhydratedFe oxideshavebeensuccessfully studied by Vempati et
ai. (1990a,b). In other cases,samplesmay deterioratebecauseof the vacuum
environmentand by localizedheatinginducedby the photon(x-rays for XPS) or
electronprobe.This is especiallytrue for clay minerals,where high vacuumcan
irreversibly damagethe surface.Theseproblemscannotbe easily solved; how-
ever, they may be betterunderstoodby monitoring the surfacechemicalchanges
with time.
There are severalbasictexts and reviews concerningsurfaceanalysisand
in particularXPS (Muilenberg, 1979; Defosse& Rouxhet,1980; Briggs & Seah,
1983; Ratner, 1983; Eland, 1984; Hochella, 1988; Payntner,1988; Perry, 1990;
Cocke et aI., 1994) that the readeris encouragedto consult for greaterdetail.
X-RAY PHOTOELECTRONSPECTROSCOPYPRINCIPLES
When a solid material is bombardedby a photon, the photon'senergy is

transmittedto an electron.Shouldthe photonbe of sufficient energy,the electron
can be ejectedfrom the atom. This processis called photoemissionand is depict-
ed in Fig. 12-1. When the excitationoccurssufficiently closeto the surface,the
photoelectroncan escapefrom the material. The photoelectronleavesthe atom
in an excited state which then must lose the energygained. The excessenergy
can be lost through various de-excitationprocessesincluding the ejection of a
secondelectron called an Auger electron (see Fig. 12-1). All elementshave a
unique set of core level electronswhose binding energiesare spreadout over a
thousandelectronvolts(eV) or more. Theseare the electronsthat are of interest
for XPS analysis.By determiningthe binding energiesof the photoelectons,the
X·RAY PHOTOELECTRON SPECTROSCOPY 359
Incident X-ray
Photon Photoelectron
I
T
Productionof Photoelectronsand Auger Electrons:
1) Absorption of X-my pholOn by core electron
2) Ejection of core-level photoelectron
3) RelMuliion or higher shell electroninto hole
4) Intern.-Ily recombinedenerg.yof relaxation
5) Ejection of Auger electron
Fig_ 12-1. Illustration of photoelectronand Auger electronprocessinducedby x-ray bombardment.
elementof origin can be determined.In fact, small changes(a few eV) in the
binding energyreflectsdifferencesin the chemicalstate(suchas oxidation state)
of the atom of origin and the number of photoelectronsproducedreflects the
quantity of the originating atomsin the excitation region.
To excite these core level electrons,photons with energiesgreater than
1000 eV should be used (x-ray photons).The most commonly used x-ray lines
for XPS analysisare the AlKa (1486.6eV) and the MgKa (1253.6 eV). These
sourceshave sufficient energyto excite the core level electronsand to penetrate
deepinto the bulk sample.However,only thosephotoelectronsoriginating from
the near-surfaceregion (top few nm) are able to escapefrom the samplewithout
energy loss. It is this fact that makes XPS a surface sensitive technique.The
depth from which a particular photoelectroncan escapewithout loss of kinetic
energy is dependenton the attenuationlength of the electron in the sample
matrix_ The attenuationlength is definedas the distancein the matrix over which
an electrontravels and suffers an energyloss only 36.8% [100(e-I )] of the time.
Escapedepth is simply the attenuationlength times the cosineof the angle (8 in
Fig. 12-1) betweenthe surfacenormal and the direction of the escapingelectron.
Aside from being dependenton the matrix, the attenuationlength shows a lop-
sided vee-shapeddependenceon the electron'skinetic energy and has a mini-
mum at about50 e V (Seah& Dench,1979). Over the energyrangeof most inter-
est (10-1200 eV), the attenuationlength varies from 2 to 10 atomic layers.
Typically, the sampling depth or what Hochella (1988) calls the "information
depth" is consideredto be equivalentto three times the attenuationlength.
Once the photoelectronhas escapedfrom the surfacewithout collision or
other energy loss, it has a certain kinetic energy(KE) that can be relatedto the
binding energy for the electron (Eb) which is characteristicfor the electronic
360 VEMPATI ET AL.
Conducting Non-conducting
Spectrometer
Sample Sample
KE KE'
Vacuum
Level
-
/
<l>s } <l>c
---.~
Conduction Uncharged
>
Band Work
Function
Fermi / Valence \
~--/r--Fe·'L L:: S)
Level Band
bv Pinned
Fermi Levels
Mismatchin
Fermi Levels
E~
Core Level
Fig. 12-2. Energy level diagram for x-ray induced photoelectronsfrom conductiveand insulating
samples.
energylevel of origin. By measuringthe kinetic energyof the photoelectron,its

binding energycan be determined.For a conductingsamplein electricalcontact
with the spectrometer,the relationship between the measuredkinetic energy
(KEsp) and binding energyis
EB =hz - KE - <l>s =hv - KEsp - <l>sp [1]
where the EB term includesthe electronbinding energy(Eb), intra-atomicrelax-

ation shift (Ea) and interatomicshift (Es) (both Ea and Es are on the order of a
few eV or less); hz is the energyof the incident x-ray photonand <l>sp is the work
function of the spectrometer(the work done on the photoelectronas it arrives at
the electronmultiplier tube to be counted).Thesetermsmay be betterunderstood
by examining Fig. 12-2. The Fermi level is defined as the highestfilled elec-
tronic state at 0 K and the binding energy, EB, is defined as zero at the Fermi
level. The work function of the sample,<l>s is the additional amountof energy
requiredto move an electronfrom the Fermi level to the vacuumlevel (the point
where the electron is no longer energeticallyassociatedwith the atom and is
often referredto as the free electronlevel). For a conductingsample,electrical-
ly coupled withthe spectrometer,their respectiveFermi levels are pinned at the
sameenergyasshownin Fig. 12-2. This meansKE + <l>s =KEsp + <l>sp and allows
Eq. [1] to be written. Now the electron'strue kinetic energy(KE) and the sam-
ple's work function need not be known. The <l>sp can be determinedusing stan-
dardswith well-defined binding energies(such as Au) and should remain con-
stantif the spectrometersystemis reasonablysafe.This allows the EB to be com-
X-RAY PHOTOELECTRONSPECTROSCOPY 361
pareddirectly from sampleto sampleif measuredon the sameinstrument;how-

ever, if the conductivity of the sample'ssurfaceor a good electrical connection
with the instrumentis in doubt then an internal standardshould be used.
Samples,such as soils, mineralsand other geologicmaterialsare not con-
ductive and so the sample'ssurfacewill be electrically isolatedfrom the spec-
trometerand their Fermi levels will not be pinned to the sameenergy (see Fig.
12-2). Equation [1] is no longer valid and a new term <l>c, the static surface
charge,must be considered.This chargeis typically positive becauseof loss of
photoelectrons.The nonconducting sample'swork function, <l>cs, now consistsof
two terms: the sample'sunchargedwork function, <l>s, and the surfacecharge,<l>c.
For the nonconductingsamplein Fig. 12-2, the kinetic energyof the photoelec-
tron (KE') is smaller than would be expectedfor an unchargedsituation by <l>c.
Consequently,one can write
EB,R = hv- KEsp - <l>sp > E' B. [2]
where the reported binding energy (EB,R), will be greater than the actual
unchargedbinding energy (E'B)' For proper analysis, a correction between
reported(EB,R) and the true binding energy (E'B) must be made. This problem
can be handledby using an internalstandardthat hasa well-known binding ener-
gy. Two of the most commoncorrectionmethodsare the: (i) adventitiouscarbon
method,and(ii) surfacegold method.Thesemethodsalongwith chargingeffects
will be discussedlater in the text.
The formation of an Auger electron (see Fig. 12-1) involves three elec-
trons (ijk) and thereforethreeenergylevels have to be considered.The impinge-
ment of a x-ray photon or an electron on an atom leadsto the ionization of an
inner shell electron(i), this leadsto the creationof a hole in the core level result-
ing in a transferof an electronfrom a higher atomic level (j) to fill the inner core
hole. This de-exitationprocessor releaseof excessenergyresultsin emissionof
a x-ray photon leading to the ejection of anotherelectron, k, called the Auger
electron.The Auger electronkinetic energyis given by
EK(ijk) = EB(i) - EB(j) - EB(k) - E(jk) + R(jk) [3]
where E(jk) is the interactionbetweenthe two holes in the final state,and R is

the total relaxation energy, i.e., the intra- and interatomic relaxation energy
betweenthe two holes in the final state.
Anotherterm that hasbeenusedto describethe chemicalstateis the Auger
parameter(Wagner& Joshi, 1980)which is definedasthe differencebetweenthe
kinetic energyof the most intenseAuger line, EK(ijk) and the most intensepho-
toelectronlines KE(P).
a = EK(ijk)- KE(P) = EB - E~ [4]
where EB and E~ are photoelectronand apparentAuger binding energies,respec-

tively. In order to keepthe Auger parameterpositive and independentof the pho-
ton excitationenergy,the modified Auger parameter,a', is used
362 VEMPATI ET AL
Table 12-1. Mg (Is) binding energies,Mg KLnLzJ Auger kinetic energiesand dRs valuesof Mg
compounds(Seyama& Soma,1988).
Compounds Mg(ls) MgKLnLzJt
Mg· F2 1306.5 1176.8 0.0
Exchangeable 1305.3 1179.0 1.0
Nonexchangeable 1303.8 1181.0 1.5
Orthopyroxene 1304.0 1180.7 1.4
MgCI2 ·6H20 1304.8 1180.2 1.7
MgO 1303.9 1181.3 1.9
MgBr2· 6H20 1305.3 1180.7 2.7
t As defined in Eq. [3].
:j: Extra atomic relaxationfactor.
a' =a + hv [5]
The advantageof using the Auger parameteris that it is independentof sample

chargingwhich is an importantconsiderationfor soil samples,mineralsandother
geologicmaterials,many of which are insulatorsand exhibit charging.
The Auger parametersare usedto understandthe chemicalenvironmentof
the minerals.The Auger parametervaluesare directly relatedto the polarizabil-
ity of the mineral. Studieson the structuralclay components,i.e., 0, Si, AI, and
Mg showthat theseelementshavelow polarizability indicating that the electrons
in silicate frameworks are delocalized (Gonzalez-Elipe et aI., 1988).
Furthermore,chemicalstateplots, plots of the Auger kinetic (eV) vs. XPS bind-
ing energy(eV) are usedcommonlyto differentiatethe exchangeable and nonex-
changeableions in the clay minerals(Seyama& Soma,1985; GonzalezElipe et
aI., 1988). Table 12-1 shows that the Mg Auger parametervalue for the
exchangeableMg is betweenMgCl2 and MgF2; whereas,the values for the
nonexchangeable ion is close to MgO. The other parameterthat is commonly
usedin the literatureis theextra-atomicrelaxationenergydifference(L1Rs) which
is calculatedas follows: {[XPS binding energy(sample)+ Auger kinetic energy
(sample)]- [XPS binding energy(standard)+ Auger kinetic energy(standard)]).
The L1Rs of the nonexchangeable Mg ion is greaterthan that of the exchangeable
ion indicating that the densityof electronssurroundingthe nonexchangeable Mg
is greater.
X-RAY PHOTOELECTRONSPECTROSCOPY
INSTRUMENTATION
The basic componentsof XPS are: ultrahigh vacuum chamber, x-ray

source,electronanalyzer,detector,datacollection and handling,and accessories
(Fig. 12-3). The most commonly usedultrahigh vacuum(UHV) pumpsare ion
pumps,vapordiffusion pumps,turbomolecularpumpsandcyropumps.The heart
of the instrumentis the analyzer.Today the most commonlyusedanalyzeris the
hemispherical-sector type. In this system,the electronsare subjectedto a retard-
ing field by a constantpassenergyand resolvedby their different pathsbetween
the two charged hemispheres.Slits having similar width are placed at the
X-ray
Electron EnergyAnalyzer
Source
: ---------t::~::~~
l:'oow~
Incident Electron Binding Energy (eV)
Sample
X-rays Detector
Ein= hv
Signal Processing
and Display
Fig. 12-3. Schematicdiagramof the ESCA spectrometer.
entranceand exit of the analyzer.The passenergy(PE) is defined as the poten-

tial difference neededbetweenthe two chargedhemispheresso that an electron
of the appropriateenergy may passthrough the analyzerwithout colliding wit~
it or the exit slit. A high PE increasesthe intensity and full width at half maxi-
mum (FWHM), and a low PE decreasesthe FWHM and intensity of the peak.
There are two basic analyzermodesthat are available: (i) fixed retarding ratio
(FRR), and (ii) fixed analyzertransmission(FAT). In the FRR mode, the sensi-
tivity (S) is proportionalto the kinetic energyof the photoelectron.Thereforethe
sensitivity at low kinetic energy is reducedto that of high kinetic energies.This
meansthat in this mode the S for transition metal elementsis reducedcompared
to Al or Si. Also, the area of sample analysisremainsconstantthroughoutthe
whole kinetic range. In caseof the FAT mode, S is inversely proportionalto the
kinetic energyof the photoelectron.Therefore,the S at lower kinetic energiesis
improved over the FRR mode and thus provides an increasedS for transition
metal elements.In the caseof FAT mode, the areaof the sampleanalyzeddoes
vary slightly with the kinetic energy.
The detectoris either a single- or multichanneldetector,the latter allows
for increaseddatacollection speed.The x-ray sourceis usually a dual anodetype
with Al and Mg as sourcemetals.Thesesourcesgive a sharp Kal,2 with a line
width of 0.8 to 0.9 eV and the intensity of theselines is half of that total emitted
intensities.The position of the main x-ray lines of theseelementsare
MgKal = 1253.6eV Ka2 = 1253.4eV

Al Kal = 1486.6eV Ka2 = 1486.3eV
Other metalsare usedon a limited basisto provide alternativeenergies.To obtain

a high resolutionXPS spectrum(-0.2 eV), a monochromaticx-ray sourceis used
at the expenseof signal intensity and excessive
chargingfor insulators.A charge
neutralizerwhich providesa flux of low-energyelectronsis usedin someexper-
364 VEMPATI ET AL.
Electron Energy
Analyzer
Charge X-ray Source

Neutralizer
"
MechanicalCleaning
Sample / and/orCleavage
Transporter
/ Sample
Transporter
Gasor
Liquid........
~ Sputteror
Vapor Coating
Chemical Physical
Main Analysis Treatment
Reaction Chamber
Chamber Chamber
Fig. 12-4. Sketchof a typical XPS spectrophotometer.
imentsto compensatefor the chargingof nonconductingsamples(seeFig. 12-4).

A sputtergun is usually presentto provide etchingof the surfaceby bombard-
mentof Argon ions, thus enablingdepthprofiling of samples.The sampleis gen-
erally placed on an tilting stagefor angle resolvedXPS studies.Since samples
often require some form of in situ treatmentbefore or after analysis,a sample
treatmentchamberis usually a part of the instrument.Additional accessories
that
may be present in the spectrometerare: in-vacuum fracturing devices, vapor
deposition,heatingand cooling stages,gas/liquid inlet valves,etc. Most modern
instrumentsmay have other surfacesensitivetechniqueslike AES, LEED, high
resolutionelectronenergyloss spectroscopy(HREEL), ultraviolet photoelectron
spectroscopy(UPS), secondaryion massspectroscopy(SIMS), and ion scatter-
ing spectroscopy(ISS).
ReferencingBinding Energy
X-ray photoelectronspectroscopyspectraof insulators,suchas soils, min-

erals and other geological materials,often exhibit considerablebinding energy
shifts and peak broadeningbecauseof surfacecharging.This is due to the fact
that thereis no electricalcontactbetweenthe sampleand instrument.In order to
correctfor thesecharge-inducedshifts, standardsare often used. Oneof the com-
mon methodsto calibratethe spectraof thesematerialsis to usethe charge-shift-
ed C(1s) XPS peak, and this C is usually referredto as an adventitiousC (ele-
ment not inherentto the sample). This C(ls) peakis consideredto be at 284.6eV
(some use 285.0 eV as well). Most of the samplesurfacescontain C which is
derivedasa contaminantfrom air and/orvacuumsystemcontamination.The sec-

ond commonmethodfor chargecorrectionis the use of a Au(4f7/2) line at 83.8
eV. In this methoda small amountof gold is depositedon the samplesurfaceto
provide a better referencematerial. The latter is the best method; however,the
disadvantageis that the Au( 4f7/2) binding energy hasbeenshownto vary with the
thicknessof the Au deposition due to extra-atomicrelaxation (Kohiki et aI.,
1983) resulting in a compositeAu peakdue to the Au/substrateinterfaceeffects.
This results in differential charging between the sample and deposited Au.
However,this is not a problemif a thicker Au film is used(Kohiki & Oki, 1985).
The matterof choosingAu depositionor adventitiousC is a matterof choiceand
care should be taken while calculatingabsolutebinding energies.
X-ray Photoelectron SpectroscopyAnalysis
X-ray photoelectronspectroscopyprobesthe core level and valencelevel

electronic structure of atoms in the near surface region. The valence electron
occursin the region of 0 to 20 eV. It is difficult to probethe bondinginteractions
at the surfaceof multicomponentmaterialsbecauseof the overlap of the many
transitions,i.e., low intensity of the peak becauseof their low photoionization
crosssection.Additional peaksfrom environmentalcontaminationmake it diffi-
cult to obtain a good spectrum.For this reason,most of the bonding interactions
at the surfacesof solids are studiedutilizing the coreelectronicstates.X-ray pho-
toelectron spectroscopyspectraare usually collected as wide or survey scans
(usually ~ 1000 eV) and narrow or high resolution scans (usually 20-50 eV
dependingon the elementof interest).
The wide scans provide information for elemental identification. Table
12-2 providesa quick referenceof binding energydataof the elementscommon
to the soil environment.A goodsourceof basicelementalreferencespectrais the
Handbook of X-ray Photoelectron Spectroscopy (also known as the "Phi" hand-
book (Wagneret aI., 1979). However, recent instrumentationhas computerized
elemental identification software. The narrow high resolution scans used for
determiningthe peak'sexactbinding energyand areaof the peakfor delineation
of the chemicalstateand quantitativeanalysis.The peakwidth, measuredas the
FWHM, also may provide additional chemicalenvironmentalinformation.
When identifying and analyzingXPS data,one shouldbe awareofa num-
ber of spectralfeaturesthat can help in the analysisprocess.It is important to
note that peaksfrom p, d, and f levels occur as doubletsand that transition met-
als from unfilled "d" shells are broad becauseof multiple splitting (seebelow).
For transitionelementsin paramagneticor high spin states,an additional peakis
observedat high binding energieswhich is referredto as a satellite peak. Also,
all the potentialAuger lines shouldbe considered.The most importantfactor that
one has to considerin insulating surfacessuch as soils is samplecharging and
this should be correctedeither by adventitiousC or gold-dot method. Some of
the processesthat are involved in XPS are discussedbelow. Thesephenomaare
useful for identification of elements,magneticstates,oxidation states,and other
chemicalinformation.
366 VEMPATI ET AL.
Table 12-2. Principal binding energiesfor the elementsin numericalorder(adaptedfrom Watneret

aI., 1979).
Spin orbit Spin orbit
Principal Core splitting or Principal Core splitting or
binding electronic x-ray binding electronic x-ray
energies Element levels source energies Element levels source
t7 Lu 4f7 [2] t128 Eu 4d
tI4 Hf 4f7 [2] 129 Sm 4d
t19 Ga 3d t130 P 2p3 [1]
23 0 2s t134 Sr 3d5 [2]
t22 Ta 4f [2] t137 Pb 4f7 [5]
25 Sn 4d tI40 Gd 4d
t29 Ge 3d5 [1] 141 As 3p3 [5]
30 F 2s t146 1b 4d
t31 W 4f7 151 Si 2s
37 V 3p t152 Dy 4d
t40 Re 4f7 [2] tI56 y 3d5 [2]
41 Ne 2s t157 Bi 4f7 [5]
t42 As 3d5 [1] t159 Ho 4d
43 Cr 3p 163 Se 3p3 [6]
48 Mn 3p tI64 S 2p3 [1]
49 I 4d5 [2] t167 Er 4d
t50 Mg 2p t175 Tm 4d
t51 Os 4f7 [3] t179 Zr 3d5 [2]
53 Fe 3p 181 Se A [AI]
t56 Li Is t182 Yb 4d
t56 Se 3d5 [1] 182 Br 3p3 [7]
60 Co 3p 186 Ga A [Mg]
t61 Ir 4f7 [3] 188 P 2s
61 Xe 4d5 [2] t189 B Is
64 Na 2s 196 Lu 4d5 [10]
67 Ni 3p t199 CI 2p3 [2]
t69 Br 3d5 [1] t202 Nb 3d5 [3]
t71 Pt 4f7 [3] 208 Kr 3p3 [8]
t73 AI 2p 211 Hf 4d5 [11]
75 Cu 3p3 [2] 226 Ta 4d5 [12]
77 Cs 4d5 [3] 228 S 2s
t84 Au 4f7 [4] t228 Mo 3d5 [3]
t87 Kr 3d5 [1] 240 Rb 3p3 [9]
89 Zn 3p3 [2] t242 Ar 2p3 [2]
89 Mg 2s 243 W 4d5 [13]
90 Ba 4d5 [3] t253 Tc 3d5 [4]
98 Er A [AI] 260 Re 4d5 [14]
t99 Si 2p3 [1] 260 Na A [Mg]
tI03 Hg 4f7 [4] 260 1b A [AI]
103 La 4d5 [3] 262 Zn A [Mg]
104 Ga 3p3 [3] 262 As A [AI]
109 Ce 4d5 [3] 270 Sr 3p3 [11]
109 Ge A [Mg] 271 Cl 2s
t111 Rb 3d5 [1] 279 Os 4d5 [14]
t112 Be Is t280 Ru 3d5 [4]
115 Pr 4d t285 C Is
117 Ho A [AI] 285 1b 4p3 [37]
t118 Tl 4f7 [4] t294 K 2p3 [3]
118 A1'2s 297 Dy 4p3 [40]
121 Nd 4d 297 Ir 4d5 [15]
122 Ge 3p3 [4] 299 Y 3p3 [12]

301 Mg A [AI] 564 Pr A [Mg]
t307 Rh 3d5 [5] 568 Cu A [AI]
309 Ho 4p3 [44] 573 Ag 3p3 [31]
315 Pt 4d5 [17] t573 Te 3d5 (10]
320 Ar 2s t574 Cr 2p3 [9]
321 Er 4p3 [47] 599 F A [Mg]
330 Zr 3p3 [14] 600 Ce A [Mg]
t333 Th 4f7 [9] 602 Gd A [AI]
333 Tm 4p3 [51) 619 Cd 3p3 [34]
t335 Pd 3d5 [5] t619 I 3d5 [12]
335 Au 4d5 [18] 634 La A [Mg]
335 Cu A [Mg] 637 Eu A [AI]
341 Yb 4p3 [48] t639 Mn 2p3 [11]
342 Ge A [AI] 641 Ni A [AI]
t347 Ca 2p3 [3] 653 Ba A [Mg]
360 Lu 4p3 [53] 665 In 3p3 [38]
361 Hg 4d5 [20] 667 Mn A [Mg]
361 Nb 3p3 [15] 669 Ne A [AI]
t368 Ag 3d5 [6] t670 Xe 3d5 [13]
369 Gd A [Mg] 676 Th 4d5 [37]
t377 U 4f7 [11] 682 Sm A [AI)
380 K 2s t685 F Is
385 Tl 4d5 [21] 685 Cs A [Mg]
394 Mo 3p3 [17] 696 Cr 2s
t398 N Is n07 Fe 2p3 (13]
t399 Sc 2p3 [5] 709 Xe A [Mg]
404 Eu A [Mg) 713 Co A [AI]
t405 Cd 3d5 [7) 715 Sn 3p3 [42)
408 Ni A [Mg] t726 Cs 3d5 (14]
412 Pb 4d5 [22] 726 Cr A [Mgl
419 Ga A [AI] 736 U 4d5 [42)
436 Ne A [Mg) 738 I A [Mg)
440 Ca 2s 745 0 A [Mg]
440 Bi 4d5 [24) 758 Nd A [AI)
t444 In 3d5 [8] 767 Sb 3p3 [46)
449 Sm A [Mg) 772 Te A [Mg)
t454 Ti 2p3 [6] t778 Co 2p3 [15)
462 Ru 3p3 [22] t781 Ba 3d5 [l51
480 Co A [Mg] 781 V A [Mgl
N85 Sn 3d5 [8) 784 Fe A [AI]
493 Na A [AI] 797 Pr A [AI]
495 Zn A [AI] 799 Sb A [Mg]
497 Rh 3p3 [24] 816 Sn A [Mg]
499 Sc 2s 820 Te 3p3 [51]
t512 V 2p3 [8] 832 F A [AI]
525 Nd A [Mg) 833 Ce A [AI]
526 Dy A [AI] 835 Ti A [Mg)
t528 Sb 3d5 [9] t836 La 3d5 [17)
t531 0 Is 843 In A [Mg]
533 Pd 3p3 [27] t853 Ni 2p3 (18]
551 Fe A [Mg] t863 Ne Is
561 Ti 2s 867 La A [AI]

368 VEMPATI ET AL.

870 Cd A [Mg] 1103 S A [Mg]
874 N A [Mg] 1103 Cd A [AI]
t884 Ce 3d5 [18] 1107 N A [AI]
886 Ba A [AI] t:j:1117 Oa 2p3 [27]
896 Ag A [Mg] 1126 Eu 3d5 [30]
900 Mn A [AI] 1129 Ag A [Mg]
916 Sc A [Mg] 1149 Sc A [AI]
918 Cs A [AI] 1154 Bi A [Mg]
926 Pd A [Mg] 1159 Pd A
t932 Pr 3d5 [20] 1161 Pb A [AI]
t933 Cu 2p3 [20] 1168 TI A [Mg]
942 Xe A [AI] 1179 Hg A [Mg]
952 Rh A [Mg] 1183 Au A [Mg]
959 Cr A [AI] 1185 Rh A [AI]
964 Ca A [Mg] 1186 Od 3d5 [32]
971 U A [Mg] 1197 Ca A [AI]
971 I A [AI] 1204 U A [AI]
978 0 A [AI] 1212 Ru A [AI]
979 Ru A [Mg] t:j:1217 Oe 2p3 [31]
t981 Nd 3d5 [21] 1223 C A [AI]
990 C A [Mg] 1239 Th A [AI]
1005 Te A [AI] 1239 K A [AI]
1006 K A [Mg] 1241 Tb 3d5 [35]
1006 Th A [Mg] 1272 Ar A [AI]
1014 V A [AI] 1296 Dy 3d5 [37]
t1022 Zn 2p3 [23] 1299 Mo A [AI]
1032 Sb A [AI] 1303 Mg Is
t1034 Pm 3d5 [26] 1304 CI A [AI]
1039 Ar A [Mg] 1310 B A [AI]
1049 Sn A [AI] 1319 Nb A [AI]
1068 Ti A [AI] t:j:1324 As 2p3 [35]
1071 CI A [Mg] 1336 S A [AI]
t1072 Na Is 1387 Bi A [AI]
1076 In A [AI] 1394 Pb A [AI]
1077 B A [Mg] 1401 TI A [AI]
t1081 Sm 3d5 [27] 1412 Hg A [AI]
1086 Nb A [Mg] 1416 Au A [AI]
t Thosecore level lines that are most useful for identifying chemicalstates.For core electroniclev-
= =
els: 3 sublevel3/2, 5 =sublevel5/2 and 7 sublevel7/2. Spin orbit splitting (in eV) is indicat-
ed in brackets.Auger lines are designatedby A and the x-ray sourceis indicatedin brackets.
:j: Core level lines that are most useful in identifying chemicalstates,and are specialfor AI source.
Spin Orbit Splitting
The XPS peakfrom p, d, and f energylevelsoccuras doublets.Thesedou-

blets, which result from spin orbit splitting, are useful for chemicalstate inter-
pretation.Table 12-3 showsthat for I = 1, whereI is the orbital angularmomen-
tum quantumnumber,the electronicstatessplit into two peakswith the peakfor
the higher j level occurringat a higher binding energies.The relative intensities
of thesepeaksare 1:2, 2:3 and 3:4 for p, d and f energylevels, respectively.
X·RAY PHOTOELECTRONSPECTROSCOPY 369
Table 12-3. X·ray nomenclaturefor the spin-orbit splitting.

Subshell Quantumnumber Area ratio
Is (i-coupling)
s 0 1/2
p 1 1/2,3/2 1:2
d 2 3/2,5/2 2:3
f 3 5/2,7/2 3:4
Satellites
Satellitesdue to multiple electronexcitation and ligand transferprocesses

are extremelyuseful in chemicalstatecharacterization.If suchinteractionoccurs
during the photoionizationprocess,the photoelectronwill lose its energyand an
additional peakwill be observedin the XPS spectrum.Thesepeaksoccur at the
high binding energy side of the main photoelectronpeak and are termed as
shake-upor shake-offsatellitesdependingon whetherthe other electronsbeing
excited are only promotedto an excited stateor to the continuum,respectively.
The transitionelementshave unpairedelectrons(paramagnetic)and producethe
most intensesatellite structures.
Multiplet Splitting
The emissionof a core level photoelectroncan be interactively coupledto

one or more valenceelectronsthrough a phenomenonknown as multiplet split-
ting. In multiplet splitting, the unpaired"hole" createdby photoemissionin the
core level interactswith the unpairedvalenceelectrons.This phenomenonis use-
ful for studying the paramagneticmetal ions. Although the multiplet splitting
phenomenoncausesbroadeningof the 2p photoelectronspectraof transition
metal ions, it also producesan useful splitting of the 3s level as well. For exam-
ple, the splitting distance, referred to as AE, betweenthe Mn(3s) peak (Fig.
12-5), hasbeenusedto identify the oxidation statesof Mn (Murray et aI., 1985).
Atomic ConcentrationsCalculations
Quantitative analyseswith accuracy as good as 10% are possible using

XPS, but care with calibration is needed.Using the relative heightsor areasof
XPS spectralpeaksalongwith suitablecalibrationfactors, it is possibleto deter-
mine the relative elementalcompositionof the near surfaceregion of a sample.
Currently,the mostwidely usedapproachfor XPS quantitativeanalysisis a com-
bination of empirically derived sensitivity factors and a basic "Three Step
Model" of photoemission.The model relatesmeasuredintensitiesto basicmate-
rial propertiesand involves: (i) photoionization, (ii) photoelectrontransportation
to the surface,and (iii) transmissionto the detector.We will presenta brief dis-
cussionof the basicaspectsof quantitativeanalysisby XPS; however,the read-
er may wish to refer to texts by Powell and Seah(1990), Briggs and Seah(1983),
Powell (1978) and Wagner(1978) for a more detaileddiscussion.
370 VEMPATI ET AL.
Mn(3s)
Region
90 85 80 75
Binding Energy (eV)
;;- 6
0
$ 1;
om
0
~is. !
Fig. 12-5. (a) Multiplet splitting in the Mn(3s)
tI)
5
•
XPS spectrum, and (b) the experimentally ~ ~
determinedrelationship between the splitting :;:" 4
distance(E) and Mn oxidation state(~) Oku et +
at. (1975); (x) Evans and Raftery, (1982); (+)
Zhao and Young, (1984); and (D) Foord et at. 2 3 4 5 6
(1984). Mn Oxidation State
If we considera clean, flat sample,homogeneousin the analysisvolume,

the intensity of a photoelectronpeak Ix (numberof photoelectronsper second)
specific to elementx is given by
[6]
where
nx = the numberof atomsof the elementx per cubic centimeter,
J = the x-ray flux in photons percentimetersquaredper second,
(J = the photoionizationcross-section in centimeterssquared/atom,
8 = an angularefficiency factor basedon the angle betweenthe x-ray
sourceand the detector,
A. = the attenuationlength of the photoelectronin the sample,
p = the probability that the photoelectronwill undergoenergylosses
other than by collision (for example,plasmonloss),
A = the areaof the samplefrom which photoelectronsare detected,
F = the instrumentaldetectionefficiency.
For a real sample,additional factors such as attenuationof the escapingphoto-

electrons by the adventitious C layer and shadowing effects due to surface
roughnessand/orparticle sizecancauseadditionalcomplications.Solving for nx ,
Eq. [6] can be rewritten as
[7]
such that the terms in the denominatorcan be consolidatedinto a single term

called the S factor, Sx' Unfortunately,the S factor involves instrumentspecific,
matrix specific as well as elementalspecific terms; consequently,researchersdo
not use Eq. [7] for quantitativeanalysis.Instead,the ratio of two elementsin the
sampleis used. For a homogenousbinary material (AB) the following equation
may be written
[8]
Equation[8] may be usedif the SA/SB ratio is matrix independent.This is possi-

ble if cr, A, and F for both elementA and elementB are assumedto changeiden-
tically or by the samefactor for different materials.Using this assumption,sets
of relative S factors may be developed(Briggs & Seah, 1983; Wagner et aI.,
1979). Sensitivity factors are usually determinedexperimentally in the same
spectrometerfor pure elementsamplesand reportedas a relative S factor, being
normalizedso the fluorine (Is) S factor is one. A generalizedform of Eq. [8] can
be written as
nx Ix/Sx
C -----
x- En
i I
- U/S·
I I I
[9]
wherethe atomicfraction of elementx, Cx, in a samplecanbe calculatedby mea-

suringthe intensity of a particularpeakfor eachelement(total of i elements)pre-
sent in the sample.
Use of S factorsas describedabovewill normally providesemiquantitative
results (within 10-20%). However, problemsmay arise becauseof inhomoge-
neoussamples,matrix effects (such as variation in A or cr), or C contamination.
To compensatefor theseproblems,additional factors such as matrix correction
factors (Briggs & Seah,1983) needto be incorporatedinto Eq. [9].
Angle ResolvedX-ray PhotoelectronSpectroscopy

and Depth Profile Studies
Angle-resolvedXPS isa nondestructivemethodto study the changesin the

chemical composition as a function of depth. Sampling depth (escapedepth
times three) is a function of the anglebetweenthe axis normal to the samplesur-
face and the electroncollection path. Henceby changingthe sampletilt angle, a
variation in the depthof analysisis achieved.The effective depth of analysis (d)
is calculatedusing the equation
d = 3Acos8 [10]
whereA the attenuationlength (measuredexperimentally)and8 is the sampletilt

angle (Fig. 12-1).
Ion sputteringtechniquesallow one to study the chemicalcompositionof
minerals along a depth profile. Mineral surfacesare erodedby bombardingthe
372 VEMPATI ET AL.
surfacewith energeticions of 0.5 to 5 keV energy.Argon ions are most com-

monly used.To determinethe amountof material removedthe rate of material
removedmust be calibrated.This is bestdone by sputteringaway a thin layer of
materialsimilar to the materialof interest.To accomplishthis, thin layersof Si02
grown on silicon often are used. From the electronicsindustry, the knowledge
and technologyis availableto grow Si02 layers of precisethicknesson silicon
wafers. Recently, comparisonsbetween untreatedand Ar-sputtered materials
have been made to interpret the chemical changes along a depth profile
(Hellmannet aI., 1990; Inskeepet aI., 1991).
SAMPLE PREPARATION
How one preparestheir sampleis strongly dependenton thesefactors: (i)

the natureof material, (ii) the vacuumconditionswhich needto be maintained,
and (iii) any desired additionalexperimentaltreatments.If UHV conditionsare
desired,the sample,its holder and anymounting material must be UHV com-
patible. This can be a problem when the samplehas relatively high vapor pres-
sureor hasa tendency tooutgas.To achievea reliable XPS spectruma basepres-
sureof 1.33 x 10-6 PA (10-8 Torr) is preferred.However,short of grosssurface
contamination(fingerprints, cutting oil, etc.), most mineralswith the exception
of somehighly porousmaterialswith poorly boundwater of hydrationare UHV
compatible(Hochella, 1988). If possible,samplesshould be relatively flat and
orientedperpendicularwith respectto the spectrometer,exceptin caseof angle-
resolvedstudies.The samplearea should be larger than the area of the beam
diameter(usually a coupleof mm2) andsinceonly the near-surface region is ana-
lyzed, samplesmay be very thin.
Most XPS systemsare only able to handle a sampleholder in a specific
way; therefore,certain designelementsfor all the samplesholdersfor a particu-
lar systemmust be the same.Once theserestrictionsare met, the sampleholder
can take on any numberof different forms to meet the needsof various sample
and experimentalrequirements.Usually, theseholders are made from an UHV
compatible materials that is conductive such as stainlesssteel, AI or 02-free
high-conductivity(OFHC) copper.The choiceof materialcan be very important
if any part of the holder is "seen" by the spectrometerbecauseof the spectral
interferences.
Samplesmay be mountedin variety of different ways. For larger single
piece samples,e.g., rocks, a mechanicalmeansof mounting often works. This
usually involves a screw or clip that holds the sample in place. Mechanical
mountingworks well as long asthe clampinghardwareis not "seen"by the spec-
trometeror doesnot interferewith the spectrallines of interest.For smallersam-
ples,silver paint or carbonpaint is often usedto glue down a sample.Gluessuch
as thesework bestbecausethey are conductiveand UHV compatible.A couple
of drawbacksto thesematerialsis that the bond may not be as secureas in the
mechanicalcase,and for the porousmaterialsthe glue may permeatethroughthe
sampleand appearon the surface.This is particulartrue for carbonpaint. Powder
samplesareperhapsthe mostdifficult to mount on to a holder. Oneof the biggest
problemsis spectralinterferencefrom the mounting material as well as prefer-

ential charging. A number of different ways of mounting powder are used.
Powder may be pressedinto pellet so they can be mounted by a mechanical
method.The drawbackhere is that pressingmay affect the surfacechemistryof
the sample.Another methodis to sprinkle the powderonto a double stick tape.
Many haveusedthis methodsuccessfullybecause the principle spectralinterfer-
ence is in the C region. Recently, a conductive double stick carbon tape has
appearedon the market and has proven very useful for mounting conductive
samplesand may help to reducechargingproblems.Hard powdersamplesalso
may be pressedinto a soft metal foil such as In. To summarize,there is no best
methodfor handling and mounting samples.The best rule is to avoid materials
that have interfering lines and that eachsamplehas to be handledaccordingto
the natureof the sampleand the experimentproposed.
SamplingHandling andTreatment
Samplehandlingis extremelyimportantbecauseone hasto protectthe sur-
facesfrom extraneouscontaminations.After the samplehasbeenplacedinto the
spectrometer,the samplemay be treatedin a specialchamber(Fig. 12-4). The
samplescan be cleanedby mechanicalmeanswhile under vacuum using metal
brushes,scrappersor samplecleavers.Referencematerialssuch as Au can be
depositeddirectly on the samplein va:cuumor a reactantcan be addedvia the gas
phaseor by depositionin an inert atmosphere.Also, the samplecan be heatedor
cooled during chemicaltreatments.
RECENT ADVANCES AND DEVELOPMENTS
Significant improvementshave been developedin the XPS instrumenta-

tion in recentyears.One of the more importantdevelopmentis the ability to map
a region of a sample,at a particularphotoelectronenergy,with a spatial resolu-
tion of lO)lm. This allows scientistsa meansof examiningchangesin the surface
composition and chemical state acrossa heterogeneoussample. Furthermore,
experimentsare underway in Siegbahn'slaboratoryto developliquid phaseXPS
(Barr, 1991; Siegbahn,1990). This would be a valuable tool for probing the
natureof interactionsat the vapor/solidand liquid/solid interfaces.In particular,
this should be extremely useful in the study of chemical interactionsbetween
clay and liquid and vapor phases.One areathat needimprovementis in decreas-
ing the analysistime so that samplesthat are sensitiveto prolongedexposureto
x-rays may be analyzed.This can be achievedby improving electronanalyzers
and detectors(Hochella, 1988).
ACKNOWLEDGMENTS
Dr. David Cocke is the Jack M. Gill Professorof Analytical Chemistry
located at Lamar University, Beaumont, Texas. Drs. Vempati and Hess are
researchassociatesworking with ProfessorCocke. Their contributionsare par-
tially supportedby the TexasAdvancedTechnologicalResearchProgramof the
374 VEMPATI ET AL.
Texas Higher Education Coordinating Board and the Robert A. Welch

Foundation.The authorsare grateful to Dr. Micheal F. Hochella, Jr., and two
anonymousreviewersfor their commentsandsuggestions.Also, part of the man-
uscript contributed by RKV was written when he was a National Research
Council postdoctoral research associateat the National Aeronautic Space
Administration-JohnsonSpace Center, Houston, TX, and he is grateful to
NASA-JSC and the National Academyof Sciences,Washington,DC, for pro-
viding this fellowship. The authorsalso are grateful to Mr. StevenJ. Pytel for
assistancewith the figures.
REFERENCES
Bancroft, G.M., I.R. Brown, and F.W. Fyfe. 1977. Calibrationstudiesfor quantitativex-ray photo-
electronspectroscopyof ions. Anal. Chern.49:1044-1048.
Barr, T.L. 1991.Advancesin the applicationof x-ray photoelectronspectroscopy(ESCA). II. New
methods.Crit. Rev. Anal. Chern.22:229-325.
Borade,R.B., A Adnot, and S. Kaliaguine. 1990. Characterizationof acid sitesin pentasilzeolites
by x-ray photoelectronspectroscopy.J. Catal. 126:26-30.
Briggs, D., and M.P. Seath.1983. Practicalsurfaceanalysis.JohnWiley & Sons,Inc., New York.
Cocke,D.L., R.K. Vempati, and R.L. Loeppert.1994.Analysis of soil surfacesby x-ray photoelec-
tron spectroscopy.p. 205-235.In J. AmonetteandI.W. Stucki (ed.) Quantitativemethodsin
soil mineralogy.SSSA,Madison,WI.
Defosse,C.P.,andP.G. Rouxhet.1980.Introductionto x-ray photoelectronspectroscopy.p. 169-203.
In I.W. Stucki and w.L. Banwart(ed.) Advancedchemicalmethodsfor soil and clay miner-
als research.D. Riedel Publ. Co., Boston,MA.
Eland,I.H.D. 1984. Photoelectronspectroscopy.Butterworth,Boston.
Evans,S., and E. Raftery. 1982. Determinationof the oxidation stateof manganesein lepidolite by
x-ray photoelectronspectroscopy.Clay Miner. 17:477-481.
Foord,J.S.,R.B. Jackman,andG.c. Allen. 1984.An x-ray photoelectronspectroscopic investigation
of the oxidationstateof manganese. Philos. Mag. A49:657-663.
Gonzalez-Elipe,A.R., I.P. Espin6s,G. Munuera,J. Sanz,and J.M. Serratosa.1988. Bonding-state
characterizationof constituentelementsin phyllosilicatemineralsby XPS and NMR.I. Phys.
Chern.92:3471-3476.
Hellmann,R., C.M. Eggleston,M.F. Hochella,Jr., and D.A Crerart.1990.The formation of leached
layers on albite surfacesduring dissolution under hydrothermal conditions. Geochem.
Hochella,M.F. Ir. 1988.Auger electronand x-ray photoelectronspectroscopies. p. 573-637.In EC.
Hawthorne(ed.) Spectroscopicmethodsin mineralogyand geology.Vol. 18. Min. Soc.Am.,
Washington,DC.
Inskeep,w.P., E.A Nater,D.S. Vanderwoort,P.R. Bloom, and M.S. Erich. 1991.Characterizationof
laboratoryweatheredLabradoritesurfacesx-ray photoelectronspectroscopyandtransmission
electronmicroscopy.GeochimCosmochim.Acta 55:787-801.
Kohiki, S., T. Ohmura,and K. Kusao. 1983.Appraisalof a new chargecorrectionmethodsin x-ray
photoelectronspectroscopy.1. ElectronSpectrosc.Relat. Phenom.28:229-337.
Kohiki, S.T., andK. Oki. 1985.An appraisalof evaporatedgold asan energyreferencein x-ray pho-
toelectronspectroscopy.1. ElectronSpectrosc.Relat. Phenom.31:85-90.
Koppelman,M.H., AB. Emerson,and I.G. Dillard. 1980. AdsorbedCr(lII) on chlorite, illite and
kaolinite: An x-ray photoelectronspectroscopicstudy. Clays Clay Miner. 28:119-124.
Muilenberg,G.E. 1979.Handbookof x-ray photoelectronspectroscopy.Perkin-ElmerCorp., USA.
Murray, J.W.,I.G.Dillard, R. Giovanoli, H. Moers,andW. Stumm.1985.Oxidationof Mn(I1): Initial
mineralogyoxidationstateand aging. Geochim.Cosmochim.Acta 49:463-470.
Oku, M., K. Hirokawa,and S. Ikeda. 1975.X-ray photoelectronspecroscopyof manganese-oxygen
systems.1. ElectronSpectrosc.Relat. Phenom.7:465-473.
Payntner,R.W. 1988.Introductionto x-ray photoelectronspectroscopy.In B.D. Ratner(ed.) Surface
characterizationof biomaterials.Elsevier,New York.
X·RAY PHOTOELECTRONSPECTROSCOPY 375
Perry, D.L. 1990. Applications of surfacetechniquesto chemicalbondingstudiesof minerals.Am.

Chern.Soc. Symp. Ser. 323:389-402.
Powell, C.J. 1978. The physical basisfor quantitativeanalysisby Auger electronspectroscopyand
x-ray photoelectronspectroscopy.p. 5-30. In N.S. Mcintyre (ed.) Quantitativesurfaceanaly-
sis of materials.ASTM STP643. Am. Soc. Test Mater., Philadelphia.
Powell, C.J.,andM.P. Seah.1990.Precision,accuracy,anduncertainityin quantitativesurfaceanaly-
sis by Auger electron spectroscopyand x-ray photoelectronspectroscopy.J. Vac. Sci.
Technol.8:735-763.
Ratner, B.D. 1983. Study of biomaterialsby electron spectroscopyfor chemical anlaysis. Ann.
Biomed. Eng. 11:313-336.
Seah,M.P., and W.A. Dench. 1979. Quantitativeelectronspectroscopyof surfaces:A standarddata
basefor electroninelasticmeanfree pathsin solids. Surf.InterfaceAnal. 1:2-11.
Seyama,H., and M. Soma.1985. Bonding statecharacterizationof the constituentelementsof sili-
cate minerals by x-ray photoelectron spectroscopy.J. Chern. Soc. Faraday Trans. 1
81:485-495.
Seyama,H., and M. Soma. 1988. Applications of x-ray photoelectronspectroscopyto the study of
silicate minerals.Res.Rep. no. 11. Natl. Inst. Environ. Stud.,Japan.
Siegbahn,K. 1990.From x-ray to electronspectroscopyand new trends.1. ElectronSpectrosc.Relat.
Phenom.51:11-36.
Stucki, J.W., C.B. Roth, and W.E. Baithinger. 1976. Analysis of iron-bearingclay mineralsby elec-
tron spectroscopyfor chemicalanalysis(ESCA). Clays Clay Miner. 24:289-292.
Vempati, R.K., R.H. Loeppert,and D.L. Cocke. 1990a.Mineralogy and reactivity of amorphousSi-
ferrihydrites. Solid Statelonies 38:53-61.
Vempati, R.K., R.H. Loeppert, D.C. Dufner, and D.L. Cocke. 1990b. X-ray photoelectronspec-
troscopyto differentiatesilicon-bondingstatein amorphousiron oxides. Soil Sci. Soc. Am.
J. 54:695-698.
Wagner,C.D. 1978. How quantitativeis electronspectroscopyfor chemicalanalysis?An evaluation
of the significant factors. p. 31-46. In N.C. Mcintyre (ed.) Quantitativesurfaceanalysisof
materials.ASTM STP643. Am. Soc.TestingMater., Pheladelphia.
Wagner,C.D., and A.J. Joshi. 1980. The Auger parameter,its utility and advantages.A review. 1.
ElectronSpectrosc.Relat. Phenom.5:259-266.
Wagner,C.D., W.M. Riggs, L.E. Davis, J.F. Moulder, and G.E. Muilenberg. 1979. Handbookof x-
ray photoelectronspectroscopy.Phys.ElectronicsDiv., EdenPrairie, MN.
Zhao, L.Z., and V. Young. 1984. XPS studiesof carbon supportedfilms formed by the resistive
depositoinof manganese. J. ElectronSpectrosc.RelatedPhenom.35:45-54.
Published 1996
Chapter13
X-Ray Absorption Fine Structure

Spectroscopy
SCO'IT E. FENDORF,University of Idaho, Moscow, Idaho
DONALD L. SPARKS,University of Delaware, Newark, Delaware
With the ever-increasingimportanceof maintainingagriculturalproductivity and

environmentalquality, it is essentialfor researchersto determinethe chemical
and physical propertiesof soils and waters,and to ascertainand predict the fate
of many substancesin thesesystems.Determining the elementalcomposition,
chemicalspeciation,and reactivity of soils directly is essentialto accuratelychar-
acterize these media. Such information provides one with a knowledge of the
bioavailability, mobility, and toxicity of addedfertilizers, pesticidesand herbi-
cides,and pollutants.
A multitude of spectroscopicand microscopic techniquesare currently
availablefor investigatingsoils and soil chemicalreactions,with the numberof
techniquesand their capabilities advancingrapidly. These techniquesprovide
detailed information on constituentscomposingsoils and their chemical state.
Unfortunately,no single techniqueis a panaceafor investigatingthesecomplex
and heterogeneoussystems,thus it is beneficial to utilize a multitude of tech-
niquesto obtain a completeand accuratedepictionof the chemicalenvironment.
One method that has recently proven to be a powerful meansfor obtaining the
speciationand local structureof elementspresentin soils is x-ray absorptionfine
structure(XAFS) spectroscopy. X-rayabsorptionfine structurehas a variety of
advantagesfor studying soils which include: elementspecificity, in situ investi-
gations, determination of oxidation states,and the local chemical and structural
environmentof an element.
X-ray absorption fine structure offers information which is somewhat
unique amongthe currently availablemethods.It probesthe local chemistryand
structureof a single elementthroughouta sample.The oxidation state, type of
nearestneighbors,coordinationnumber,bond distances,and orbital symmetries
of the x-ray absorbingelementcan be accuratelydeterminedin an array of media
(Eisenberger& Lengeler, 1980). Becausethe information obtainedwith XAFS
differs from that of other spectroscopiesand microscopies,when used in con-
Book Seriesno. 5.
377
378 FENDORF& SPARKS
junction with them XAFS offers a complementarymeansfor detailing the prop-

ertiesof a soil and studyingreactionstherein.
Vibrational spectroscopies(infrared and Raman)give information on the
molecularaspectsand interactionsof a compound.Magneticspectroscopies such
as nuclear magnetic resonance(NMR) and electron paramagneticresonance
(EPR) can be usedto determinethe chemicalenvironmentof a species.Both of
thesespectroscopies can be conductedundernoninvasivesampleconditions,and
studiescan be performedon solids, surfaces,or solutions. They are, however,
subjectto the necessityof having an elementor compoundwhich is activatedby
the respectiveperturbation,a magneticfield or a low-energy photon induced
excitation, and vibrational spectroscopiesprovide no direct electronicinforma-
tion. Electronicspectroscopies, e.g., ultraviolet (UV), visible (VIS), or lumines-
cent, rely on the excitation of an element'svalanceelectrons.In so doing they
give oxidation stateinformation, and undersomeconditionslimited information
on the structuralenvironmentof the adsorbingelement.However,detailedstruc-
tural information, such as bond distancesand the type and numberof coordinat-
ing atoms,is not derivedfrom thesespectroscopies. Scatteringtechniquescan be
employedon soils to determinestructural information on a sample; however,
most conventionaltechniques,e.g., x-ray diffraction, only probe the long-range
order of a sampleand not the local structureof a given element.Additionally,
they do not provide chemicalstateinformation.
Microscopic techniquesalso can be usedto study soils and soil chemical
reactions, providing a wealth of information. Scanning electron microscopy
(SEM) provides three-dimensionalparticle morphology and local particle sur-
face featuresgenerally on a micrometerscale (although high-resolutionSEMs
are capableof betterthan lO-nm resolution). Higher resolutioncan be obtained
with transmissionelectron microscopy(TEM) so that details about the atomic
orderingof a samplecan be obtained.Both TEM and SEM can be equippedwith
energydispersivespectrometersfor elementalanalysis,and electronenergyloss
spectroscopy(EELS) is availableon many TEMs yielding information similar to
the edgestructurein XAFS.
Recently introducedsurfaceprobing microscopies(SPM), such as scan-
ning force (SFM) and scanningtunneling microscopies(STM), yield detailed
surface information. Their application should provide exciting details on the
microtopologyand reactivity of soil surfaces.All of the microscopictechniques
provide a meansfor obtaining the spatial proximity of surface modification;
however,with the exceptionof EELS and in somecasesSTM, they do not pro-
vide electronicinformation nor the local chemicaland structuralenvironmentof
a given elementor compoundthroughoutthe sample.Additionally, most of the
electronmicroscopiesmust still be conductedundera vacuumenvironmentand
introduce the sample to electron bombardment-whichcan severely damage
many samples.
X-ray absorptionfine structurecircumventsmany of theselimitations and
thereforeis an extremely useful tool for investigatingsoils and their reactions,
especiallywhen employedin conjunctionwith complimentarytechniques.The
attributesand limitations of XAFS are the focus of this chapter.The aim is to
provide the readerwith an understandingof the physical bases,experimental
X·RAY ABSORPTION FINE STRUCTURESPECTROSCOPY 379
considerations,and dataanalysisproceduresof XAFS. We give a brief overview

of the principlesof XAFS followed by a summaryof dataanalysistechniquesfor
XAFS that provide structural parameters.The final portion of this chapter
describesthe experimentalproceduresthat are employedwith XAFS, application
of XAFS to soils is addressedthroughoutthis chapter.
PHYSICAL BASIS OF X-RAY ABSORPTIONFINE STRUCTURE
When x-raysare impingedupon a samplethey can be elasticallyor inelas-

tically scattered.As they passthrougha sampleof thicknessx they are exponen-
tially attenuated.This attenuationis proportional to the sample thickness,the
incident x-ray intensity (10), and a constantof the material at a particular energy
(p, the material'sabsorptioncoefficient)
It =10 exp (-px) [1a]
which upon rearrangementcan be expressedas,
In Ie/It =fIX [lb]
where It is the transmitted intensity. Photoelectricabsorption is one possible

inelasticprocessaccountingfor this attenuationand is the basisfor XAFS spec-
troscopy.
The absorptioncoefficientdescribesthe ability of the materialto absorbx-
rays at a given energy.This parameter,p, is a function of energyso that an ele-
mentsability to absorbx-rays at different energiesis reflectedby the value of p.
Sometimesa convenientway to expressthis quantity is with a massabsorption
coefficient,pip, to accountfor the quantity of matter through which the photon
beamhastraveled.Simple dividing p by the densityof the material,p, yields this
parameter.Absorption coefficientsare additive so that a material'stotal absorp-
tion coefficient, (pip)m, is the summationof the absorptioncoefficientof its con-
stituents:(p/P)m =l:{p,/p k The valuesof the absorptioncoefficientsfor most ele-
ments over a wide range of energieshave been tabulatedby McMaster et a1.
(1969).
Measuringchangesin p with changesin x-ray energyprovidesinformation
on the chemicalandstructuralstateof the absorber(Sternet aI., 1975).The infor-
mation containedin XAFS results from the x-ray excitation of an atom's core
electrons.With sufficient energy input the core electronscan be completely
ejectedfrom the atom, the energyrequiredfor this processis the electronbind-
ing energy (BE). Quantum mechanicalselection rules specify that for energy
absorptionto occur a transition exactly equal to the incident energy must take
place.Thus, if an unoccupiedvalenceorbital is present,and the transitionof the
core electron to this unoccupiedstate is symmetry allowed (which is governed
by dipole selectionrules), then absorptionmay take place at a defined energy
below that of BE. When the incident photon energy equalsthe BE of the core
electron,absorptiontakesplace ejecting the electronfrom the atomic shell into
380 FENDORF& SPARKS
O PhotoeleClrOn :} .
4> : conttnuum
..
> ;
;
:
o (,,~.:~.:..:..:~:_:...:~~.~__ ~,._::,".,:< } "bound" valencestateS

, , -: -:« ",' _ . _' .:
: .:. ::,: :: ::~~ ~:~:~:~::... :.~.'<

T
j \~ ~;)\,",.c L shell
..
: : ~ ~. .~ ~: ~ ~ ~ l
\. f.
h"~~ " j K shell

i. •••••••• ;.:.;.:.;.:.;.;.::.:..:.:;.::.::.:..:.::.::••::•••.• .•.••••••••••••••
Fig. 13-1. An x-ray photon inducedexcitation of an atom'score-electron.The excited electroncan

either enter unoccupiedorbitals or be ejectedfrom the atomic shell into the continuum.An elec-
tron which is ejectedfrom the shell is termeda photoelectron
.
the continuumcreatinga photoelectron.Figure 13-1 gives a schematicdepiction

of theseprocesses.
In XAFS, an energyrangeis chosento encompassthe BE for the coreelec-
trons of the atom of interest.Since every elementhas a specific core configura-
tion, the binding energiesare unique to each element.Thus, XAFS is element
specific. For example,if one wished to analyzeeu in a sample,which has a Is
electron binding energy of 8979 eV, the incident energy would start slightly
below 8979 e V and then be increasedprogressivelybeyondthis energyvalue.
The energy balancefor photoelectricabsorptionis such that the kinetic
energy(KE) of the photoelectroncreatedby x-ray absorptionis equalto the inci-
dent energy(Ie) minus the BE of the excitedelectron:KE = Ie - BE. The abrupt
increasein photon absorptionat BE producesan absorptionedge (Fig. 13-2).
BeyondBE x-ray absorptioncontinueswith the photoelectronscreatedhaving a
greaterand greaterKE. Figure 13-2 depictsan absorptionedgeillustrating three
general spectral regions: the pre-edge,near-edge,and extendedportion. The
near-edgeregion is denotedXANES (x-ray absorptionnearedgefine structure)
while the region extendinggenerallyfrom 50 eV abovethe edgeand beyondis
termedEXAFS (extendedx-ray absorptionfine structure).
Becausethe absorptionprocessesare due to an electronictransitionof the
core electron to an unoccupiedorbital or into the continuum,XAFS inherently
possesses electronic information on the absorbingelement.This information is
primarily locatedin the pre-edgeand XANES spectralregions.Electronic tran-
sitions are governedby dipole selectionrules. Theserules dictate that electron
transitionsfrom one orbital to another(state-to-statetransitions)can only occur
X-RAY ABSORPTION FINE STRUCTURESPECTROSCOPY 381
X-ray absorption spectral regions
<l--I~ t>
PRE-EDGE NEAR EXTENDED
EDGE (EXAFS)
(XANES)
-1+---~----'---~-----'----~---r----~---r----~--~
5750 6000 6250 6500 6750 7000
Energy (eV)
Fig_ B-2. When the x-ray energyis sufficient to eject a core electron,a dramaticincreasein absorp-
tion occurs.The threespectralregionsresultingfrom this phenomenonare depicted:the pre-edge,
the nearedge (XANES),and the extendedportion (EXAFS).
with unit changesin the orbital angularmomentumquantum number(Le., 61 =

1); s~p and p~d transitionsare allowed, but s~d or p~f are not. However,
mixing of orbitals often occurs,thus transitionsthat would otherwisebe forbid-
den are observed.The excitation of a core electronto an unoccupiedorbital is
thereforegovernedby the numberof unoccupiedstatesas well as the symmetry
correlation betweenthe core and empty valence orbitals. These state-to state
transitionsoften result in intense,sharpabsorbances that are termedwhite lines.
This is exemplified by the strong edgefeature in Cr(VI) which resultsfrom the
s~d transition(for examplesee,Davenport& Isaacs,1991; Manceau& Charlet,
1992; Bidoglio et aI., 1993); the tetrahedralsymmetryof CrOl- resultsin mixed
p-d orbitals thus allowing for the otherwiseforbidden transition. This intense
white line is absentin Cr(lll) due to the lack of p-d orbital mixing and fewer
unoccupiedd-orbitals.
Furtherinformation on the oxidation stateof the absorbingelementis pro-
vided by the energyposition of the absorptionedgeand white line. The binding
energyof a core electronis a function of the electronconfigurationof the atom-
ic shell. Sincethe numberof protonsis fixed, a varying numberof electronshave
to compensatefor this charge. When valence electronsare removed from an
atomic shell (oxidation), the remaining electronsare drawn closer to the posi-
tively chargednucleusto increasechargeshielding.As a consequence, the BE is
increasedand a greateramountof energyis necessaryfor photoelectricabsorp-
tion--shifting of the absorptionedgeand white line(s) to higher energies.Thus,
the absorptionedgeor white line energyposition can be used to determinethe
oxidation state of an absorbingelement.In fact, for ionic compoundsa linear
relationshipbetweenthe energyof the absorptionedge,or white line, and oxida-
tion stateresults(Kunzl, 1932).
382 FENDORF & SPARKS
a)
hv
A
=>
Energy
b)
~-----~:-\~~
Energy
mulLiple scattering
Fig. 13-3. If an atom was not coordinatedby other atomsthen a smoothmonotonicdecayin x-ray
absorptionwould occur at energiesgreaterthan BE (a). However, atomsare always in a coordi-
nated environmentwhich results in the interaction of backscattered(in-coming) photoelectron
wavesin the emanatingwaves(b). This producesa fine structurein the x-ray absorptionspectraat
energies greater than BE.
As can be seenfrom electronstate-tostatetransitions,the x-ray absorption

processesare dependenton the fmal-state wave function (Stem, 1974; Lee &
Pendry, 1975), e.g., the overlap of the core and empty valenceorbital strongly
determinesthe white line intensities.A further result of the fmal statewavefunc-
tion is that when theincidentenergyis greatenoughto createa photoelectron,its
interactions with the atoms coordinating the absorberaffect the absorption
process.If the excited atom were in an imaginaryvacuumenvironmentwith no
other atomssurroundingit, then onewould observea smoothmonotonicdecay
in the amplitudeabovethe absorptionedge(Fig. 13-3a).However,this is never
the casein reality. All atomshave somekind of coordinatingenvironmentand
thus the photoelectronis affectedby the interactionswith its surroundings.
Electrons have both a particle and wave nature. Consideringthe wave
propertiesallows one to glean insight as to the interactiveprocessesof the pho-
toelectronand the neighboringatoms that producethe oscillations beyond the
edge. When a photoelectronis created,the electronwave will propagateaway
from the central atom (the absorber),it may scatteroff neighboringatoms,and
then finally return to its point of origin. This scenariois schematicallyillustrat-
ed in Fig. 13-3band 13-4a.Undersuchconditionsthe outgoingwavesthen will
interact with incoming waves that resultedfrom the backscattering.When the
a)
100cractjnc WaVes
b)
c)
-+-----
Fig. 13-4. A schematicdepictionof (a) the outgoingand incoming wave interactions,(b) the resul-
tant amplitudeof in-phaseinteractionand (c) out-of-phaseinteraction.
outgoing and incoming waves are in-phasewith each other, constructiveinter-

ferenceoccursenhancingabsorption(Fig. 13-4b). As the KE changes,the pho-
toelectron wavelength changesuntil the waves are 1800 out-of-phase.This
resultsin destructiveinterferenceand a loss in absorptionintensity (Fig. 13-4c).
The consequence of thesescatteringphenomenaand wave interactionsis that the
intensity of the x-ray absorption,at energiesgreaterthan BE, oscillate with a
dependenceon the structuralenvironmentof the absorber.Mathematicallymod-
eling theseoscillations,basedon physical considerations,providespreciselocal
structuralinformation on the absorber.
The length that the photoelectrontravels, or its survival pathlength,simi-
lar to the ion meanfree path (IMFP), is dependentupon its KE. The pathlength
is greatestat low KE, decreasesto a minimum, and then graduallyincreaseswith
384 FENDORF& SPARKS
150
100
0<
~
'-"
...c::
.....
~
0..
Q)
10
~
~
t::
~
Q)
~
1
2 10 100 1000 2000
Energy (eV)
Fig. 13-5. The ion meanfree path (IMPF) of an electronas a function of its KE (from Teo, 1986).
increasingKE (Fig. 13-5). The pathlengthof the photoelectronhas important

consequences on the analysisof theseoscillations.If the pathlengthis long then
multiple scattering(scatteringoff more than one atom before returning to the
central atom; seeFig. 13--3b) can significantly contributeto the oscillatory por-
tion of the spectra.Undersuchconditionsone must considerthesemultiple scat-
tering pathsin a mathematicalrepresentationof the modificationson the absorp-
tion intensity inducedby the structuralenvironmentof the absorber.In contrast,
if the pathlengthis short suchthat only electronsof single scatteringtrajectories
survive, then one can make a simplifying assumptionthat a mathematicalrela-
tionship of single scatteringcan describethe oscillatory function (Stem et aI.,
1975). This is generally the casefor EXAFS and is the key to structural infor-
mation obtainedfrom this spectralregion.
core-holeproduction
t///
e·
Fig. 13-6. 1\'10 relaxation processesare viable after a

core-hole is produced:fluorescencex-ray or Auger
Auorescence Auger electronemission.
X-RAY ABSORPTIONFINE STRUCTURESPECfROSCOPY 385
After the ejection of a core electron,there are two pathsfor relaxationof

the excited state: fluorescent x-ray or Auger electron emission. Figure 13-6
showstheserelaxationprocesses.Auger and fluorescenceemissionare inverse-
ly related, electron production decreasesand x-ray production increaseswith
atomic number.Fluorescentand Auger yields are tabulatedin many texts (e.g.,
CRC Handbook of Chemistry and Physics, Lide, 1993; E-196). Becauseboth
emission processes are relatedto the core-holeproduction,their measurements
can be utilized to acquireXAFS spectra.Due to the measurementof a core-hole
productthe relationshipof the fluorescentto the incident intensity is not one of
exponentialdecayand can be expressedas
[2]
Fluorescencedetectionturns out to be very beneficial for studiesof dilute sys-

tems and is commonly practiced(Stem & Heald, 1979). However, for concen-
trated samplesthe fluorescentx-rays may be absorbedby other processes(self-
absorption),which removesthe relation presentedin Eq. [2]. That is, fluores-
cenceis only proportional toabsorptionfor dilute samples.Auger electrondetec-
tion is often beneficial for surfacestudies.The application of these detection
techniqueswill be discussedfurther in later sectionsof this chapter.
SYNCHROTRONSAND BEAM LINE HARDWARE
Synchrotronfacilities produceextremelyintensephoton fluxes, which are

greaterthan a million times thoseof conventionalstationaryor rotating anodex-
ray tubes.Due to the high intensitiesof synchrotronx-ray sources,XAFS can be
performedon soils, suspensions, andsolutionswithout the needfor samplealter-
ations, i.e., in situ analysis. This is a great advantagesince no artifacts are
inducedby the analytical environmentnecessaryfor measurement,e.g., drying,
heating,or evacuation.High intensity x-ray sourcesnow usedfor experimenta-
tion are actually storagerings rather than synchrotrons.With synchrotrons,par-
ticles are acceleratedevery 0.1 to 1 ms and then the electronor positron is dis-
charged.Storagerings, on the other hand,acceleratethe particlesfor hours pro-
ducing stable x-ray sources.The storagering is an elliptical structurethat is a
polyhedronwith bendingmagnetsat its apices.Electronsor positronsare accel-
eratedat nearly the speedof light and then theyare injected into the ring. The
bendingmagnetsthen pull the particlesaroundthe ring maintaininga constant
speedwhile changing their trajectory, which is in essencean acceleration.A
physicalconsequence of particle accelerationis the productionof light in the x-
ray energy. In the ring, x-rays are given off tangentialto the curvature,and so
beam lines are installed on the ring to capture and collimate these photons.
Figure 13-7 gives a representationof a synchrotronstoragering.
From Fig. 13-7 it can be seenthat there are multiple beamlines at a syn-
chrotron.In somecaseseachline is split into variouscomponents.The synchro-
tron providesthe collimated high intensity x-rays, but eachbeamlinemust then
condition the photonsfor the type of experiment.Somebeamlinesare dedicated
386 FENDORF& SPARKS
to scattering,othersto microprobeanalysis,and a few to x-ray absorptionexper-

iments. Each of the techniquesmay require different instrumental features and
beamconditioning.
The physics of the storagering determineat what energy the maximum
intensity will occur. When planning an experimentit is thus important to con-
sider which facility will offer the best x-ray sourcefor the study. Studyingvery
heavy or very light elementswill be most fruitful at the developingthird gener-
ation x-ray sources.The advancedphoton source (APS), Argonne National
Laboratory,will provide high intensitiesat very high energiesmaking the study
of heavyelementalK-edgespossible.High energiesare currently availableat the
Cornell High EnergySynchrotronSource(CHESS),but theAPS will havemuch
greater intensities. In contrast, the advancedlight source (ALS), Lawrence
BerkeleyLaboratory,is beingdevelopedfor the study of light elementscommon
to biological systems.Midrangeenergies,optimal for studyingfirst- and second-
row transition elements,is offered by The National SynchrotronLight Source
(NSLS), Brookhaven National Laboratory, running since the mid-1980s.
Stanfordis homeof the oldestsynchrotronin the USA, the StanfordSynchrotron
Radiation Laboratory (SSRL), which first ran strictly in parasitic mode, and
offers a host of energyranges.
Before XAFS analysiscan take place, the x-ray white light must be nar-
rowed to a finite wavelengthwith the capacity to changethe selectedwave-
length. The smallerthe rangeof energiesimpinging upon the samplethe better
the resolution. Energy selection is accomplishedby passingthe incident light
through a crystal monochromator.The most frequently used monochromator
arrangementis two Si or Ge crystals; for Si the (111), (220), or (400) facesare
photons
"
Fig. 13-7. A depictionof a synchrotronstoragering and the associatedbeamlines for conductingx-
ray experiments.
X-RAY ABSORPTIONFINE STRUCTURESPECTROSCOPY 387
employed.The wavelengthof light passingthroughthe monochromatoris deter-

mined by Bragg'slaw (nA =2d sin 8). The principle energyis obviously n = 1,
with higher energiespassingat larger n values-higher-orderharmonics.The
higher-harmonicscan be a sourceof experimentalconcern,as discussedin the
following section.Grazing incident mirrors or detuningthe monochromatorcan
be usedto reject unwantedhigher-orderharmonics.The higher-orderharmonics
havea narrowerenergyrangethan the principle so that if the two crystalsare off-
set slightly the unwantedhigh energy will be severely limited relative to the
selected energy. Detuning to half of the maximum principle intensity will
decreasethe harmonicsby at least 100 times (Heald, 1988b). Since the energy
must be varied during the courseof an XAFS experiment,the monochromators
are rotated with a goniometer,changingthe diffraction angle and thus wave-
length (energy) of the passingx-rays. Defects in the crystals can causehigh
intensity, but narrow, spurious peaks in a spectra.Although data analysis can
remove many glitches it is always best to have them absent to begin with.
Sometimesjust rotating the crystal slightly will removesuch artifacts.
The facilities providedto the uservary from beamlineto beamlinein addi-
tion to the synchrotronfacility. Generally,ionization chambersare always pro-
vided alongwith genericsample stages. Extendedx-ray absorptionfine structure
equipmentpools have beenestablishedat most locationswhich can be reserved
upon request. Included in thesepools are Lytle ionization and 13-elementGe
solid-state fluorescent detectors.Additionally, equipment for sample cooling
may be availablein the pooledequipment.Specialexperimentalset-upsare often
constructedby the user.
EXPERIMENTAL AND DATA ANALYSIS
ExperimentalConsiderations
To provide the intensity necessaryfor performingXAFS measurements in

a reasonabletime periodand in solvatedsystems,high intensity x-ray sourcesare
necessary.As a consequence,XAFS experimentsare conductedat synchrotron
facilities. Becauseof the competitive time allocation at these facilities, it
becomesimperative to have well-thought out experiments.In this section, we
will provide someinsightson considerationsneededfor fruitful experiments.
The different synchrotronfacilities in the USA are summarizedin Table
13-1. Thesehigh-intensity x-ray sourcespermit in situ investigations,but only
for elementsheavierthan Sc. Lighter elementsare subjectto a greaterintensity
Table 13-1. Synchrotronfacilities in the USA.

Name and location Operation
National SynchrotronLight Source(NSLS), Brookhaven,NY In operation

StanfordSynchrotronRadiationLaboratory(SSRL), Stanford,CA In operation
Cornell High Energy SynchrotronStation (CHESS),Ithaca,NY In operation
AdvancedPhotonSource(APS), Argonne, IL In start-up (1996)
AdvancedLight Source(ALS). Berkeley,CA In operation
388 FENDORF& SPARKS
a)
tro"'n_s" 'to:ra:_ge. :.:R~i! .lng:.:!. . jI:~4- Is.r

Elec..... • 6
":i~:t~w,~
Monochromator Controlled
eiwiron ment
Absorption= j.lX = In (f)
b)
. I,
Absorpllon= j.lX:= (r.)
Fig. 13·-8. A schematicillustration of an experimentaldesignfor XAFS experiments.After leaving

the storagering, a specific energy is obtainedby passingthe high intensity x-rays through a crys-
tal monochromator.The beamis then further monochromizedby passingthroughmetal slits, enter-
ing the incident intensity detector,and impinging on the sample.The sampleis rotated45 0 to the
incident beam for fluorescencedetection.If only transmissiondetection is performed,then the
samplemay be turnednormal to the incident beam. A referencematerial in the beampath is desir-
able for conductingXANES experimentswhereoptimal energycalibration is neededfor the edge
or white line positions.
loss by the sample environmentand must therefore be investigatedin a low

absorbingmediasuchas a vacuumor He(g). Unfortunately,this limits the in situ
study of many importantsoil constituents,such as AI or Si, with XAFS.
A further point is the considerationof the absorberin the sample.This will
dictate whetheran experimentis feasible, and if so what mode of detection is
most appropriate.Measuringchangesin the incident vs. the transmittedx-ray
intensity is the most historic meansof obtaining XAFS measurements . It is
achievedby simply placingtwo detectorsin the beampath,one prior (10) and one
post (It) to the sample(Fig. 13-8). Ionization chambersare the most common
detectorsfor 10 and It. Neglectingfor the momentinterferenceeffects, samples
with concentrationsexceedingabout 3 to 5% work very well with this mode of
detection.Model compounds(with high absorberconcentrations)are particular-
ly suitablefor this design.In fact, any samplewith high absorberconcentrations
needeitherto be diluted or run in transmissionmodesincefluorescenceemission
is only proportionalto absorptionin dilute systems.
The expectedattenuationof a samplecan be determinedby rearrangingEq.

[lb].
[3]
in which Wi is the weight of the absorberin the sample,and A is the areaexposed

to the beam.Additionally, this expressioncan be used to determinethe optimal
samplethicknessesfor a given experimentalarrangement.Optimal statisticsfor
transmissionrequire that about0.20 to 0.25 of the incident beambe absorbedby
10,0.50to 0.60 by the sample,and the rest by It (Heald, 1988a).For experiments
where a third detectorbeyondIt is needed,the proportionsof absorptionby the
10, I" and the sample shouldadd to less than one.
Detectionby fluorescenceemissionis desirablefor dilute samplesand for
reducingharmonicexcitations(Jaklevicet aI., 1977). In a dilute samplevery lit-
tle of the incident photonswill be attenuated,leading to the comparisonof two
large and similar numbersin the 10 and It detectors.This leads to the need for
very accuratemeasurements, or a very high signal-to noise(SN) ratio will result.
This restriction can be overcome by measuringa phenomenonrelated to the
core-holdproduction,e.g., fluorescenceor Auger emission,which thus enhances
the SN. Fluorescencedetection is very important for measurementof natural
materialssince often the concentrationsof the elementof interestare very low.
Additionally, higher-energyharmonicsoften are not attenuatedgreatly by the
sampleso that they impinge on the It detectorcausingdistortions in absorbance
measurements.With fluorescencedetectioll, higher-order harmonics passing
through the monochromatorresult in less distortion since these nonattenuated
photonsdo not reachthe Ir detector.
Fluorescentexperimentaldesignsare somewhatmore complex than trans-
mission experiments(Fig. 13-8). The sampleis usually turned 45° to the inci-
dent beamwith Ir positioned45° from the sample,thus being 90° off the inci-
dent beam.The emissionoccursover a full 41t solid angle, i.e., in all directions,
so that collecting the maximum intensity is arduous.Accordingly, it is desirable
to position a wide-angledetectoras close to the sampleas possible.Even so, it
is operationallyonly possibleto collect from a single side; thus, already making
maximumcollection only half of the emission.One quickly seesthat only a por-
tion of the emittedintensity is detectedunderthe bestof circumstances.An addi-
tional complication results from elastic and Compton scatteringof the incident
beam.Someof thesescatteredx-rays can reachthe detector,adding erroneously
to the detectedsignal if an energydiscriminatingdetectoris not employed.Since
fluorescentionization chambersare still the most commonly used, this can be a
seriousproblem.
There are two means commonlyemployedfor diminishing the scattered
primary beam intensity reachingI r. The first is to position an energy-discrimi-
nating filter betweenthe sampleand the detector.In practicesucha filter is made
of a thin elementalfilm (3-5 absorbancelengths thick) which strongly absorbs
the primary beamenergy,but not the fluorescentenergy.That is, the materialhas
an absorptionedgeat an energybelow that of the primary beam,but greaterthan
the fluorescent energy. For K-edge analysis, Z-l element filters are the best
390 FENDORF& SPARKS
choice,but the Z-1 filter is only appropriatefor 23 < Z < 44; for the study of ele-
mentsaboveZ = 44, Z-2 filters are appropriate(Lytle et aI., 1984). For Ledges
(commonly employedfor heavier elements)one must find an elementwith a
strongabsorptionbetweenthe excitation andemissionenergies.Although the fil-
ter reduceselasticor Comptonscatteringfrom reachingI r, the filter will emit flu-
orescentx-rays from this absorption.To limit the influencesof the filter emis-
sions a linear set of metal slits, Soller slits, are placedin betweenthe filter and
the detector(Heald, 1988a). Energy discriminatingalso can be achieved with
solid-statedetectorssuch as those made from Ge (Cramer et aI., 1988). With
thesedetectorsselectedenergiescan be acquiredrather than the full spectrum
emitted.
In conjunctionwith concentration,interferencesmust be evaluatedin the
experimentaldesign.Interferencescan arise from three sources:(i) absorbance
of the incident beam by an elementother than that specified, (ii) overlapping
edges,or (iii) absorbanceof the fluorescentx-rays. The latter is only of concern
if the experimentis performedwith fluorescencedetection.Interferenceeffects
(i) and (iii) originate from elementalconstituentswith edgesslightly below the
intended absorber,while effect (ii) is due to absorbancesat energiesslightly
greaterthan the absorber's.
Incident intensity lossesoccur if another element in the sample has an
absorbanceat an energynot much less than the elem~nt of interest.This effec-
tively reducesthe concentrationof the absorber,making detectionof low con-
centrationsdifficult. Unfortunately,there is no meansto overcomethis compli-
cation other than to removethe interfering elementfrom the sample.Becauseof
this problem it is difficult, or in somecasesimpossible,to study Siwith XAFS
in a matrixabundantin Al. The converseis not true: Al can be studiedby XAFS
with Si presentsincethe higherbinding energyof Si will not be exciteduntil 280
eV abovethe Al K-edge. However, the presenceof Si in an Al study leadsto a
complicationof the secondtype. When scanningthe Al edgeone would normal-
ly acquirethe spectraapproximately1000 eV beyondthe edge.However,the Si
edgewill occurat 280 eV abovethat of Al, making measurements at higher ener-
gies meaningless.Although this doesnot completelyinhibit sucha study, it does
decreasethe resolutionof the bond distance(L\r), which is determinedby L\r =
n/(L\k), where.1 is the k-rangeevaluated.
Solid-statedetectorsnot only provide energydiscrimination,but they also
can havemultiple channelsin a single unit, i.e., the detectoris actuallycomposed
of multiple singledetectors(Crameret aI., 1988).Multichanneldetectorsalso are
referred to as "multi-element" detectorsbecauseeach solid-statecomponentis
termedan elementof the detector.Thesedetectorsthus acquiremany signalsat
one time which can be averagedto producea single scan.Consequently,if one
employs a 13-elementdetector,13 signals are obtainedin the sameamount of
time as a normal single spectra.Obviously, this greatly enhancesonesresolution
(by the "';13).
Although fluorescencedetectionhasmany advantages,especiallyfor soils,
it is not without hindrances.Self-absorptioneffects may alter the spectralfine
structure, leading to erroneousstructural parameters.The sample should be
either sufficiently dilute or thin so that the changein optical density is less than
0.1 betweenenergiesbelow and above the absorptionedge.The comparisonof

fluorescent spectra,that are in question, with transmissionspectraallows an
assessment of self-absorptioninfluences.An additional disadvantagewith fluo-
rescenceis that for elementsin the soil having absorptionedgeswithin -1000e V
of the edgeof interest,the emittedfluorescentx-rays are absorbedby the sample
and thus inhibit detection. For example,it would not be possible to study Ni
sorbedon Fe-oxideswith fluorescentdetectionsincethe Ni Ka x-rays (7480eV)
emittedwould be absorbedby the Fe (with a K-edge of 7111 eV). This compli-
cation can only be circumventedby detectingin transmissionor electroncapture
modes.
In addition to the desired energy, the monochromatorcrystals can pass
higher energiescorrespondingto additionalwavelengthsthat satisfy Bragg'slaw
and are allowed by the structureof the crystal. For Si(111) monochromators,the
third-order harmonicis of concern.This implies that for the selectionof a given
energy,an energy3x this amountalso may exist. The result of this phenomenon
is that the 3rd-harmonicenergywill passthrough the samplewith little attenua-
tion, creatingdifficulties for absorbancemeasurements (It will not be an accurate
measurement).Furthermore,at selectedenergiesbelow the edgeof interest,the
higher harmonicwill havean energymore than sufficient to excite the absorber.
This can dramaticallydiminish the edge-jumpand distort information contained
in. the EXAFS. Therefore,one should always insure that harmonicrejection is
maximized.Either detuningthe monochromatoror employinga harmonicrejec-
tion mirror can decreasehigher energythroughout.Detuningalso can be usedas
a checkfor harmonicinfluences.
Effectively, only the harmonicsshould excite the sampleand influence If
at selectedprinciple energies belowthe edge.Thus,if 10 is unchanged,a decrease
in Ifllo must result from a decreasein If (Lytle et aI., 1984). At theseenergies,
when If is not decreasedwith further detuningthen higher-orderharmoniceffects
should be negligible. This method for monitoring harmonicsis summarizedas
follows. The energyis tunedso that the principle is below the edge(maybe-50
to -100 eV) and Ifllo recorded,then the monochromatoris detunedto reduce10
by 10% and Ifllo is again read.This procedureis continueduntil Ifllo (or log lofl)
is unchangedwith detuning,at this point harmoniceffects should be minimal.
Samplescan be in the form of solids, suspensions,or solutionsfor XAFS
analysis.For solutions,concentrationsof about 5 mM are requiredfor measure-
ment of most species,but this dependson the constituentscomposingthe solu-
tion. Employmentof 13-elementsolid-statedetectorsallows one to measurecon-
centrations10 to 100 times less. Suspensionsare commonly investigatedin soil
science.If one wishesto discern information on the solid comprising the sus-
pension,it is often advantageousto consolidatethe solid phasewithout remov-
ing it from the sampleenvironment.This can be performedusing filtration while
keepingthe filtrate moist, or through centrifugation.Powderedsolids can easily
be studied,while larger fractions can either be groundand analyzedas a powder
or cut to fit the experimentalset-up. For fluorescencedetection,large fractions
can be analyzedprovided the surfacedoesnot obstructthe emitted x-rays from
reachingthe detector.If a large solid is to be measuredby transmissionit should
be cut into thin section-thethicknessof which can be determinedby Eq. [3].
Cooling a sampleis often conductedfor XAFS experimentsto reducethe ther-

mal disorderand thus the Debye-Waller factor (which is an amplitudereducing
componentof the EXAFS). Liquid N temperatures(77 K) are desirablefor this
purposesincefurther cooling may be costly and only modestlydiminishesther-
mal motion.
Time is alwaysof the essence at a synchrotronfacility. Therefore,it is pru-
dentto optimizethe availablebeamtime while acquiringthe spectralquality nec-
essaryfor accuratestructuralanalysis.Accordingly, the energyregion for scan-
ning, the monochromatorstepsize (Le., the energywidth moved), and the inte-
gration time at each point should be adjustedfor the desiredexperiment.For
XANES analysis,one may not wish to scan a large spectralregion, but rather
limit the energiesto within 50 to 100 e V of the edge. One also would want a
small step size to resolvesharp featureswhich reside in this region. However,
sincethe maximumresolutionsetby the monochromatoris usuallyabout0.2--0.3
eV, it is not useful to makestep sizessmallerthan this. If EXAFS analysisis to
be the thrustof the experimentthen a large scanregion is necessary;starting200
eV below the edge(-200 eV) and scanningto about 800 to 1000 eV is a com-
mon procedure,but the specific experimentshould be considered,e.g., if an
interfering excitation residesat energiescloserto the edge,then unlessan ener-
gy discriminatedetectoris employedscanningbeyondthat energydoesnot pro-
vide useful data. The region capturedfrom -200 to -50 eV provides for esti-
matesof the background.Becauseno significant featuresshould arise here, a
large steppingrate (10 eV) and low integrationtime (1 s) can be used. Nearing
the edge,the stepsize shouldbe narrowed(1-2 eV steps)to give the resolution
necessaryfor XANES features.The integrationtime neednot be increaseddue
to the intensity obtainedin this spectralregion. Although variable,at about50 to
100 eV beyondthe edgethe EXAFS amplitudebeginsto decaywhile the peri-
odicity increases.Hence,the integrationtime shouldbe enlarged(anywherefrom
2-16 s is typical), while the step sizescanbe increased.In our depiction,we have
usedthreespectralregionswith differing parametersto acquirean EXAFS spec-
tra. Certainly, greater(or fewer) regionscould be used,we mustagainstressthat
it is dependentupon the intent of the given experiment.
SpecialExperimentalDesigns
For certain applications, experimental set-ups may be altered. X-ray

absorptionfme structureis not inherently surfacesensitive,yet many studiesin
soil and environmentalscienceswish to discern surfacestructures.The easiest
way to do this is to limit the absorberto the surface.Then, XAFS would be sur-
face sensitive since the element being examinedresides only at the surface.
When this can not be done a special experimentaldesign must be employed.
SurfacesensitiveEXAFS experimentsare commonlynamedSEXAFS.
Surfacesensitivitycan be obtainedby limiting to the surfacethe absorbing
element, incident photons, or detected signal. A product of the excitation
process,as earlier discussed,are Auger electrons.The short IMFP of electrons
meansthat thosedetectedmust haveoriginatedfrom the surfaceregion (30-50A
deepdependingupon the energyand the samplecomposition).Hence,measure-
ment of the absorptionprocessesthroughelectroncaptureis necessarilysurface

sensitive.Unfortunately,becauseof the short IMFP of electrons,theseexperi-
mentscannotbe conductedin situ, they must be conducted invacuumor at least
a He(g) environment orthe electronswill be attenuatedprior to detection.
Total external reflection of x-rays impinging on a smooth surface will
result at glancing incident anglesbecausethe index of reflection is slightly less
than one. Surfacesensitivity resultsas a consequence, with the x-ray penetration
depth less than about 30 A. These experimentsare termed "glancing angle"
XAFS. While they are not restrictedto vacuumenvironments,they do require a
specialalignmentof the sampleand the incident beam-withpreciseand vari-
able anglesavailable.
Determinationof oxidationstatescanbe obtainedfrom the energyposition
of white lines or the absorptionedge.Most beamlines can provide about0.15 to
0.20 eV resolution,which can yield accurateoxidationstateinformation because
a unit changein valenceresults in about a 1 to 2 eV shift in energy. But, the
experiment must be accurately calibrated to a standardto determine energy
shifts. This can be doneby runningthe standardbetweeneachsample,and recal-
ibrating. A better meansfor obtaining the necessarycalibration, however, is to
employ an internal standard.This doesnot require introducingthe standardizing
materialinto the unknownsample,but ratherhaving two spectraacquiredsimul-
taneously.Figure 13-8 showsa diagramof the detectorarrangementproviding
for thesemeasurements. A third ionization chamber,I r, is placedbeyondIt and
the referencematerialmountedin betweenIt and I r . That makesIt the equivalent
of an 10 detectorfor the referencematerial.For this procedure,in calculatingthe
thicknessof the unknownsample,evenfluorescentexperimentsshouldnot be so
thick as to completely attenuatethe incident beam, i.e., it is necessaryto have
someintensity left to reachthe calibratingstandardand I r •
Electron,Symmetry,and ChemicalEnvironmentDetermination
A unique feature of XAFS is that both detailedspeciationinformation is

provided inaddition to accuratestructuralmeasurements. The pre-edgeand, pri-
marily, the XANES regionsof XAFS containinformation on the electronicstate
of the absorbingelement,its symmetry,and thus its local chemicalenvironment.
Spectralfingerprints of XANES allow for the identification of a speciesevenin
complex matrices,and undersomecircumstancesprovide quantitativemeasure-
ments. Detailed electronic information on the absorbingelement can be dis-
cerned using quantum mechanical models of state-to state transitions (e.g.,
Sham,1985; Zaanenet aI., 1985; Petiauet aI., 1987).
Multiple or complexunknownchemicaland structuralenvironments,such
asthoseoften found in soils, often precludethe useof EXAFS analysisfor deter-
mining the structurein which an elementresides.The XANES spectrado, how-
ever, provide an opportunity to discern structuraland chemical information on
the elementof interestunder the aboveconditions.Although detailed,complex
calculationscan be performedon XANES featuresto determineaccuratestruc-
tural or electronicinformation,employmentof a fingerprint methodis often most
useful to determine elemental speciationin unknown samples.This is usually
accomplishedby comparingthe XANES featuresof an unknownto known com-

pounds.The useof statitisticalfitting methodsimprovesthe accuracyand details
obtainedusing a fmger print type method.A further advantageof XANES mea-
surementsis that the relatively intensefeaturesin this region allow for the mea-
surementof low elementalconcentrations,i.e., measurementof dilute systems.
Generally,the intensityof the XANES featurespermitsthe determinationof oxi-
dationstatesthroughwhite line or edgeposition at concentrations100x less than
those neededfor EXAFS analysis,coordinationenvironmentsthrough spectral
finger prints can be determinedat concentrationslOx lower than thoseneeded
for EXAFS analysis.
The oxidation stateof an elementis easily determinedby the energyposi-
tion of white lines or the absorptionedgesince,aspreviouslystated,the binding
energy of the core electronwill be influencedby the valenceshell occupancy.
Generally,for eachchange inoxidation stateabouta 1to 2 e V shift occursin the
edgeposition. By taking careto standardizeto a known compound,usinga metal
foil or an elementalsampleof the absorbingelement,an energy resolution of
approximately0.15 eV can be obtained,allowing for the determinationof oxi-
dationstates.A useful linear relationshipbetweenthe edgeenergyand oxidation
stateoccursfor ionic compounds.This was first postulatedby Kunzl (1932), but
is ideally only valid for isolatedionic compounds.Significant covalentbonding
or othercomplicatingfactors may removethis linear relationship.However,the
strong electronwithdrawing capacity of speciessuch as 0 2- in oxyanionspre-
serve the linear relationshipbetweenthe edge position and valencestate-as
demonstratedby the linear relationshipexhibited in S compounds[Fig. 13-9]
(Franket al., 1987; George& Gorbaty,1989; Huffman et aI., 1991;Waldo et aI.,
1991; Vairavamurthyet aI., 1993,1994; Morra et ai., 1994,unpublisheddata).In
fact, the intensity and energydependenceof white line featureshasallowed for
a quantitative determinationof S speciesin complex matrices such as heavy
petroleums,petroleumsourcerock, and coal (George& Gorbaty,1989;Waldo et
ai., 1991; Huffman et al., 1991; Vairavamurthyet ai., 1994).
In addition to chemicalspeciationinformation, XANES featuresalso can
be usedto discernsymmetryinformationon the x-ray absorbingelement.Again,
this is most easily approachedby identifying features in model compounds
which representa coordinationenvironment,and using this information to deter-
mine the coordinationsymmetryof an elementin an unknown.Waychunaset ai.
(1983) and Waychunas(1987) usedsuchan approachto ascertainthe site geom-
etry of Fe and Ti, respectively,in various minerals.Furthermore,the polarized
propertiesof synchrotronradiation permittedthe elucidationof bound and con-
tinuum state symmetriesof sulfate, chlorate, thiosulfate, and dithionate with
XANES (Tyson et ai., 1989).
Rather rigorous calculations can be conductedon XANES features to
model the state-tostate transitionsand determinethe electronicsymmetriesof
the absorbingspecies.Commonly,experimentsare conductedin which energies
are selectedto excite different electronshells.This providessymmetryinforma-
tion by comparingtransitionsfrom core electronsof differing characterto the
empty orbitals; for example,the Lz,3 excitationswere usedto measurethe ionic-
ity and solid-state effects in oxides of 3-d transition metals (Grunes, 1983;
X-RAY ABSORPTION FINE STRUCTURE SPECTROSCOPY 395
a) sulfide
elemental sulfur
2460 2465 2470 2475 2480 2485 2490 2495

Energy (eV)
7
6 b)
II.)
e;; 5
U'l 4
-=: 3
0
.~ 2
"'01
·R 0
°_1
CIl
-2
-3
2465 2470 2475 2480 2485
Energy (eV)
Fig. 13-9. Often XANES has advantagesfor determiningstructuresin unknown compounds,and
provideschemicalstate information. The fingerprint utility of XANES has many advantagesfor
studying sampleswith great heterogeneityor with very low concentrationsof the absorber.The
XANES spectrafor varoiusS-bearingcompounds(a) illustrates theability of XANES featuresfor
identifying different local chemical and structuralenvironments.The linear relationshipbetween
the energyposition of the white-line peakposition and S oxidation stateallows for the identifica-
tion of S oxidation statesin unknown samples(b).
396 FENDORF& SPARKS
Waddingtonet aI., 1986). Muller and Wilkins (1984) presenta formalism to cal-
culate x-ray spectra from core excitations and illustrate this approach with
detailed computationsof the K, L, and M absorptionspectraof palladium. A
recent publication by Stohr (1992) provides an excellent review of electronic
considerationof XANES spectra.
Sulfur and Metal Speciationin Soils
The utility of XANES for speciatingelementswithin soils is exemplified

with S (Morra et aI., 1994,unpublisheddata).In the K-edgeanalysisof S ( (2472
eV), a whiteline feature is apparentdue to the transition of the Is electroninto
the empty p orbitals. The peakposition or inflection point of the white line can
be usedto determinethe oxidationstateof S. Using organicand inorganicS com-
poundsof known oxidation statesa relationshipbetweenthe white-line energy
position and S oxidation statewas developed(Fig. 13-9). This relationshippro-
vides the opportunityto discernthe oxidation stateof S in soils and soil materi-
als. It is important to realize that the "apparent"oxidation statesof organic S-
containingcompoundssuchas cysteineand methionineare not necessarilyinte-
gers. For example, we confirmed oxidation statesof +0.4 to +0.6 previously
reportedfor cysteine,cystine, and methionine(Frank et aI., 1987; Waldo et aI.,
1991).
Humic materials obtained from the International Humic Substances
Societywere placeddirectly into the sampleholder of the spectrometerwithout
any additional treatment.All spectrafrom the humic materialsproduced white
line peaksindicating the presenceof S in multiple oxidation states.Soil and peat
humic acids producedwhite lines indicative of S in oxidation statesof +6.0 and
lessthan +1.0 (Fig. 13-10). Fulvic acidsproducedwhite lines indicative of S in
oxidation statesof +5.0 and speciesbetween0 and +2.0 (Fig. 13-10). We con-
cluded that soil humic acids containedS in the +6.0 oxidation state,indicating
the presenceof ester-bondedsulfates.In contrast,fulvic acids containedS pre-
dominately in the +5.0 oxidation state. Specieswith this oxidation state likely
exist as sulfonic acids. However, reducedforms of S also are presentin both
humic and fulvic acids. ReducedS in soil and peat humic acids with oxidation
statesbetween0 and +1.0 representsamino acid S.
Previousto the use of XANES, S speciationin soil, although critical to
agricultureand environmentalquality, was approachedusing indirect techniques
that resultedin crude fractionation. Additionally, theseearlier techniquesmost
often relied on the differential reductionof S speciesto HzS using hydriodic acid
or RaneyN. As a result of using thesetechniques,there remaineda tremendous
amount of uncertainty about the precursor molecules. Using XANES spec-
troscopy an accuratedepiction of the chemicalstateof S in soils was obtained
directly and nondestructively.
In addition to the speciationof S, XANES providesa meansfor defming
the oxidation stateof metalsin the solid-phase.By noting the inflection point or
peakposition of the metal absorptionedge,the oxidation stateof U in sediments
(Bertschet aI., 1994) and Mn (Schultzeet aI., 1995) in soil mineralswas deter-
mined. In somecases,the pre-edgewhite line featurescan be used to note the
':<
2482.2
a) ~
2480.4
b)
* i
* ~
.c- Suwannee River
....... ::.l
00 FU~ z
$:I .c-
....... l'"l
00
$:I
E
~ B
(1) 2474 $:I
~
::>
....... (1) ~
~ ~ ::>
....... ~
!;Il
~
~ ~
~ Soil Fulvic Acid
Soil Humic
;
o
~
2465 2470 2475 2480 2485 2490 2495 2465 2470 2475 2480 2485 2490 2495
Energy (eV) Energy (eV)
Fig. 13-10. White-line positionsin the XANES spectraof S from humic materials(a) indicatesoxidation statesof +6.0 (2482.2-2482.5eV) and less than +1.0 (2474
eV). In fulvic acids (b), the white-line positionsof the S XANES spectraindicate oxidation S specieswith oxidation statesof +5.0 (2480.4 eV) and between0 and ~
+2.0 (2473-2475eV). ~
398 FENDORF& SPARKS
oxidation stateof the absorber.This is clearly exemplified in the caseof Cr, in

which Cr (VI) hasa distinct pre-edgefeaturewhich is absentin Cr(III). However,
more subtlepre-edgefeaturescan be usedto glean information on the electron-
ic stateof metals.Recently,Bajt et aI. (1994) usedthe XANES pre-edgefeature
of Fe to deducethe presenceof Fe(II) or Fe(III) in primary minerals.The used
of pre-edgefeaturesas opposedto main-edgepeakpositionsis that the former is
generally not influenced by the bonding environmentof the absorber.That is,
multiple scatteringeventsmay skew the peakenergyposition of the absorption
edgemaking its energyposition dependenton the bonding environmentas well
as the electronicstateof the absorber.If electronicinformation is desired,it may
be beneficial to use a spectralfeature that is not influenced by the scattering
events,suchas pre-edgefeatures.In addition to pre-edgefeatures,the inflection
point of the main-edgealso is useful for determiningthe oxidation stateof the
absorbersince its position also will be influencedonly minimally by scattering
events.
Structural Analysis
Detailed structural parameterscan be extractedfrom both XANES and

EXAFS spectra.PresentlyEXAFS is utilized more extensivelyto deducenear-
est neighbor numbers, types, and distancesbecauseof the simpler physical
descriptionof the processesoccurringin this spectralregion. In this chapterwe
will only review the proceduresfor determiningprecisestructural parameters
from EXAFS analysis. Those readersseeking more extensivediscussionson
quantitativeXANES structuralmeasurements are referredto Petiauet aI. (1987),
Durham(1988), and StOhr (1992).
Due to the shorter IMFP above about 30 to 50 e V from the edge (Fig.
13-10),electronswhich scatteroff coordinatingatomsand do not return direct-
ly to the absorberwill have negligible influenceson absorption,i.e., only single
scatteringis significant. This allows one to employ a simplified, relative to mul-
tiple scatteringconsiderations,mathematicaldescription of the backscattering
interferenceson the x-ray absorption.In so doing onecan deducethe local struc-
ture in which the absorberresides.This simplification is the basisfor the single
scatteringsingleelectrontheory commonlyemployedfor EXAFS analysis(Stem
et aI., 1975).
The following discussionon EXAFS dataanalysisassumesthat good-qual-
ity spectrahave beenacquired,experimentalconsiderationsfor obtainingquali-
ty data are presentedin the next section.Subsequentto data acquisitionvarious
runs of an experimentare often averagedto further improve the spectralquality
and minimize glitches inducedby instrumentalfluctuations. Even with the best
of data, however, there still may be extremely sharp anomalousfeaturesas a
result of unwantedreflectionsin the crystal monochromator.Becausethesefea-
turesaregenerallyof a very narrow energywidth, it is possible,and desirable,to
removetheseglitches. Simply removing the data points completelyor fitting a
function from both sidesof the glitch is usually satisfactoryfor removing such
artifacts.This procedureis appropriatefor the large periodicity observedat ener-
a)
-1~--~--r-~---.--~--_r--~--.---r-~
5750 6000 6250 6500 6750 7000
0.5,-------
_ _ _ _ _ _ _ _ _ ---,
_
b)
O+---~--,_---_,--~--_r------._----~
5750 6000 6250 6500 6750 7000
Energy(eV)
Fig. 13-11. X-ray absorption spectrafor (a) y-CrOOH collected in transmissionmode, and (b)
K2Cr207 measuredby fluorescencedetection.One should note the decreasingabsorptionbeyond
the edgewhen collectedin transmissionmode which is on contrastto the increasingbackground
observedby fluorescencedetection.
gies about100 e V abovethe edge,but shouldbe appliedwith caution in the near

edgeregion where "true" sharpspectralfeaturesmay reside.
Examplesof deglitchedspectracollected in transmissionand fluorescent
modes are depicted in Fig. 13-11. Spectracollected by transmissionhave a
decreasingbackgroundwhile an increasingbackgroundresults when they are
collectedby fluorescence,the backgroundneedsto be subtractedin either case
before further analysis.The backgroundsare usually approximated,to a good
degree,by a pre-edgeregion from approximately-200 to -50 eV below the
edge.To remove the pre-edgecontributions,a low-order (often linear) polyno-
mial function is typically fit to this region (Fig. 13-12a);deviationsresult, how-
ever, with solid-statedetectorswhere Gaussianfunctions are typically needed
(Crameret aI., 1988).Following backgroundremoval,the absorptionintensity is
normalizedto a unit jump heightby dividing the magnitudeof the spectraby the
intensity changeat the absorptionedge(Fig. 13-12b).A unit jump height curve
with the pre-edgesubtractedis shown in Fig. 13-12c.
400 FENDORF& SPARKS
0.5
s::::::
.8
...... 0
fr
o
15 -0.5
<
-1
-1.5+----,---,-----,---.---,---..---'--,--'
5750 5900 6050 6200 6350 6500 6650 6800
Energy (eV)
I
subtraction
1.4,--------------------,
V
b)
s::::::
.....
o
1
......
~0.7
CZl
~ nonnalizing
point
5750
1
O~~~~_.----r_--._--_.--_.--~~
5900 6050 6200 6350 6500 6650 6800
Energy (eV)
Fig. 13-12. Whethercollectedin fluorescenceor transmission,the backgroundneedsto be subtract-
ed from the spectra.This is accomplishedby fitting a low-order polynomial equationto the pre-
edge region (a), and then subtractingthis equationfrom the spectralcurve (b). Additionally, the
jump-heightof the curve is usually nonnalizedto unity, as shown in (c).
The oscillatory portion of the EXAFS function arisesfrom its structural

environment.Therefore,one wishes to removethe contributionsof the atomic
type absorption(the absorptionwhich would occur if the atom were not coordi-
natedby other atoms)leaving only the oscillatory contributionin order to glean
detailedstructuralparameters.To a good approximation,the atomic absorption
can be modeledby the smoothlyvarying envelopeof the EXAFS. Slight varia-
tions in the methodfor isolating the oscillatoryportion of the spectrahavearisen,
I
normalization
1.5,-----------------------,
V
c)
s:::::
o
......
.....
e-
O
[/J
,.0
--< 0.5
o '-
5750 5900 6050 6200 6350 6500 6650 6800
Energy (eV)
Fig. 13--12. Continued.
but the basic stepsare the same.The envelopeportion of the data are fit with a
spline function, the spline function is then subtractedfrom the total absorption,
and finally the difference is divided by a normalizing function (Fig. 13-13).
Including the normalizationfunction, the isolating of the oscillationsis
1.1
Q)
u
§
f:
~ 0.95
~
'. ·-x
0.8+----,----,----,--...,------,-----;
5950 6100 6250 6400 6550 6700 6850
Energy (eV)
Fig. 13--13. After the backgroundhas beensubtractedfrom the EXAFS spectrathe portion arising
from scatteringneedsto be isolated.A spline function is thus usedto model the atomiclike absorp-
tion that occurs,subtractionof the spline function leavesonly contributionsarising from the coor-
dination environmentof the absorber.
14.-------------------____________________ ~
a) spline too floppy

14,----------------------------------------. s
c)
] ~
"2 .a
.....
s::
~ ~
~
tz t
2 345 6 7 8 9
Distance(A) 2 3 4 5 6 7 8 9
14.---------------________________________ ~
Distance(A)
b) spline too rigid
]
.....
s::
~ Fig. 13-14. Optimization of the spline function (the number of knots and the
~ ~
weighting) is determinedfrom the derivative curve and the Fr. The derivative Z
curve should not follow featuresin the X(k) function. The Fr of y-CrOOH is =
o
ti: illustrated.In (a) a splinewith too many knots with too high of a weightinggave
~
a spline that was too flexible to model the atomic-like absorption.This results r<.o
in a diminished peak intensity and broadeningof the feature. If the spline is
excessivelyrigid, the Fr will have a spuriouspeakat the low-energyside and
2 3 4 5 6 7 8 9 also will have reduced intensity.A good choice for a spline function is shown
Distance(A) by the Fr in (c). i
X·RAY ABSORPTIONFINE STRUCTURESPECTROSCOPY 403
x = Vtrlata - Ilspline)/#norm [4]

whereX is the oscillatory contributions,Ildata is the total absorption,Ilspline is the
assumedatomic contributions,and#norm is the normalizingfunction.
The spline function has a variable number of fixed points, knots, which
determineits flexibility. Additionally, it is weightedto provide greaterflexibili·
ty at different energies;the higher the weighting the greaterthe flexibility at
higher energies.Both the numberof knots andweighting are varied to obtain the
best representationof the absorptionenvelope.Optimization of the spline func-
tion can be achievedby monitoring featuresin the Fourier transform(Ff) func-
tion, and the shapeof the derivativeof X. One should seeka derivative function
which is independentof the oscillationsin X. If the derivativecurve follows fea-
tures in X then the flexibility of the spline function is too great-eitherthe
weighting and/orthe numberof knots needsto be decreased.Additionally, a low
Ff magnitudewill result if the spline is excessivelyflexible (Fig. 13-14a). In
contrast,a spline fit which is too rigid will producespuriouspeaksat unrealisti-
cally low distancesin the Ff (Fig. 13-14b).When the Ff magnitudeof known
real peaks(usually those arising from first shell contributions)are maximized
and spuriouspoints onthe low distanceside, and noiseon the high distanceside,
are minimized then a good spline function hasbeenacquired(Fig. 13-14c).
Finally, the starting and endingknot positionsof the spline function must
be addressed.Generally,the starting point can be taken at the midpoint on the
low energyside of the initial peak.The endpointcan be chosenat various loca-
tions, and often it is desirableto try different ones,but the choice should corre-
spond to the inflection point of a peak at the high energy end of the spectra.
Figure 13-14 presentsthe fitting of a spline function and an exampleof choos-
ing the endpointsof a spline function.
The EXAFS function needsto be normalizedto some consistentunit to
allow for a comparisonbetweendifferent spectra.Two choicesfor this purpose
often usedin data analysisare the spline function itself or a Victoreen function
(Teo, 1986).Although the splinefunction is often usedas the normalizingfactor,
it may containspecific interactionsof an atomic environmentand thereforemay
not always be the best choice, but nonethelessit is commonly used. Victoreen
functions are devoid of atomic environmentdependencies and are a convenient
meansfor normalizationsincethey havebeentabulatedfor most elements(Teo,
1986).
With the oscillatory portion of the EXAFS isolated,the energy units are
transformedfrom electronvoltsto angstroms-I;the equation in angstroms- 1 is
denotedas k space-kbeing the photoelectronwavenumber.
In Eq. [5], E is the energy(eV), Eo is the binding energy(eV), Ae is the photo-

electronwavelength,m is the massof an electron, and h is Planck'sconstant
divided by 21t. Defining the true Eo is experimentallyand theoreticallyarduous,
in practicea referenceEo is chosenwith the crucial point being to make a con-
sistentchoicebetweensamplesand referencematerials.The first inflection point
Fig. 13-15. Isolation of the scatteringinfluenceson the x-ray absorptionyields the X(k) function,
which is derived from the subtractionof the spline function modelingthe atomic absorptionfrom
the experimentalspectraas shown in Fig. 13-11. The contributionof the total amplitudeand total
phaseparametersin Eq. (5) are illustratedfor the secondshell of y-CrOOH.
in the absorptionedge is often used as the practical Eo, althoughother choices

include a distinct featurein the edgestructureor the half-heightpoint of the edge.
In k space,the X function is physically expressed
f Nj 2(r· -
X = k rf S~ (k) Fj(k) exp(-2Foj) exp L {
r- A)] sin [2krj + cj>(k)] [6]
which providesfor the determinationof the structuralparameters:coordination

number(Nj) and interatomicdistance,r. The spectralcontributionof thesecom-
ponentsis shownin Fig. 13--15. The coordinationnumberaffectsthe amplitude,
with greaterN increasingthe amplitude,while r influencesboth the amplitude
and phase.The disorderin the sampledue to slight differencesin structure,stat-
ic effects,and thermalvibrationsis reflectedin the Debye-Wallerfactor, o. Since
the photoelectriceffect is more rapid (on the order of 10-15 s) than vibrational
motions(10-13 s) the atomic positionsdo not reflect the averagevibrational posi-
tion but rather capturethe atoms in differing statesof their vibrational motion,
producinga thermalvibrational disorder.The disorder(static and thermal) caus-
es an energy-dependent exponentialdecreasein the amplitude.
The type of the backscatteris expressed bythe magnitudeof the backscat-
tering amplitude,Fj(k), which representsthe jth shell coordinatingatom. Many
body effects, the excitation of electronsother than the intendedcore-electron,
such as shake-up(excitation) and shake-off(ejection), result in an amplitude

reductionfactor, S6. The finite core-holelifetime and interactionsof the photo-
electron with valenceelectronsinduce further reductionsin the amplitude; the
meanfree path, A, representstheselosses.The core-holelifetime is a function of
the atomic number(Z) and the core electronthat is excited(i.e., it differs for K,
L" L", L"" etc., shells).It decreases as Z increasesand is shortestfor the Is elec-
tron, increasingwith further distantelectrons.Consequently,one must be careful
of experimentson K-shells of high Z elementssince the shortnessof the core-
hole lifetime may severelylimit resolvingany structuralinformation.
Inelastic processescan be double counted in the core regions; thus, the
parameter~ in Eq. [3], the core radius, is employedto eliminate this error. The
core radiuscan usually be approximatedby r of the nearestneighbor(Sternet aI.,
1980). The periodicity of X is determinedby r and a frequencyaltering affect, <1>,
from Coulombic interactions.As the photoelectronis ejectedit experiencesthe
absorber'sCoulombic potential, upon scatteringfrom a neighboring atom it
experiencesthat atom'sCoulombicpotential,finally as it returnsto the absorber
atom it once againexperiencesthat Coulombicpotential; therefore,ultimately it
experiencesthe neighboring atom's once and the absorber'sfield twice. This
alters the phase by a defined amount, <1>, dependenton the backscatterand
absorbertype. Although Eq. [3] may appearsomewhatcumbersome,the true
complexity of this event is hidden in the parametersF(k) and<1>(k).
A crucial factor in the predictedcurve is the calculationof the phaseshift.
The phaseshift can be determinedfrom model compounds,theoreticalcalcula-
tions, or a combinationof these.Model compoundshave beenthe most preva-
lent, and apparentlythe most accurate(Eisenberger& Lengeler, 1980), for the
past decade. However, with advancementin the theoretical calculations of
EXAFS, recentdevelopmentshaveprovideda meansfor accuratelydetermining
phaseshifts from theory (McKale et aI., 1988; Rehr et aI., 1992). Additionally,
these developmentshave allowed for the considerationof multiple scattering
paths.
When model compoundsare employed,their choice is partially dependent
on the methodusedfor determiningphaseshifts, but thosewith structuressimi-
lar to that expectedfor the unknown are always the most desirable(Eisenberger
& Lengeler, 1980). In fact, some fitting routines require that the structuresbe
similar, e.g., the ratio method,while othersonly needhavethe samecentralatom
and neighbor.Becausethe phaseshift is dependentonly on the core stateof the
atom, they are transferablebetweencompoundsof similar elementalconstituents
but differing structures.However, similar structuresare always most desirable
and should be employedwhen available.
The phasefunctions between periodic neighboring elementsare within
experimentalerror, providing the opportunity to use model compoundsof Z ± 1
of the absorberand neighboringatoms if no other model compoundsare avail-
able. One should note that this also makesit inherently difficult to distinguish
betweenneighboringatom types if they are periodically close.Finally, all model
compoundschosenshouldhavea high symmetryto reducethe ambiguity result-
ing from a plethoraof bonddistances.A limitation in using model compoundsto
define phaseshifts and amplitude parametersis that it may be difficult to find
406 FENDORF& SPARKS
a)
3 5 7 11 13 15
b)
3 5 7 11 13 15
Fig. 13 -16. Transformationof the isolatedoscillatory portion of the spectrainto k-space(A-I) pro-
ducesthe X(k) function. The X(k) function for y-CrOOH is shown in (a) unweightedand(b) for 12.
suitable materials. In addition, further problems arise if shells overlap.

Nonetheless,oncethe phaseshift functions havebeenproperly definedit is pos-
sible to ascertainthe type and numberof neighboringatoms,their distancefrom
the absorber,and their disorder.
Returningto our discussionon the analysisproceduresfor extractingstruc-
tural information, the oscillatory spectralcomponentshave been isolated pro-
ducing the X(k) function, an exampleof which is presentedin Fig. 13-16. The
amplitude of X(k) decayswith increasingenergy. This is unfortunatesince the
determinationof interatomic distancesdependson the frequency and not the
amplitude, and yet the larger amplitudes may dominate the smaller ones.
Therefore,unlessone wishesto emphasizea particularregion of the XAFS spec-
trum, obtaining approximatelyequal amplitude over the entire energy range is
most appropriate.Weighting schemesof k!', with n equal to 1,2, or 3, have been
suggestedfor Z > 57, 36 < Z < 56, and Z < 36, respectively(Teo & Lee, 1979).
Fourier transformationof the X(k) yields a spectrumin real spacewith

peaksproportionalto the distancesof the atomic shells,i.e., the Fr gives a radi-
al structurefunction (RSF). Various routinescan be usedfor the Fr, but for prac-
tical purposestheseare incorporatedinto data analysispackagesand therefore
will not be discussedhere. For a more thoroughdiscussionon Fr routines the
readeris referred to Sayersand Bunker (1988). Although phaseshifts can be
incorporatedinto the Fr, generally they are not so that the peaksin the spectra
are offset by the phaseshift, i.e., the peakscorrespondto atomic distancesuncor-
rectedfor phaseshifts. However,the Fr doesgive a first approximationof atom-
ic distances,and it allows for the isolation of variousshell contributions(Sayers
et aI., 1971). Figure 13-14 showsa Fr spectraand Fig. 13-17 illustratesthe iso-
lation of the first shell.
The isolated shell in the FT must then be transformedback to k-spaceto
determineaccuratestructuralparameters.The backtransformedfunction is often
referredto as a Fourier-filtered(FF) spectra.One shouldalwaysjudiciously pre-
sent raw spectrain addition to the FF since the latter are smoothedextensively,
prohibiting evaluation of the general data quality. In fact, the XAFS research
community often requiresthat raw spectraalways be presentedwith any further
EXAFS analysis.
A curve-fitting routine producingthe best fit betweena predictedcurve,
from varied atomic dimensionsand parameters,with that of the experimental
curve is implementedto define the structureof the specimen.A backtransformed
spectraand a predictedcurve for y-CrOOH are illustrated in Fig. 13-18. The
phaseshift, backscatteringamplitude, mean free path, core radius, and many
body effectsare fixed by the atomic constituents:they are eitherdeterminedfrom
model compoundsor from theory. With theseparametersdefined,r, N, and cr can
be varied to obtain the best fit betweenthe predictedand experimentalspectra.
Under the bestof circumstancesthis can yield valuesof r :!: 0.01 A and N ± 20%.
SinceEo is a distant dependentparameter,and in practiceis experimental-
ly defined,it is often usedas a variable.This partially offsetsany errorsin its def-
inition. Thus, during a curve-fitting routine four parametersare varied to opti-
mize the fit of the predictedto the experimentalcurves:Eo, r, N, and cr. Varying
theseparametersmust be done with careso as not to have parametersoffsetting
errors in the other, which could lead to grossly inaccurate(and often physically
impossible)structures.Variablesthat are correlatedwith eachothershouldnot be
floated together: cr and N are strongly correlated,and r and Eo are moderately
correlated.
The resultsthus far shouldprovide detailedstructuresof the first shell con-
tributions, but the spectralcontributionfrom more distantshellsstill remainto be
determined.Going back to the RSF (the Fr spectra),one must now isolate the
secondpeak, backtransformit, and fit the experimentalcurve with a predicted
one to obtain its structuralcomposition.This procedureis continueduntil all the
prominentFr peakshave beensufficiently analyzed.Now one should have an
accuratedepictionof the unknown'sstructure.However,this compositestructure
shouldbe further testedfor accuracy.By using all the structuralcomponents,the
full experimentalX function can be predicted.An exampleof this is presentedin
Fig. 13-18 along with the resulting local structural model derived from the
14 ~
QC
I a) I b) 1\
mfirSIShell
<L)
I
"0
.-
.-.s ~
s:::
b.O .,Becondshell
~ / \I X
--
::E
~
v
U\r ~J t /\1\_ -I I I I I I
3 5 7 9 11 13
2 3 4 5 6 7 8 9 A-I
0
Distance (A)
c)
~
z
Fig. 13-17. To accuratelydetermine the structural componentsof a
~~\ ~ ~ IV t:l
0
given shell, the shell needsto be isolated in the FT and then back-
V 2!;
transformedinto k-spacefor modeling. The isolation of the first- P.o
VV
VV en
shell and second-shellcontributionsin y-CrOOH are shown in (a).
The backtransformedspectraof the first-shell is exhibitedin (b) and
~ 7
~~ 9 11 13
~
3 5
that of the secondpeak in (c). A-I ~
en
b) ~
a)
~ ~
S~
-.
~
X
O"'-S'T°"-
I
~
/ ~"Cr-O
o I
4 6 8 10 12 14 ~ OH
A-I Si I I
""0 •• '
3.39'~'/"'" c\r.::-;;-0, n
.'
' " S1-0
2.9 '."
---":Cr-o
I
o
~
~ ° I
s~O jH
Fig. 13-18. Determiningpreciseneighboringatom types, number,and
distanceis dependenton fitting the backtransformedspectrawith a
predictedcurve.The full backtransformedX(k) for y-CrOOH with the
predictedspectrais shown in (a). A schematicstructuralmodel of y-
CrOOH from thesevaluesis illustratedin (b).
f OH
/'''-OH $
410 FENDORF& SPARKS
EXAFS results.If one cannotobtain a good fit betweenthe predictedand exper-

imental curvesfor the full spectra,then the structuralparametersare suspect.A
secondmeansrecentlyavailablefor testingthe validity of the acquiredstructure
is to derive a model spectrawith thesevaluescompletelyfrom theory. The theo-
ry presentedby Rehret al. (1992) allows for this and is presentedin the computer
programsFEFF.
For energiesgreaterthan approximately50 eV abovethe absorptionedge
the single-scatteringtheory usually works quite well. However, for somestruc-
tures multiple-scatteringeffects can be influential. Such is the caseif atoms,in
successivelyclose shells, reside in a co-linear array. In this structural arrange-
ment the wave is focused by a first neighbor onto the second, thus greatly
enhancingthe latter'samplitude.Fortunately,in most casesthis phenomenonhas
a limited effect due to the IMFP restrictionsfor this trajectory,but nonethelessit
can be significant in many structures.One shouldbe awareof thesepossibilities
in predictedstructuresor model compounds.Under such conditions, multiple
scatteringeffects must be considered,which again can be accomplishedusing
recentcodesdevelopedby Rehr et al. (1992).
ExtendedX-ray Absorption Fine StructureAnalysis of Soil Materials
The EXAFS can provide much information on the structureof soil con-
stituentsand on structuralalterationinducedby reactions.The majority of stud-
ies utilizing this techniqueso far were conductedon rathersimple, homogeneous
systemsratherthan naturalsoils or sediments.Although EXAFS hasnot yet been
applied extensively to soils, it does offer many prospectsfor enhancingour
knowledgeof soils and soil reactions.
One of the primary usesof EXAFS is to elucidatethe surfacestructureof
sorbedmetal ions on surfacescommonto soils or waters,the referencesand gen-
eral subjectsof many suchstudiesare provided in Table 13-2. This information
is essentialfor determiningthe stability of a sorbate,which dictatesthe potential
for desorptionfrom a surfacephase.Furthermore,this information is neededfor
the developmentof accuratemechanisticmodels that can predict the fate of
metal ions in soils.
Whethera metal ion binds as an outer-sphereor inner-spherecomplexcan
be discernedwith EXAFS. The absenceof backscatterers beyondthe first shell
in the RSF (derived from the Fourier transformeddata as discussedearlier) is
evidencefor an outer-spheresurfacecomplex(Hayeset aI., 1987). In the caseof
an inner-spheresurfacecomplex, the coordinationenvironment(i.e., the inter-
atomic distancesand coordinationnumber)of the sorbate-sorbentcomplex can
be obtained(seefor exampleHayeset aI., 1987; Chisholm-Brauseet aI., 1990a,
b; Waychunaset aI., 1993).
RecentEXAFS studieshave shownthe presenceof multinuclearsorbates,
confirming and elaborating upon electron spin resonance(McBride, 1982;
McBride et aI., 1984; Bleam & McBride, 1986)and x-ray photoelectron(Tewari
& Lee, 1975; Crowther et aI., 1983) spectroscopicevidencefor nucleatedsor-
bates. Multinuclear chromium hydroxide surface phaseswere identified on
goethite (Charlet & Manceau,1992) and silica (Fendorf et aI., 1994b). While
Table 13-2. SelectedXAFS studieson soil mineralsand surfacecomplexes.

Subject Reference
Manganesestructures Arrenhiuset al. (1979)

Structuresof glasses Calas& Petiau(1983)
Local chemicalenvironmentsin Fe minerals Waychunaset al. (1983)
Structureof 0 in SiOz glasses Marcelli et al. (1985)
Local structuralenvironmentsin Fe minerals Waychunas(1986)
Seleniteand selenateon goethite Hayeset al. (1987)
Sulfur and Zn environmentsin zincblende Sainctavit etal. (1987)
Site characteristicsof Ti in minerals Waychunas(1987)
Local structureof Fe and Mn oxides Manceau& Combes(1988)
Local Fe structuresin hydrousferric hydroxides:
Fe structuresduring the formation of ferric gels Combeset al. (1989)
Fe structuresin hematiteformation from ferric gels Combeset al. (1990)
Co(II) sorption on rutile, alumina, and kaolinite Chisholm-Brause(1990a)
Pb(II) sorbedon y -A1 z0 3 Chisholm-Brause(1990b)
Cr(III) pillared smectites Corker etal. (1991)
Sulfur forms in coal Huffman et al. (1991)
Structuresof Pb(II) on a-FeOOH Roe et al. (1991)
Sulfur speciationin heavy petroleusm Waldo et al. (1991)
Cr(III) sorption on Fe(OH)3(am) and goethite Charlet& Manceau(1992)
NP(V) retentionon goethite Combeset al. (1992)
Uranyl structureson silica and montmorillonite Dent et al. (1992)
Cr(III) reactionswith birnessite Manceau& Charlet (1992)
Incorporationmechanismsof Srz+ in calcite Pingitore et al. (1992)
Chromium oxidation stateson ferrous hydroxides Bidoglio et al. (1993)
Multinuclear Cu speciesand Zn-sulfides Helz et al. (1993)
ReducedCr statesin olivine Sulten et al. (1993)
As sorption on ferrihydrate Waychunaset al. (1993)
Fe oxidation statesin minerals Bajt et al. (1994)
Speciationof U in soils and sediments Vairavamurthyet a\. (1994)
Surfacestructureof Cr(lll) on silica Fendorfet al. (1994b)
Sulfur speciationin marine sediments Vairavamurthyet a\. (1994)
Sulfur speciationin soils Morra et al. (1994, unpublisheddata)
Mn oxidation statesin soils Schulzeet a\. (1995)
nucleatedCr(III) sorbateswere observedon both materials,the local structureof

the sorbatesdiffered; the 'Y-CrOOH type local structureoccurredon silica while
a-CrOOH was presenton goethite.However, at high surfacecoveragesa phase
transitionfrom the a to the 'Y-CrOOH phasewas observed.
In addition to sorptionexperiments,XAFS hasbeenusedto determinethe
structureof minerals. The complex structureof various Mn-oxides was deter-
mined utilizing EXAFS, in one of its first applications to natural materials
(Arrenhius et aI., 1979). The site vacanciespresentin many Mn-oxides were
determinedas was the structuralenvironmentof interlayercations(e.g., Cu and
Zn). Furtherinvestigationsusing XAFS spectroscopyhave revealedmany of the
structuraldetails of Mn-oxides(Manceau& Combes,1988).
The precipitationof Fe-(hydr)oxidegels and their transformationinto crys-
talline (hydr)oxides is an important factor affecting soil chemical reactions.
However,detailsof thesereactionsare difficult to obtainevenin simple systems.
By employing EXAFS spectroscopyto theserelatively simple systems,in situ
investigationswere conductedon the structureof the amorphousFe-(hydr)oxide
gels and their progressivedevelopmentinto crystalline forms (Combeset aI.,

1989, 1990).
Data Analysis Programs
Numerouscomputer programs have been written for EXAFS analysis.

Many of these are free to general users and can be accessedthrough public
domain accounts.Othersare proprietaryand may be very expensive.Here, we
will provide a brief discussionon only a few of thesepackageswith an empha-
sis on thosethat are free or of nominal cost to the would-be user.
At various points in this text we have alluded to the advancementin the
application of theory to XAFS basedon the results of Rehr et ai. (1992). The
product of these advancementsis available in a relatively easily employed
computerpackage,FEFF. Version 6.0 is the most recently availableform of this
computer code, and it can explicity account for multiple scattering events.
Additionally, it provides an opportunity to accuratelycreate a spectrastrictly
from atomic coordinatesalthough it can be restricted to calculateonly single
scatteringeventsfor comparativepurposes.Comparisonsbetweenexperimental
and FEFF output have proven accuratefor many systems.It is not, however,a
data analysispackagefor data reduction;this routine is for comparingtheoreti-
cal to experimentalresultsor for generatingneededamplitude and phasefunc-
tions. The computercode is availablefor a small userfee and can run in a vari-
ety of operatingsystems.Interestedusersin FEFFshouldcontact:FEFFProject,
PhysicsDepartmentFM-15; University of Washington;Seattle,WA 98195.
A public domain accessibleprogram for data reduction and analysis is
available from the University of Washington.The University of Washington
packageis availablefor a multitudeof operatingsystemsincluding Macintosh.It
provides a convenientand user friendly meansfor data reduction, isolation of
atomic shells, and back transformationof thesecontributions.The fitting rou-
tines in this packagehavebeensuccessfullyappliedto a variety of systems.The
ratio methodis availablein addition to a FITT routine that useslinear compila-
tions of model compoundsto obtain a predictive curve. Another public domain
packageis now availableat Stanford University-EXAFSPAK. This code also
hasa userfriendly environment,and providesthe userwith a strongand flexible
meansfor dataanalysis.The fitting procedureis very powerful and can be cou-
pled with FEFF for computation of phase and amplitude functions.
Unfortunately,it is only currently compatiblefor VAX-VMS operatingenviron-
ments. Both EXAFSPAK and the University of Washingtonprogram can be
accessed electronically. The address for EXAFSPAK is
EXAFSPAK@BIOGG.SSRLOl.SLAC.STANFORD.EDU,and that for the
Macintosh version of the University of Washington package is
XAFSDB.IIT.EDU. Many other routinesare availableand new onesare current-
ly beingdeveloped.Someusershaveconstructedtheir own routinesfor their spe-
cific uses.Becausethis text is designedprimarily for the unfamiliar experimenter
we haveonly provideda brief overviewof a few ofthe easily accessibleand low-
cost (but not low-quality) computerpackagesavailable.
SOURCESOF FURTHERINFORMATION
Other excellentsourcesof information on XAFS spectroscopyare avail-
able. Books devotedto XAFS include thoseby Teo (1986), Koningsbergerand
Prins (1988), and Stohr(1992). In additionsto thesefine booksthereare numer-
ous review articleson this subject;someof thosethat are particularly relevantto
XAFS applied to soil scienceinclude thoseby Brown and Parks(1989), Brown
et al. (1989), Charletand Manceau(1993),and Fendorfet al. (1994a).Therealso
are a plethora of researcharticles involving XAFS spectroscopy;some of the
more pertinentstudieswith respectto soil scienceare listed in Table13-2.
REFERENCES
Arrenhuis,G., K. Cheung,S.E. Crane,M. Fisk, J.Z. Frazer,J. Korkisch, T. Mellin, S. Nakao,A Tsai,
and G. Wolf. 1979. Counterionsin marine manganates.lnC. Lalou (ed.) La genesedes nod-
ules de manganese.Coil. Int. CNRS (Paris)289:333-356.
Bajt, S., S.R. Sutton, and J.S. Delaney. 1994. X-ray microprobeanalysisof iron oxidation statesin
silicates and oxides using x-ray absorption near edge structure (XANES). Geochim
Bertsch,P.M., D.B. Hunter, S.R. Sutton,S. Bajt, and M.L. Rivers. 1994. In situ chemicalspeciation
of uranium in soils and sedimentsby micro x-ray absorption spectroscopy.Environ Sci.
Technol.29:980-4.
Bidoglio, G., P.N. Gibson, M. O'Gorman,and K.J. Roberts. 1993. X-ray absorptionspectroscopy
investigationof surfaceredox transformationsof thallium and chromium on colloidal miner-
ai oxides. GeochimCosmochimActa 57:2389-2394.
Bleam, W.E, and M.B. McBride. 1986.The chemistryof adsorbedCu(II) and Mn(lI) in aqueoustita-
nium dioxide suspensions. 1. Colloid InterfaceSci. 103:124--132.
Brown, G.E., Jr., and G.A Parks.1989. Synchrotron-based x-ray absorptionstudiesof cation envi-
ronmentsin earth materials.Rev. Geophys.27:519-533.
Brown, G.E., Jr., G.A Parks,and C.J. Chisholm-Brause.1989. In situ x-ray absorptionspectroscop-
ic studiesof ions at the oxide-waterinterfaces.Chimica 43:248-256.
Calas,G., and J. Petiau. 1983. Structureof oxide glasses:Spectroscopicstudiesof local order and
crystallochemistry:Geo-chemical implications. Bull. Miner. 106:33-55.
Charlet,L., andA Manceau.1992. X-ray absorptionspectroscopicstudy of the sorptionof Cr(III) at
the oxide-water interface: II. Adsorption, coprecipitation, and surface precipitation on
hydrousferric oxide. J. Colloid InterfaceSci. 148:443-458.
Charlet, L., and A Manceau.1993. Structure,formation, and reactivity of hydrousoxide particles:
Insights from x-ray absorptionspectroscopy.p. 117-164.In J. Buffle and H.P. van Leeuwen
(ed.) Environmentalparticles.Vol. 2. Lewis Publ.
Chisholm-Brause,C.J., P.A O'Day, G.E. Brown, Jr., and G.A Parks. 1990a.Evidencefor multinu-
clear metal-ion complexesat solid/solution interfacesfrom x-ray absorptionspectroscopy.
Nature (London) 348:528-530.
Chisholm-Brause,C.J., AL. Row, K.E Hayes,G.E. Brown Jr.,G.A Parks,and J.O. Leckie. 1990b.
XANES and EXAFS study of aqueous Pb(lI) adsorbed on oxide surfaces. Geochim.
Combes,J.M., A Manceau,G. Calas,and J.Y. Bottero. 1989. Formationof ferric oxidesfrom aque-
oussolutions:A polyhedralapproachby x-ray absorptionspectroscopy:I. Hydrolysis and for-
mation of ferric gels. Geochim.Cosmochim.Acta 53:583-594.
Combes,1.M., A Manceau,and G. Calas.1990. Formationof ferric oxidesfrom aqueoussolutions:
A polyhedralapproachby x-ray absorptionspectroscopy:II. Hematiteand formation of fer-
ric gels. Geochim.Cosmochim.Acta 54:1083-1091.
Combes,1.M., C.J. Chisolm-Brause,G.E. Brown, Jr., G.A. Parks,S.D. Conradson,P.G. Eller, I.R.
Triay, D.E. Hobart, and A Meijer. 1992. EXAFS spectroscopicstudy of neptunium(V)sorp-
tion at the a-FeOOH/waterinterface.Environ. Sci. Technol. 26:376--382.
Corker, J.M., J. Evans,and J.M. Rummey. 1991. EXAFS studiesof pillared clay catalysts.Mater.
Chern. Phys. 29:201-209.
Cramer,S.P.,O. Tench,M. Yocum, and G.N. George.1988. Nuc\. Inst. Meth. 586-588.
Crowther,D.L., J.G. Dillard, andJ.W. Murray. 1983.The mechanismof Co(II) oxidationon synthetic
birnessite.Geochim.Cosmochim.Acta 47:1399-1403.
Davenport,AJ., and H.S. Issacs.1991.Application of x-ray absorptionspectroscopyto the study of
corrosionand inhibition. Corrosion91, no. 74. Nat\. Assoc.CorrosionEng., Cincinnati, OH.
Dent, AJ., J.D.F. Ramsay,and S.W. Swanton.1992. An EXAFS study of uranyl ion in solution and
sorbedonto silica and monotmorilloniteclay colloids. J. Colloid InterfaceSci. 150:45-60.
Durham, P.J. 1988. Theory of XANES. p. 52~6. In D.C. Koningsbergerand R. Prins (ed.) X-ray
absorption. Principles, applications,techniquesof EXAFS, SEXAFS, and XANES. John
Wiley & Sons,New York.
Eisenberger,P., and B. Lengeler. 1980. Extendedx-ray absorptionfine-structuredeterminationsof
coordinationnumbers:Limitations. Phys. Rev. B22:3551-3562.
Fendorf,S.E., D.L. Sparks,G.M. LambIe, and M.J. Kelley. 1994a.Applicationsof x-ray absorption
fine structurespectroscopyto soils. Soil Sci. Soc. Am. J. 58:1583-1595.
Fendorf,S.E., G.M. LambIe, M.G. Stapleton,M.J. Kelley, and D.L. Sparks.1994b. Mechanismsof
chromium(I1I) sorption on silica. I: Cr(lII) surface structure derived by extended x-ray
absorptionfine structure(EXAFS) spectroscopy.Environ. Sci. Techno\.28:284-289.
Frank, P., B. Hedman,R.M.K. Carlson,T.A. Tyson, AL. Roe, and K.O. Hodgson. 1987. A large
reservoir of sulfate and sulfonate residues within plasma cells from Aseidia eeratodes,
revealedby x-ray absorptionnearedgestructurespectroscopy.Biochemistry 26:4975-4979.
George,G.N., and M.L. Gorbaty. 1989. Sulfur K-edge x-ray absorptionspectroscopyof petroleum
asphaltenes and model compounds.J. Am. Chem.Soc. 111:3182-3186.
Grunes,L.A. 1983. Study of the K edgesof 3d transition metalsin pure and oxide form by x-ray-
absorptionspectroscopy.Phys.Rev. B27:2111-2131.
Hayes,K.F., A.L. Roe, G.E. Brown, K.O. Hodgson,J.O. Leckie, and G.A. Parks.1987.In situ x-ray
absorption study of surface complexes: selenium oxyanions on a-FeOOH. Science
(Washington,DC) 238:679-682.
Heald,S.M. 1988a.Designof an EXAFS experiment.p. 87-118.In D.C. Koningsbergerand R. Prins
(ed.) X-ray absorption. Principles, applications, techniques of EXAFS, SEXAFS, and
XANES. JohnWiley & Sons,New York.
Heald, S.M. 1988b.EXAFS with synchrotronradiation. p. 119-161.In D.C. Koningsbergerand R.
Prins (ed.) X-ray absorption.Principles,applications,techniquesof EXAFS, SEXAFS, and
XANES. JohnWiley & Sons,New York.
Helz, G.R., J.M. Charnock,D.J. Vaughan,and C.D. Gamer. 1993. Multinuclearity of aqueouscop-
per and zinc bisulfiile complexes:An EXAFS investigation. Geochim. CosmochimActa
57:15-26.
Huffman, G.P.,S. Mitra, F.E. Huggins,N. Shah,S. Vaidya, and F. Lu. 1991. Quantitativeanalysisof
all major forms of sulfur in coal by x-ray absorptionfine structurespectroscopy.Energy &
Fuels5:574-581.
Jaklevic, J., J.A. Kirby, M.P. Klein, AJ. Robertson, G.S. Brown, and P. Eisenberger. 1977.
Fluorescencedetectionof EXAFS: Sensitivity enhancement for dilute speciesand thin films.
Soil StateCommin. 23:679-682.
Koningsberger,D.C., and R. Prins. 1988. X-ray absorption.Principles,applications,techniquesof
EXAFS, SEXAFS,and XANES. John Wiley & Sons,New York.
Kunzl, V. 1932. A linear dependenceof energy levels on the valency of elements.Collect. Czech.
Commun.4:213-224.
Lee, P.A, and J.B. Pendry.1975. Theory of the extendedx-ray absorptionfine structure.Phys.Rev.
B1:2795-2811.
Lide, D.R. (ed.). 1993. Handbookof chemistryand physics.CRC Press,Ann Arbor, Ml.
Lytle, F.W., R.B. Greegor,D.R. Sandstrom,E.C. Marques,J. Wond, C.L. Spiro, G.P. Huffman, and
F.E. Huggins. 1984. Measurementof soft x-ray absorptionspectrawith a fluorescent ion
chamberdetector.Nuc\. Instr. Method. 226:542-548.
Manceau,A, and L. Charlet. 1992.X-ray absorptionspectroscopicstudy of the sorptionof Cr(I1I) at
the oxide-waterinterface: I. Molecular mechanismsof Cr(I1I) oxidation on Mn oxides. J.
Colloid InterfaceSci. 148:425-442.
Manceau,A, and J.M. Combes.1988. Structureof Mn and Fe oxidesand oxyhydroxides:A topop-
logical approachby EXAFS. Phy. Chem. Mineral. 15:283-295.
Marcelli, A, I. Cavoli, A. Bianconi,J. Garcia,A Gargano,C.R. Natoli, M. Benafatto,P. Chiaradia,
M. Fanfoni, E. Fritsch, F. Calas,and J. Petiau.1985. Local structurein SiOz glassesby oxy-
gen Kedge XANES. J. Phys.46(C8):107-112.
McBride, M.B. 1982. Cuz+-Adsorptioncharacteristicsof aluminum hydroxide and oxyhydroxides.

Clays Clay Miner. 30:21-28.
McBride, M.B., AR. Fraser,and W.J. McHardy. 1984. Cu2+ interactionwith microcrystalolinegibb-
site. Evidencefor orientedchemisorbedcopperions. Clays Clay Miner. 32:12-18.
McKale, AG., B.w. Veal, AP. Paulikas,S.K. Chan,and G.S. Knapp. 1988. Improved ab initio cal-
culationsof ~mplitude and phasefunctions for extendedx-ray absorptionfine structurespec-
troscopy.J. Am. Chern. Soc. 110:3763-3768.
McMaster,W.H., N. Kerr del Grande,1.H. Mallet, and J.H. Hubell. 1969. Compilationof x-ray cross
sections.III. UCRL-50174.U.S. Atom. Energy Comm.
Morra, M.J., S.E. Fendorf,and P.R. Brown. 1994. Speciationof sulfur in soil organicmaterial using
x-ray absorptionnearedgestructure(XANES) spectroscopy.Soil Sci. Soc. Am. J.
Muller, J.E., and 1.w. Wilkins. 1984. Band-structureapproachto the x-ray spectraof metals. Phys.
Rev. B29:4331-4348.
Petiau,1., G. Calas, and P. Sainctavit. 1987. Recent developmentsin the experimentalstudiesof
XANES.1. Phys. 48(C9):1085-1096.
Pingitore, N.E., Jr., F.W. Lytle, B.M. Davies, M.P. Eastmann,P.G. Eller, and E.M. Larson. 1992.
Mode of Incorporationof Sr2+ in calcite: Determinationby x-ray absorptionspectroscopy.
Geochim.Cosmochim.Acta 56:15311538.
Rehr,1.1., R.C. Albers, and S.1. Zabinsky. 1992. High-ordermultiple-scatteringcalculationsof x-ray
absorptionfine structure.Phys. Rev. Lett. 69:3397-3400.
Roe, AL., K.F. Hayes,C. Chisholm-Brause,G.E. Brown, Jr., G.A Parks, K.O. Hodgson,and 1.0.
Leckie. 1991. In situ x-ray absorptionstudy of lead ion surfacecomplexesat the goethite-
water interface.Langmuir 7:367-373.
Sainctavit,P., J. Petiau,G. Calas,M. Benfatto, and c.R. Natoli. 1987. XANES study of sulfur and
zinc K-edges in zincblende: Experimentsand multiple-scatteringcalculations.J. Phys. 48,
C9:1109-1112.
Sayers,D.E., and B. Bunker. 1988. Data Analysis. p. 211-253.In D.C. Koningsbergerand R. Prins
(ed.) X-ray absorption. Principles, applications, techniques of EXAFS, SEXAFS, and
XANES. John Wiley & Sons,New York.
Sayers,D.E., E.A Stem,and F.W. Lytle. 1971. New techniquefor investigatingnoncrystallinestruc-
tures: Fourier analysis of the extended x-ray absorption fine structure. Phys. Rev. Lett.
27:1204-1207.
Schulze, D.G., T. McCay-Buis, S.R. Sutton, and D.M. Huber. 1995. Determinationof manganese
oxidation statesin soils using x-ray absorptionnear-edgestructure(XANES) spectroscopy.
Soil Sci. Soc. Am. 1. 59:1540-1548.
Sham,T.K. 1985. L-edge x-ray absorptionsystematicsof the noble metals Rh, Pd, and Ag and the
main-group metals In and Sn: A study of the unoccupieddensity of statesin 4d elements.
Phys. Rev. B31:1888-1902.
Stem,E.A 1974. Theory of extendedx-ray absorptionfine structure.Phys. Rev. 810:3027-3037.
Stem, E.A, and S.M. Heald. 1979. X-ray filter assemblyfor fluorescencemeasurements of x-ray
absorptionfine structure.Rev. Sci. lnstrum. 50:1579-1582.
Stem, E.A, B.A Bunker, and S.M. Heald. 1980. Many-body effects on extendedx-ray absorption
fine structureamplitudes.Phys. Rev. B21:5521-5539.
Stem, EA., D.E. Sayers,and F.W. Lytle. 1975. Extendedx-ray absorptionfine-structuretechniques.
III. Determinationof physical parameters.11:4836-4846.
Stohr, J. 1992. NEXAFS spectroscopy.Springer-Verlag,New YorklBeriin.
Sutton, S.R., K.W. Jones,B. Gordon, M.L. Rivers, S. Bajt, and 1.V. Smith. 1993. Reducedchromi-
um in olivine grainsfrom lunar basalt15555: X-ray absorptionnearedgestructure(XANES).
Teo, B.K. 1986. Basic principles and dataanalysis.Springer-Verlag,New YorklBerlin.
Teo, B.K., and P.A Lee. 1979. Ab initio calculationsof amplitudeand phasefunctions for extended
x-ray absorptionfin estructurespectroscopy.J. Am. Chern. Soc. 101:2815-2832.
Tewari, P.H., and W.J. Lee. 1975. Adsorption of Co(II) at the oxide-water interface. J. Colloid
InterfaceSci. 52:77-88.
Tyson, T.A, A.L. Roe, P. Frank, K.O. Hodgson,and B. Hedman.1989. Polarizedexperimentaland
theoreticalK-edge x-ray absorptionstudiesof S042-,CI03-, S2032-,and SZ06 2-. Phys. Rev.
B39:6305-6315.
Vairavamurthy,A, B. Manowitz, G.W. Luther, III, and Y. Jeon. 1993. Oxidation state of sulfur in
thiosulfate and implications for anerogic energy metabolism.Geochim. Cosmochim.Acta
57:1619-1623.
Vairavamurthy,A., W. Zhou, T. Eglinton,andB. Manowitz. 1994.Sulfonates:A novel classof organ-

ic sulfur compoundsin marine sediments.Geochim.Cosmochim.Acta 58:4681-4687.
Waddington,w.G., P. Rez, I.P. Grant, and CJ. Humphreys.1986. White lines in the Lz,3 electron-
energy-lossand x-ray absorptionspectraof 3d transition metals.Phys. Rev. B34:1467-1473.
Waldo, G.S., R.M.K. Carlson,J.M. Modowan,K.E. Peters,and J.E. Penner-Hahn.1991. Sulfur spe-
ciation in heavy petroleums: Information from x-ray absorption near-edge structure.
Waychunas,G.A. 1986. X-ray K-edge absorptionspectraof Fe mineralsand model compounds:II.
EXAFS. Phys. Chern. Miner. 13:31-47.
Waychunas,G.A. 1987. Synchrotronradiation XANES spectroscopyofTi in minerals:Effects ofTi
bonding distances,Ti valence,and site geometryon absorptionedge structure.Am Miner.
72:89-101.
Waychunas,G.A., MJ. Apted, and G.E. Brown, Jr. 1983.X-ray K-edgeabsorptionspectraof Fe min-
erals and model compounds:Near-edgestructure.Phys. Chern. Miner. 10:1-9.
Waychunas,G.A., B.A. Rea, C.C. Fuller, and J.A. Davis. 1993. Surface chemistryof ferrihydrite:
Part 1. EXAFS studiesof the geometryof coprecipitatedand adsorbedarsenate.Geochim.
Zaanen,J., G.A. Sawatzky,J. Find, W. Speier,and J.e. Fuggle. 1985. Lz,3 absorptionspectraof the
lighter 3d transition metals.Phys. Rev. B32:4905-4913.
Published 1996
Chapter 14
Salinity: Electrical Conductivity

and Total Dissolved Solids
J. D. RHOADES, U.S. Salinity Laboratory, Riverside, California
INTRODUCTION
The term salinity refers to the presenceof the major dissolvedinorganicsolutes,

essentially Na+, Mg2+, Ca2+, K+, Cl-, SOl-, HCO:3 and C01-, in aqueoussam-
ples. As applied to soils, it refers to the soluble plus readily dissolvablesalts in
the soil or, more usually, in an aqueousextractof a soil sample.Salinity is quan-
tified in termsof the total concentration(or, occasionally,the content)of suchsol-
uble salts. The diagnosis,assessment, managementand needfor reclamationof
salinesoils and the suitabilityof watersfor variouspurposes,induding irrigation,
are evaluatedusing information of soil and water salinity.
For certain soil salinity considerations,it would be desirableto know the
individual concentrationsof the major solutesin the soil water over the rangeof
water contentsthat occur in the field and to obtain this information in the field,
without the taking of soil samplesand the carryingout of laboratoryanalyses.No
practicalmethodsare availableat presentto permit suchdetaileddeterminations,
but total salinity can be measuredin situ using electrical signals from various
typesof sensors(Rhoades,1978, 1990; Rhoades& Oster,1986).The latter deter-
minationsare often sufficient for purposesof diagnosing,surveying,and moni-
toring soil salinity, and for assessingthe adequacyof leachingand drainage,even
though they only give information of total soluble salt concentration,and hence
supplantthe needfor carrying out conventionallaboratoryanalytical procedures;
in other casesthey greatly minimize the numberof samplesrequiring composi-
tional analysesbecausecorrelationsfrequently exist betweensalinity and the con-
centrationsof individual solutesand their ratios within the samegeneralareaof
the landscape(Skarieet aI., 1987).
The total soluble salt concentration(or content) of a water can be deter-
mined from either measurementof its electrical conductivity (EC) or of its
residue-weightupon evaporationto drynessafter filtration. Likewise, soil salini-
ty canbe determinedfrom either measurementmade:(i) on an aqueousextractof
the soil sample,(ii) or on a sampleof displacedsoil solution. Alternatively, soil
Book Seriesno. 5.
417
418 RHOADES
salinity can be measuredin situ using buried porouselectrical conductivity sen-

sors that imbibe soil water or it can be estimatedfrom EC measurements made
on watersaturatedsoil pastes.It also can be estimatedfrom measurement of bulk
soil EC made:(i) in situ using electrical probes,or (ii) remotely using electro-
magnetic(EM) systems.The appropriatemethodof measuringsalinity shouldbe
selectedbasedon the specific conditions, needsand equipmentand manpower
availability.
Water salinity is measuredmore accurately,quickly and simply from EC;
however, for some purposes,total dissolvedsolids may be a more appropriate
methodof appraisal.If only a practical measureof soil salinity is needed,mea-
surementof bulk soil EC made using either EM induction or four-electrode
devicesis recommended.To monitor soil water salinity as the soil dries between
irrigations, either the imbibition-or the time domain reflectrometric(TDR)-
type of salinity sensoris recommended.When determinationof the concentration
of a particularsolute is needed,then either extractionof soil samplesor acquisi-
tion of soil solutionsamples isrequired,along with appropriatelaboratoryanaly-
sis. Collection of watersamplesfrom soils using suction-cupextractorsis useful
for field monitoring needs,but this procedureis usually limited to relatively wet
soil conditions (Rhoades,1978; Rhoades& Oster, 1986). Displacementof soil
solution from soil samplescollectedand kept at field moisturecontentprovides
for a meaningfulsoil-water sample,but the laboratoryproceduresfor acquiring
the latter samplesare more demanding.Soil sampleextractsgive "relative salin-
ities" only, since the soils are adjustedto unnaturallyhigh water contentsduring
extraction.
A combinationof the varioussalinity sampling/measurement methodsmin-
imizes the needfor the more difficult and time-consumingcollection, extraction
and chemical analysisprocedures.This combinationapproachis recommended
when monitoring salinity changes,or when characterizingthe salinity of large
fields and projects.For the latter, extensiveuseof portable(betteryet of mobile)
EM and four-electrodeequipmentand techniquesfor measuringbulk soil EC and
for assessingsoil salinity in the field are recommended,with the supplemental
use of the other methodsonly as neededfor calibration purposesor for charac-
terizing solute composition. Field methods for measuring bulk soil EC are
described in Methods of Soil Analysis, Part I (Physical and Mineralogical
Methods)of the SSSABook Series(see Rhoades& Oster, 1986) and elsewhere
(Rhoades,1990), mobile versions of these systemsare describedby Rhoades
(1992).
This chapterdescribesprevalent laboratorymethodsfor determiningsalin-
ity (soil and water) basedon measurements of EC or total dissolvedsolids after
evaporationat 180°C. It also describeslaboratory methodsfor making and for
obtainingaqueousextractsof soil samples1. Various methodsfor determiningthe
I Methodsfor collecting samplesof soil water per se are not providedin this chapter,but the fol-
lowing provides some useful referencesin this regard. Samplesof soil solutions may be obtained
from soils by meansof displacement,compaction,centrifugation,molecularadsorptionand vacuum
or pressureextraction methods(Richards,1941). Displacementmethodsare describedby Adams
(1974); combinationdisplacement/centrifugation methodsby Gillman (1976); Mubarak and Olsen
(1976, 1977) and Elkhatib et al. (1986); a combinationvacuum/displacement method by Wolt and
SALINITY, ELECTRICAL CONDUCTIVITY & DISSOLVED SOLIDS 419
concentrationsof individual inorganic solutes (i.e., Na+, Ca2+, Mg2+, K+, CI+,
SOi-, HCO), NO), COJ-,H3B03, etc.) in watersandsoil extractsin commonuse
in laboratorieshaving modern instrumentationare describedelsewherein this
publication. Analogousmethodologysuited to laboratorieswithout such conve-
niencesare given in an earlier publication (seeBower & Wilcox, 1965).
SATURATION AND OTHER AQUEOUS EXTRACTS
Principles
Becausepresentmethodsof obtaining soil water samplesat typical field

watercontentsare not very practical,aqueousextractsof the soil samplesare usu-
ally madeat higher than normal water contentsfor routine soil salinity diagnosis
and.characterizationpurposes.Becausethe absoluteand relative amountsof the
varioussolutes areinfluencedby the soil/waterratio at which the extractis made
(Reitemeier,1946), the soil/waterratio usedto obtain the extractshould be stan-
dardizedto obtain resultsthat can be appliedand reasonablyinterpretedgeneral-
ly. Soil salinity is mostgenerallydefinedand measuredon aqueous extracts of so-
called, saturatedsoil-pastes(U.S. Salinity Laboratory Staff, 1954).This water
content(and water/soil ratio) varies with soil texture but is usedbe~ause it is the
lowest one for most soils for which sufficient extractcan be practically removed
from a soil sample for the compositional analysis of major constituentsand
becauseit is betterrelatedto soil-watercontentsunderfield conditions.For these
samereasons,crop toleranceto salinity also is expressedin terms of the EC of
the saturationextract(ECe, Mass & Hoffman, 1977; Maas, 1986, 1990).
Apparatus
1. Plasticcontainerswith snaptightlids of 250-mL capacityor greater.

2. Vacuumline, suctionapparatus(Richardsfilter funnel2, Buchnerfunnel,
or commercialvacuum manifold) filter paper (medium grade, such as
Whatmanno. 50) and samplebottles [28.4 g (1 oz.) or larger, to collect
and store extracts]with sealablecaps.
3. Balanceor scaleaccurateto at least 1 g.
4. Extractionbottlesfor soil suspensions.
5. Mechanicalshaker.
Graveel (1986); a simple field-pressurefiltration method by Ross and Bartlett (1990); adsorption
techniquesby Shimshi(1966) and by Tadrosand McGarity (1976) and centrifugationtechniquesby
Davies and Davies (1963), Yamasuki and Kishita (1972), Gilman (1976), Dao and Lavy (1978),
Kinniburgh and Miles (1983) and Elkhatib et at. (1987). Comparisonsof the various methodshave
been made by Adams et at. (1980); Kittrick (1983); Wolt and Graveel (1986); Menzies and Bell
(1988) and Rossand Bartlett (1990). The different suction-typesamplersand other methodsfor sam-
pling soil solution and variouserrorsassociatedwith them havebeencritically reviewedby Rhoades
(1978), Rhoadesand Oster(1986), Litaor (1988) and Grossmanand Udluft (1991).
2 Richards,L.A. (1949).
420 RHOADES
Reagent
1. Sodium hexametaphosphate [(NaP03)6] solution, 0.1%. Dissolve 0.1 g

of (NaP03)6 in water, and dilute the solution to 100 mL.
Procedure
SaturationExtract
Weigh 200 to 400 g of air-dry soil of known watercontentinto a taredplas-
tic containerhaving a snaptightlid. Weigh the containerplus contents.Add dis-
tilled water to the soil with stirring until it is nearly saturated.Allow the mixture
to standcoveredfor severalhours to permit the soil to imbibe the water and the
readily solublesaltsto dissolve,and then add more waterwith stirring to achieve
a uniformly saturatedsoil-water paste.At this point, which is generally repro-
ducible to within ±5%, the soil pasteglistens as it reflects light, flows slightly
when the container istipped, slides freely and cleanly off a smoothspatula,and
consolidateseasily by tappingor jarring the containerafter a trench is formed in
the pastewith the side of the spatula.After mixing, allow the sampleto stand
(preferably for another2 h), and then recheck the criteria for saturation.Free
water should not collect on the soil surface,nor should the pastestiffen marked-
ly or lose its glisten. If the paste is too wet, add additional dry soil of known
amount(weight) to the pastemixture. Upon attainmentof saturation,weigh the
containerplus contents.Recordthe increasein weight due to the amountof water
added.Transferthe saturatedsoil pasteto a filter funnel fitted with highly reten-
tive filter paper.Apply vacuum,and collect the filtrate in a test tube or bottle. If
the initial filtrate is turbid, refilter or discardit. Terminatethe filtration when air
beginsto passthroughthe filter. Add 1 drop of 0.1% (NaP03)6 solution for each
25 mL of extract.
Extractsof Soil/WaterRatiosof One:Oneand One:Five

Weigh a sampleof air-dry soil of appropriatesize, and transferit to a flask
or bottle. Add the requiredamountof distilled water (an equal weight for a 1:1
extract,5times the weight for a 1:5 extract), stopperthe container,and shakeit
in a mechanicalshakerfor 1 h. If a mechanicalshakeris not available,shakethe
containervigorously by hand for 1 min at least four times at 3D-min intervals.
Filter the suspensionusing highly retentive filter paper. (Discard or refilter the
initial filtrate if it is turbid.) Add 0.1% (NaP03)6 solution at the rate of 1 drop/25
mL of extract.
Calculations
Calculatethe saturationwater percentage(SP) of the saturatedpastefrom
the weight of oven-drysoil (Ws) and the weight of water(W) added(Ww), includ-
ing that initially presentin the soil sampleas,
[1]
Comments
The extraction ratios(1:1, 1:5, etc.) are easierto make than that of satura-
tion, but they are less well relatedto field soil water compositionand content.
More importantly, salinity and compositionalerrorsfrom dispersion,hydrolysis,
cation exchange,and mineral dissolutionincreaseas the water/soil ratio increas-
es(Reitemeier,1946).As a compromise,Sonneveltandvan den Ende(1971) rec-
ommendeda 1:2 (1 part soil = 2 partswater) volume extract, since it is closerto
the saturationextractratio but quickerto make.Whenrelative changes ratherthan
absolutesolute concentrationsare needed,thesewider extractionratios have the
advantagesof speedand greatervolume.
The EC of extractsof gypsiferoussoils decreasesless, as the water/soil
ratio increases,due to the dissolutionof gypsum.This gypsumdissolutionexag-
geratesthe concentrationof soluble salts presentin the soil at lower water con-
tents,especiallythoseexisting underfield condition~.
Soil samplesshould not be oven-driedbefore extractingfor determination
of soluble salts,becauseheatingto 105°C convertsat leasta part of the gypsum
(CaS04• 2 H20) to plasterof paris (CaS04• 1/2 H20). The latter hydratehas a
highersolubility in waterthan doesthe former. The solubilitiesof other saltsand
mineralsalso may be affected.
The weight of soil requiredwill dependon the number.and kind of deter-
minationsto be made on the extract, the analytical methodsemployed,and the
salt contentof the soil. In general,from one-fourthto one-third of the water in
saturatedsoil pastescan be practically removedby vacuumfiltration.
In determinationsof extractwater contents,especiallywhen a high ratio of
soil to water is used,it is desirableto correctfor the water contentof the soil sam-
ple. For example,an air-dry samplecontaining2% water on an oven-dry basis
can be adjustedto a soil/waterratio of 1: 1 by adding98 mL of water to 102 g of
air-dry soil. At soil/waterratios of 1:5 or greater,no correctionis ordinarily made
for water in the air-dry sample.
Specialprecautionsshouldbe takenin preparinga saturatedsoil pastewith
peat and muck soils or very fine or very coarse-texturedsoils (Prichard et aI.,
1983). If possible,peatand muck soils should not be allowedto dry appreciably
following collection becausetheir saturationwater content changeswith dehy-
dration. Peatand muck, especiallyif coarseor woody, requirean overnightimbi-
bition periodto obtain a definite endpointfor the saturationpoint. After the first
wetting, pastesof these soils usually stiffen upon standing. Adding water and
remixing then yields a mixture that usually retains the characteristicsof a satu-
rated paste.With fme-texturedsoils, enoughwatershouldbe addedimmediately,
with a minimum of mixing, to bring the samplenearly to saturation.This mini-
mizes the formation of clumpsof soil during stirring, speedsthe mixing process,
and helpsattain a more definite endpoint.Carealso shouldbe takennot to undu-
ly overwetcoarse-textured soils. The presenceof free water on the surfaceof the
pasteafter standingis an indication of oversaturationin the caseof coarse-tex-
turedsoils. Evensmall amountsof free watercanlead to appreciableerrorsin sat-
uration pastewater contentsfor thesematerials.However,the effect on the value
422 RHOADES
of the EC of the extract (ECe) is small and usually does not significantly affect
salinity diagnoses.
Sodiumhexametaphosphate is addedto the extract to preventthe precipi-
tation of CaC03 from the extract upon standing.The quantity of (NaP03)6 solu-
tion addedincreasesthe Na concentrationabout 0.5ppm, or 0.02mmolJL, which
is inconsequentialcomparedwith the possibleloss of CaC03. Alternatively, a
subsampleof the extractshouldbe analyzedimmediatelyor immediatelydiluted
twofold and usedfor the Ca and alkalinity determinations.
Alternative methods of preparing the saturated soil paste have been
describedby LongeneckerandLyerly (1964),who proposedwetting the soil sam-
ple on a capillary saturationtable, by Beatty and Loveday (1974) and Loveday
(1972), who recommendedpredeterminingthe amountof water at saturationon
a separatesoil sampleusing a capillary wetting technique,and by Allison (1973),
who recommendedslowly addingsoil to water (oversaturationmethod).Similar
results are obtainedwith thesemethods.The choice of method is primarily one
of personalpreference.
Thymol can be addedto the pasteto minimize the effect of microbial activ-
ity on saturationextractcompositionduring equilibration(Carlsonet aI., 1971).
The extractsshould be storedat about4°C until analyzed.
ELECTRICAL CONDUCTIVITY OF WATERS

AND AQUEOUS EXTRACTS
Principles
Electrical conductivity is a numericalexpressionof the ability of an aque-

ous solution to carry an electric current. It is generallyrelatedto the total solute
concentrationand can be usedas a quantitativeexpressionof dissolvedsalt con-
centration,eventhough it is also affectedby the mobilities, valencesand relative
concentrationsof the individual ions presentin the solution.
The determinationof EC generally involves the physical measurementof
resistance(R) in ohms.The R of a conductingmaterial(suchas a salinesolution)
is inversely proportionalto its cross-sectionalarea(A) and directly proportional
to its length(L). The magnitudeof the R measuredthereforedependson the char-
acteristics(dimensionsand spacings)of the conductivity cell and electrodes.
Specific resistance(Rs) is the R of a cubeof the sample1 cm on edge.Practical
conductivity electrodesare not of this dimensionand measureonly a given frac-
tion of the specificR; this fraction (RlRs) is commonlyreferredto as the cell con-
stant(K).
The reciprocalof R is conductance(C). It is expressedin reciprocalohms,
or mhos. When the cell constantis applied, the measuredconductanceat a spec-
ified temperatureis convertedto specificC, the reciprocalof the specificR (here-
in called EC)
1
EC=-=K/R. [2]
Rs
Electrical conductivity has been customarily reported in micromhos per

centimeter (Ilmho/cm) or in millimhos per centimeter (mmho/cm). In the
InternationalSystemof Units (SI), the reciprocal of the ohm is the siemen(S)
and, in this system,EC is reportedas Sim or as dS/m. Therefore,IS =1 mho or
1 dS/M =1 millimho/cm.
Electrolytic conductivity (unlike metallic conductivity)increaseswith tem-
peratureat a rate of approximately1.9% per degreeCentigrade.Since each ion
has a different temperaturecoefficient, for precise work, conductivity ideally
should be determinedat 25°C. However, EC can be measuredat other known
temperaturesand correctedto the 25°C referenceusing appropriatetemperature
coefficients(usually basedon NaCl). Sodiumchloride solutionshave a tempera-
ture coefficientthat closelyapproximatesthat of mostsurface-andgroundwaters.
Potassiumchloride solutionshave a lower temperaturecoefficientof conductivi-
ty than typical water aqueoussolutions.
Apparatus
ConductanceMeter
Any instrument capable of measuring conductancewith an error not
exceeding1%, or IllS, whicheveris greater,is suitable.The instrumentmay be
an automaticor manually operatedwheatstonebridge, a deflection meter or a
self-recordingrecorderor indicator.The preferredinstrumentis one which mea-
suresthe ratio of the alternatingcurrent(1000 Hz) throughthe cell to the voltage
acrossit, sinceit gives a more linear readingof conductance,and one which also
includesautomatictemperaturesensingand compensation.
Thermometer
For high-precisionmeasurements, sampletemperatureis critical, and spe-
cial electricalthermometersare desiredin the instrumentalong with circuitry to
compensatefor temperature.When the instrumentis not providedwith automat-
ic temperaturecompensation,sampletemperature(t) should be manually deter-
mined using a thermometerand the EC values measuredat temperaturet then
adjustedto the referencevalue of 25°C usingtemperature-correction relations(or
tables). For high-precisiondeterminations,use a thermometeraccurateto the
nearestO.I°C and coveringthe rangeof 20 to 30°C. For routine or field determi-
nations,a thermometeraccuracyof 0.5°C is adequate.If the measuringinstru-
ment is providedwith a manualtemperaturecompensator,adjustthis to the mea-
suredsampletemperature.If it hasautomaticcompensation,allow sufficient time
to permit equalizationof sampleandconductivitycell temperatures.If it hasnei-
ther, measureand recordthe temperatureof the samplesat which the conductance
is measuredand adjust the value using an appropriatetemperature-correction
relation (seetext below).
Conductivity-Cell/Electrodes
Selecta conductivity-cell having a cell constantappropriateto the EC of
the sample(seeTable 14-1). Flow-throughcells should be usedfor measuring
424 RHOADES
Table 14-1. Recommended

cell constantsfor variousconductivity ranges.
Rangeof conductivity Cell constant
dS/m
0.OOOO5~.020 0.0001
0.001~.2 0.001
0.01-2 0.01
0.1-20 0.10
1-200 0.50
solutions of EC values lower than 0.01 dS/m, in order to avoid contamination

from the atmosphere.Recommendedrate of flow through the cell is 0.3 mls. In
all other cases,pipet type- or dip-type cells can be used.The cell systemshould
assurethe retentionof calibrationunderexpectedconditionsof flow, pressureand
temperaturechangesand should be resistantto corrosion. Ideally, it should be
equippedwith a meansfor the accuratemeasurementof sampletemperatureas
discussed above. Either platinum-ornonplatinumtype electrodes-can be used,
dependingupon accuracyrequirements.Platinumtype electrodesare more accu-
rate. Electrodesmade of durable common metals can be used for continuous
monitoring or field use.
Reagents
1. Platinizing solution. One gram chloroplatinic acid (H2PtCI6 • 6H20)

plus 12 mg lead acetatein 100 mL of pure water (EC < 0.001 dS/m).
2. Cleaningsolution. Mixture of one part by volume of isopropyl alcohol,
one part of ethyl ether,and one part of HCI (1 + 1).
3. Standardpotassiumchloride (KCI) solution, 0.010M. Dissolve 0.7456
g of reagentgradeanhydrousKCI in pure water (EC < 0.001 dS/m) and
add more water to make to 1 L at 25°C. Store in a glass-stoppered
borosilicate glass bottle. This referencesolution has an EC of 1.413
dS/m at 25°C and is suitablefor most solutionswhen the cell hasa con-
stant betweenone and two. For other cell constantsuse stronger or
weakerKCI solutions,as listed in Table 14--2.
Procedure
Preparationof Platinum Electrodes

Clean,new platinum-electrodes with chromic-sulfateacid solution and pla-
tinize the electrodesbeforeuse.Immersethe electrodesin the platinizing solution
and connectboth to the negativeterminalof a 1.5-V dry cell battery.Connectthe
positive side of the battery to a piece of platinum wire and dip the wire into the
solution. Continuethe resulting electrolysisreactionuntil both cell electrodesare
coatedwith platinum black. Use a currentsuch that only a small quantity of gas
is evolved.Subsequently,cleanand replatinizethe electrodeswheneverthe read-
ings becomeerraticor a sharpendpointcannot be obtainedor whenvisual exam-
ination showssome platinum black has flaked off. Rinse electrodesthoroughly
after useand keepimmersedin distilled water when not in use.
Table 17-2. Conductivityof KCI solutionsat 25°C.

Concentration Conductivity
M dS/m
0.001 0.147
0.01 1.413
0.02 2.767
0.05 6.668
0.1 12.900
0.2 24.820
0.5 58.640
1 111.900
Calibration of Cell
Rinseconductivitycell with at leastthreeportionsof 0.01 M KCL solution.
Adjust temperatureof a fourth portion to 25.0 ± O.l°C. MeasureR (or C) of this
portion and note the temperature.Repeatthe measurementon additionalportions
of the referenceKCL solution until the R value obtainedremainsconstantwith-
in the requiredlimits of precision.
Computecell constant
K = (0.001413)(RKCd/[l + 0.019(25- t)], [3]
where RKCL is the measuredR (ohms) and t is measuredtemperatureCC).
Measurement of Sample Conductivity

Rinsecell with one or more portionsof sample.Ideally, adjustt of the final
portion of sampleto 25 ± O.l°C and measureR or C; if not practicalor necessary
to adjust the t of the sampleto 25°C, measuret and R, or e, at the samet. The
techniqueusedshouldbe the sameas that employedto calibratethe cell. Record
thesevalues.
Calculation
When sampleR is measured,conductivity (dS/m) at 25°C is
EC Z5 = 1,000K[1 + 0.019(25- t)]/Rx, [4]
where Rx is measuredR of the sample(ohms), t is the measuredt and K is the

cell constant(m-1).
When sampleconductanceis measured,conductivity (dS/m) at 25°C is
EC25 = Cx (1,000)K[1 + 0.0191(25- t)], [5]
whereex is measurede of the sample(mhos).

Reportthe conductivity at 25°C in termsof dS/m to the nearest1% of deter-
mined conductivity.
426 RHOADES
Comments
Laboratorymeasurements of EC are relatively accurate.Errors are usually

relatedto electrodefouling and inadequatesamplecirculation.
Besidesbeing usedto define and measuresoil and water salinity and crop
tolerancesto salinity, EC is usedto estimatealiquot size for chemicalanalysis,to
checkreliability of chemicalanalyses,to estimatetotal dissolvedsolids in a sam-
ple (by multiplying conductivity, in dS/m, by an empiricalfactor varying between
5.50 to 9.00 dependingon the solublecomponentsand temperature),to evaluate
and expresscorrosionlikelihood, and to assessthe purity of demineralizedwater
(freshly distilled water has a conductivity of 0.0005 to 0.002 dS/m, increasing
after a few weeksof storageto 0.002 to 0.004 dS/m due to absorptionof atmos-
pheric CO2 and NH3).
Exposureof a sampleto the atmospheremay causechangesin conductivi-
ty, due to the loss or gain of dissolvedgasesor water. This is quite important in
the caseof waterswith low concentrationsof dissolvedelectrolytes.Carbondiox-
ide, normally presentin the air, can drastically changethe EC of "pure" waters,
as can atmospherecontainingammoniaor acid gases.Contactwith air shouldbe
avoidedby using flow-through type conductivity-cells.
Becauseof markeddifferencesin the equivalentweights, equivalentcon-
ductivities, and proportionsof major solutesin soil extractsand water samples,
the relationshipsbetweenEC and salt concentrationor betweenEC and osmotic
pressureare only approximate.They are still quite useful, however.Theserela-
tionshipsare:
1. Total cation (or anion concentration),mmolescharge/liter== lOx EC, in
dS/m,
2. Total dissolvedsolids, mg/liter == 640 x EC, in dS/m,
3. Osmoticpressure,kPa at25°C == 0.36 x EC, in dS/m.
Pure water of low and known conductivity should be usedin the prepara-
tion of the standardKCI solutions.The EC of this water shouldbe addedto that
given in Table 14-2 and usedin Eq. [3] to determinethe cell constant,since the
conductivity of the referencesolution is that of the water plus that of the added
KCI.
The temperaturecoefficient varies with concentrationand upon the nature
and compositionof the dissolvedelectrolytes.The lower the concentration,the
higher the coefficient, due to the effect of temperatureupon the dissociationof
water. The best way to correct for the temperatureeffect on conductivity is to
hold the temperatureof the sampleand cell constantat 25 ± 0.5°C. If this cannot
be done, the next best way is to make determinationsof sampleEC at several
temperatures,to plot the paired readingsand to interpolatethe EC at 25°C from
the smoothedcurve. For less-accurateneeds,the EC at 25°C (in dS/m) canbe cal-
culated by multiplying the EC measuredat the measuredtemperatureby an
appropriatecoefficient ft. The value of f t can be estimatedas [1 + 0.0019 (25 -
t)].
For routine field or laboratorytesting, the cell constantof the conductivity
cell should be determinedand periodically checkedby comparing instrument
readings taken with referenceconductivity solutions at a known temperature,

preferably25°C.
For otherdiscussionsof EC seeAmericanSocietyfor Testingand Materials
(1986) and Helrich (1990).
ELECTRICAL CONDUCTIVITY OF SATURATED SOIL PASTES

AND DETERMINATION OF SOIL SALINITY
Principles
Soil salinity, in terms of ECe, may be estimatedfrom measurementof the

EC of the saturatedsoil-paste(ECp) and estimatesof saturationpercentage(SP).
The measurement of ECp and the estimateof SP can be madeusing an EC-cupof
kI).own geometry and volume. The method is suitable for both lal?oratory and
field applications,especiallythe latter, becausethe apparatusis inexpensive,sim-
ple and rugged and becausethe determinationof ECp can be made much more
quickly than of ECe.
Rhoadeset al. (1989a)haveshownthat the following relation describesthe
EC of saturatedsoil pastes,
[6]
where ECp and ECe are as definedpreviously,8w and8s are the volume fractions
of total water and solids in the paste,respectively,8ws is the volume fraction of
water in the pastethat is coupled withthe solid phaseto provide aseries-coupled
electrical pathwaythrough the paste,ECs is the averagespecific EC of the solid
particles, and the difference (8w - 8ws) is 8we, which is the volume fraction of
water in the pastethat providesa continuouspathwayfor electricalcurrent flow
throughthe paste(a parallel pathwayto 8ws). Assumingthe averageparticle den-
sity (Ps) of mineral soils to be 2.65 glcm3 and the density of saturationsoil-paste
extracts(Pw) to be 1.00, 8s and 8w are directly relatedto SP as follows
Pw 100
8w = SP + [ -P-s- + SP 1 , [7]
[81
As shown by Rhoadeset al. (1989a,b),the SPof many mineral soils can be ade-
quately estimatedin the field for purposesof salinity appraisalfrom the weight
of the paste-filledcup. Figure 14-1 may be usedfor this purpose,for details of
the relationsinherentin this figure seeWilcox (1951).
Electrical conductivity of the saturationextract can be determinedfrom
measurementof ECp and SP (using Eqs. 6-8), if values of Ps' 8ws and ECs are
known. These parameterscan be adequatelyestimated, as demonstratedby
428 RHOADES
100
90
80
70
W 60 FOR BUREAU OF SOILS CUP
r~
OF 50 cm 3 VOLUME
Z 50
W
~
~ 40
W
t-
en
~ 30
~
~
a::
~
20
!;i
en
GRAMS PASTE
Fig. 14-1. Theoreticalrelation betweensaturationpercentage(SP) and weight (g) of 50 cm3 of satu-
rated paste,assuminga particle density of 2.65 g/cm3•
Rhoadeset al. (1989a). For typical arid land soils of the southwesternUSA, Ps
may be assumedto be 2.65 glcm3; ECs may be estimatedfrom SPas ECs =0.019
(SP) - 0.434, and the difference(9w - 9ws) may be estimatedfrom SP as (9w -
9ws) =0.237 (SP)O.6657.
Apparatus
1. Conductancemeter.Almost any laboratoryC metercan be used,prefer-

ably a temperaturecompensatingtype as describedin "Apparatus."An
exampleis shownin Fig. 14-3.
2. Largecup-typeconductivity cell.The 50-cm3 volume "Bureauof Soils"
cup is suitable (U.S. Salinity Laboratory Staff, 1954). An exampleis
shownin Fig. 14-2.
3. Portablebalance.Capableof weighing accuratelyto the nearest0.5 g.
An exampleis shownin Fig. 14-2.
Reagents
1. Standardpotassiumchloride solutions, 0.010 and 0.100 M solution.

Sameas describedin "Reagents."
Fig. 14-2. Picture of portablebalanceusedin the field to determinethe weight of the saturatedsoil-
pastefilling the "Bureauof Soils" cup.
Procedure
Rinse and fill the conductivity cup with KCl solution. Adjust the conduc-
tivity meterto read the standardconductivity. Rinse and fill the cup with the sat-
uratedsoil-paste,tap the cup to dislodgeany air entrappedwithin the paste.Level
off the pastewith the surfaceof the cup. Weigh the cup plus paste,subtractthe
cup tare weight to determinethe gramsof paste(W p) occupyingthe cup. Connect
the cup electrodesto the conductivity meterand determinethe ECp' correctedto
25°C, directly from the meterdisplay.
Calculation
Obtain the SP value from Fig. 14-1 correspondingto Wp. Obtain ECe from
Fig. 14-4 given ECp and SP, using the curve correspondingto the SP value, or
calculateECe using the following equation
ECc = (-b ± ~ b2 - 4ac)/2a, [9]
wherea =[8s (8w - 8ws)], b =[(8s + 8wsf (EC s) + (8w - 8ws) (8wsECs) - (8s) EC p]
and c =-(8ws )(ECs)(ECp).
Comments
Sensitivity analysesand tests have shown that the estimatesused in this

methodare generally adequatefor salinity appraisalpurposesof typical mineral
430 RHOADES
Fig. 14-3. Pictureof "Bureauof Soils Cup" filled with saturatedsoil pasteconnectedto conductance
meter.
arid-landsoils of the southwesternUSA (Rhoadeset aI., 1989b).For organicsoils

or soils of very different mineralogyor magneticproperties,theseestimatesmay
be inappropriate.For suchsoils, appropriatevaluesfor Ps, ECs and Ows will need
to be determinedusing analogoustechniquesto thoseof Rhoadeset ai. (1989a).
It should be noted that (ECeOe) is not equivalentto (ECwOw) becausedif-
ferent amounts of soil are involved in the two measurements.The relation
betweenthesetwo productsis
[10]
Data to supportthis is given in Rhoadeset ai. (1989b, 1990).

SALINITY, ELECfRlCAL CONDUCTIVITY & DISSOLVED SOLIDS 431
o
.. "
6
w
.:
-
4
u
...
IV
)(
W
I
c o
o
:;
--:
-...IV::3 .. ~-------- ---'- --~---:
sp, 20 30 40 50 60 70 80
-
IV
40 90
VJ 100
35
-
o
>- .,."
';
:; 25
U
:::II
~ 20
C
o
o ,a
-.,
ii 10
u
1(1
.~
u 5
iii 0
o 2 4 6 8 10 12 14 16 18 20
Electrical Conductivity of Saturation-

Paste, EC p , dS/m
Fig. 14-4. Relations between electrical conductivity of saturatedsoil-paste(ECp), electrical conduc-

tivity of saturationextract(Eee) and saturationpercentage(SP), for representativearid-landsoils.
TOTAL DISSOLVED SOLIDS IN WATERS

AND AQUEOUS EXTRACTS
Principles
The amount(concentration)of total dissolvedsolids in a sampleis deter-

mined by weighing the residueobtainedafter evaporatinga samplethat hasbeen
filtered to remove particulate matter. A sample is filtered through a standard
membrane(usually 0.45 Ilm in pore size) filter and the filtrate is evaporatedto
drynessin a weigheddish and dried to a constantweight of 180°C. The increase
in weight representsthe total dissolvedsolids. The definition is operationalsince
certaincolloids may not be removedby filtration and the evaporationresidueusu-
ally differs in compositionfrom the dissolvedmatterinitially presentin the water.
432 RHOADES
Apparatus
1. Samplereservoir.A chemicallyresistantcontainerof 1- to 4-L capacity.

2. GlassPetri dish. 150-mmdiam.
3. Evaporatingdish. A straight-wall or round bottom dish of SO- to 100-
rom diam. and approximately 200-mL capacity made of platinum,
porcelainor high-silica glass.
4. Heater. A controlled electric hot plate, infrared lamp or steam bath
which is capableof maintainingthe t of the evaporatingsamplenearthe
boiling point.
5. Muffle furnace.Capableof operationat 550 ± 50°C.
6. Desiccator.Large sealablechamberprovidedwith a desiccantcontain-
ing a color indicator of moistureconcentration.
7. Drying oven. Capableof operationat ISO ± 2°C.
S. Analytical balance.Capableof weighing to an accuracyof 0.1 mg.
9. Filtration apparatus.Any suitablecommercialmembranefiltration sys-
tem capableof removing particulates >0.45 rom in size.
Procedure
Heat cleanevaporatingdish to 180 ± 2°C for 1 h in oven, storein desicca-

tor until needed,weigh just before use. Choosesample volume to yield upon
evaporationapproximately25 mg of residue,if only the amountof residueis to
be determined,or 100 mg of residueif it is to be analyzedfor composition.Pass
the measuredvolumeof samplethroughthe filter membraneandthenfollow with
three successivelO-mL volume incrementsof distilled water. Quantitatively
transferthe filtered samplealiquot to a samplereservoir.Fill an evaporatingdish
(that previouslyhasbeenignited at 600 ± 25°C for 1 h, cooledin a desiccatorand
weighed)to nearly full with a portion of the sample.Heat the dish to evaporate
this portion, but do not allow it to boil or dry. Periodically add more of the sam-
ple from the reservoirto the dish until the reservoiris empty. Rinse the reservoir
severaltimes with pure water, adding the rinsings to the contentsof the evapo-
rating dish. Then evaporatethe remainderof the materialin the dish to neardry-
ness.Transferto a 103°Coven and completethe evaporation.Dry the dish + con-
tentsfor 1 h at ISO ± 2°C, cool in a desiccator,and then weigh. Repeatthe cycle
of drying (1 h periods),cooling, desiccatingandweighinguntil a constantweight
is obtainedor until loss in weight is no more than 4% of the previousweight or
0.5 mg, whicheveris less.Recordthe weight of residueas total dissolvedmatter.
Calculation
Calculatethe concentrationof total dissolvedsolids in milligrams per liter

(essentiallypartsper million), as follows
total dissolved solids, mg L-l =(A - B)l000/sample volume, mL, [11]
whereA is weight of dried residueplus dish (mg) and B is weight of dish (mg).
Comments
Highly mineralized waters with a considerableCa2+, Mg2+, Cl- and/or

SO~-content may be hydroscopicand require prolongeddrying, proper desicca-
tion, and rapid weighing.
Sampleshigh in HCO)" require careful drying at 180°C to insure complete
conversionof HCO)" to COJ-. The temperatureat which the residueis dried has
an important bearing on results, becauseweight lossesdue to volatilization of
organicmatter, mechanicallyoccludedwater, water of crystallizationfrom heat-
induced chemical decomposition,as well as weight gains due to oxidation,
dependon temperatureand time of heating.Residuesdried at 103 to 105°C may
retain not only water of crystallization but also some mechanically occluded
water. Loss of CO2 will result in conversionof HCO)" to COJ-. Loss of organic
matterby volatilization usually will be very slight. Residuesdried at 180°C will
lose almost all mechanicallyoccluded water. Organic matter may be lost by
volatilization, but not completelydestroyed. Some carbonatesmay be converted
partially to oxides.
The results may not agreewith the theoreticalvalue for solids calculated
from the chemical analysis of the sample. In general, evaporatingand drying
watersamplesat 180°Cyields valuesfor dissolvedsolidscloserto those obtained
throughsummationof individually determinedmineralspeciesthan the dissolved
solids valuesobtainedthrough drying at the lower temperature.
REFERENCES
Adams, F. 1974. Soil solution. p. 441-481.In E. W. Carson(ed.) Theplant root and its environment.
Univ. PressVirginia, Charlottesville.
Adams,E, C. Burmester,N.V. Hue, and EL. Long. 1980.A comparisonof column-displacement and
centrifugemethodsfor obtainingsoil solutions.Soil Sci. Soc. Am. J. 44:733-735.
Allison, L.E. 1973.Oversaturationmethodfor preparingsaturationextractsfor salinity appraisal.Soil
Sci. 116:65-69.
AmericanSociety for Testingand Materials. 1986.Annual book of ASTM standards.Section11. Vol.
11.01.ASTM, Philadelphia,PA.
Beatty, HJ., and J. Loveday. 1974. Soluble cations and anions. p. 108-117. In J. Loveday (ed.)
Methodsfor analysisof irrigatedsoils. Tech. Commun.54. Commonw.Bur. Soils, Commonw.
Agric. Bur., FarmhamRoyal, Bucks, England.
Bower, CA., and L.v. Wilcox. 1965. Solute salts. p. 933-951.In c.A. Black et al. (ed.) Methodsof
soil analysis.SSSABook Ser. 5. ASA and SSSA,Madison,WI.
Carlson,R.M., R. Overstreet,and DJ. Naylor. 1971. Effect of microbial activity on saturationextract
composition.Hilgardia 40:553-564.
Dao, TH., and T.L Lavy. 1978. Extraction of soil solution using a simple centrifugationmethodfor
pesticideadsorption-<lesorptionstudies.Soil Sci. Soc. Am. J. 42:375-377.
Davies, B.£., and R.I. Davies. 1963. A simple centrifugationmethodfor obtaining small samplesof
soil solution. Nature (London) 198:216-217.
Elkhatib, E.A., O.L Bennett,V.c. Baligar, and RJ. Wright. 1986. A centrifugemethodfor obtaining
soil solution using an immiscible liquid. Soil Sci. Soc. Am. 1. 50:297-299.
Elkhatib, B.A., 1.L. Hem, andTE. Staley.1987.A rapid centrifugationmethodfor obtainingsoil solu-
tion. Soil Sci. Soc. Am. J. 51:578-583.
Gillman, G.P. 1976.A centrifugemethodfor obtainingsoil solution. Div. Rep. 16. Div. Soils, CSIRO,
Townsville, Queensland,Australia.
Grossman,1., and P. Udluft. 1991. The extractionof soil water by the suction-cupmethod:A review.
J. Soil Sci. 42:83-93.
434 RHOADES
Helrich, K (ed.). 1990. Official methodsof analysis. 15th ed. Association of Official Analytical
Chemists,Inc., Arlington, VA
Kinniburgh, D.G., and D.L. Miles. 1983. Extraction and chemicalanalysisof interstitial water from
soils and rocks. Environ. Sci. Technol. 17:362-368.
Kittrick, J.A 1983. Accuracy of several immiscible displacementliquids. Soil Sci. Soc. Am. 1.
47:1045-1047.
Litaor, M.1. 1988. Review of soil solution samplers.Water Resour.Res. 24:727-733.
Longenecker,D.E., and PJ. Lyerly. 1964.Making soil pastesfor salinity analysis:A reproduciblecap-
iIlary procedure.Soil Sci. 97:268-275.
Loveday,1. 1972. Moisture contentof soils for making saturationextractsand effect of grinding. Div.
Soils Tech. Pap. 12a. Commonw. Sci. Industr. Res. Organ., CanberraCity, ACT 2601,
Australia.
Maas,E.Y. 1986. Salt toleranceof plants.Appl. Agric. Res. 1:12-26.
Maas,E.V. 1990. Crop salt tolerance.p. 262-304.In KK Tanji (ed.) Agricultural salinity assessment
and management.ASCE ManualsRep. Eng. 71. ASCE, New York.
Maas,E.Y., and GJ. Hoffman. 1977. Crop salt tolerance--currentassessment. J. Irrig. DrainageDiv.,
ASCE 103(IR2):115-134.
Menzies,N.W., and L.c. Bell. 1988.Evaluationof the influenceof samplepreparationand extraction
techlliqueon soil solutioncomposition.Aust. J. Soil Res. 26:451-464.
Mubarak,A, and R.A Olsen. 1976. Immiscible displacementof the soil solutionby centrifugation.
Soil Sci. Soc. Am. 1. 40:329-331.
Mubarak,A, and R.A Olsen. 1977.A laboratorytechniquefor appraisingin situ salinity of soil. Soil
Sci. Soc. Am. J. 41:1018-1020.
Prichard,T.L., GJ. Hoffman, andJ.L. Meyer. 1983. Salinationof organicsoils in the Sacramento-San
JoaquinDelta of California. Irrig. Sci. 4:71-80.
Reitemeier,R.F. 1946. Effect of moisturecontenton the dissolvedand exchangeableions of soils of
arid regions.Soil Sci. 61:195-214.
Rhoades,1.D.1978. Monitoring soil salinity: A review of methods.p. 150-165.In L.G. Everett and
KD. Schmidt (ed.) Establishmentof water quality monitoring programs.Am. Water Resour.
Assoc.,Minneapolis,MN. .
Rhoades, J.D.1990. Determining soil salinity from measurementsof electrical conductivity.
Rhoades,J.D. 1992. Instrumental field methods of salinity appraisal.In G.c. Topp et al. (ed.)
Advancesin measurementof soil physical properties:Bringing theory into practice. SSSA
Spec.Publ. 30. ASA, CSSA, and SSSA, Madison,WI.
Rhoades,J.D., and J.D. Oster. 1986. Solute content.p. 985-1006.In A Klute (ed.) Methodsof soil
analysis.2nd ed. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Rhoades,J.D., N.A Manteghi,P.J. Shouse,and w.J. Alves. 1989a.Estimatingsoil salinity from sat-
uratedsoil-pasteelectrical conductivity. Soil Sci. Soc. Am. J. 53:428-433.
Rhoades,J.D., B.L. Waggoner,PJ. Shouse,and WJ. Alves. 1989b.Determiningsoil salinity from soil
and soil-pasteelectrical conductivities:Sensitivity analysisof models. Soil Sci. Soc. Am. J.
53:1368-1374.
Rhoades,J.D., PJ. Shouse,WJ. Alves, N.A. Manteghi,and S.M. Lesch.1990. Determiningsoil salin-
ity from soil electrical conductivity using different modelsand estimates.Soil Sci. Soc. Am.
J. 54:46-54.
Richards, L.A. 1941. A pressure-membrane extraction apparatusfor soil solution. Soil Sci. Soc.
51:377-386.
Richards,L.A. 1949. Filter funnels for soil extracts.Agron. 1. 41:446.
Ross,D.S., and R.I. Bartlett. 1990. Effects of extractionmethodsand sample storage on propertiesof
solutionsobtainedfrom forestedspodosols.J. Environ. Qual. 19:108-113.
Shimshi,D. 1966. Use of ceramicpoints for the samplingof soil solution. Soil Sci. 101:98-103.
Skarie, R.L., J.L. Richardson,GJ. McCarthy, and A Maianu. 1987. Evaporite mineralogy and
groundwaterchemistry associatedwith saline soils in EasternNorth Dakota. Soil Sci. Soc.
Am. J. 51:1372-1377.
Sonnevelt,c., and 1. van den Ende. 1971. Soil analysisby meansof a 1:2 volume extract. Plant Soil
35:505-516.
Tadros,V.T., and J.W. McGarity. 1976.A methodfor collectingsoil percolateand soil solution in the
field. Plant Soil 44:655-667.
U.S. Salinity LaboratoryStaff. 1954. L.A. Richards(ed.) Diagnosisand improvementof saline and
alkali soils. U.S. Dept. of Agriculture Handbookno. 60. USDA, Washington,DC.
Wilcox, L.v. 1951. A methodfor calculatingthe saturationpercentagefrom the weight of a known

volume of saturatedsoil paste.Soil Sci. 72:233-237.
Wolt, 1., and J.G. Graveel. 1986. A rapid methodfor obtainingsoil solution using vacuumdisplace-
ment. Soil Sci. Soc. Am. 1. 50:602-{)05.
Yamasuki,S., and A. Kishita. 1972. Studieson soil solution with referenceto nutrient availability. I.
Effect of various potassiumfertilizer on its behaviorin the soil solution. Soil Sci. Plant Nutr.
18:1-{).
Published 1996
Chapter 15
Carbonate and Gypsum
RICHARD H. LOEPPERT, Texas A&M University, College Station, Texas
DONALD L. SUAREZ, USDA-ARS, U.S. Salinity Laboratory,

Riverside, California
The carbonateminerals and gypsum exert a dominating influence on soils in

which they are presentbecauseof their relatively high solubility, and in the case
of the carbonates,their alkalinity and pH buffering properties.In this chapter,
proceduresfor the determinationof quantity, reactivity and equilibrium relations
of thesemineralswill be discussed.
CARBONATE
Inorganic carbonatein soil occurspredominantlyas the sparingly soluble

alkaline-earthcarbonates,calcite (CaC03) and dolomite (CaMg(C03)2). Calcite
is usually the dominantform in active pedogenicenvironments(Doner & Lynn,
1977; Nelson, 1982). There are only a few reported occurrencesof aragonite
(CaC03) and vaterite (CaC03) in soils. Sodium carbonateand magnesiumcar-
bonateand hydroxycarbonatearecommonin evaporatesor in regionsof high-salt
depositionin soil. Concentrationof dissolvedcarbonateis controlledby equilib-
rium relationsof the solid-phasecarbonatesand gas-phaseCO2. Concentrationof
dissolvedcarbonateis likely to be higher in systemswith high partial pressures
of CO2, e.g., in flooded soils or in microenvironmentsof high microbial activity,
or in sodic soils, becauseof the high solubility of Na2C03. Also, calcite and
dolomite usually control the activities of Ca2+(aq) and Mg2+(aq) in soils contain-
ing theseminerals.
Calciumcarbonatecontentsof carbonate-influenced soils rangefrom traces
to greaterthan 80%. Calcite existsin a variety of forms, from nodulesof 1 cm or
greaterdiameterto submicrometerparticlesto well-formed rhomboids.The pres-
enceof carbonatesis usually associatedwith neutral to alkaline soils, but solid-
Book Seriesno. 5.
437
438 LOEPPERT& SUAREZ
phasecarbonatesin the form of nodulesare known to exist in some acid envi-

ronments.
Soils differ considerablywith respectto particle-sizedistribution of the car-
bonatephase.Calcite in active pedogenicenvironmentsexists predominantlyin
the fine-silt and coarse-clayparticle-sizefractions (Bui et aI., 1990); alluvial ba-
sins in the USA containcalcite predominantlyin the silt fraction (Suarez,1977).
Well-formed rhombohedralcrystalsof calciteare not commonin soils with flour-
ishing plant and microbial populations,due to the presenceof chemical agents,
e.g., phosphateand organicacids,that are readily adsorbedon the calcite surface
and can retardor preventnormal crystal growth processes.Calcite is more com-
monly observedas spheroidalaggregatesof microcrystallineparticles(Bui et aI.,
1990). Dolomite is observedpredominantlyin the silt and fine-sandparticle-size
fractions. The largerparticle size of dolomite, comparedto calcite, is attributable
to its nonpedogenicorigin and its slower rate of dissolution.Soils vary consider-
ably with respectto the relative reactivitiesof the carbonatecomponent,due to
differences in carbonatemineralogy, particle size and morphology. Pedogenic
carbonates,due to their aggregatedmicrocrystallinemorphology,are usually ob-
servedto have relatively high reactivities.Calcite has a considerablyhigher dis-
solution rate than dolomite, approximately100-fold.
The carbonateminerals, due to their relatively high solubility, reactivity
and alkaline character,act as pH buffers; the pH valuesof most calcareoussoils
are within the rangeof 7.5 to 8.5. It is becauseof thesepropertiesthat carbonates
play an important role in pedogenic,chemical and rhizosphereprocessesin the
soil.
TOTAL CARBONATE ANALYSIS
Soil carbonateis usually quantified by acid dissolution as summarizedin

the reactionsbelow (Allison & Moodie, 1965; Nelson, 1982):
[1]
and
with the determinationof eitherW consumptionor Ca (and Mg) or CO2 produc-

tion. Alternatively, a dry combustionprocedure,basedon the precombustionof
organic matter at 575°C in an O2 streamand the subsequentcombustionof car-
bonateat 1000°Cand collection of CO2 (Rabenhorst,1988) has beenutilized.
Methods involving determinationof CO2 have usually been preferred,
sincein the absenceof decompositionof organicmatter,the measurement of CO2
production provides an absolutemeasureof carbonate;however, it is essential
that precautionsbe takento ensurethat thereis no interferencefrom organicmat-
ter oxidation. Carbondioxide releasedcan be measuredgravimetrically(Allison,
1960; Allison & Moodie, 1965), titrimetrically (Bundy & Bremner,1972),mano-
metrically (Martin & Reeve, 1955; Presley, 1975), volumetrically (Dreimanis,
1962), spectrophotometricallyby infrared spectroscopy,or by gas chromatogra-
CARBONATE & GYPSUM 439
phy. Methodsinvolving H+ consumption(V.S. Salinity Lab. Staff, 1954; Moore

et aI., 1987)and Ca (orMg) production(EI Mahi et aI., 1987)also havebeenuti-
lized, but specialprecautionsare required,since neitherH+ consumptionnor Ca
release are specific for the carbonate dissolution reaction (e.g., the cation
exchangecomplex can be a sink for H+ and a sourceof Caz+). The methodsof
H+ consumptionthat involve reactionwith a strong acid, such as HCI addition
and back titration of the unreactedacid, are usually not suitabledue to the prob-
lem of consumptionof W by other soil componentsat high W activities; how-
ever, methodsinvolving the reactionof a weak acid suchas aceticacid (Loeppert
et aI., 1984; Moore et aI., 1987) for the determinationof total soil carbonateand
of pH 4.0 sodiumacetate(Bloom et aI., 1985)for the determinationof carbonate
in the clay-sizefraction have beensuccessfullyutilized.
Manganesedioxide in the soil can interfere with the acid dissolutionpro-
ceduresdue to its influence on oxidation of organic matter (Allison & Moodie,
1965). In HCI solution, Cl- reactswith MnOz to produceMnz+ and Clz. The Clz
or one of its reactionproducts,HOCl, can oxidize organic matterand thus result
in the releaseof CO2. Since Fe2+ or Snz+ are more easily oxidized than Cl-, their
presenceensuresthe reductionof MnOz without the formation of Cl z and COz.
The releaseof CO2 from organic matter can be minimized by the addition of
FeClz or FeS04to the acid (Martin & Reeve, 1955; Allison, 1960; Allison &
Moodie, 1965).
Methods have been utilized for the simultaneousquantitativedetermina-
tion of calcite and dolomite basedon: (i) the relative ratesof calcite and dolomite
dissolution upon reactionwith HCI (Skinner & Halstead,1958; Skinner et aI.,
1959; Dreimanis, 1962; Evangelouet aI., 1984); (ii) the selectivedissolutionof
calcite by a citrate buffer and the total carbonatedissolutionby HCI (Peterson&
Chesters,1966; Petersonet aI., 1966);(iii) the relative intensity ratio of x-ray dif-
fraction (XRD) peaks(Tennant& Berger, 1957; Diebold et aI., 1963; Runnells,
1970; VIas & Sayin, 1984); or (iv) differential thermalanalysis(DTA) (Warne&
Mitchell, 1979). Each of thesemethodsrequire the use of finely and uniformly
groundsoil samples.
Methodswhich have beenutilized for the determinationof total carbonate
are summarizedin Table 15-1. The authorsof this chapterprefer the acid disso-
lution procedurefollowed by the manometricdeterminationof COz, due to its
simplicity, though the other methodspresentedin the text also will give reliable
results.The choice of procedurewill dependto a large extent on the equipment
availableto the researcher.Eachof the procedureshasseveralsourcesof error of
which the analyst should be aware (as discussedin the appropriatesections
below). In all cases,the soil samplemust be finely ground. Also, CaC03 or soil
standardsshould be utilized to checkthat accuratequantitativeresultsare being
obtained. Previous reviews by Allison and Moodie (1965) and Nelson (1982)
containadditionalvaluableinformation. The XRD and DTA methods,utilized by
someresearchers, havefound their greatestutility in the qualitativeidentification
of mineral phasesand are not discussedin this chapter.Readersare referred to
other sources(Table 15-2) for discussionsof thesemethodologies.Methodsfor
the quantitativedeterminationof calcite and dolomite are summarizedin Table
15-2.
440 LOEPPERT & SUAREZ
Table 15-1. Methodsfor total carbonatedeterminationof soils.

Component Determination
Principle analyzed method References
Acid CO2 Gravimetric Allison, 1960; Allison & Moodie, 1965
dissolution (NaOH absorbent)
Titrimetric Tinsley et aI., 1951; Bundy & Brumner,
(NaOH absorbent) 1972
Manometric Martin & Reeve,1955; Presley,1975;
Nelson, 1982; Suarez& Wood, 1984;
USDA-SCS,1984
Volumetric Dreimanis,1962
Gravimetric Allison & Moodie, 1965; U.S. Salinity
(C02 loss) Lab. Staff, 1954
HOAc consumed pH (HOAc/OAc ratio) Moore et aI., 1987; Loeppertet aI.,
(from HOAc) 1984
HOAc consumed Titration (difference Bloom et aI., 1985
(from pH 4 method)
NaOAc)
H+ consumed Titration (difference U.S. Salinity Lab. Staff, 1954
(from HCI) method)
Ca,Mg Atomic absorption EI Mahi et aI., 1987
Dry CO2 Gravimetric Rabenhorst,1988
combustion (NaOH absorbent)
Table 15-2. Methodsfor quantitativedeterminationof calcite and dolomite.

Component Determination
Principle analyzed method References
Differential CO2 Manometric Skinneret aI., 1959; Skinner& Hal-
kinetics stead,1958; Evangelouet aI., 1984
Volumetric Dreimanis,1962
Selective Ca,Mg Citrate, HCI Petersonet aI., 1966; Peterson& Ches-
dissolution (atomic absorption) 1966
X-ray Mineralogy XRDt Tennant& Berger,1957; Diebold et aI.,
diffraction 1963; Runnells,1970; VIas & Sayin,
1984
Differential Mineralogy DTAt Warne& Mitchell, 1979
thermal
t XRD =x-ray diffraction, DTA =differential thermalanalysis.
Pressure Calcimeter Method
Principles
The carbonatecontentof a samplecan be determinedby reactionwith acid,
in a closedsystem,to form CO2. At constanttemperature,the increasein pressure
is linearly relatedto the quantity of carbonatepresentin the sample.The slope, a,
of the equation,
CaC03 = a (H) + b [3]
whereH is the pressureand a and b are empirically determinedconstants,is less

than one, becauseof the equilibrium relationshipbetweenCO2 partial pressure
and dissolved CO2• That is, as the pressureincreases,the quantity of CO2

remaining in the liquid phase also increases.Since the equilibrium of CO2
betweensolution phaseand gasphaseand the pressureof the gasare both depen-
dent on temperature,the temperatureof the closedreactionvessel(pressurecal-
cimeter) must be controlledwith either a controlledtemperaturewater bath or a
constanttemperatureroom. Pressureis measuredwith either a manometeror a
pressuretransducer.It is more convenientto run pure calcite standardsat the
given temperatureand barometric pressureand construct a calibration curve,
ratherthan to makespecificcorrectionsfor temperatureandbarometricpressure.
As with the other acid dissolution methods, ferrous iron, as either FeCl2 or
FeS04,can be addedto limit the oxidation of organicmatterand the subsequent
evolution of CO2 from this source.The use of an oxidation inhibitor is especial-
ly importantfor any procedureusing strong acids that may result in the genera-
tion of elevatedtemperatureswithin the soil.
Variations of pressurecalcimeters,especiallywith regardto the designof
the system for the initial mixing of sample and acid, have been described
(Williams, 1949; Martin & Reeve,1955; Skinner et aI., 1959; Evangelouet aI.,
1984). For example,Evangelouet al. (1984) useda magnetoutsidethe pressure
chamberwhich could be manipulatedto dispensethe soil samplefrom a metal
cup within the chamber.
A modification of the pressure-calcimeter procedurebasedon the relative
ratesof reaction of calcite and dolomite has been used to quantitatively deter-
mine calcite and dolomite in mixed-phasesystems(Skinner & Halstead,1958;
Skinner et aI., 1959; Evangelouet aI., 1984). An estimateof the calcite and
dolomite contents canbe obtainedby taking measurements after 1 to 2 min (cal-
cite) and after 1 to 2 h (calcite + dolomite). More accuratedeterminationsof cal-
cite and dolomite require developmentof time-pressurecurves, using either a
mercury manometer(Skinner et aI., 1959; Turner & Skinner, 1959, 1960) or a
pressuretransducer(Evangelouet aI., 1984). Skinner and Halstead(1958) and
Skinneret al. (1959) plotted log (P~-PI) vs. time during reactionof soils with 4
M HCI in a pressurecalcimeter,took the approximatelylinear portion of the
curveoccurringafter 1 min and extrapolatedthis line to time 0 (to). The intercept
value at to, PD' representsthe CO2 derived from dolomite, and P~-PD represents
the CO2 derivedfrom calcite. The pressuretransducer,suchas that describedby
Evan-gelouet al. (1984), is more highly suited than the Hg manometerfor this
purposedue to its more rapid responseand higher sensitivity and the easeof
obtainingdata.
Method
Apparatus
1. For use of a Hg manometer:
A. Pressurecalcimeter,reactionvessel(Fig. 15-1). Use a 90-mL (3-
oz) glass bottle (A), approximately 92-mm height and 40-mm
diam., with plastic screwcap (B). Drill a 3-mm hole in the plastic
cap.
B. Hypodermicneedle(El ), size 26.
442 WEPPERT& SUAREZ
em
F-'>
\1 3.
2'
I'
e
g B-'> I'
N
1,~
~
E,
20
G-4 3.
A -- Reaction bottle
B -- Plastic cap D
C -- Rubber gasket
D -- Soil + Water I -- Plastic leg cemented to
E1 , Ez -- Hypodermic needles bottom of vial
F -- Rubber tubing J -- Via l, plastic
G -- Hg manometer K -- Syringe, plastic, 1 cm 3, with
H -- 6 M HCI / FeClz reagent size 26 hypodermic needle
Fig. 15-1. Pressurecalcimeterapparatus(Nelson, 1982).
C. Gasket(C), rubberor plastic. Cut gasketsfrom rubbersheeting,to

fit just insidethe plasticscrewcapsof the pressurecalcimeter.The
gasketmust form a sealwith the top of the bottle.
D. Vials (1), heavy wall polystyreneor high density polypropylene,
18-mL capacity,22-mm diam., 46-mm height. Cut vials down to
2S-mmheight and attachvial to side bottom of the bottle.
E. Construct a 90-cm Hg manometer(G) by bending 3- to 4-mm
internal diameter(i.d.) glasstubing or by connectingtwo piecesof
glasstubing with Tygon tubing. Attach rubber tubing to one side
of the manometer.To the other end of the rubber tubing attacha
syringe barrel (K), with the upper lip removed,polypropylene,1
cm3, with hypodermicneedle(size 26).
2. For usewith pressuretransducer:
A. Use a SOO-mLglassbottle (clean,usedreagentbottle) with plastic
cap.
B. Sameas with manometer.
C. Sameas with manometer.

D. Sameaswith manometer.
E. Pressuretransduceraccuratein the 0 to 20 kPa (aboveatmospher-
ic) pressurerangewith a 0 to 1 V, or less,full-scale output.
F. Strip chartrecorderor analog-to-digitalboardfor personalcomput-
er.
Reagents
1. Hydrochloric acid (HC1), 6 M, with 3% (by weight) ferrous chloride
(FeC12 • 4 H20): Add 500 mL of concentratedHCl to 400 mL of deion-
ized water,then add30 g of FeC12 • 4 H20 and dilute to a total volume
of 1 L.
2. Sand, acid-washedand rinsed, and ground to passa 200 mesh in.-1
(75J..lm nominal pore size) sieve.
3. Calciumcarbonate(CaC03), reagentgrade.
4. Carbon-dioxide(C02) free deionizedwater: Boil deionizedwater for
10 min, cool rapidly in an ice bathto approximatelyroom temperature,
and stopperto preventcontactwith atmosphericCO2•
Procedure
Calibration.Prepare0.5, 1.0, 2.0, 5.0, 7.5, 10, 15 and 20% CaC03 stan-
dards by adding dry reagentgrade CaC03 to acid-washedand rinsed sand.
Transfer2.000g of eachmixture to the bottomof a dry reactionbottle. Add 5 mL
of freshly prepared,CO2-free deionizedwatergently down the side of the bottle;
avoid splatteringthe standardsampleonto the reactionvesselwall. Pipette5 mL
of 6 M HCI-FeC12 reagentinto the plastic vial and insert, with the aid of tweez-
ers or forceps,onto the platform of the reactionvessel.Wipe the rim of the reac-
tion bottle with glycerol. Insertgasket(C) into the plasticcup (B), and fastenthe
plasticcap tightly onto the reactionbottle (A).
Insertthe hypodermicneedle(E1) throughthe 3-mm hole of the plasticcap
(B) and allow 10 s for pressurein the containerto equilibratewith atmospheric
pressure.Removethe hypodermicneedle,then mix the HCI-FeCI2 reagentwith
the sampleby tilting the vial (J) and bottle (A). After the acid and samplehave
reactedfor 1 min, fully immersethe reactionbottle (A) in water at room temper-
atureto checkfor leaks.If no leaksare evident,removebottle (A) from the water
bath. After 2 min, tilt and rotatethe reactionbottle (A) to mix the acid with soil
particlesthat are on the sidesof the bottle. After 1 h, swirl the bottle without
splatteringsolution on the gasket(C).
Insert the hypodermicneedle(E2) throughthe 3-mm openinginto the bot-
tle (A). If a manometeris being used, record the difference in height of Hg
betweenthe two armsof the manometer.Use the sameprocedureon two blanks
(Hb) (reagentsonly). Subtractthe blank (Hb) readingsfrom eachof the standards
to obtainHe corrected.Calculatethe regressionequationusinga linear regression
procedure
[4]
wherea and b are the regressionparameters.
If a pressuretransduceris being used, read the pressurepeaks off the

recorderor enter pressurevaluesdirectly into the personalcomputerfor subse-
quentprocessing.
Samples.Weigh 2.00 ± 0.01 g of air-dry soil. Transferthe sampleto a reac-
tion flask (A) and proceedas describedabove for the standards.If pressureor
manometerreadingsexceedthe value recordedfor the higheststandard,repeat
the samplewith an appropriatelyreducedmassof soil (i.e., 0.5-1.0g of soil).
Calculations
Calculatethe massof CaC03 (g) in the soil sampleusing the linear regres-
sion betweencarbonatemassand pressure.
Calculatethe calcite equivalent
g CaC03
calcite equivalent(massfraction) = ·1 I
g sOl sampe
Comments
Estimatesof CaC03 and CaMg(C03h (dolomite) contentscan be obtained
by taking measurementsafter 2 min (calcite) and after 1 to 2 h (calcite +
dolomite). More accuratedeterminationsrequiredevelopmentof a time-pressure
curve, as describedby Turner and Skinner(1959, 1960) for use with a manome-
ter and Evangelouet al. (1984) for usewith a pressuretransducer.
GravimetricMethod
Principles
In this methodthe inorganiccarbonatesare decomposedby treatingthe soil
in a reactionflask with 1 M H2S04 containingFeS04as an antioxidantto prevent
the releaseof CO2 from organicmatter(Allison, 1960; Allison & Moodie, 1965).
The CO2releasedduring the dissolutionof carbonateis carriedby meansof a pre-
purified CO2-free air or N2 streamthrougha seriesof trapsto removeinterfering
gasesas follows: (i) concentratedH2S04 to removewater vapor, (ii) Zn metal to
remove tracesof H2S04, and (iii) Mg(CI04)2 to remove the last tracesof water
vapor. The gasstreamthen passesthrough a Nesbitt bulb which containsNaOH,
a CO2 absorbent.The CO2 originatingfrom the carbonatedecompositionreaction
is determinedby weighing the Nesbitt bulb before and after the absorptionof
CO2• The chemicalreactionsinvolved in the variousstepsof the processare sum-
marizedbelow:
1. purification of purgegas
[5]
2. carbonatedecomposition
[6]
3. removal of water vapor
[7]
4. removalof tracesof H2S04
[8]
5. removalof tracesof water vapor
[9]
6. absorptionof CO2
[10]
Method
Apparatus
A modified versionof the apparatusfor digestion,trappingof undesirable
gasesand CO2 absorption that was originally describedby Allison (1960) is
shown in Fig. 15-2. If the sameapparatusalso is to be usedfor determinationof
total organicC, additionaltrapsare neededto removethe N-, S- and halogen-con-
taining gasesfrom the gasstream,as describedin Chapter36 (Sawhney,1996).
The original apparatusas describedby Allison (1960) can be constructed
from the following parts (Nelson & Sommers,1982): (A) Hoke needlevalve to
control air flow; (B) 25-cm high soda-limetower; (C) 100-mL Kjeldahl flask to
fit a no. 2 stopper;(D) Allihn four-bulb condenserfitted with a no. 2 stopperat
the bottom end and a two-hole no. 2 stopperat the top end; (E) 60-mL open-top
G H
Concentrated
H2S0 4
Fig. 15-2. Modified Allison apparatusfor the gravimetric determinationof carbonate(Nelson &
Sommers,1982).
separatoryfunnel; (F) 25- by 90-mm shell vial with no. 4 stopper;(G and H) 15-
cm long U-tube; and (I) Nesbitt absorptionbulb. Use neoprenestoppersandgum
rubbertubing for all connections,coat all glass-to-rubbertube connectionslight-
ly with silicone lubricant.
ItemsC throughE (Fig. 15-2) canbe replacedwith glasswarewith ground-
glassjoints (standard-taper24/40) (Nelson & Sommers,1982). The following
parts are needed:(C) 100-mL round-bottomflask (Coming 4320); (C-l) distill-
ing adaptertube (Coming 9421),which containsan inlet tube for bubbling CO2-
free air into the digestionacid mixture; (D) Allihn condenser,with approximate-
ly 30-mm jacket length (Coming 2480); (E-1) distilling tube with suction side
arm (Coming 9420) (side arm is connectedto purifying traps); (E) graduated
separatoryfunnel (Coming 6382A).
The carrier gas streamcan be supplied by either a laboratory source of
compressedair or bottled air or N2. Carbon dioxide in the carrier streamis re-
movedby passingit throughthe soda-limetowerB or othersuitableCO2 absorb-
ing system.In the original (Allison) apparatus,the outlet of the soda-limetower
is connectedto a glasstube of 4-mm outsidediameter(o.d.) that extendsthrough
the upper stopperof condenserD, downward through the condenserand dips
about 1 cm below the acid in digestionflask C. The stemof the funnel E should
extend into condenserD to at least 5 cm below the stopperto avoid contact
betweenthe acid and the stopper.In the ground-glassapparatus(Fig. 15-2), the
stem of the graduatedseparatoryfunnel should extendthrough the entire length
of the condenserD. The stopcockof the graduatedfunnel may be lubricatedwith
stopcockgrease;however,if the sameapparatusis to be usedfor determination
of total organic C by concentratedH2SOJH3P04digestion, regular stopcock
lubricant shouldnot be usedon the stopcock.
The purifying trapsare usually mountedon a panelwith attachedbase.The
vial for the H2S04 trap is fitted with a no. 4 stopperthat has approximately0.6
cm cut off to provide a tight sealwith the vial. Fill the trap not more than one-
third full with concentratedH2S04, Preparethe inflow tube from the end of a 5-
mL pipettewith the tip extendingnot more than 1.3 cm into the acid. The outlet
tube from the H2S04 trap connectsto the U-tube. Placea glass-woolplug in the
bottomof the U-tube. Fill the inlet sidewith 30 meshin.-1 (600-llm nominalpore
size) granularZn and the outlet side with anhydrousMg(CI04)2' Place a loose
plug of glass woolin eachend of the U-tube and stopperimmediatelyto prevent
hydration of the anhydrousMg(CI04)z.
The Nesbitt bulb shouldbe layeredsuccessivelywith a plug of glasswool,
a 3-cm layer of 1.15- to 2.13-mm (8- to 14-mesh)in.-1 CO2 absorbent,a 2-cm
layer of 0.84- to U5-mm (14- to 20- mesh) in.-1 CO2 absorbent,a l-cm layer
of anhydrousMg(CI04)z, and a layer of glasswool.
During the digestionstep, heat must be applied to rapidly bring the reac-
tion mixture to a boil. This can be most easily accomplishedby useof a Bunsen
or Meekerburner(Allison, 1960).
Reagents
1. Digestion acid for carbonates:Dissolve 56 mL of concentratedsulfu-
ric acid (H2S04) and 92 g of ferrous sulfate heptahydrate(FeS04•
7H20) in 600 mL of deionizedwater. Cool the solution, dilute it to

approximately1 L, and storein a well-stopperedcontainer.This solu-
tion is approximately1 M H2S04 and contains5% FeS04,to act as an
antioxidant.
2. Calcium oxide, Ascarite II or other suitable CO2 absorbentfor the
soda-limetower.
2. Sulfuric acid, concentrated.
3. Granularzinc, 30 meshin.-l.
4. Magnesiumperchlorate[Mg(CI04)2], anhydrous.
5. Absorbent for carbon dioxide (C02): Ascarite II or other suitable
absorbent,approximately8- to 14-mesh in.- l and 14- to 20-mesh
in.- l ; an indicating absorbentis preferred.
Procedure
Transfera soil sample,previously ground to passa 30 mesh in.-l sieve,
containingnot more than 250 mg of CaC03 equivalentinto a 100-mL digestion
flask, and connectto the condenser(Fig. 15-2). Weigh the Nesbitt bulb, attachit
to the system,and openthe valve at the top of the bulb. Pour 25 mL of the diges-
tion acid into the burette at the top of the condenserwith the stopcockclosed.
Allow the acid to enter the digestionflask, and immediatelyclose the stopcock
to preventloss of CO2, Be sure that the air delivery tube extendsat least 5 mm
below the acid level in the digestionflask. Tum on the cooling water to the con-
denser.Adjust the carrier streamto a flow rate of about 2 bubblesS-l, and main-
tain this rate during digestion.Apply heat slowly, and bring the contentsof the
flask to a boil in about 4 min. Continuegentle boiling for exactly 3 min more.
Removethe flame, adjust the carrier streamto 6 to 8 bubbless-l, and continue
the aerationfor 10 min. Shutoff the air streamand disconnectthe digestionflask
from the condenser.Close the stopcock on the Nesbitt bulb, disconnectthe
Nesbitt bulb from the system,and weigh it immediately.Make a blank determi-
nation using the identical procedurebut without a sample.Before determining
the carbonatecontentof soil samples,the procedureshouldbe checkedwith fine-
ly ground CaC03 standardsto be sure that a quantitativedigestionand determi-
nation of carbonateis being obtained.
Calculations
gC
InorganiccarbonateC, massfraction,
g soil
= (g CO2 from sample- g CO2 from blank ) ( atomic weight of C )

C g water-freesoil molecularweight of CO2
=( g CO2 from sample- g CO2 from blank)

• (0.2727)
g water-freesoil
Calcium carbonate,%
= (g CO2 from sample- g CO2 from blank ) •

g water-freesoil
molecularweight of caC03)
( • (100)
molecularweight of CO2
g CO2 from sample- g CO2 from blank)

=( . • (2.274) • (100)
g water-freesoIl
Comments
The H2S04 trap should be preparedanew at the beginning of each day's
operation,or more often if frothing occurs.One or more reactionswith standard
CaC03 should be madeat the beginningof the day and a single standarddeter-
mination for at least every 10th sample during the day to determinewhether
quantitativeanalysesof CaC03 are beingobtained.Low CO2 valuesmay indicate
incompletedecompositionof carbonateor incompletepurge of CO2 (digestion
and purge time shouldbe increasedby a few minutes;longer reactiontimes may
be neededif samplesare known to contain dolomite), leaks in the apparatus
(checkcarefully using an aqueousleak detector),or a depletedCO2 absorbentin
the Nesbitt bulb (repack the Nesbitt bulb). High CO2 values may indicate a
depletedsoda-limetower, a depletedwater trap [reprepareboth the H 2S04 and
Mg(CI04h traps], or a poorly conditionedapparatus(run 1 or 2 blank samples
until systemequilibrium is attained).
When the apparatusis idle overnightor for longerperiods,the Nesbitt bulb
should be detachedand stored in a desiccator,and the tubes connecting the
Mg(CI04h trap should be clampedto preventhydration of the desiccant.
Since strongacids are usedin this procedure,essentialprecautionssuchas
propereyewearand clothing and protectivebarriersshouldbe usedto shield the
operator.
SimpleTitrimetric Procedure
Principles
This procedureis basedon the dissolutionof soil carbonateas describedby
Eq. [1] and [2] and the subsequentreactionof CO2 with aqueousKOH or NaOH
by the following reaction
[11]
An aliquot of the aqueousKOH or NaOH absorbentis first titrated with standard
HCI to the phenolphthaleinendpoint,which involves thefollowing reactions,
[12]
and
+- Rubber Stopper
~:f=I==;?
Glass Tube
2 M KOH
Soil Sample
Fig. 15-3. Digestion vesselfor the titrimetric determinationof carbonate(Bundy & Bremner,1972).
[13]
and then to the bromcresolgreenendpoint
[14]
The inorganic carbonatecontentis proportionalthe HCI consumedin this latter

reaction.
Method
Apparatus
The digestionapparatusshouldbe preparedas illustratedin Fig. 15-3. The
digestionchamberis a wide mouth, 8 oz (approximately240-mL) Frenchsquare
(or similar) bottle. The chamberis fitted with a rubberstopperwith a single hole
(6-mm diam.) to tightly hold a glasstube (length, 110 mm; i.d., 5 mm). The upper
end of the glass tube is sealedwith a sleeve-typerubber septum(plug diam., 5
mm; cap diam., 9 mm). A 5-mL beaker(to hold the KOH solution) is attachedto
the glassrod by meansof Epoxy cementor a rubberband, so that the bottom of
the beakeris about5 mm abovethe lower end of the tube.
Reagents
1. Potassiumhydroxide (KOH), 2 M. Dissolve 112.2 g of reagentgrade
KOH into CO2 free deionizedwater (which hasbeenboiled to remove
dissolvedCO2), and dilute the solution to 1-L total volume. Store in a
tightly stopperedpolypropylenebottle.
2. Hydrochloric acid (Hel), 2 M. Add 167 mL of concentratedHCI to
about 700 mL of deionizedwater, and dilute the solution to 1-L total
volume.
3. Hydrochloric acid (HCI), 1 M. Add 83 mL of concentratedHCI to

about 700 mL of deionizedwater, and dilute the solution to 1-L total
volume.
4. Hydrochloricacid (HCI), 0.1 M. Take 100 mL of the 1 M HCI solution
and dilute to exactly 1-L total volume. This reagentshould be stan-
dardizedagainsta standardtris-hydroxyaminomethane (THAM) solu-
tion.
5. Phenolphthaleinindicatorsolution. Dissolve0.05 g of phenolphthalein
in 50 mL of 95% ethanol,and add 50 mL of deionizedwater.
6. Bromcresol green indicator solution: Dissolve 0.1 g of bromcresol
greenin 250 mL of 0.0006M NaOH.
7. n-octyl alcohol.
Procedure
Weigh a sampleof finely ground (to pass 100 mesh in.-1 sieve; 150-llm
nominal pore size) soil (not more than 8 g) containingup to 30 mg of inorganic
C into an 8-oz (240-mL) squarebottle, add 1 drop of n-octyl alcohol,placeexact-
1y 5 mL of 2 M KOH in the 5-mL beakermountedon the stopperassembly,and
stopperthe bottle tightly. Remove50 mL of air from the bottle by insertinga 50-
mL gassyringeinto the septum,then inject 20 mL of 2 M HCI into the bottle with
a hypodermicsyringe. Swirl the bottle gently for a few secondsto mix the con-
tents, taking care to minimize the splatteringof the soil onto the walls of the jar
and to preventthe loss of NaOH from the beakeron the stopperassembly.After
allowing the bottle to standat room temperaturefor 16 to 24 h, gently unstopper
the bottle and quantitatively transfer the contentsof the beakeron the stopper
assemblyto a 125-mL Erlenmeyerflask (markedto indicatea volume of 50 mL)
with the aid of C0z-free deionized water, to give a final volume of 50 mL.
Stopperthe Erlenmeyerflask if any time is to elapsebefore titration.
Add 0.3 mL of phenolphthaleinindicator to the flask, titrate with 1 M HCI
until the pink color beginsto fade, and continuethe titration with 0.1 M HCI until
the phenolphthaleinendpoint(colorless)is reached.Then add 0.8 mL of brom-
cresolgreenindicator solution and titrate with standard0.1 M HCI to the brom-
cresol green end-point(the color changeat the endpoint is from blue to bright
yellow). Alternatively, the samplecould be titrated to the HCO)" pH endpoint
(approximately8.2) and then to the H2C03 pH endpoint(approximately4.5).
Calculations
gC
InorganiccarbonateC, massfraction,
g soil
(mL HCls - mL HCld (mmolc HCI mL-l) (0.012 g C mmol.,--l)

= g soil
Calcium carbonate,%
(mL HCls - mL HCld (mmolc HCI mL-l) (0.050 gCaC03 mmolc)

= .
g soIl
(100)
where, mL HCls = mL of standard0.1 M HCl required to titrate the sample

from the phenolphthaleinendpointto the bromcresolgreen
endpoint;
mL HClc = mL of standard0.1 M HCI required for this titration in a
blank analysisperformedexactly as describedfor the sam-
ple analysisbut with no soil sampleaddedto the bottle; and,
concentrationof the standardHCI is expressedin mmole
HCI mL-l.
Volumetric CalcimeterMethod
Principles
In the volumetriccalcimetermethod,the carbonatesare treatedwith excess
acid, as illustrated by Eqs. [1] and [2], and the CO2 is determinedvolumetrical-
ly (Allison & Moodie, 1965). Under conditionsof constantpressureand temper-
ature, the increasein volume of the systemis a direct measureof massof CO2
evolvedand henceof soil carbonatedecomposed.The apparatususedto measure
the increasein volume is calleda volumetric calcimeter.The apparatusdescribed
herein is the Chittick apparatus,as utilizedby Dreimanis(1962). Since the vol-
ume of a given massof CO2 is dependenton both pressureand temperature,cor-
rections must be made to adjust for deviations in CO2 volume from those
observedat standardtemperatureand pressure.Also, the solubility of CO2 in
water or acid is dependenton temperatureand PCO2' as well as on the achieve-
ment of equilibrium betweengaseousphaseand solution phaseCO2, It is very
difficult to make solubility correctionsbasedon physical constantsdue to the
uncertaintyof CO2 equilibrium. The problemof evaluatingCO2 equilibrium (or
disequilibrium) isusually overcomeby calibratingeachspecificcalcimeterappa-
ratus under the exactconditionsto be utilized in the analysis.
The Chittick apparatushasbeenusedto determinecalcite and dolomite in
mixtures,basedon the relative ratesof reactionof theseminerals.
Method
Apparatus
1. Volumetric calcimeter(Dreimanis, 1962). The volumetric calcimeter
is shown in Fig. 15-4. The principle componentsare: (A) 250-mL
Florenceflask with two-hole rubberstopper,(B) 25-mL addition tube
with stopcock,(C) three-waystopcock,(D) manometer,and (E) level-
ing bulb. Tubing connectionscan be madewith Tygon tubing.
2. Magneticstirrer.
Reagents
1. Hydrochloric acid (HCI), 6 M, with 5% (by weight) ferrous chloride
(FeCI2 • 4H20): Add 500 mL of concentratedHCI to 400 mL of deion-
ized water, then add 50 g of FeCl2 • 4H20, and dilute to a total volume
of 1 L.
2. n-amyl alcohol.
ML
E
B
20
200
Fig. 15-4. Volumetric calcimeterfor the determination

of carbonate(Dreimanis,1962).
Procedure
1. Calibrationof calcimeter:
A. Weigh dry fine-grained(small enoughparticle size to passthrough
a 100 meshin.-1 sieve; ISO-11m nominal pore size) reagent-grade
CaC03 to the nearest0.1 mg into separatedecompositionflasks.
Samplesof approximately10, 20, 30, 50, 75, 100, 150, 200, 300,
400, 600 and 800 mg are recommended.
B. Placea stirring bar in the flask, and add two dropsof amyl alcohol.
C. Install the sampleflask in the system,and fill the graduatedfunnel
(B) to the 25-mL mark with HCI-FeCI2 solution.
D. Openthe three-waystopcock(C) to the atmosphere,and adjustthe
liquid level of the measuringburette(D) to exactly0 mL by adjust-
ing the height of the leveling bulb (E).
E. Close the systemto the atmospherewith the three-waystopcock
(C) (180° rotation), and lower the leveling bulb about 2 cm.
F. Simultaneouslybegin to add HCI-FeCI2 solution from the graduat-
ed funnel(B) to the sampleand begin lowering the leveling bulb.
The leveling bulb liquid level should be kept 1 to 2 cm below the
liquid level in the measuringburette(D).
G. After the sampleis moistened,tum on the magneticstirrer (slow
stirring rate).
H. Closethe stopcockof the graduatedfunnel (B) after 20 mL of acid
has beendispensed.
I. When the level in the gas buretteceasesto drop (usually less than
3 min), equalizeliquid levels in the leveling bulb (E) and the mea-
suringburette(D), and readand recordthe volume of CO2 that has
been evolved. Also record the temperature(1) and barometric
pressure(P).
2. Determinationof total carbonatein soils:
A. Add 0.5 to 5.0 g ± 1 mg of soil which has beenground to passa
100 meshin- 1 sieve (150-~m nominal pore size) to the decompo-
sition flask (A). The sample shouldcontain no more than 600 mg
CaC03 equivalent.
B. Performsteps(b) through (i) as describedabove.Longer reaction
times will be requiredif the soil containsdolomite.
3. Determinationof calcite and dolomite in soils:
A. Performsteps(b) through(i) as for the determinationof total car-
bonate;however,in this case,two readingswill be taken.The first
readingshouldbe taken at 30 s and the secondreadingat 30 min.
B. Exactly 30 s after the addition of acid, equalizeliquid levels in the
measuringbulb and the measuringburetteand readand recordthe
volume of CO2 that has beenevolved. Also, record the tempera-
ture (1) and barometricpressure(P).
C. Turn off the magneticstirrer exceptfor a 15- to 30-s stirring peri-
od every 5 to 10 min. Maintain the liquid level in the leveling bulb
1 to 2 em below that in the measuringburette.
D. At 30 min following the addition of acid, repeatthe measurements
as madeat the 30-s reading.
Calculations
1. Calibrationof calcimeter:
A. Correctthe CO2 volume for the standardsby subtractingthe aver-
age CO2 volumesfor the reagentblanksas follows
VC02(corr) = VCOz(sld) - Vc02(blank)"

B. Reduceall correctedCO2volumesto thoseat standardtemperature
and pressure(STP) using the following equation:
273 K) ( P mm Hg )
VC02(STP)= Vco2(corr) ( T OK 760 mm Hg
C. Determinethe calculatedCaC03, WCaC03(cal)' from the VC02(STP)

valuesusing the following equation:
( 100 g CaC03 mol- 1L )

1 ) (
Weaco3(cal) = [Vco2(corr)l 22.414Lmol- 1 1000mL
D. Plot actualCaC03, WCaC03'on the y-axis vs. WCaC03(cal) on the x-

axis. The plot shouldbe closeto a straightline. The slopeC is the
correction factor betweenthe actual and calculatedCaC03. The

value of C shouldbe less than 1 and is relatedto the actual quan-
tity of CO2 remainingdissolvedin the HCI digestionagentunder
the conditionsof analysis.
W CaC03 =(C) [W caC03(cal)]·

2. Determinationof carbonatein soils:
A. The V C02(STP)is calculatedas in (a) and (b) above.
B. The weight of CaC03 is calculated asfollows
100 g CaC03 mOl-I) ( 1L )

(
WCaC03 = (C) [V C02(corr)] 22.414L mol-1 1000 mL
C. Calcium carbonateequivalentis calculated asfollows
.
CaC03 eqUIvalent,% = (Wcac03g ) (100).
Wsoilg
3. Determinationof calcite and dolomite:

A. The V C02(STP)for the first and secondCO2 determinationsis cal-
culatedas in (A) and (B) above.
B. The weight of CaC03 is calculated asfollows
( 100 g CaC03 mOl-I) ( 1L )

Wcalcite =(C) [V 1-C02(corr)] 22.414L mol-l 1000 mL
Wdolomite = (C) [V 2-C02(corr) - V 1-C02(corr)]·
( 100 g dolomite mOl-I) ( 1L )

22.414L mol-1 1000 mL
C. Calcium carbonateequivalentis calculatedas follows
Calcite, % = (Wcalcite g ) (100)

Wsoilg
Dolomite, % = (Wdolomite g) (100).

Wsoilg
Comments
The major source of error for this procedureresults from the degreeof
degassingof CO2 from the digestionmixture. Uniformity of procedure,includ-
ing volume of acid, agitationof the digestionmixture and time of measurement
of CO2 volume, is essential.
For the determinationof calcite and dolomite, corrections are usually

requiredsincenot all of the calcite and a portion of the dolomite is dissolvedat
the 30-s reading.Also, the dolomite may not be totally dissolvedat the 30-min
reading. Therefore,correctionsare utilized to more closely reflect the actual
quantitiesof calcite and dolomite. Theseconstantsmust usually be calibratedin
each laboratory for the specific conditions of analysis,by the use of soils of
known calcite and dolomite contentsor of reagentgradecalcite and dolomite.
GravimetricMethod for Loss of CarbonDioxide
Principles
When carbonatesare decomposedwith acid as describedby Eqs. [1] and
[2] in an opensystem,CO2 is releasedto the atmosphere.The decreasein weight
resultingfrom CO2 loss is proportionalto the carbonatecontentof the soil. The
method is adoptedfrom U.S. Salinity Lab Staff (1954) and Allison & Moodie
(1965).
Method
Reagents
1. Hydrochloric acid (HCI), 3 M. Transfer250 mL of concentratedHCI
to 500 mL of deionizedwater and dilute to a total volume of 1 L.
Procedure
Weigh (to the nearest0.1 mg) a stoppered,50-mL Erlenmeyerflask con-
taining 10 mL of 3 M HCI. Alternatively, a 70-mL (20-dram)snap-lidpolypropy-
lene vial with 2-mm diam. holesin the caps canbe utilized. Transfera 1- to lO-
g air-dried soil sample(containing0.1-0.3 g of CaC03 equivalent)to the con-
tainer, a little at a time, to preventexcessivefrothing. After effervescencehas
subsided,replacethe stopperloosely on the flask and swirl the flask occasional-
ly for about 15 min. At intervalsof about 15 min, removethe stopperand swirl
the flask for 10 to 20 s to displaceany accumulatedCO2 with air. Replacethe
stopper,and then weigh the flask and its contentsto the nearest0.1 mg. Repeat
the agitation and weighing procedureuntil the weight of the containerdoesnot
changeby more than 1 to 2 mg. The reactionis usually completewithin 1 h.
Calculations
Weight of CO2 =Differencebetweeninitial andfinal weights(flask + stop-
per + acid + soil)
g CO2 lost) ( g C mol-1 )

C03-C, % = ( (100)
g soil g CO2 mol-1
=(g CO2!ost) (0.2727)(100)

gsod
( g CO
CaC03, % = .
2 lost ) (g CaC03 mOl-I)
(100)
g sod g CO2 mol-1
I
= (g COs.lost (2.273) (100)
g sod ")
Comments
The major sourcesof error with the weight loss procedureare evaporation
of water and failure to quantitativelydegasCO2• Errors due to decompositionof
organic matter can be reduced by the addition of 3% FeCl2 to the HCI.
Reasonableresults can be achievedwith proper adherenceto uniform experi-
mental procedure; however, precision is usually lower than by the methods
involving direct quantification of evolved CO2. This method is not suitable for
soils with low CaC03 contents.
Acetic Acid Dissolution Method
Principles
A procedurefor the routine determinationof soil carbonatewhich requires
only a pH meteris basedon the following reaction(Loeppertet aI., 1984; Moore
et aI., 1987)
CaC03 + 2 HC2H30 2 ~ Ca2+ + 2 C2H30i + H20 + CO2 [15]
The neutralizationof acetic acid in the abovereactionis then expressedas
[16]
An equilibrium expressionfor the reactionis
[17]
which may be written in terms of CaC03 contentas
pH = a log lCaC03 )
T- CaC03
+ b, [18]
in which T equalsthe total amount of CaC03 which could be completely neu-

tralized by the quantity of acetic acid addedto the system;a and b are constants
for the empirically determinedequationfor a seriesof CaC03 standards.
The experimentalprocedure involves addition of a known quantity of
acetic acid to a given quantity of soil. The pH of the reaction mixture is then
determinedfollowing the complete dissolution of CaC03. Calcium carbonate
contents are determined from a standard curve of pH vs. log[CaC03/(T -
CaC03)] for known quantitiesof CaC03.
Method
Apparatus
1. pH meter,which gives a digital output of 0.01 pH unit or better.
Reagents
1. Calcium carbonate,fine-grained,reagent-grade.
2. Acetic acid, 0.4 M (H~H302); weigh 24.02g of glacial aceticacid and
dilute to 1 L total volume.
Procedure
To preparea standardcurve, weigh fine-grainedreferencecalcite samples
to the nearest0.1 mg and place in 70-mL (20-dram) snap-lid vials with 1-mm
holes in the capsto allow escapeof CO2 during the dissolutionreaction.The 1-
mm holesare large enoughto allow exchangeof CO2, yet small enoughto mini-
mize the volatilization of HC2H30 2. Sampleweightsof 10, 30, 50, 100, 200, 300
and 400 mg are suggested.
Weigh soil samples(previouslygroundto passa 100-meshin.-1 sieve; 150-
Ilm nominal pore size) to the nearest0.001 g and place into 70-mL (20-dram)
snap-lid polypropylenevials with 1-mm diam. holes in the caps. Soil weight
shouldbe less than 2 g for soils with less than 20% CaC03 equivalent(CCE), 1
g for soils with 20 to 40% CCE and 0.5 g for soils exceeding40% CCE.
Add 25.0 mL of 0.40 M CH3COOH to calcite standardsand soils in the
snap-lidvials. Placethe vials on a rotary shakerovernight(approximately16 h).
After overnight equilibration, measurepH of suspensionsof standardsand soils
to the nearest0.01 pH unit. If the pH exceeds5.00, the analysisshouldbe repeat-
ed with a lower quantity of soil.
Calcu/ations
Plot a standardcurve of pH vs. mg CaCOy'(T - mg CaC03), whereT rep-
resentsthe quantity of CaC03 that would reactstoichiometricallywith the acetic
acid added.As describedabove(with 25 mL of 0.4 M CH3COOH added),T is
500 mg. Determinethe slopeand interceptof the linear equation
mg CaC03 )
pH =a log ( +b [19]
T-mgCaC0 3
CalculateCCE from the experimentallydeterminedpH values of the soil sus-

pensionsusing the following equation
CCE % = 100 (T) [20]

, S (1 + lO(b - pH)la)
CCE, g kg-1 = 10 (CCE, %) [21]
where,a and b are constantsderived empirically from the standardcurve, and S

is the oven-drysoil weight in milligrams.
458 WEPPERT& SUAREZ
Comments
This procedureprovidesa meansof obtainingquantitativedeterminations
of soil carbonatewith readily available equipment,but it is subjectto errors to
which the CO2evolutionproceduresare not subject.Possiblesourcesof error are:
(i) consumptionof W by the soil cation- exchangecomplex, (ii) dissolutionof
soil componentsotherthan CaC03, (iii) incompletedissolutionof the solid-phase
carbonate,(iv) volatilization of acetic acid, and (v) errors in pH determination
(Loeppert et aI., 1984). Errors due to nonspecific interactionsof W with clay
minerals,which are most prevalentat low CaC03 contents(Le., at the low pH
valuesat high HC2H302/~H302 ratios), can be reducedby standardadditionsof
Ca (e.g., 0.1 M CaCI2) to all samplesand standards.Errors due to specific inter-
actions of H+ with soil organic matter, which are most severeat low carbonate
and high organic matter contents,cannot be totally eliminated;for example,at
2% actual CaC03 and 4% organic matter content,the measuredCaC03 content
was 2.2%, at 20% CaC03 and 4% organic matter the measuredCaC03 content
was 20.2% (Loeppertet aI., 1984). Under the experimentalconditionsdescribed
above,errors due to decompositionof layer-silicatemineralsare minimal; how-
ever, samplescontaining readily decomposableminerals,e.g., zeolites, may be
subjectto errorsin carbonatedetermination.On the otherhand,contacttime must
be sufficient to allow for the complete dissolution of solid-phasecarbonate.
Longer reactiontimes may be requiredfor samplescontainingdolomite, due to
the slower reactionrate of this mineral. The preciseand reproducibledetermina-
tion of pH is essentialfor the successof the procedure;therefore,the care and
conditioningof the pH electrodeis very important.
CARBONATE REACTIVITY
Particle-sizedistribution, surfaceareaand reactivity are importantproper-

ties of soil carbonateswhich influencesoil pedogenic,chemicaland rhizosphere
processes.Calcium carbonate providesa reactivesurfacefor adsorptionand pre-
cipitation reactions,for example,of phosphate(Boischotet aI., 1950; Talibudeen
& Arambarri, 1964; Arner et aI., 1985)., trace metalsand organic acids. Carbo-
nate reactivity can influence the rateof volatilization of ammonia(Ryan et aI.,
1981). Carbonatereactivity also influences rhizosphereprocesses,especially
thoseprocesses in which acidificationis an importantfactor. For example,the Fe-
deficiencystressresponseof dicots involves the exudationof protonsand acidif-
cation of the rhizosphere;the effectivenessof Fe-deficiencystressresponseof
dicots is thereforenegativelyinfluencedby the neutralizationof plant-produced
acidity, which is influencedby the reactivity of the carbonatephase(Loeppertet
ai., 1988; Morris et ai., 1990).
Severalmethodshave been used to assesseither carbonatereactivity or
quantity of reactivecarbonate.A widely usedprocedureis the "active" carbonate
method(Drouineau,1942; Boischot and Hebert, 1947), which involves reaction
of the soil with ammoniumoxalateor oxalic acid for a predeterminedtime. Other
scientistshave used quantity of clay-sizecarbonateas a measureof active car-
bonate.A radio-tracertechniqueinvolving the useof 45Ca has beenproposedto
measurethe specificsurfaceareaof CaC03 in the soil (Talibudeen& Arambarri,

1964; Abedi & Talibudeen,1974; Holford & Mattingly, 1975); however,results
by this proceduremay not be reliable in soils which containappreciablequanti-
ties of layer silicates. Glover (1961) demonstratedthe utility of ethylenedi-
aminetetraaceticacid (EDTA) for the evaluationof soil carbonatereactivity.
The evaluationof carbonatereactivity involves the reactionof the soilwith
either (i) a complexingagentthat reactsspecifically with the carbonatephaseor
(ii) a standardacid. In the former case,the rate of depletionof complexingagent
from the solution, or the rate of releaseof Ca (andMg) is monitoredas an index
of carbonatereactivity; in the latter caseeither rate of consumptionof H+ is mon-
itored with a pH-stat titrator (Moore et aI., 1989; Morris et aI., 1990; Hartwig &
Loeppert,1991)or rate of productionof CO2 is monitoredwith a Hg manometer
or a pressuretransducer(Suarez& Wood, 1984). The active carbonateand car-
bonatereactivity proceduresare kinetically basedmethods,and the results are
strongly influencedby factorswhich influencethe rate of reactionat the carbon-
ate mineral surface,e.g., samplepretreatment,agitation procedureand tempera-
ture. Therefore, for each of these procedures,the experimentalmethodology
mustbe adheredto carefully to enablecomparisonsbetweensamples.Table 15-3
providesa summaryof the availablereactivity methods.
CarbonateReactivity by pH Stat
Principles
The rate of reactionof H+ with CaC03 is directly relatedto surfaceareain
a well-stirred system,and within the pH rangeof three to five is approximately
first-order with respectto H+ activity (Berner & Morse, 1974). In the pH-state
procedure,standardacid is titrated into an aqueoussuspensionof a calcareous
soil at a rate to maintain pH constantat a predeterminedvalue during the car-
bonatedissolutionreaction(Eq. [1 D. A plot of cumulativeacid additionvs. time
is obtained,for which the slopeat a given time is equalto the reactionrate at that
point in time. Moore et aI. (1989) developeda procedureto determinereactive
surfaceareaof soil carbonatesusing pH-stattitrations at pH three and five. This
procedureutilizes rate constantsfor the dissolution of Iceland spar calcite and
cumulativesoil carbonatedissolutiondata,which are incorporatedinto simulta-
neousmultiple linear equationsemployingthe equal diameter-reduction hypoth-
Table 15-3. Methodsfor evaluationof carbonateparticle-sizedistribution or reactivity.

Reacting Determination
Principle species method References
Acid H+ pH-stat Morris et aI., 1990; Moore et aI., 1989;

dissolution Hartwig & Loeppert,1991, 1992
kinetics CO2 (manometric) Suarez& Wood, 1984
Surface Oxalate MKn04 titration of Drouineau,1942; Boischot & Hebert,
complexation 1947
45ea 45eaanalysis Talibudeen& Arambarri, 1964; Abedi
& Talibudeen,1974
EDTA ea, Mg (atomic Glover, 1961
absorption)
460 WEPPERT& SUAREZ
esis (Swartzendruber& Barber, 1965). The particle-sizedistribution of the soil

carbonateis then determinedby a least squarestechnique.The calculatedparti-
cle-sizedistribution actuallyrepresentsthe hypotheticalparticle-sizedistribution
of an equivalentquantity and reactivity of Icelandsparcalcite.
The rate of reactionof H+ with soil carbonateis highly dependenton acces-
sibility of carbonatesurfacesitesto reactionwith H+; therefore,samplepretreat-
ment and degreeof dispersionof the soil strongly influencesthe resultsof the
pH-stattitration procedure(Hartwig & Loeppert,1991).If the researcher's inter-
est is in assessingthe relative reactivity of the carbonateparticlescomposingthe
soil, then the soil aggregatesshould be totally dispersedby Na saturationand
either sonicationor vigorous agitation of the sample,prior to pH-stat titration
(Hartwig & Loeppert,1991).If, on the other hand,the interestis in assessingthe
relative reactivity of the carbonatephasein an aggregatedsoil, then minimal dis-
persionof the aggregatesis desirable.In this case,the only pretreatmentis usu-
ally to wet the samplegently with a dilute CaCl2 solution, to minimize disper-
sion prior to titration. .
Method
Apparatus
1. Automatic pH-stat titrator with a graphicalor digital output of cumu-
lative titrant volume vs. titration time. A stirrer assemblywith paddle
stirrer ratherthan a magneticstirring bar is preferred,to minimize the
grinding of soil particlesat the bottom of the titration vessel.
Reagents
1. Hydrochloric acid (HC1), 0.5 M: Transfer 41.7 mL of concentrated
HCI to 600-mL deionized water and dilute to l-L total volume.
Standardizethis reagentagainsta standardsolution of tris-hydroxy-
aminomethane(THAM).
2. Calcium chloride (CaCI2), 0.01 M: Weigh 1.47 g of reagentgrade
CaCl2 • 2H20, dissolvein 600 mL deionizedwater, and dilute to l-L
total volume.
3. Sodium chloride (NaCl), 1 M: Weigh 58.4 g of reagentgrade NaCl,
dissolvein 600 mL deionizedwater, and dilute to l-L total volume.
Procedure-Titration of Dispersed Soils for Determination of Effective

Particle-size Distribution
Sample Pretreatment.Place an accuratelyweighed 0.5000-g air-dried
soil sample(previously crushedto passa 2-mm pore-sizesieve) into a 40-mL
polypropylenecentrifuge tube, and saturatewith Na by three successivetreat-
mentswith 20 mL of 1.0 M NaCI. Following eachNaCl treatment,centrifugethe
suspensionat approximately3000 x G for 10 min and discard the supernate.
Following the final treatmentwith NaCl, centrifugethe sampleat approximate-
ly 3000 x G for 10 min, discardthe supernate,washonce with20 mL of deion-
ized water, centrifuge, and discard the supernate.Following each addition of
NaCI or water, shakethe samplevigorously, sonicatefor 30 s with a 0.22-cm
diam. probe at a power output of 7 W [a power settingof approximately5 with
the Bransonmodel 350 sonifer (Heat Systems,Farmingdale,NY)], and shake

vigorously againto dispersethe sample.If the sampleremainsdispersedfollow-
ing centrifugationof the final water treatment,then totally dispersethe sample
by vigorousagitationand transferquantitativelyto a titration vessel.If the sam-
ple is not adequatelydispersed,then washwith deionizedwater one more time
to ensureadequatedispersion.If a sonicatoris not available,then instead,shake
the samplevigorouslyfor 10 min during eachstepof the dispersionand washing
process.The sonicationtreatmentis severeenoughto promotedispersionof the
silicate clays, yet mild enoughto preventappreciablegrinding of the carbonate
aggregates.The washingstepis minimized in order to reducethe dissolutionof
carbonateandto ensureat leasta partial Na saturation ofthe soil cation-exchange
complex.
Titration. Transferthe soil samplefrom step 1, above,quantitativelyinto
an appropriatetitration vessel.Adjust the pH stat titrator to the desiredtitration
pH. Allow an air purge of approximately200 cm3 min-1 to passover the sus-
pension,but avoid the bubblingof air in the suspension.The sampleis stirredfor
exactly30 s on the titration assemblybeforeinitiation of titration. Separatesam-
ples shouldbe titrated at pH 3 and 5 for 28 min using 0.5 M HCI. Obtain plots
of cumulativevolume of acid consumedvs. time at eachpH value. Titrations at
both pH 3 and 5 are utilized, sincethe pH 3 titration enablesa betterevaluation
of sand-sizeparticlesand the pH 5 titration allows the evaluationof clay-size
particles.
Calculations.The rate of dissolutionof soil carbonateis describedby the
following equation
( Kt)2
p
+ ... + 41t qn K' rno - [22]
where
dQ
- rate of dissolution,milligram/minute,of the soil carbonate,taken
dt directly from the slopeof the pH stat titration curve at time t
qa to qn = numberof particlesin particle-sizefractions a throughn for the
initial particle radii, rao to rno
rao to rno = initial particle radii; thesevaluesrepresentthe various particle-
size classes;the recommendedinitial particle-radiiare 5 x 10-6,
2.5 x 10-5, 2 x 1Q-4, 10-3, 5 x 10-3 cm to representthe fine-clay,
coarse-clay,fine-silt, coarse-siltand sandparticle-sizefractions,
respectively.
K = dissolutionrate constantfor Icelandsparcalcite, milligrams per
minute per centimetersquare.The reactionrate constantof cal-
cite can be determinedby the titration of crushedand thorough-
ly washedIcelandsparcalcitethat hasbeenseivedto obtainnar-
row-rangesof particle size (Hartwig & Loeppert, 1991). Usually

in the determinationof rate constant,the Iceland spar calcite is
pretreatedto remove5% of the crystal massto ensurethat surface
adsorbedcrystalliteshave beenremoved.The reactionrate con-
stantis takenasthe initial dissolutionrate of the mineral. The sur-
face areafor the rate constantdeterminationcan be either calcu-
lated using assumptionsof sphericalgeometryof the particle-size
separatesor experimentallydeterminedusing a BET N2-adsorp-
tion adsorptionprocedure.
p = densityof calcite = 2.71 g cm-3
The variablesqa to qn are calculatedfrom the multiple linear regressionequations

generatedusing reactionrates,dQ/dt, at time intervals along the pH-stattitration
curves atpH 3 and pH 5. The SAS Institute (1985) GeneralLinear Model (GLM)
procedureor other suitable linear regressionmodel would provide a meansof
solving the multiple linear regressionequations.
The CaC03 equivalentweight of the soil is the sum of the CaC03 equiva-
lent weight of the individual particle size classes
where, Vao to Van are the volumesof the individual particlesof the particle-size
classesa through n, and Wa to Wn are the total weightsof particle in the particle-
size classesa through n. The proportion of the total carbonatein eacheffective
particle-sizeclassis calculatedby dividing the total effective weight of carbon-
ate in that classby the total weight of carbonatein the soil, e.g., WafWt for parti-
cle classa.
The total effectivesurfaceareaof the carbonatephasemay be calculatedas
follows
[25]
Procedure-Titration ofAggregated Soils for Determination

of Relative Carbonate Reactivity
SamplePretreatment.Placean accuratelyweighed0.500-gair-dried soil
sampleinto a 40-mL polypropylenetitration vessel.Carefully add 20 mL of 0.01
M CaCl2 solution,while beingcareful not to agitatethe soil sample.Gently rotate
the vesselat an angle for a few rotations to ensurethat the soil aggregatesare
thoroughly saturatedwith water, allow the sampleto standfor 30 min, and gen-
tly rotate the vesselagainfor a few rotationsprior to titration.
Titration. Adjust the pH-stat titrator to pH 5.0. Allow an air-purge of
approximately200 cm3 min-1 to passover the suspension,but avoid bubbling.
Initiate the titration immediately following initiation of agitation of the sample
with the paddlestirrer on the titration assembly.Allow the titration to proceedfor
20 min, during which time a plot of cumulativevolumeof acid consumedvs. time

is obtained.
Calculation.Calcium carbonatereactivity as determinedby this procedure
is usually expressedas milligram per gramper minute,which is determinedasthe
slope of the linear regressionof the dissolutioncurve over the first 20 min or as
the net dissolutionrate
mg CaC03 dissolved
CaC03 reactivity at pH 5.0 =
(soil g) (time min)
50 mg CaC03 )
(HCI mL) (HCI mmolc mL- 1) (
= _________________________
m
__m_o_lc_____
[26]
(soil g)(time min)
Comments
The pH-statprocedurefor carbonatereactivity providesan excellentmeans
of comparingsoils with respectto relative carbonatereactivity. The pH-stat titra-
tion has most commonly beenperformedat pH 5, though other pH values(e.g.,
4,6 and 7) havealsobeenutilized. Morris et al. (1990)determinedthat pH 5 titra-
tion was the most effective pH for assessment of the role of carbonatereactivity
on a rhizosphereprocess[Fe-deficiencystressresponseof soybean(Glycine max
L.)] that is influencedby the relative reactivity of the carbonatephase.The pH-
statcarbonate-reactivity titration is highly dependenton pH andsoil pretreatment
procedure.The choiceof pretreatmentand reactionpH is influencedby the objec-
tive of the experiment.
CarbonateReactivity by a ManometricProcedure
Principles
In the pressuremethod,a fixed acid concentrationis reactedwith an aque-
ous suspensionof a calcareoussoil in a closedreactionvessel(Suarez& Wood,
1984). The solution is preferably buffered to maintain constantpH during the
measurementtime. Addition of acid results in dissolutionof CaC03 and release
of CO2 as shownin Eq. [1]. A pressuretransducerand a strip chart recorderallow
measurementand display of the pressurevs. time relation. The releaseof CO2 is
proportional to the dissolution rate (reactivity) which is proportional to the
CaC03 surfacearea,though results are highly dependenton samplepreparation
(Suarez& Wood, 1984). As with the pH-stat methods(seeCarbonateReactivity
by pH-Stat),the choiceof pretreatmentproceduredependson whetherthe results
are to be used for evaluationof reactivity in the aggregatedsoil or reactivity of
the dispersedparticles. Reactivesurfacearea is calculatedfrom comparisonof
measuredsoil rates to measuredrates of well crystallizedcalcite of known sur-
face area.
To Valve and Pressure

Clomp Transducer
Reaction Vesse I
Wot.r Both
.--...u--- Magnet in Stopper
w tt - - - HOAc Solution
Wire Stand
Stir Bar -~:!5~r- Sample - NoOAc Solution
--+---- Insuloted Stir Plate
Fig. 15-5. Modified pressurecalcimeterfor the determinationof carbonatereactivity (Suarez&

Wood, 1984).
Method
Apparatus
1. 500-mL wide-mouthjar (reaction vessel),rubber gasketring, plastic
screwcap and a three-wayglassstopcock(Fig. 15-5). Glassstopcock
is insertedand sealedinto the plastic cap using epoxy glue.
2. Small wire tripod with a plastictable (tofit into the jar asshownin Fig.
15-5).
3. No.4 polypropylenestopper(cup shaped)attachedto the table with a
wire loop on one side (stopperis hinged to allow it to tip and empty
contents).
4. Small Teflon-coatedstir bar wedgednearthe top of the stopper.
5. Large stir bar.
6. Insulated pressure transducer (e.g., Kistler Model 314D, Kistler
InstrumentCorp., Amherst, NY) with 0- to 2-kPa sensitivity and 0.1-
V output.
7. Water bath and submergedstirrer, or insulatedstirring plate and 1 L
glassjar with deionizedwater.
8. 1.5-mmi.d. thick wall polypropylenetubing.
9. 0.1-V full-scale deflection chart recorder.
10. Magnet.
Reagents
1. Sodiumacetate(NaC2H30 2), 1.0 M; Weigh 136.08g of NaC2H30 2 • 3
H20, dissolvein 600 mL deionizedwater, and dilute to 1 L total vol-
ume.
2. Acetic acid (HC2H30 2), 2.0 M; Weigh 120.1 g of glacial acetic acid,
dissolvein 600 mL deionizedwater, and dilute to 1 L total volume.
Method
The experimentalsetup is shown in Fig. 15-5. The rubbergasket,plastic
screw cap and 3-way glass stopcockare attachedto the glass jar. The system
shouldbe pressurizedto checkfor leaks.
1. A measuredamountof calcite standardor soil (0.1-10.0 g dry weight)

is placedinto the reactionvessel.
2. Add 100 mL of deionizedwater and 5.0 mL of 1.0 M Na acetateto the
reactionvessel.
3. Add 5.0 mL of 2.0 M acetic acid to the polypropylenestopper.
4. Insert the large stirring bar, wiretripod and polypropylenestopperinto
the reactionvessel.
5. Cap the jar, and place in the 1 L water bath on top of an insulatedstir-
ring plate in a temperature-controlled room.
6. Connectthe tubing to the pressuretransducer.
7. After 5 min of stirring (300 rpm), closethe stopcockfrom the open-to-
air position to the open-to-transducer position.
8. Tip the plastic stopperby manipulatingthe external magnetnear the
plastic stopper,and initiate the chart recorder.
9. Stop the reactionafter 5 min.
10. Examine pressurevs. time curve and calculate the maximum slope
(rate). This will ordinarily be the straight line portion of the curve.
11. Repeat the procedurewith standardsof known surface area (from
0-0.02 m2).
12. Constructa calibration curveby plotting standardsurfaceareaagainst
measuredreactionrates(Pa S-I).
13. Use calibrationcurve to determinereactivecalcitesurfacearea(calcite
equivalent).
Comments
The procedurecan be usedto determinetotal carbonateby reactingfor 1 h
and measuringtotal pressure.Use of strongeracid is not recommendedsince
reactionratesincreaseand do not allow sufficient time to ensurethat the acid is
well mixed when the maximum slope of the pressure-timecurve is determined.
Surfaceareasare measuredat the low Pcoz pressure(0.4-1.6 kPa aboveatmos-
pheric) and under conditionsof buffered pH. As with other procedures,FeS04
can be addedto samplesto preventoxidation of organicmatter and formation of
COz, but decompositionof organic matter is ordinarily not a problem due to the
use of weak acid and the short reaction times. Surfaceareaof calcite standards
should be determinedby the BET methodusing N2 •
Active Carbonate
Principles
The active carbonatemethod, originally developedby Drouineau(1942),
involves reactionof the soil with 0.1 M ammoniumoxalatefor 2 h, followed by
the determinationof unreactedoxalateby titration with 0.1 M KMn04. Boischot
and Hebert (1947) evaluatedthe procedureand recommendedthat the pH of the
ammoniumoxalateextractantbe adjustedto 9.0. Somescientistshave utilized a
pH 7.0 bufferedoxalate(Carter, 1981; Ryan et aI., 1981),and othershaveuseda
0.2 M ammonium oxalate with no further pH adjustments.The reaction of
ammoniumoxalatewith calcium carbonatecan be written as follows
WEPPElU& SUAREZ
The oxalatealso reactswith dissolvedand exchangeableCa to form Ca oxalate;

therefore,the procedureshouldmore appropriatelybe called an active Ca proce-
dure ratherthan an active carbonateprocedure.The reactionof unreactedammo-
nium oxalate(following acidification with H2S04) is summarizedby the follow-
ing equation
The active carbonateprocedureprovides a rapid approximation of the

quantity of clay and fme-silt size carbonatesin the soil and can be usedto obtain
a comparativebut not an absolutemeasureof reactivecarbonatecontentof the
soil. Resultsby this procedurehavebeenshownto correlatehighly with compar-
ative estimatesof soil-carbonatereactivity by the pH-statprocedure(del Campil-
10 et aI., 1992).The activeoxalateprocedureis a kinetically basedprocedure,the
results of which are highly dependenton sample pretreatmentand handling.
Precautionsshouldbe takento ensureuniformity of samplehandlingandshaking
procedures.Major sourcesof error include solution and exchangeable Ca and the
inability of oxalateto reactwith Mg in Mg-substitutedcalcite.
Method
Reagents
1. Ammonium oxalate [(NH4hCz04], 0.1 M, adjustedto pH 9.0. Place
12.61 g of H2Cz04 • 2 H20 into a 1000-mL beaker,adjust to pH 9 by
addition of 2 M Nf40H, and make to alL final volume.
2. Potassiumpermanganate (KMn04), 0.02M. Add 3.16 g of KMn04 to
1 L of deionizedwater. Keep the solution at a gentleboil for about1 h,
cover, and let standovernight.Filter througha fine-sinteredglassfun-
nel. Storethe solution in a amberglassbottle.
3. Sulfuric acid (H2S04),3 M. Transfer167 mL of concentratedH2S04 to
600 mL deionizedwater, and dilute to 1 L total volume.
4. Sulfuric acid (H2S04), 1 M. Transfer56 mL of concentratedH2S04 to
600 mL deionizedwaterand dilute to 1 L total volume.
Procedure
1. Standardizationof potassiumpermanganate. Dry about1.5 g of primary
standardammoniumoxalate(NaC20 4) at 110°Cfor at least 1 h. Cool in a desic-
cator,weigh threeindividual 0.2 to 0.3 g samplesto the nearest0.1 mg into 400-
mL flasks. Dissolvein 250 mL of 1 M H2S04, Heatto 80 to 90°C, andtitrate with
0.02 M KMn04 at this temperature.The pink color resultingfrom addition of an
aliquot of titrant should be permittedto disappearbefore further titrant is intro-
duced.Finely divided KMn04 will be formed along with Mn2+ if the KMn04 is
addedtoo rapidly and will causethe solution to acquire a faint brown discol-
oration. The first persistentpink color (30 s) should be taken as the endpoint.
Determinea blank by titrating an equalvolume of 1 M H2S04,
2. Reactionwith ammoniumoxalate.Weigh 1 g of air-dried soil which has

beencrushedto passa 20 meshin- I (850-llm nominal pore size) sieve,and place
in a 40-mL snaptop polypropylenevial (with a 2-mm diam hole in the top). Add
25 mL of 0.1 M ammoniumoxalate,stopper,and shakefor exactly 2 h on a rotary
shakerset to oscillate at approximately240 cycles min-I. Immediately transfer
the suspensionto a 40-mL polypropylenecentrifugetube, and centrifugefor 20
min at 3000 x G.
3. Titration with potassiumpermanganate. Transfera lO-mL aliquot of the
supernateto a 50-mL Erlenmeyerflask, and add 5 mL of 3 M H 2S04. Determine
the quantity of oxalateremainingin solution by titration with 0.02 M KMn04 as
describedabove.
Calculations
1. Calculatethe concentrationof the KMn04 titrant as follows
Concentrationof KMn04, M
[29]
The final concentrationof KMn04 is calculatedas the averageof repli-

cates.
2. Calculatethe active carbonatecontentas follows
Active carbonatecontent,%
5 mol H 2C20 2 ) (100 g CaC03 )

(Yo - Y s) (C) (dilution factor) (
2 mol KMn04 mol
= (100)
soil g
[30]
Active carbonatecontent,g kg-1
5 mol H2C20 z ) (100 g CaC03 )

(Yo - Vs) (C) (dilution factor) (
2 mol KMn04 mol
= ------------------------------------------------
soil kg
[31 ]
where
Vo = volume (L) of KMn04 requiredto titrate 10 mL of oxalateextractant.
Vs = volume (L) of KMn04 requiredto titrate 10 mL of oxalateextractantfol-
lowing treatmentwith sample
C = concentrationof KMn04 in mol L- 1
468 WEPPERT& SUAREZ
Comments
The maximumquantityof active CaC03 that canbe dissolvedby the above
procedureis 200 g kg-1 soil. If active Caco3 contentsgreaterthan about 150 g
kg-1 are obtained,then the analysisshouldbe repeatedwith a smalleramountof
soil and/ora largervolume of ammoniumoxalateextractant.
CARBONATE EQUILmRIA-SOLID PHASE
Principles
Carbonateequlibria can be characterizedby the reactionof dissolvedCO2
and the dissociationof the resultantweak acid with carbonate-containing miner-
als, principally calciteand dolomite. Dissolutionof CO2 gasinto water resultsin
the speciesCO2(aq). Upon hydrationthe speciesH2C03° is formed, (referredto as
H2C03). The weakacid dissociatesinto HCO)" and CO~- with pK1 and pK2 val-
ues of 6.35 and 10.33, respectively,at 25°C. Typically the terms HC03* or
H2C03 areusedto denotethe sumof C02(aq)and H2C03 0 • The solubility of CO2
is given by the equation
[32]
whereKc~ =10-1.47 at 25°C when Pc~ is expressedin atmospheres. The solu-

bility of CO2 increaseswith decreasingtemperature,with a Kc~ of 10-1.11 at O°C
and 10-1.64at 40°C.
The dissociationof carbonicacid is represented
by the following equations
(H+)(HCO)")
K1 = [33]
(H2C03)
(H+)(CO~-)
K2 = [34]
(HCO)")
where parentheses representactivities. At 25°C, K1 = 10-6.35 and K2 = 10-10.33,

while at 40°C, K1 = 10-6·30andK2 = 10-10.22•
The total amountof inorganicC decreaseswith increasingtemperatureat
ftxed Pc~, wheretotal C is deftnedby IC = C02(aq)+ H2C03a + HCO)" + CO~
+ CaHCOj + CaC03° .... The solubility of CO2 decreasesonly slightly with
increasingionic strength,while the total dissolvedinorganicC contentincreases
substantially(primarily dueto decreases in the HC03- activity coefficientandion
pair formation of RC03- and CO~- with divalent metals).
Reactionof a carbonatesolid phasewith a solution free of net alkalinity
resultsin an increasein pH dueto dissolutionof C~- which protonatesto HCOl .
Typically the dissolutionof calcite is written as
[35]
Although this a good overall representationof the major species,it obscuresthe

shift in minor species.Increasedconcentrationor activity of HCOj" requiresthat
there be a decreasedconcentrationor activity of H+, since H2C03° is fixed by
Pc02. A small but significantcomponentof calcitedissolutionresultsin the reac-
tion
CaC03(s) + W ~ Ca2+ + HCOj" [36]
The pH of water in equilibrium with atmosphericCO2 is pH =5.65, while at cal-

cite equilibrium the solutionis at pH =8.4. The reactionshownin Eq. [35] results
in approximately5 x 10--4 moles of calcite dissolvedwhile the reactionrepre-
sentedin Eq. [36] resultsin 2 x 10-6 molesof dissolvedcalcite. With increasing
PC02' the pH for calcite-equilibriumis shifted downward;at PcC}z =100 kPa (1
atm), pH =6.01.
Solving speciationproblems in the carbonatesystem requires that one
know at leasttwo of the following variables:(i) total dissolvedinorganicC, (ii)
inorganicC alkalinity, (iii) pH, and (iv) partial pressureof CO2. Typically inor-
ganic C alkalinity and H are determinedsince they are the easiestto determine
experimentally.Useof total dissolvedinorganicC may be conceptuallypreferred
since total concentrationsare usually measuredfor other dissolvedspecies,but
the measurementis unstablewithout specialprecautions,due to the undissociat-
ed dissolvedCO2• The determinationof pH and PC02 are also unstable,but pH
can be rapidly measuredusing electrodes.In contrastto total dissolvedC, inor-
ganic carbonatealkalinity is stablein the absenceof dissolutionor precipitation.
ReactionRates
The two importantkinetic processesin the carbonatesystemare equilibra-

tion of the gasphasewith the dissolvedspeciesand equilibrationof the dissolved
specieswith the solid phase.Reactionsamongthe dissolvedspeciesin the bulk
solution are sufficiently fast that reactionratesneednot be consideredand equi-
librium canbe assumed.In contrast,disequilibriumbetweengasand liquid phase
must be consideredwheneverprecipitationor dissolution is occurring. This is
particularly importantat low CO2 partial pressuresand/orhigh rate of precipita-
tion of dissolutionwhere the relatively slow ratesof diffusion of CO2 and con-
versionto H2C03* can result in disequilibrium.
It is generally recommendedthat determinationsof equilibria with solid-
phasecarbonatesin the laboratorybe doneat high P CO2' especiallyfor soils. Dis-
equilibrium betweenthe solid carbonatesand liquid phasecan almostalwaysbe
expectedfor soil-watersystems,especiallywhen equilibrium is approachedfrom
supersaturation,due to the presenceof dissolvedorganicswhich inhibit precipi-
tation. An additionalproblemis the decompositionof organicmaterialwhich can
result in a large increasein total dissolvedalkalinity. The combinationof these
processes makedeterminationof the stability of soil carbonatesdifficult. The best
procedurefor determinationof CaC03 stability is likely by reactionat high CO2
with gentlestirring. Dissolutionreactiontime for soil calcitein solutionmay gen-
erally be on the order of 0.5 to 3 d, assumingthat gas-solutionequilibrium is
maintained.
470 WEPPEKf& SUAREZ
In most circumstances,carbonatedissolution is a diffusion controlled

process.This is readily demonstratedby the reaction dependenceon stirring.
Under acidic conditions(pH < 5) in a stirredsystem,calcitedissolutionrate can
be representedby the expressionR = k1aH+' At 25°C, Plummeret al. (1979) list
a kl of 0.05 mmol calcite cm2s-1 at a stirring rate of 2000 rpm. At pH values
above6, the dissolutionreactionrate is relatedto PCOz whenPC02 is greaterthan
10 kPa, while the rate is constantwhen the PcOz is below 10 kPa (at constant
pH).
Extractionof Soil Waterfor CarbonateEquilibrium Determinations
The calcitesaturationstatusof a soil watersystemdependson the soil PcOz

aswell ason the watercomposition.Becauseof the dependence on the gasphase
composition,the saturationstatusis bestdeterminedin the field by extractionof
the soil water and immediatedeterminationof pH and subsequentpreservation
of the sample for further laboratory analysis. Extraction of the water phase
requireseitherapplicationof a vacuum,useof a squeezingtechnique(press)or
immiscible displacementwith an organicsolvent.Installationof vacuumextrac-
tors is recommendedfor ongoingmonitoring of a site.
Useof conventionalvacuumextractorswill resultin CO2 degassingandan
upwardshift in pH (Suarez,1987).Use of a multichamberedextractordescribed
by Suarez(1986) minimizesthe air phasein the sampleand allows for flushing
of the samplechamber.This extractordesignreducesthe pH error to below 0.05
pH units, when the extractoris operatedat the minimum vacuumlevel required
to obtain the sample.Upon obtaining the samplethe pH must be immediately
measuredin the field with a metercalibratedwith pH standardsat the tempera-
ture of the sample.After measurementof the pH, the sampleshouldbe tightly
cappedand placedin an ice chestor refrigeratoruntil determinationof alkalini-
ty, preferablywithin a few hours.Subsequently,the sampleis filtered/dilutedfor
analysisof the othermajorion components,typically Ca, Mg, Na, K, S04,Cl and
N03• The chemical speciation program should have temperaturedependent
expressionsfor calculationof ionic activity coefficientsand activities.
GYPSUM
Gypsum(CaS04• 2H20) is almostalways the calcium sulfatemineral in

soils (Nettletonet aI., 1982). Anhydrite (CaS04)is rarely found, as is bassanite
(CaS04• 0.5H20), exceptundersurfacesoil conditionsof high temperatureand
very low humidity or at very high salinity. In microcrystallinesize,theseminer-
als will transforminto gypsumupon rewettingthe soil.
In the subsurface(rock formations)anhydriteis somewhatmore common
thanin soils, existingin associationwith marineevaporates.Gypsumin arid land
soils is usuallypedogenic,asmostsurfacewatersareundersaturated with respect
to gypsum,and it dissolvesrelatively easily and is rarely transportedin fluvial
processes.Gypsummay occur as white surfacecrustsin arid environments,but
also readily producessilt-sizedcrystalsto severalcentimetersin length.
Method
Principles
Determinationof gypsumhasgenerallybeendoneby dissolutionand mea-
surementof Ca and/orSOi- in a dilute extract.In order to usethe procedurein a
quantitativemanner,correctionmustbe madefor dissolvedsulfates.In addition,
some estimate of soil gypsum is desired in order to select the appropriate
soiVwaterratio for extraction.This is requiredto ensureboth completedissolu-
tion of the gypsumand yet have sufficient sulfate for analysis.The previously
recommendedmethod(Nelson, 1982) involved an initial semiquantitativedeter-
minationof gypsumby measuringweight lossdue to dehydration.This wasdone
by subtractingthe weight lossof soil underP20 5 from the weight lossafter being
placed in an oven overnight at lO5°C. After this calculation, the appropriate
soil/waterratio is determinedand the total sulfateis analyzed.
A soil often containsgypsumif the Ca and S04concentrationsof the satu-
ration extract exceed20 mmolc L-l (Nelson, 1982). A quick test for the likely
presenceof gypsumcanbe madeby preparinga dilute waterextract(1:2 or 1:5),
andseparatingand treatingthe supernatantwith an equalvolume of acetone.The
formationof a white precipitateindicatesgypsum(U.S. Salinity LaboratoryStaff,
1954).
Alternative methodsinclude the infrareddehydrationmethod(El Prince&
Turjoman, 1983), the resin method (Frenkel et aI., 1986) and XRD (Khan &
Webster,1986).
Multiple Dilution Procedure

Preparea saturationextractwith 25 g of soil asdescribedin Thomas(1995,
seeChapter16), andextractthe water. Grind 50 g of air-dry soil until it passesan
80 meshin- 1 (180-~m nominal pore size) sieve. Split and weigh 25-g portions
into 1oo-mL and 1-L flasks. Add 100 mL and 1 L of deionizedwaterto the 100-
mL and 1-L flasks, respectively.Mix the suspensions overnighton a reciprocat-
ing shaker.
Filter eachof the suspensionsthrough Whatmanno. 42 filter paper,and
collect the filtrate. Determinesulfate as recommendedin Sawhney(1996, see
Chapter36) on thesetwo extractionsand on the saturationextract.
Calculations
S04' mmol = ( S04mmol) ( L ) (mL deionizedwater) [37]

L lOoo mL
S04(g) =S04(DE)- S04(SE) [38]

where
S04(g) = sulfatefrom gypsum,mmol
S04(DE)= sulfatein dilute extract,mmol
S04(SE)= sulfatein saturationextract,mmol
472 WEPPEKI' & SUAREZ
g gypsum ( 0.172g CaS04• 2H20) ( 1 )

g soil = (S04(g)' mmol) mmol 25 g soil [39]
Comments
Heating the soil at 105°C overnight before grinding convertsgypsum to
bassanite(Rivera et ai., 1982) which is more soluble and dissolvesfaster than
gypsum,and ensuresa more completedissolutionreaction.
REFERENCES
Abedi, M.J., and O. Talibudeen.1974. The calcareoussoil sof Azerbaijan. I. Catenadevelopment
relatedto the distribution and surfacepropertiesof soil. J. Soil Sci. 25:357-372.
Allison, L.E. 1960.Wet combustionapparatusandprocedurefor organicand inorganiccarbonin soil.
Allison, L.E., and C.D. Moodie. 1965. Carbonate.p. 1379-1400.In C.A. Black et aI. (ed.) Methods
of soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA, CSSA, and SSSA,Madison,WI.
Amer, F.A., A. Mahmoud,and V. Sabel. 1985. Zeta potentialand surfaceareaof calcium carbonate
as relatedto phosphatesorption.Soil Sci. Soc. Am. J. 49:1137-1142.
Berner, R.A., and J.W. Morse. 1974. Dissolution kinetics of calcium carbonatein seatwater.Iv.
Theory of calcite dissolution.Am. J. Sci. 274:108-134.
Bloom, P.R., K. Meter, and J.R. Crum. 1985. TItration methodfor determinationof clay-sizedcar-
bonates.Soil Sci. Soc. Am. J. 49:1070-1073.
Boischot,P., M. Coppenet,and J. Hebert. 1950. Fixation de I'acide phosphoriquesur Ie calcairedes
sols. Plant Soil 2:311-322.
Boischot, P., and J. Hebert. 1947. Determinationof available calcium in soil by the ammonium
oxalatemethodand its use to determinethe readily assimilablecalcium of liming materials.
Ann. Agron. 17:521-525.
Bui, E.N., R.H. Loeppert,and L.P. Wilding. 1990. Carbonatephasesin calcareoussoils of the west-
ern United States.Soil Sci. Soc. Am. J. 54:39-45.
Bundy, L.G., andJ.M. Bremner.1972.A simple titrimetric methodfor determinationof inorganiccar-
bon in soils. Soil Sci. Soc. Am. Proc. 36:273-275.
Carter,M.R. 1981.Associationof total CaC03and active CaC03with growth of five tree specieson
chernozemicsoils. Can. J. Soil Sci. 61:173-175.
del Carnpillo, M.C., J. Torrent,and R.H. Loeppert.1992.The reactivityof carbonatesin selectedsoils
of southernSpain.Geoderma52:149-160.
Diebold, F.A., J. Lemish, and C.L. Hiltrop. 1963. Determinationof calcite,dolomite, quartzand clay
contentof carbonaterocks. J. Sediment.Petrol. 33:124-139.
Doner,H.E., andW.C. Lynn. 1977.Carbonate,halide,sulfateandsulfide minerals.p. 279-330.In J.B.
Dixon and S.B. Weed (ed.) Minerals in the soil environment.SSSA, Book Ser. 1. SSSA,
Madison,WI.
Dreimanis,A. 1962. Quantitativegasometricdeterminationof calcite and dolomite by using Chittick
apparatus.J. Sediment.Petrol. 32:520-529.
Drouineau,G. 1942. Dosagerapidedu calcaireactif de sols. Ann. Agron. Vol. 12.
EI Mahi, H.E., I.S.lbrahim, M. Abdel, H.M. Magid, and A.M.A. Eltilib. 1987.A simple methodfor
the estimation of calcium and magnesium carbonatesin soils. Soil Sci. Soc. Am. J.
51:1152-1155.
EI Prince, A.M., and A.M. Turjoman. 1983. Infrared dehydrationmethod for determininggypsum
contentof soils. Soil Sci. Soc. Am. J. 47:1089-1091.
Evangelou,v.P., L.D. Whittig, and K.K. Tanji. 1984. An automatedmanometricmethodfor quanti-
tative determinationof calcite and dolomite. Soil Sci. Soc. Am. J. 48:123&-1239.
Frenkel,H., Z. Gerstel,andJ.v. Renger.1986. Determinationof gypsumand cationexchangecapac-
ity in arid soils by resin method.Geoderma39:67-77.
Glover, E.D. 1961. Method of solution of calcareousmaterial using the complexingagent,EDTA. J.
Sediment.Petrol. 31:622-626.
Hartwig, R.C., and R.H. Loeppert.1991. Pretreatmenteffect on dispersionof carbonatesin calcare-
ous soils.Soil Sci. Soc. Am. J. 55:19-25.
Hartwig, R.C., and R.H. Loeppert.1992.A pH-statprocedurefor evaluatingreactivity of agricultur-

allimestone.Soil Sci. Soc. Am. J. 56:302-308.
Holford, I.C.R., and G.E.G. Mattingly. 1975.Surfaceareasof calciumcarbonatein soils. Geoderma
13:247-255.
Khan, S.U., and G.R. Webster.1986. Determinationof gypsumin solonetzicsoils by an x-ray tech-
nique.Analyst 93:400-402.
Loeppert,R.H., S.C. Geiger,R.C. Hartwig, and D.E. Morris. 1988. A comparisonof indigenoussoil
factors influencing the Fe-deficiencychlorosis of sorghumand soybeanin the calcareous
soils. J. PlantNutr. 11:1481-1482.
Loeppert,R.H., C.T. Hallmark, and M.M. Koshy. 1984. Routine procedurefor rapid determination
of soil carbonates.Soil Sci. Soc. Am. J. 48:103~1033.
Martin, S.E., and R. Reeve.1955. A rapid manometricmethodfor determiningsoil carbonate.Soil
Sci. 79:187-197.
Moore, J.J.,R.H. Loeppert,L.T. West, and C.T. Hallmark. 1987.A routine methodfor calcium car-
bonateequivalentof soils. Commun.Soil Sci. PlantAnal. 18:265-277.
Moore, T.J., R.H. Loeppert,and R.C. Hartwig. 1989. Steadystateprocedurefor studyingthe effec-
tive particle size distribution of soil carbonates.Soil Sci. Soc. Am. J. 54:55-59.
Morris, D.R., R.H. Loeppert,andT.J. Moore. 1990.Indigenoussoil factorsinfluencing Fe chlorosis
of soybeanin calcareoussoils. Soil Sci. Soc. Am. J. 15:1329-1336.
Nelson,D.W., andL.E. Sommers.1982.Total carbon,organiccarbonandorganicmatter.p. 539-579.
In A.L. Page(ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA,Madison,WI.
Nelson,R.E. 1982.Carbonateandgypsum.p. 181-197.In A.L. Page(ed.) Methodsof soil analysis.
Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Nettleton, W.D., R.E. Nelson, B.R. Braskerand P.S. Derr. 1982. Gypsiferoussoils in the western
United States.p. 147-168.In J.A. Kittrick (ed.) Acid sulfateweathering.SSSASpec.Pub\.
10. SSSA,Madison,WI.
Peterson,G.W., andG. Chester.1966.Quantitativedeterminationof calciteand dolomite in pure car-
bonatesand limestones.J. Soil Sci. 17:317-327.
Peterson,G.w., G. Chesters,andG.B. Lee. 1966.Quantitativedeterminationof calcite anddolomite
in soils. J. Soil Sci. 17:328-338.
Plummer,L.N., D.L. Parkhurst,and T.M.L. Wigley. 1979. Critical review of the kinetics of calcite
dissolutionand precipitationchemicalmodeling-speciation,sorption,solubility, and kinetics.
p. 537-573.In E.A. Jenne(ed.) Chemicalmodelingin aqueoussystems.ACS, Washington,
DC.
Presley,BJ. 1975. A simple method for determiningcalcium carbonatein sedimentsamples.J.
Sediment.Petro\.45:745-746.
Rabenhorst,H.C. 1988.Determinationof organicandcarbonatecarbonin calcareoussoils usingdry
combustion.Soil Sci. Soc. Am. J. 52:965-969.
Rivera,E.D., C.T. Hallmark, L.T. Westand L.R. Drees.1982.A techniquefor rapid removalof gyp-
sum from soil samples.Soil Sci. Soc. Am. 1 46:1338-1340.
Runnells,D.D. 1970. Errors in x-ray analysisof carbonatesdue to solid-solutionvariation in com-
position of componentminerals.J. Sediment.Petro\.40:1158-1166.
Ryan, 1, D. Curtin, and I. Safi. 1981. Ammonia volatilization as influencedby calcium carbonate
particle size and iron oxides.Soil Sci. Soc.Am. J. 45:338-341.
SAS Institute. 1985. SAS user'sguide: Statistics.Version 5 ed. SAS Inst., Inc., Cary, NC.
Sawhney,B.L. 1996. Extraction of organic chemicals.p. 1071-1084.In D.L. Sparkset al. (ed.)
Methodsof soil analysis.Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and ASA,
Madison,WI.
Skinner,S.I.M., and R.L. Halstead.1958. Note on rapid methodof determinationof carbonatesin
soils. Can. J. Soil Sci. 38:187-188.
Skinner,S.l.M., R.L. Halstead,andlE. Brydon. 1959.Quantitativemanometricdeterminationof cal-
cite and dolomite in soils and limestones.Can. J. Soil Sci. 39:197-204.
Suarez,D.L. 1977.Ion activity productsof calciumcarbonatein watersbelow the root zone.Soil Sci.
Soc.Am. J. 41:31~315.
Suarez,D.L. 1986. A soil water extractor that minimizes C02 degassingand pH errors. Water
Resour.Res.22:876-880.
Suarez,D.L. 1987. Predictionof pH errors in soil water extractorsdue to degassing.Soil Sci. Soc.
Am. J. 51:64-67.
Suarez,D.L., and J.D. Wood. 1984. Simultaneousdeterminationof calcite surfaceareaand content
in soils. Soil Sci. Soc. Am. 1 48:1232-1236.
474 WEPPERT & SUAREZ
Swartzendruber,D., and S.A. Barber. 1965. Dissolution of limestone particles in soil. Soil Sci.
100:287-291.
Talibudeen,0., and P. Arambarri. 1964.The influenceof the amountand the origin of calcium car-
bonateson the isotopically-exchangeablephosphatein calcareoussoils. J. Agric. Sci.
62:93-97.
Tennant,C.B., and R.w. Berger. 1957. X-ray determinationof dolomite-calciteratio of a carbonate
rock. Am. Miner. 42:23-29.
Thomas,G.W. 1996.Soil pH and soil acidity. p. 475-490.In D.L. Sparkset al. (ed.) Methodsof soil
analysis.Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Turner, R.C., and S.1M. Skinner.1959.An investigationof the interceptmethodfor determiningthe
proportionof dolomite andcalcite in mixturesof the two. II. Experimentalrate of solution of
dolomite and calcite in samplesconsistingof a numberof crystals.Can. 1. Soil Sci. 40:232
242.
Turner,R.c., and S.I.M. Skinner.1960.An investigationof the interceptmethodfor determiningthe
proportionof dolomite and calcite in mixturesof the two: II. Experimentalratesof solution
of dolomite and calcite in samplesconsistingof crystals.Can. J. Soil Sci. 40:232-241.
Ulas, M., and M. Sayin. 1984. Quantitative determination of calcite and dolomite in soils by x-ray
diffraction. J. Soil Sci. 35:685-691.
U.S. Departmentof Agriculture-Soil ConservationService.1984. Proceduresfor collectingsoil sam-
plesand methodsof analysisfor soil survey.Soil Surv. Invest. Rep. 1. (Rev.) U.S. Gov. Print.
Office, Washington,DC.
U.S. Salinity LaboratoryStaff. 1954. Diagnosisand improvementof saline and alkali soils. USDA
Handb.60. U.S. Gov. Print. Office, Washington,DC.
Warne,S.S.1.,and B.D. Mitchell. 1979. Variable atmosphereDTA in identification and determina-
tion of anhydrouscarbonatemineralsin soils. J. Soil Sci. 30:111-116.
Williams, D.E. 1949.A rapid manometricmethodfor the determinationof carbonatein soil. Soil Sci.
Soc. Am. Proc. 13:127-129.
Published 1996
Chapter 16
Soil pH and Soil Acidity
G. W. THOMAS, University of Kentucky, Lexington, Kentucky
Soil pH is probably the single most informative measurementthat can be made

to determinesoil characteristics.At a single glance,pH tells much more about a
soil than merely indicating whetherit is acidic or basic. For example,availabili-
ty of essentialnutrientsand toxicity of other elementscan be estimatedbecause
of their known relationshipwith pH.
The term pH was "invented" by the SwedishscientistSorensen(1909) in
order to obtain more convenient numbers and the idea quickly caught on.
Gillespie and Hurst (1918) seemto havebeenamongthe earliestto determinepH
(or PH, as it was then called) electrometricallyusing a platinum-palladiumblack-
hydrogengas electrode,a calomel referenceelectrodeand a fairly cumbersome
potentiometerand galvanometersystem.At that period, it was still much more
commonto use colorimetric methodswith indicator dyes than the electrometric
method.
This changedrapidly, however.Sharpand Hoagland(1919) useda similar
but lessinvolved methodthan Gillespieand Hurst (1918) and Healy and Karraker
(1922) used a commercially available platinum-hydrogengas electrode,poten-
tiometerand galvanometerwhich had beendesignedby Clark (1920).
The decadeof the 1920ssaw the developmentof the quinhydroneelectrode
which was lessfragile and much lessexpensivethan the hydrogen-platinumelec-
trode. But, it was the developmentof the glasselectrodein the 1930sthat brought
the determinationof pH very rapidly to its presentimportanceand convenience.
The BeckmanModel G pH meter(circa 1931) was practically indestructibleand
could be used as a portableas well as a laboratory instrument.Although it was
cumbersomeby today'sstandards,it was virtually foolproof (exceptfor the con-
stantly failing batteries)and many are still capableof operatingif not actually
operatingtoday.
As recentlyas two decadesago, the useof the small, handheldportablepH
metersthen availableto determinepH in the field was a very impreciseand haz-
ardousundertakingbecauseboth electrodesand meterswere subject to sudden
failures but this haschangedratherabruptly in the last few years.Microcircuitry
and plastic have contributedto rugged pH metersand electrodesthat withstand
Book Seriesno. 5.
475
476 THOMAS
considerableabuseand which, in terms of real cost, are very inexpensivecom-

paredto the pH metersand electrodesof just a few yearsago.
The value of pH as an indicator of soil conditions and the quality of the
equipmentnow available at reasonablecost make the acquisition of pH data a
fairly simple chore at present.This chapterdiscussesthe methodsused, their
strengthsand weaknesses and someaspectsof interpretationof the data.
DEFINITION OF pH
The conceptof pH was derived from the ion productof water, which dis-
sociatesvery slightly
H20 ... H+ + OH- [1]
Kw =[W][OH] =1 x 10-14
at 25°C temperaturewhereH+ and OH- in bracketsare activities. When [H+] and
[OW] are equal, eachhas an activity of (10- 14 )1/2 or (10-7). pH was defined by
Sorensen(1909) as the negativelogarithm to base10 of the hydrogenion con-
centration,but is now defined in terms of hydrogenion activity
1
pH = or - log [W] [2]
log [W]
Thus, the pH of pure waterwould be -log of 1 x 10-7 or 7. As defined,any solu-

tion with a pH below sevenis consideredacidic, and one with a pH greaterthan
sevenis defined as basic.
Becauseonly dissociatedhydrogenions affect pH, the type of acid that is
presentis important in interpreting pH values. Acids are roughly divided into
strong and weak basedon their degreeof dissociation.Strong acids arealmost
completely dissociated(e.g., hydrochloric acid) while weak acids are slightly
dissociated(e.g., aceticacid). In the caseof weak acids,the total amountof acid
in solution can be calculatedfrom the pH and the dissociationconstantusing the
relationship
[3a]
or in a form that is simpler to use
[A-]
PH - pK. = log-- [3b]
a [HA]
(whereKa is the dissociationconstantfor the acid, [A-] is the activity of dissoci-

ated anion and [HA] is the activity of undissociatedacid in solution. Becausein
pure acid systems(A-) = (H+), where ( ) refers to concentrations,the negative
log of a weak acid concentrationis given by
SOIL pH & SOIL ACTIVITY 477
pHA = 2pH - pKa [4]
At a concentrationof 1 M, pHA = 0 and pH = 1/2 pKa' so that pH also can be

usedto determinethe value of pKa in a molar concentrationof acid.
In mixed acid-saltsystems(buffers), Eq. [3b] can be usedto estimatede-
greeof dissociation.A modificationof that equation,the Henderson-Hasselbalch
equation, has been used to estimatedegreeof dissociation in organic matter
(Hargrove& Thomas,1981) or other pH-dependentexchangersin soil
a
pH = pKap = ~log - - [5]
1- a
(whereKap is the apparentdissociationconstant,a is the degreeof dissociation,

and ~ is a constantwhich, for organicmatter, and other polyacidsgenerallyhas
a value of aroundtwo). This equationgives the pKap at half neutralization(a =
0.5) since the log portion disappearsand pH = pKap.
SIGNIFICANCE OF SOIL pH VALUES
When usedwith soils, pH can aid the investigatorgreatly in determining

major soil characteristics.Although not adequateas a lime requirementindica-
tor, pH can be usedto make a rough estimateof lime requirementand the rela-
tive availability of both P and many of the minor elements(Zn, Cu, B, Mn, Fe,
and Mo, for example).In a more specific way, particularpH valuesin water can
be usedto predict ratherwell the dominantcationson soil cation exchangersat
the time of soil sampling and analysis. These characteristicpH values are
describedbelow.
Presenceof FreeAcids
Generally,soil pH valuesin the neighborhoodof two to three indicate the
presenceof free mineral acid, usually H2S04 • A pH value much below 4.0 is
impossibleto achievewith Al3+ -saturationso that acidity 10 to 100 times more
intense (pH 2-3) indicates not only the presenceof H+ but also a continuing
sourceof them. A H-saturatedsoil, preparedfrom H2S04 will not persistbut will
rather quickly form an AI-soil due to dissolution of the clay minerals by
exchangeableW (Coleman& Craig, 1961). Therefore,a soil with pH of two to
threealwaysis found to havea bountiful supply of free acid. The usualsourceof
this acid is pyritic mineralswhich, upon oxidation,form the H2S04. Likely occa-
sions where this excessof acid is found are in mine spoils and recently drained
marinesediments,both of which are frequently well-suppliedwith pyrites.
The presenceof free acid indicatesthree seriousproblems:(i) plants will
not grow, (ii) the soil minerals are slowly (or rapidly) being dissolvedand (iii)
there will be a high cost of amelioratingsoil acidity using lime. In commercial
agriculturethis cost most often cannotbe met but, for land restoration,where it
must be paid, it can equal the value of the land itself. For example 100 Mg of
478 THOMAS
CaC03 per hectareis more typical than not of the lime requiredto control acid-
ity in thesecases.
Presenceof Aluminum Ions
At pH valuesof four to five, the presenceof exchangeable, trivalent Al will

be encounteredin mineral soils and, at times even in certain organicsoils. The
hydrolysisof both exchangeable and solutionAl in Al-saturatedmontmorillonite
and hectorite,
gives a minimum pH value of 3.84 in montmorillonite(Frink & Peech,1963)and

asthe Al3+ becomessmall, the pH risesto 4.89 in hectorite(hencethe rangefrom
pH 4-5).
Values of pH in this range practically always indicate trouble aheadfor
most crop plants becauseof the negativeeffects that Al has, especiallyon plant
root growth (Foy, 1984). Nevertheless,the caseis not nearly so seriousas that of
free H+ whereno plant growth is possibleand wherecostsarevery high. In most
cases,the problemsof exchangeable Al3+ can be resolvedwith a few megagrams
of CaC03, a treatmentthat usually lasts severalyears and produceseconomic
increasesin crop yields.
Presenceof Hydroxy-Aluminum
At a pH of 5.5 and above,exchangeable Al3+ is no longer present.Instead,

Al chemistry is dominatedby a complex mixture of hydroxy-Al ions, many of
them highly polymerizedand virtually nonexchangeable (Rich, 1960).
The smaller particleshave a high net positive chargeand are true solutes
(Hsu, 1989).Thesemay be involved directly in suppressionof root growth (Alva
et aI., 1986). The larger polycationshave little direct effect on plant growth but
may strongly affect both P and K availability (Rich, 1964).
In many soils, the major part of the "buffering" region involved in practi-
cal liming is controlled by the hydroxy-Al ions adsorbedon both clay minerals
and on organic matter. From the .standpointof acidity control in soils, these
hydroxy-Al compoundsare very well buffered, indeed, (Coleman & Thomas,
1964) and while resistingthe effect of liming materials,they also resist equally
well, the tendencyof acidifying agents(such as NH4- containingfertilizers) to
reacidify the soil. They act aboutequally well as "sinks" for WandOH- and, of
course, are a major source of the so-called pH-dependentcharge (Thomas &
Hargrove,1984).
Presenceof Calcium Carbonate
At the other end of the scale,pH is invaluable as an indicator of excess

CaC03 in the soil. Although the effect of the partial pressureof CO2 on pH of
calcareoussoils is strong, normally soils with pH valuesof 7.6 to 8.3 are found
to be calcareous.ExcessCaC03 in the soil immediatelyindicatesthat no money
needbe spenton lime, that soil acidity will not be a problemand, unfortunately,
that minor elementssuch as Zn and Fe could prove to be troublesome.On the
whole, the conditionsare good but there are somepotentialproblems.
Knowledge of whethera soil is calcareousor not can be especiallyvalu-
able in areaswhere both calcareousand noncalcareoussoils are interspersedon
the landscape.Both soil treatmentsand expectationswill be likely to vary
accordingto the presenceor absenceof CaC03.
Presenceof SodiumCarbonate
When pH values stray towards nine, it can be inferred that CaC03 is no
longercontrolling the systemand that Na2C03is assumingcontrol with dire con-
sequences for the long-termhealthof the soil. When Na2C03becomesdominant,
not only is Na an importantcationon the soil exchangersbut Ca ceasesto be very
important becauseof its precipitationas CaC03. In a calcareoussoil at normal
pH valuesof 8.3 or less,Ca in solution is relatively abundantbut, as the pH rises
towardsnine, CaC03 becomes soinsolublethat Na2C03'not CaC03, buffers the
soil. Being a soluble salt, the systemis swampedwith Na at the expenseof Ca.
When the concentrationsof the normal saltsof Na, such as NaCI or Na2S04are
relatively low, the combinationof exchangeableNa and Na2C03 with low Ca
causesdispersalof the clay and of the organicmatter; the result is called black
alkali soils. Fortunately,these soils usually exist as only portions of the land-
scapebut, even so, their appearanceis a warning of further degradationof the
soil in the future.
FACTORS WHICH AFFECT SOIL pH
The Effect of Dilution

The ratio of water to soil in the suspensionhasthe effect of increasingpH
as the ratio increases.However, the effect is not as straightforwardas might be
expected.For example,increasingwater content 10 times will not give one pH
unit increaseas it would in a solution of acid. Instead,the increasein pH will be
only about0.4 as shown in Fig. 16-1 from Davis (1943). What is the reasonfor
the relative insensitivity of suspensionpH to dilution?
It appearsthat, eventhoughpH is a measureof the H+ activity in solution,
in a suspension,the dilution of the soil sampletendsto increasethe dissociation
of H+ from the soil surfaceand, in addition, to increasethe amountof hydroly-
sis from whateverforms of AI might be present.Both effectshelp to "buffer" the
solutionand, therefore,to maintainthe pH at a relatively stablevalue over a wide
rangeof dilution in acid soils. In soils with higher pH values,hydrolysisof basic
cationstendsto maintain a stablepH with dilution. In a practicalway, this effect
leadsto the conclusionthat water/soil ratios are not a highly important factor to
consider in interpreting soil suspensionpH values as long as interpretationis
basedon a consistentmeasurementand experiencewith that measurement.
480 THOMAS
0.60
0.50
0.40
1
I
<l
Q. 0.30 1 -----
_/--- - /--1-------- ----------·l
0.20
0.10
,
0.00
1: 1 1:5 1:10
Soil:Water Ratio
Fig. 16-1. The effect of dilution on pH valuesof California soils showingstandarddeviationat each
dilution (datafrom Davis, 1943).
Salt Content
A major factor influencing pH of soils is the salt contentof the soil solu-
tion. These salts may be a natural part of the soil such as NaCI, Na2S04 or
Ca(N03)2, or they may be addedin the form of fertilizers. In any event,the usual
tendencyof the saltsis to lower the pH value of the soil progressivelyas the salt
concentrationincreases.In acid soils the effect apparentlyis due to both dis-
placementof AI3+ from the exchangecomplex and to increasedhydrolysis of
various kinds of AI speciesin the presenceof salt (Ragland& Coleman,1960).
In calcareoussoils, an effect of aboutthe samemagnitudealso is observed
which ratherobviouslycannotbe explainedby the samemechanisms.Moore and
Loeppert (1987) relate this reduction in pH to the displacementof Ca2+ from
exchangesites by the salt. At a constantpartial pressureof CO2 of 60.6 Pa
(0.0006atm), their equationfor the pH of a salt-affectedcalcareoussoil is
pH = 6.48 - 0.54 log [Ca2+]

where [Ca2+] is the activity of Ca in the soil solution.
It is worth pointing out also that in tropical soils, there is frequently a ten-
dency for pH valuesto rise in the presenceof salts (Van Raij & Peech,1972).
This behavioris an indication that salt is releasingmore OH- than H+ from the
soil or, in other words, the soil has more sourcesof positive than of negative
charge.This behavioroffers one possibility for the useof both salt and water pH
values for the chemical characterizationof the soil. This characterizationhas
value for predicting the behavior of both sulfate and phosphateas well as the
cationsthat may be present.
There are two practical problemswhich have to do with the effect of salt
on pH: the first problem is the seasonaleffect first noted by Baver (1927), and
the secondproblem is the interpretation(or reinterpretation)of pH valuestaken
SOIL pH & SOIL ACfIVITY 481
7
y-O.8269x-O.2936
r'=O.7S24 •
6
•
•
•• •
•• •
U 5
• •
~
c:
•• • •
•• •
J:
Cl. 4 •• •
•••••••• ••
•• •• • •• •••
3 •
•
2
4 5 6 7 8
pH in Hp
Fig. 16-2. A comparisonof pH in water and in 1 M KCI for Nigerian soils (data from Okusamiet
al.,1987).
in 0.01 M CaClz or M KCI vs. thosetaken in water. The seasonaleffect is basi-

cally a reflection of the loss, formation or accretionof saltsduring varioustimes
of the year. Under humid conditions,the soil is most nearly free of saltsin early
springdue to winter leachingand lack of nitrification. At spring planting season,
the soil is likely to be high in saltsdue to rapid nitrification and to applicationf
fertilizer. As cropsgrow, salts are taken up gradually, lowering the salt content.
But, soonafter harvest,the salt level in the soil can be quite high becauseof the
water deficit, absenceof plant uptake, nitrification and mineralizationof nutri-
entsin crop residues.Baver (1927) showedthat the lowest soil pH occurreddur-
ing June,was nearly equaledin Septemberand that the highestsoil pH occurred
in April, generallyconfirming the scenariogiven above.
Attempting to deal with problemsof variable saltby addition of saltsdoes
not necessarilyresolve the problem. As shown in Fig. 16-2, (Okusami et aI.,
1987) the addition of 1 M KCllowers pH aboutone unit at pH 4 and more than
two units at pH 8, giving still anotherset of numbersof calibratementally with
observedsoil conditions.
The approachof Schofieldand Taylor (1955) which hasbeenwidely used
in Englandand Canada(Turneret aI., 1963) is to determinepH in 0.01 M CaClz
The resultingdifferencepH - 1/2pCahasbeencalled the lime potentialbecause
it correspondsto the meanactivity of Ca(OH)z.It is somewhat,but not entirely
invariablewith naturally changingsalt concentrationin soils.
Websterand Harward (1959), using both water pH and pH - 1/2 p (Ca +
Mg), showedthat the ,.2 (,.2 = proportion of variation explainedby the simple
482 THOMAS
regressionequation)for the relationshipbetweenwaterpH and percentageCa +

Mg saturationwas slightly betterthan the ,.z for a similar relationshipbetween
pH - 1/2 P (Ca + Mg) andpercentageCa + Mg saturation.Perhapsmore impor-
tantly, their valuesof l/2p(Ca+ Mg) obtainedusing0.001M CaCl2showedprac-
tically no variationover a rangeof basesaturationsfrom 15 to 150%,good evi-
dencethat Ca2+from CaCl2essentiallycoveredup any soil differences.This is a
clear indication that the extra trouble involved in determiningpH in 0.001 M
CaCl2was not worth the effort.
In summary,even with the problemsinvolved, a soil-water suspension
gives valueswhich can be usedwith considerableconfidence,if not precision,
for making practicalfield decisions.The use of saltsof one kind or anotherto
correctfor variablesaltsin the soil doesnot, in general,lead to betterinterpreta-
tion.
CarbonDioxide Content
The effect of CO2on the pH of calcareoussoils is very largeand,given the

variability of the partial pressureof CO2 in the soil atmosphere,it can be con-
sideredthe single largestfactor affecting the measuredpH of calcareoussoils.
Bradfield (1942) publisheddata on the pH of CaC03 over a rangeof Pc~ and
Whitney and Gardner(1943) determinedpH on a numberof westernU.S. soils
as affected by Pc~. Their data for soils are plotted on a graph drawn from
Bradfield'sdatafor CaC03(Fig. 16-3) and the agreementis excellent.Two soils
dominatedby Na2C03were not plotted becausethe pH valueswere so much
higher, but eventhey had the sameslope as the other soils. Figure 16-3 shows
clearly that the reaction
is the major controlling factor which determinespH in calcareoussoils.

Naturally, in soils which are dried and groundprior to determiningthe pH, the
effect of CO2shouldnot be so importantbut wherepH is determinedon a fresh
sampleor in situ, the effect can be very large.
SuspensionEffect
The reductionin pH (and the occasionalrise) that occurswhen the elec-

trodesareplacedin the soil suspensionratherthanthe supernatantsolutionabove
the suspensionhas beencalled the suspensioneffect. The interpretationsof the
suspensioneffectbasicallyhavebeentwo: first, Marshall(1964)believedthat H+
nearthe clay surfacedissociatedsufficiently to affect pH whenthe electrodewas
placednearthem. In otherwords, he believedthat the pH was truly different in
the soil paste as comparedto the supernatantsolution. Second,accordingto
Colemanet aI. (1951),the suspensioneffect is primarily causedby the effect of
the electricalchargeof the soil on the mobilities of K+ andCl- from the calomel
electrode,ratherthan the glasselectrode.For example,a strongly negativesoil
would tend to promoteK+ mobility and impedethat of Cl whereas,a positively
9.0 .---------------------------------------~
.,...
8.5
•
8.0
7.5
•
:r:a. 7.0 •
•
6.5
•
6.0
•
5.5
5.0 L - -_ _ _ _ _ _ _ _"---_ _ _ _ _...l.-_ _ _ _ _ _- - ' -_ _ _ _ _ _ _ _--'
-4.0 -3.0 -2.0 -1.0 0.0
log Pco,
Fig. 16-3_ The effects of PC02 on pH of calcareoussoils (curve from Bradfield, 1942; points from
Whitney & Gardner,1943).
chargedsoil (an oxisol horizon, for example)would have exactly the opposite
effect. A soil with very low chargewould havevery little effect at alL
Although it is generally accepted(Olsen & Robbins, 1971) that the sus-
pensioneffect is more likely the result of the secondexplanation(that is, it is
largely spurious),there are some indications that Marshall (1964) was correct
about relative H+ activity on the soil surfaceas comparedwith those in the soil
solution. For example,Swobodaand Kunze (1968), basedtheir conclusionson
ionization of weak organic bases,concludedthat the surfacepH of clays had to
be lower than that of the bulk suspensionin order for ionization of the basesto
occur.
Nevertheless,distancesa few nanometersfrom the clay surfacehavevery
little to do with those obtained with a large electrode in a soil suspension.
Perhaps,the emergingfield of microelectrodesfor pH measurementwill shed
further light on the subject.
FUNDAMENTALS OF pH MEASUREMENT
Colorimetric Determinations of pH
Colorimetric methodsfor pH determinationare basedon weak acids or

weak bases,the colors of which changewith undissociatedor dissociatedforms.
For example,the dissociationof an indicator weak acid gives
acid alkaline [6]

color color
484 THOMAS
The overall color observedis a function of the relative concentrationsof

HI and 1-, which varieswith pH accordingto the following equation
pH = pKHI + log -
W] + log fI- [7]
[HI]
where pKHI is the negativelogarithm of the dissociationconstantof HI andf r- is

the activity coefficientof the indicatorion. The color achievedwill vary with the
concentrationof solutions used becauseof its effect on f I-. For example,with
bromcresolgreen,chlorophenolred andbromothymolblue, pH valuesin a 0.5 M
NaCI concentrationhavea positive error of between0.3 and0.4 pH units (Peech,
1965).
Another error in colorimetric methodsis causedby the use of ethanol to
dissolve some indicators. The dissociationconstantis decreased,changingthe
pH of color changeto higher values.This is especiallyimportant when indica-
tors are useddirectly on the moist soil but not so importantin soil suspensions.
ElectrometricMeasurements
In the vast majority of cases,pH is determinedusing a glass electrode-
calomelelectrodesystem.This is representedby the following diagram
Ej
HgIHgClz, KCI(sat) I test solutionIglass10.1M Hel, AgClIAg
wherethe single barsrepresentphaseboundariesand the doublebar representsa

liquid junction potential markedEj. The observedvoltage of the cell, E, is pro-
portional to the H+ activity of the solution on suspensionand is given by
(E -E')F
pH - -'----'--- [8]
2.303RT
whereR is the gasconstant,T, the absolutetemperatureandF is the Faradaycon-

stant.At 25°C, this reducesto
(E -E')
pH - -'----"- [9]
0.0591
The term E' includesthe potential of the reference(calomel) electrode,the Ag-

AgCI portion of the glasselectrode,the asymmetrypotential of the glasselec-
trode and the liquid junction potential,E j • When the pH meter is standardized
with a buffer of known pH, the value of E' has beenevaluated,and effectively
canceledout. Therefore,
pH = pHs + (E - Es) [10]

0.0591
wherepH~ is the pH of the standardbuffer solution,Es is the voltageat pHs, and

E is the voltage of an unknown solution or suspension(Peech, 1965). This
assumesthat the junction potentialwill not changefrom the standardbuffer to a
soil suspensionwhich, of course,it will as hasbeendiscussedearlier.
Equation[10] signifiesthat, in practicalterms,the voltagechangefor 1 pH
unit is 0.0591 V (or 59.1 mY) at 25°C, and all commercialpH metersare cali-
bratedon this basis.
ELECTROMETRICMEASUREMENT OF SOIL pH
Equipment
There is currently such a proliferation of equipmentfor determiningsoil
pH that it is impractical to discussthem all. In general,the electrometeritself
varies in quality with price and the cheaperelectrometerscannot be used for
readingsmore precisethan ±0.1 pH unit. For field work they are entirely ade-
quate.For more preciselaboratorywork they probably are not.
Combinationelectrodes,which usually have a Ag-AgCI referencerather
than the usual calomel referenceelectrode,do not give results as accurateas
those obtainedwith the standardtwo-electrodesystem. Nevertheless,for field
work the combinationelectrodesmay be preferablebecausethey are easierto
use. Similarly, plastic-encasedelectrodesare clearly preferable for field use
becausethey resist breakage.On the other hand, standardglass electrodesare
much easierto clean betweensamplesand should be more useful in laboratory
applications,unlesscareless operators are a problem.The new "Ross" electrodes
which use a platinum wire and a redox filler solution to replacethe traditional
Ag-AgCl and Hg-HgCI2/KCI systemshave the advantagethat the referenceis
not sensitive to temperaturechangesand readings can be made much more
quickly. Their disadvantageis that they cost two to three times as much as con-
ventionalelectrodes.
Although digital readoutis now much more commonthan the traditional
analogand has many advantages,particularly with indecisiveoperators,the lat-
ter hasthe advantageof visually determiningthe approachof pH to an equilibri-
um value. This can be a very useful feature in certain kinds of studies,such as
titrations.
It shouldbe stated,in general,that the move to solid-stateinstrumentshas
eliminatedmany of the problemsof the older machines.On the other hand,many
of those problems(shielding, grounding) could be handledby an experienced
operator.When the new machinesfail, they tendto fail completely a require a
return trip to the manufacturer.
Standardizationof the Meter

Most modem pH meters are equippedwith a meter testing program to
checkthe meterseparately.This is done with a shorting plug installed to deter-
mine whetherthe meter is stable internally. Having done this check-upaccord-
ing to the operatingmanual,the meterand electrodesmay be standardized.
486 THOMAS
For soils, a two-buffer standardization,at pH 7 and pH 4, should be per-

formed. This will determinewhether the electrodesare working properly and
also will store the slope value (0.0591 V per unit of pH) in the memory of the
electrometer.
In general,with solid-statepH meters,the procedureis to use a buffer of
pH 7.0 and allow the machineto display the number.If the number is 7.0, the
value is approvedby pressingthe yes button and the electrodesare cleanedand
placedin the pH 4.0 buffer. The secondbuffer readingshould be very close to
4.0. If it is, that numberalso can be approvedby pressingthe "yes" button and
the pH meterand electrodesare now ready for use. If the metercannotbe stan-
dardized,it indicateselectrodeproblemswhich are coveredin the next section.
On older pH meterswithout programmedcheck sequencesand without
automatic temperature compensation(ATC), the procedurefor standardizationis
as follows: the pH 7 buffer is introduced,the manualtemperaturecontrol is set
to the temperatureof the buffer and the meter is set at 7.0. The electrodesare
rinsed and blotted and the pH 4.0 buffer is introduced.If the buffer value reads
4.0 :t 0.1, the instrumentis now readyfor use.If not, adjustthe readingto pH 4.0
usingthe temperaturecompensationknob. Repeatthis with the pH 7.0 buffer and
againwith the pH 4.0 buffer until both readingsaresatisfactory.If this cannotbe
done,one or both electrodesprobably are faulty.
ElectrodeProblems
Glassusedin the H+-sensitiveelectrodeis characterizedby low electrical

resistance,low melting point, and high Na content(Dole, 1941). In addition, the
glassmust have the tendencyto hydrateeasily if it is to be predictablyrelatedto
the activity of H+. That the adsorptionof water by the pH-sensitiveglasswas of
overwhelmingimportancewas known as early as the 1920s(Dole, 1941).
The most commonproblemswith electrodesare causedby allowing them
to dry out. In the caseof the glass electrode,there is a hydratedlayer of glass
which facilitatesthe movementofH+ (Westcott,1978).Hydration doesnot occur
immediatelywhen the glasselectrodeis placed in water or dilute acids so that
there will be a delay of somehours as proper hydration occurs. In addition, the
surfaceof the electrode,upon drying, may becomecoatedwith carbonatesor
other compounds,rendering it somewhatimpermeableto H+. The electrode
shouldbe given alternate5-min soakingsin 0.1 M HCI followed by 0.1 M NaOH
and finally in 0.1 M HCI again prior to use.This treatmentwill do a goodjob of
removingany accumulatedcarbonateswhich restrict operationof the glasselec-
trode.
If the rejuvenationprocedureoutlined abovedoesnot resolvethe problem,
the electrodemay be lightly etchedin 20% ammoniumfluoride solution for 10
to 30 secand then rinsedin water. This is a "last resort" treatmentthat cannotbe
repeatedregularly because the glassis alreadyvery thin.
The calomel referenceelectrodealso suffers from drying but the causeof
potential problemsis not due to the glassbut, rather,the lack of flow of internal
filling solution from inside the electrodeto the soil suspension.The electrode
shouldbe full of solution beforeuseand the vent capshouldbe openso that flow
can occur. For soils measurements, it usually is preferredto have relatively slow
leakageto minimize contaminationof the soil suspension.Howeverif leakageis
too slow, the reading will be in error. The asbestosor glass fiber, or ceramic
opening in the referenceelectrode(all three are used) may be pluggedby KCI
crystals(usually from inside the electrode)or by soil particleson the outside.It
is necessarythat somemeasurableflow occur.This can be checkedby filling and
cleaning theelectrodeand checkingthat slight wetting occurs.
PROCEDUREFOR SOIL pH MEASUREMENT
Equipmentand Reagents
1. pH meterequippedwith glassand referenceelectrode,or combination

electrode.
2. 50- or 100-mL beakers.
3. Pipet or automaticpipet of 10 mL.
4. Standardbuffers, pH 7 and pH 4.
5. Deionizedwater.
6. 0.01 M CaCl2 solution.
7. 1 M KCl solution.
Determination-pHin water
1. Weigh out 10 g of air-dry soil in a 50- or 100-mL beaker.

2. Add 10 mL of deionizedwater to the soil in the beakerand mix well. A
stirring stick, or stirring machinecan be usedbut care should be exer-
cised to minimize contamination.(For large-scaledeterminations,a
shakingmachinecan be employedas is done in most soil testing labo-
ratories.)
3. Let standfor 10 min.
4. Swirl the suspensionin the beakerand insert the electrodes intothe sus-
pension. Electrodesmay be placed in the clear supernatantabove the
soil, directly in the sedimentedsoil, or the entire suspensionmay be
stirred during the pH determination.The important thing is that the
measurements be carried out ina consistentway. In water pH determi-
nation, valuestaken in the supernatantgenerallywill be slightly higher
than in the stirred suspension.With a salt pH, the differencesbetween
the threetechniques practically disappear.For exposedglasselectrodes,
it is useful to have a stop of sometype so that the bulb will not contact
the bottom of the beaker.McLean (1982) suggesteda glassrod, slight-
ly longer than the electrode,mounted onthe electrodeholder that makes
contactwith the beakerbefore the electrodedoes.
5. ReadpH and record as pHw.
6. BetweenpH readings,rinse the electrodeswith distilled water. Blotting
is not necessary.
488 THOMAS
pH in One-OneHundredthMolar Calcium Chloride
1. Repeatthe above procedure(1-4) but use 0.01 M CaCl2 insteadof

water to make the soil suspension.
2. ReadpH and recordas pHeacl2.
pH in One Molar PotassiumChloride
1. Repeatsteps1 to 4 but use 1 M KCI insteadof water to make the soil

suspensions.
2. ReadpH and recordas pHKo .
ALTERNATIVE MEmODS FOR pH MEASUREMENT
Use of Microelectrodes
The possibility of taking microsite pH values in intact soil systemshas

been facilitated by the use of microproceduresin plant cells (Felle & BertI,
1986). Conkling and Blanchar(1989) have usedhomemademicroelectrodesfor
this purposeand haveobtainedrelatively stableand reproducibleresults.It is not
yet clear,however,that startlingnew findings aboutthe variability of pH in a soil
matrix will be forthcoming.
Basically, the techniqueof Conkling and Blanchar, (1989) is to melt a
small cap of pulled H+ ion-sensitiveglassover a pulled glass capillary,to install
a silver wire, coatedwith AgCI and to fill it with a solution.This requiresmicro-
scopicobservationand manipulationand must be thoughtof as tediouswork.
The referenceelectrodeis a calomel electrodeconnectedto a gel-filled
KCI salt bridge to assurethat a completecircuit is made. Readingson suspen-
sions have shown close to 1:1 slopeswith conventionalelectrodesso it is clear
that the microelectrodeshavesomepromise.
It is quite interestingto note that the pioneersof microelectrodesare fol-
lowing the path of the earlier pioneers (Sharp& Hoagland,1919), making the
equipmentas they go.
Use of Test Kits
Test kits have always suffered somewhatby comparisonwith electrodes

because(i) thereare only certainpH valueswhereionic dyeschange,and (ii) the
color of the soil somewhatobscuresthosecolors. Summingup (i) and (ii), it is
clearthat using kits is partly an art and dependsconsiderablyon the pair of eyes
that is observingthe color. Masonand Obenshain(1939) found good agreement
betweencolorimetersand electrodepH values,but agreementwas in the range
of 0.3 to 0.4 pH units which is not very satisfactoryfor many applications.
Certainly, in many cases,thesefield kits can be useful in roughtly deter-
mining soil pH categoriesbut theyare more time-consumingthan a portablepH
meterand, probably,more expensivein the long run, and not as good.
pH Papers
The newer pH papersare reasonablyuseful for field work is accuracyof

closerthan 0.5pH units is not critical. Thesepapershavemaximumpracticaluse
as a first approximationin the field when no pH meteris available.As such they
are helpful in progressivesoil mapping programsor in trouble-shootingsitua-
tions. Nevertheless,the values usually have to be recheckedusing a pH meter
beforecompleteconfidencein the resultscan be attained.
REFERENCES
Alva, AK., D.G. Edwards,C.S. Asher, and EP.C. Blarney. 1986. Relationshipsbetweenroot length
of soybeansand calculatedactivities of aluminum monomersin nutrient solution. Soil Sci.
Soc. Am. 1. 50:959-962.
Baver, L.D. 1927. Factorsaffecting the W-ion con~entration of soils. Soil Sci. 23:399-414.
Bradfield, R. 1942. Calcium in the soil: I. Physico-chemicalrelations. Soil Sci. Soc. Am. Proc.
6:8-15.
Clark, W.M. 1920. The determinationof hydrogenions. Williams and Wilkins, Baltimore, MD.
Coleman,N.T., and D. Craig. 1961.The spontaneous alterationof hydrogenclay. Soil Sci. 91:14-18.
Coleman,N.T., and G.w. Thomas.1964. Buffer curvesof acid clays as affected by the presenceof
ferric iron and aluminum. Soil Sci. Soc. Am. Proc. 28:187-190.
Coleman,N.T., D.E. Williams, T.R. Nielsen, and H. Jenny. 1951. On the validity of interpretations
of potentiometricallymeasuredsoil pH. Soil Sci. Soc. Am. Proc. 15: 1O~110.
Conkling, B.L., and R.W. Blancher. 1989. Glassmicroelectrodetechniquesfor in-situ pH measure-
ments.Soil Sci. Soc. Am. J. 53:58-52.
Davis, L.E. 1943. Measurements of pH with the glasselectrodeas affectedby soil moisture.Soil Sci.
56:405-422.
Dole, M. 1941. The glasselectrode.John Wiley & Sons,New York.
Felle, H., and A BertI. 1986. The fabrication of H+-selectiveliquid-membranemicro- electrodesfor
use in plant cells. 1. Exp. Bot. 37:141~1428.
Foy, C.D. 1984. Physiological effects of hydrogen,aluminum, and managementtoxicities in acid
soils. p. 57-97. In F. Adams(ed.) Soil acidity and liming. Agron. Monogr. 12. 2nd ed. ASA,
CSSA, and SSSA, Madison, WI.
Frink, C.R., and M. Peech.1963. Hydrolysis and exchangereactionsof the aluminum ion in hectorite
and montmorillonitesuspensions.Soil Sci. Soc. Am. Proc. 27:527-530.
Gillespie,L.J., and L.A Hurst. 1918. Hydrogenion concentration-Soiltype-Commonpotatoscab.
Soil Sci. 6:219-236.
Hargrove, W.L., and G.W. Thomas. 1981. Effect of organic matter on exchangeablealuminum and
plant growth in acid soils. p. 151-166.In R.H. Dowdy (ed.) Chemistry in the soil environ-
ment. ASA Spec. Publ. 40. ASA and SSSA, Madison, WI.
Healy, 0.1., and P.E. Karraker. 1922. The Clark hydrogen-electrodevessel and soil measurements.
Soil Sci. 13:323-328. .
Hsu, P.H. 1989. Aluminum hydroxides and oxy-hydroxides. p. 331-378. In J.B. Dixon and S.B.
Weed (ed.) Minerals in the soil environment.SSSABook Ser. 1. SSSA, Madison, WI.
Marshall,C.E. 1964. Thephysicalchemistryand mineralogyof soils. Vol. I. JohnWiley & Sons,Inc.,
New York.
Mason,D.O., and S.S. Obershain.1939. A comparisonof methodsfor the determinationof soil reac-
tion. Soil Sci. Soc. Am. Proc. 3:129-137.
Mclean, E.O. 1982. Soil pH and lime requirement.p. 199-224.In AL. Pageet al. (ed.) Methodsof
soil analysis.Part 2. 2nd ed. Agron Monogr. 9. ASA and SSSA,Madison, WI.
Moore, T.1., and R.H. Loeppert. 1987.Significanceof potassiumchloride pH of calcareoussoils. Soil
Sci. Soc. Am. J. 51:908-912.
Okusami,T.A, R.H. Rust, and AS.R. Juo. 1987. Reactive characteristics of certain soils from south
Nigeria. Soil Sci. Soc. Am. J. 5:1256-1262.
Olsen, R.A, and J.E. Robbins. 1971. The causeof the suspensioneffect in resin-watersystems.Soil
Sci. Soc. Am. ProC. 35:26G--265.
490 THOMAS
Peech,M. 1965. Hydrogen-ionactivity. p. 914-926.In C.A. Black (ed.) Methodsof soil analysis.
Part 2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Ragland,J.L., and N.T. Coleman.1960. The hydrolysis of aluminum salts in clay and soil systems.
Rich, c.1. 1960.Aluminum in interlayersof vermiculite. Soil Sci. Soc. Am. Proc. 24:26-32.
Rich, c.1. 1964. Effect of cation size and pH on potassiumexchangein Nason soil. Soil Sci.
98:100-106.
Schofield, R.R., and A.W. Taylor. 1955. The measurementof soil pH. Soil Sci. Soc. Am. Proc.
19:164-167.
Sharp, L.T., and D.R. Hoagland. 1919. Notes on recent work concerning acid soils. Soil Sci.
7:197-200.
Sorensen,S.P.L. 1909. Enzymestudies:II. The measurementand importanceof the hydrogenion
concentrationin enzymereaction.Comp!. Rend.Trav. Lab. (Carlsberg).8:1.
Swoboda,A.R., and G.W. Kunze. 1968. Reactivity of montmorillonite surfaceswith weak organic
bases.Soil Sci. Soc. Am. Proc. 32:806-811.
Thomas,G.W., and w.L. Hargrove. 1984. The chemistryof soil acidity. p. 3-56. In F. Adams (ed.)
Soil acidity and liming. Agron. Monogr. 12. 2nd ed. ASA and SSSA,Madison,WI.
Turner, R.c., W.E. Nichol, and J.E. Brydon. 1963. A study of the lime potential. Soil Sci.
95:186-191.
Van Raij, B., and M. Peech.1972.Electrochemicalpropertiesof someoxisolsand alfisolsof the trop-
ics. Soil Sci. Soc. Am. Proc. 36:587-593.
Webster,G.R., and M.E. Harward.1959.Hydrogenand calciumion ratios in dilute equilibriumsolu-
tions as relatedto cation saturation.Soil Sci. Soc. Am. Proc. 23:446-451.
Westcott,C.c. 1978. pH measurements. Acad. Press,New York.
Whitney, R.S., and R. Gardner. 1943. The effect of carbon dioxide on soil reaction. Soil Sci.
55:127-141.
Published 1996
Chapter 17
Lime Requirement
J. THOMAS SIMS, University of Delaware, Newark, Delaware
INTRODUCTION: DEFINITION AND IMPORTANCE

OF LIME REQUIREMENT METHODS
Lime requirement is definedas the amountof agriculturallimestoneor otherbasic

material neededto increasethe pH of the soil from an unacceptablyacidic con-
dition to a value that is consideredoptimum for the desireduseof the soil. It also
hasbeenreferredto as the capacity factor of soil acidity becauseit representsthat
fraction of total soil acidity (soluble and exchangeable)that must be neutralized
to achieve a desiredsoil pH (McLean, 1982). Hence it is an indication of the
capacity of the soil to resist a changein pH as lime is added.Soil pH, usually
referredto asthe intensity factor of soil acidity, reflectsthe amountof acidity pre-
sent in the soil solution and servesas an index of the acid-basestatusof the soil.
Optimum soil managementfor any purposerequiresthat soil pH be adjustedto
an acceptablerange; thereforeaccurate,rapid methodsto assessthe amount of
liming material neededto effect a soil pH changeare essential.Among the con-
ditions associatedwith an adequatelylimed soil are an adequatebasesaturation
of the soil's cation exchangecapacity (CEC), the neutralizationof potentially
phytotoxic elements(e.g., AI, Fe, Mn), the reduction in solubility of hazardous
traceelementsin waste-amended soils (e.g.,Cd, Cr, Cu, Ni, Pb, Zn) and enhanced
microbial activity (McLean, 1971).
A variety of techniqueshave beendevelopedto measurethe lime require-
ment of soils and severalexcellentreviews on this topic are available (Alley &
Zelazny, 1987; McLean, 1982; Peechet aI., 1965; van Lierop, 1990). Ail lime
requirementmethods,however,are basedon similar underlying principles. First
is the obviousfact that the measuredlime requirementmust accuratelyreflect the
amountof liming material neededto raise the pH of the soil to the desiredor tar-
get pH when the lime is appliedunderfield conditions.Historically, the targetpH
has been defined as the soil pH value associatedwith optimum plant growth,
henceit will vary with type of plant to be grown and also will be influencedby
economicand logistical considerations.For example,a soil pH of 6.0 is usually
Book Seriesno. 5.
491
492 SIMS
consideredoptimum for most agronomiccrops,suchas com (Zea mays L.), soy-

bean (Glycine max L. Merr.), and wheat (Triticum aestivum), although some
leguminouscrops,suchas alfalfa (Medicago sativa L.), often respondfavorably
to applicationsof lime up to a pH of 6.5 to 7.0. However,a lower targetpH (e.g.,
5.~5.5) may be acceptableor desired for acid-tolerantplants such as potato
(Solanum tuberosum L.), blueberry(Vaccinium spp.), ornamentalshrubs,and a
large numberof grassesusedin the revegetationof highly disturbedsoils. Other
studieshave shown that liming soils to a pH of 5.5-whereexchangeablealu-
minum (Al3+) is neutralized-isoften an acceptableapproachto achievemaxi-
mum crop yields, particularly in highly weatheredsoils (Evans & Kamprath,
1970; Kamprath, 1970; McLean, 1970; Reeve& Sumner,1970; Webberet aI.,
1982). Target pH valuesof 5.2 to 5.5 are often used for peat, muck, and other
high organicmattersoils becauseeconomiccrop responsesto lime rarely occur
above these pH values (McLean, 1973; van Lierop, 1983). Soils also may be
limed for purposesother than plant growth, such as to enhancethe biodegrada-
tion of an organic pollutant by soil microorganismsor to convert a potentially
toxic trace elementinto an insolubleform. Target pH valuesfor thesesituations
may be quite different than thosebestsuitedfor crop growth.
A rangein target pH valuesmay exist for different soil managementcon-
ditions, even in limited geographicareas.Becauseof this, it is apparentthat the
most important characteristicof any lime requirement technique is that it mea-
sure only that fraction of soil acidity that must be neutralizedto increasethe
existing soil pH to the target pH under the environmentalconditions and time
intervalsthat will be encounteredin the field. Most studieshaveshownthat lime
requirementvaluesare typically intermediatein magnitudebetweenthe amount
of exchangeable soil acidity, defined as that extractedby a neutral, unbuffered
salt [e.g., 1 M KCl; (Thomas,1982)], and the amountof total potential soil acid-
ity, usually defined asthat which reactswith a bufferedsolution adjustedto a pH
of 8.2, such as BaClz-triethanolamine(Mehlich, 1953; Peech et aI., 1962;
Thomas,1982).
A secondunderlyingprinciple importantto the developmentand selection
of a lime requirementtest is the fact that lime requirementvaluesincrease astar-
get pH values increasebecauseof the contribution of pH dependentacidity to
total soil acidity (Thomas& Hargrove,1984; van Lierop, 1990). For soil pH val-
ues greaterthan 5.5 in mineral soils, lime reactsprimarily with pH dependent
acidity from clays, oxides, and organic matter. Lime requirementtests must
thereforeaccuratelymeasureall forms of acidity presentin a soil, both dissoci-
ated and undissociated,that must be neutralizedto attain the desiredpH.
A third factor to consideris that the suitability of a lime requirementtest
must be verified by comprehensivecalibration studiesthat reflect the intended
useof the soil and the variation in soil propertiesexpectedin the geographicarea
where the test will be used.Calibration studiescomparesoil pH valuespredict-
ed to result when soils are limed in accordancewith a lime requirementtestwith
pH valuesactually obtainedwhen soils are limed in field or greenhousestudies.
Thesestudiesare vital stepsin an overall liming programand must be conduct-
ed to insure that a lime requirementmethod accuratelypredicts the amount of
lime neededto eliminatethe deleteriouseffectsof excessivesoil acidity prior to
LIME REQUIREMENT 493
initiation of a soil managementprogram. Accurate calibration of lime require-

ment testsis equally importantto preventoverliming of soils with low buffering
capacities(e.g., sandy,low organicmattersoils) becausethe availability of some
importantplant micronutrients,suchas Fe, Mn, and Zn, can be reducedto yield-
limiting levels if thesesoils are limed too far abovethe targetpH.
Finally, most lime recommendationsare basedon the use of high-quality
limestone(CaC03 or Ca • MgC03) becausethis is the most common,effective,
and economicliming material available.The developmentof lime requirement
testsand associatedrecommendations, therefore,also must considerthe needfor
calibration studies that determine conversion factors between limestone and
otherliming materialsthat havedifferent neutralizingabilities. Examplesinclude
both thosethat are more reactivethan limestone(e.g., Ca oxidesand hydroxides)
and those that may be less reactive(e.g., wasteliming materialsfrom industrial
and municipal sources).
FACTORSAFFECTING LIME REQUIREMENT
A number of complex and interrelatedfactors affect the measuredlime

requirementof a soil, including the nature of soil acidity, the neutralization
sequence (PH rangebetweeninitial and desiredsoil pH), soil physicaland chem-
ical properties, and any analytical considerationsrelated to the accurateand
reproducibledeterminationof lime requirementeither by rapid Soil test method-
ology or by more time-consuminglaboratory, greenhouseor field methods
(McLean, 1982). Each of thesefactors is reviewed briefly below, emphasizing
their relevanceto the rapid soil testingmethodsthat are most commonly usedto
determinethe lime requirementsof soils.
Natureof Soil Acidity
The dominantsourcesof acidity presentin most soils include: (i) dissoci-

atedand undissociatedhydrogenions (H+) associatedwith layer silicate clays or
AI and Fe oxides; (ii) ionizable H+ that originates from the acidic functional
groups of soil organic matter such as carboxyls and phenols; (iii) the various
forms of soil AI, including exchangeablealuminum (AI3+), hydroxy-AI
([AI(OH)2+] and polymeric AI hydroxides;(iv) soluble H resulting from acidic
precipitation,soluble organic acids,or from acid-forming reactionsin soils such
as the oxidation of ammonium-based fertilizers, the hydrolysis of monocalcium
phosphatefertilizers, and the oxidation of pyrite or other sulfide-bearingminer-
als (Thomas& Hargrove,1984).
The generalgoal of liming is to neutralizeenoughof the total acidity in a
specifiedvolume of soil (e.g., the "plow layer") to increasepH to a desiredvalue.
Becauseof this a lime requirementtest must react with an appropriatepercent-
age of eachform of soil acidity to quantify the amountof limestonerequiredto
raise the soil pH to the value appropriatefor the intended use of the soil.
Obviously, a test that only measureddissociatedH+ (active acidity) would great-
ly underestimatethe amountof lime neededbecauseit ignored the contribution
494 SIMS
of exchangeableH+ and all forms of soil AI and Fe. This is why soil pH alone
cannotbe usedto determinelime requirement.Measuringexchangeable acidity
(H+ and AI 3+) would better approximatethe lime requirementof a soil, but in
many casesalso can be an underestimatebecauseit doesnot include the nonex-
changeable acidity found as hydroxy-AI coatingson the surfacesand interlayers
of clays, the AI that is complexedwith soil organicmatter, and someof the pH
dependentacidity that is expressedat soil pH values>5.5. Any techniquethat is
usedto estimatelime requirement,therefore,must be able to predict the amount
of acidity originating from all sourcesthat will react when lime is addedto the
soil.
NeutralizationSequence:Initial and DesiredpH

The amountof lime requiredquite naturally dependson how acidic the soil
is relative to the pH that is desired.The initial pH is usually determinedeitherby
naturalconditionsrelatedto soil development(weathering)and/oranthropogenic
activities such as fertilization, acidic deposition, construction, mining, and
drainage.As mentionedearlier,the desiredpH is basedon the intendeduseof the
soil andtypically rangesfrom 5.5 to 6.5. The lime required toattainthe targetpH
also dependsgreatly on thosephysicaland chemicalpropertieswhich contribute
to the buffer capacity of the soil, definedin this contextas the inherenttendency
of a soil to resist a changein soil pH. Buffer capacity,and thus the amount of
lime neededto alter soil pH from its initial to the desiredvalue (neutralization
sequence), increaseswith the amountof clay, oxides,and organicmatterpresent
in the soil. As soil pH is increasedby liming, the natureof soil acidity changes.
In soils that are highly acidic (pH < 5.0), lime reactsprimarily with soluble and
exchangeable AI3+ andsomeexchangeable H+. As theseforms of acidity are neu-
tralized and soil pH increasesto approximately5.5, the dominantforms of soil
acidity becomehydroxy-AI monomersand polymers, and soil organic matter.
Successfullime requirementtests,therefore,must first be capableof measuring
the dominantreactiveform of soil acidity presentat the initial pH in a neutral-
ization sequence.Beyond this, as noted in more detail below, a successfultest
must accuratelyquantify the different lime requirementsof soils with similar ini-
tial pH valuesbut different buffer capacities.
Soil Properties
ParentMaterial and Weathering
The nature of the original parent material from which a soil was formed
and the extent of weatheringthat has occurredwill greatly influence both the
degreeof soil acidification and the forms of soil acidity now present(McLean,
1982). Lime requirementsare normally greaterfor soils that developedfrom
more acidic parentmaterialsunderconditionsof intenseweathering.Soil acidi-
fication occurswhenthe removalof basiccations(Ca, Mg) from soils due to ero-
sion, leaching,and plant uptakeoccursmore rapidly than the naturaldissolution
of rocksandminerals(weathering)containingthesebasescan replenishthem. As
the basic cationsare removedfrom the soil, acidic cationssuchas H+, AI3+, and
Fe3+ replacethem on cation exchangesites and in the soil solution. The rate at
which this occurs depends on the compositionof the original parentmaterialand
the extent of weathering. Rocks and minerals dominated by carbonatesand
oxidesor aluminosilicatesthat containhigher percentagesof basiccationsacid-
ify more gradually,particularly in less humid regions.In arid regions,soil acid-
ification may not occur at all; here the accumulationof soluble cations often
resultsin greaterproblemswith salinity and sodicity, and lime requirementtests
are of little relevance.Conversely,parentmaterial where quartz and silica pre-
dominatecontainlower amountsof basiccationsand soils developedfrom these
materialsacidify rather rapidly, particularly in humid regionswhere rainfall and
temperaturesare more conduciveto dissolution and leaching of basic cations.
Soil acidification also is enhancedin areaswith greaterbiological activity, due
to the formation of carbonicacid (H 2C03) when CO2 originatingfrom the micro-
bial decompositionof organicresiduesdissolvesin the soil solution.
The net effect of the interactionsbetweenparentmaterial and weathering
is to create soils that will have different forms and degreesof soil acidity.
Furthermore,with time and continuedweathering,soils that initially are domi-
natedby the more permanent(exchange)acidity of 2:1 clays gradually convert
into thosedominatedby 1: 1 clays and oxidesof Fe and AI, where pH dependent
acidity is more important.The needfor lime, and the selectionof an appropriate
lime requirementtest is thus closely associatedwith well-understoodsoil forma-
tion processes.
Soil Textureand OrganicMatter Content

The ability of an acid soil to resist a changein pH when a liming material
is appliedis a measureof its buffer capacityand is most strongly influencedby
the percentageand type of clay mineralsand the amountof soil organic matter
present.Soils high in clay and/ororganicmatterare said to be well-buffered,will
have higher CECs,and will thus requiremore lime to effect a changein pH than
poorly buffered soils. The type of clay mineralspresentaffects soil acidity and
lime requirementas much or more as the amountof clay present.Soils where2: 1
clays predominatehave interlayer CEC sites and are thus more buffered than
soils dominatedby 1:1 clays. Positively chargedhydroxy-AI polymersare often
held at theseinterlayerexchangesites,further contributingto the amountof soil
acidity presentand the amountof lime required to reacha desiredpH. Texture
also can contribute to soil acidification by its interaction with leaching. Soils
with finer textureshave higher water-holdingcapacitiesand slower percolation
rates,and thus generally have slower ratesof loss of basic cationsthan coarse-
textured,easily leachedsoils. However,with time evensoils that havevery high
clay contentscan be leachedof basiccationsand becomeextremely acidic and,
becauseof their high buffer capacity,have very high lime requirements.
The organicmattercontentof soils also contributesto soil acidity and lime
requirement.Becausethere are differencesin the natureof soil acidity associat-
ed with organicmatterand clays(or Fe and AI oxides),the need forlime and the
meansto assesslime requirementdiffers as well. Ail of the soil acidity associat-
ed with organic matter originateson pH dependentCEC sites of organic func-
496 SIMS
tional groups such as carboxylic acid and phenols.Consequently,most studies

have shown that not only doeslime requirementin soils increasewith organic
mattercontentin general,but that in soils with high organic mattercontents,lime
requirementincreasesmore dramatically as target pH is raised than occurs in
high clay soils. One important implication of this is the much lower target pH,
usually in the rangeof pH 5.2 to 5.5, and thus lower lime requirement,normally
usedfor organicsoils. Thereare a numberof reasonsfor selectinga lower target
pH for organic soils. One practical considerationis that the amount of lime
required to changethe pH of high organic matter soils to "standard"target pH
valuesof 6.0 to 6.5 usedfor most plants can be substantialbecauseof the high
pH buffering capacity of these soils. These high lime rates have often been
shown to be unnecessaryand uneconomicalbecausecrop yields are often not
increasedin thesesoils by raising the pH beyond5.5 (McLean, 1973; McLean &
Brown, 1984; van Lierop, 1983). Causesfor this include the lower clay content
and thus lower amountof exchangeableAI3+ in organicsoils; thereforea major
reasonfor liming organicsoils to greaterthan 5.5 is minimized. Further,organic
soils often have adequateexchangeableCa, Mg, and K for most crops, even at
rather low pH values,due to their high CEC, and are also well-known to form
complexeswith solubleAI and Fe species(ionic and polymeric hydroxy AI, Fe)
that are less toxic to plants; hencethe needfor further liming (>5.5) to neutral-
ize hydroxy AI and Fe is reduced.
The importanceof clays,AI and Fe oxides,and organic matterto buffering
of soil acidity is so greatthat somelime requirementtestsare basedon soil pH
and eithersoil series(as an indicatoror clay and organicmattercontent)or some
rapid estimateof clay (hand texturing) or organic matter content(loss on igni-
tion). While theseapproacheshave beensuccessfulin many situations,they are
rather time-consumingand thus have usually beenreplacedby one of the rapid
chemicalmethodsdescribedin more detail below (e.g., buffer solutions).
Analytical Considerations
Lime RequirementMethod
A variety of techniqueshavebeenusedto estimatethe lime requirementof
acid soils (Adams, 1984; McLean, 1982; Soil & Plant Analysis Council, 1992;
van Lierop, 1990). The most commonapproachesnow usedare summarizedin
Table 17-1 and discussedin detail in "Lime RequirementMethods." In brief,
lime requirementmethodsconsistof either: (i) soil-lime incubations(ii) soil-base
titrations, (iii) soil-buffer eqUilibrations,(iv) exchangeableacidity (or AI), or (v)
estimatesbasedon soil pH and either soil seriesor somereadily measuredsoil
propertythat is well-correlatedwith soil buffer capacity.
Selectionof the most appropriatetechniqueto determinelime requirement
must begin with an understandingof the nature and variability in the forms of
soil acidity found in the dominantsoils of the geographicareawherethe test will
be used. Considerableresearchand practical experiencehas clearly shown that
different lime requirementtestsmust be usedin different physiographicregions
to attain the most accuraterecommendations. This is clearly shownin Fig. 17-1
Table 17-1. Summaryof lime requirementmethodscommonly usedin the USA (basedon a survey conductedof soil testing laboratoriesin 1995).
Lime requirement i
t"l
method Stateswhere methodis used Comments
SMP single buffer ~~~~u~~~~~~~~~~~~ Numerouslabs stressedthe importanceof properstor- ~
~
many regional, private soil testinglaboratories age of the SMP to protectagainstCO2 and water
vapor. Most midwesternstatesuse the SMP method
publishedin the NCR-13 regional soil testing man- i
ual (NCR-13, 1988)
Adams-Evanssingle buffer AL, DE, FL, GA, SC, TN DE, FL use "buffer index" basedon soil pH in water, ~
target pH of crop, and Adams-Evansbuffer pH
Mehlich single buffer NC,WV
Other single buffers 10, MS, MO-Modified Woodruff buffer Woodruff buffer also must be protectedfrom CO2 and
water vapor
Soil pH and measuredor AR-Soil water pH and CEC estimatedfrom exchangeableCa
estimatedsoil property CT-Soil pH, texture and organicmatter
IL-Soil water pH, soil type, and croppingsystem
LA-Soil pH, incrementaltitration of soil with CaC03
MD, NJ, RI-Soil pH and estimatedtexture
TX-Soil pH, crop, and texture (routine test); soil pH and 2X KCI exchange-
able AI in completesoil test
VA-Soil water pH and estimateof soil texturebasedon soil type Soil type or texture estimatedby individual submitting
sample,county agent,or consultant
VT-Soil pH in water and ammoniumacetatereactiveAI Soil pH actually measuredin 0.01 M CaCI2, then
equatedto water pH by adding0.6 pH units
Someprivate labs--Soil water pH and % Ca + % Mg saturationof CEC
Other methods NY-BaCI2-TEA titration method Lime requirementdeterminedbasedon titration to pH
8.2 and relationshipbetweensoil pH and basesatur-
ation; titrant also must be protectedfrom CO2 and
water vapor
None usedroutinely AZ, MT, NO, NM, NV, SO, UT, WY Calcareoussoils predominateand lime is rarely
needed,exceptionsmay include severelydisturbed
soils such as minespoil revegetationprojects
~
498 SIMS
.SMP o ADAMS-EVANS
c::J MEHLlCH OTHER
o NONE USED ROUTINELY
Fig. 17-1. Summaryof lime requirementmethodscommonly usedin the USA in 1995. Map based
on information collected in surveysand available in regional soil testing publications(NCR-13,
1988; NEC-67, 1995; SRlEG-18,1988).
which illustrates the predominanceof the Shoemaker-McLean-Pratt(SMP)

buffer solution (Shoemakeret aI., 1961) in the Northeastand north centralUSA,
while the Adams-Evansbuffer solution, which was developedfor low CEC, low
organic mattersoils, is most commonly usedin the Southeastand Mid-Atlantic
regions.Sincebuffer solutionsvary in chemicalcompositionby design,signifi-
cantover-or underestimates of the amountof lime requiredcan result if the inap-
propriatesoil-buffer test is selected.
Practicalconsiderations,suchas the amountof time availableto conducta
test and return a lime requirementrecommendation,the space available and
equipmentor suppliesrequired,any safetyor health hazardsthat may arise,and
the cost per sample also must be carefully evaluatedwhen selecting a lime
requirementtest. Generallyspeaking,this meansthat for routine soil testingpur-
posesa soil-buffer solution is preferred(or required),although severallarge soil
testing laboratoriesobtain acceptableresultsby basinglime requirementon soil
pH and soil seriesor soil properties.
LaboratoryProtocols
Strict adherenceto standardizedlaboratoryprotocolsis essentialto obtain
accurate,reproducibleresults with any soil testing method and is particularly
importantfor lime requirementmeasurements. Severalregional publicationsare
available that provide detailed protocols for the most commonly used lime
requirement methods in the USA [NEC-67, 1995 (Northeast Regional
CoordinatingCommitteeon Soil Testing); NCR-13, 1988 (North Central Re-
gional Soil Testing Committee); SERA-IEG-18 (Southern Extension and
ResearchActivity Information ExchangeGroup), 1988, 1992]. Analytical fac-
tors to considerwill vary with the type of lime requirementtest. In the caseof
buffer solutions,the most commonerrorsare: (i) failure to correctly prepareand
store the buffer solution; (ii) inconsistentor incorrect adherenceto the required
soillbuffer solution ratio; (iii) inconsistentor inadequatetime of reactionof the
buffer with the soil; and (iv) failure to allow adequateequilibration time for the
actual buffer pH measurement. Titration, colorimetric, or spectroscopicproce-
duresusedto measureexchangeableacidity or Al shoulduse appropriateextrac-
tion procedures(e.g., extractant,soil/solution ratio, shaking time and method,
type of filter paper),carefully standardizedtitrating solutionsand accuratelypre-
pared standards for atomic absorptionor inductively coupledplasmaspectrome-
ter (ICP) analysesof extracts.
Methodsused to determinelime requirementby soil-lime incubation and
soil-basetitrations have varied ratherwidely and a thoroughreview of the liter-
ature on this approach(Alabi et ai., 1986; Brown & Cisco, 1984; Fox, 1980;
McLean et ai., 1978; Mohebbi & Mahler, 1988; Sims & Dennis, 1989; Tran &
van Lierop, 1982) is strongly recommendedprior to initiating thesetime-con-
suming and costly studies.For lime requirementmethodsthat are basedon soil
pH and anothermeasuredsoil property (most commonly organic matter), it is
essentialthat the analytical methodsselectedreflect the proceduresused in the
original calibration studies that determinedthe relationship between the soil
propertiesand the amountof lime required(Keeney& Corey, 1963; Tran & van
Lierop, 1981). For example,if lime requirementrecommendations are basedon
soil pH and organic matter content estimatedby the Walkley-Black method, it
would be inappropriateto use a loss-on-ignition (LOI) method to determine
organicmatterunlessadditionalcorrelationor calibrationresearchwas conduct-
ed to verify the reliability of LOI as a similar indicator of soil buffer capacity.
LIME REQUIREMENT METHODS
Field Estimationof Lime Requirement
The most accuratemeansto determinethe lime requirementof a soil is

through a field study. Typically, field studiesinvolve adding increasingratesof
the desiredliming materialto the soil using commercial-scaleapplicationequip-
ment, allowing thelime to react foran appropriateperiodof time underthe envi-
ronmentalconditionsrepresentativeof the geographicareaof interest,and then
measuringthe changein soil pH at eachlime rate (Doerge& Gardner,1988).The
lime requirementof the soil can then be directly determinedfrom the resulting
lime response curve (plot of final soil pH vs. rate of liming materialadded).The
time and expenserequiredto conductfield studiesnormally precludesthe useof
a wide rangeof soils, liming materials,and liming situations(e.g.,crop rotations,
tillage and soil managementvariations). Becauseof this, field studiesof lime
requirementare obviously not suitable for routine soil testing programswhere
the goal is to rapidly predict the lime requirementfor a large and diversenumber
of soils. In general,field studiesof lime responsehave been conductedrather
infrequentlyand are usually doneto evaluatethe relative efficiency of a new lim-
ing material,to quantify the effect of somesoil or crop managementpracticeon
the overall effectivenessof liming, or to serve as final verification for lime
500 SIMS
requirementtestsdevelopedby someof the morerapid methodsdescribedbelow.

Field studies,however, are an important componentof lime and soil pH man-
agementand shouldbe conductedwheneverpossible,particularly as significant
changesin soil managementoccur (e.g., widespreadconversionto no-tillage
from conventionaltillage) or if alternativeliming materials(e.g., lime stabilized
sewagesludge)begin to be usedwidely throughouta region.
Soil-Lime Incubations
Soil-lime incubation studiesare the most commonmethodusedto charac-

terize the lime requirementsof soils with differing physical and chemicalprop-
erties and, to a lesserextent, to assesshow long-term changesin soil manage-
ment may affect lime requirement.The accuracy of most of the rapid lime
requirementtestsdescribedbelow (e.g.,buffer solutions)was initially verified by
using soil-lime incubationstudies.The methodologyusedto conducta soil-lime
incubation study is similar to that in a field study, although the smaller scale
(greenhouse,laboratory)involved allows for the reasonablyrapid evaluationof
many more soils. In brief, thesestudiesinvolve mixing increasingratesof the
liming materialwith a fixed weight or volume of soil, equilibratingthe soil-lime
mixture in a moist state for severalweeks or months either in a laboratory or
greenhouse,and developing a lime-responsecurve basedon the resultantpH
changes.The amount of lime required to effect the desiredpH changeis then
relatedback to the propertiesof the soils or to other soil measurements,suchas
the depressionin pH of a chemical buffer solution to which the soil has been
added,thus providing a quantitative basis for lime recommendationsby these
more rapid methods.
While incubationstudiesare ratherstraightforward,there are a numberof
importantfactors to considerwhen designinga soil-lime incubationexperiment.
Among theseare the type andfinenessofliming materialto be used(e.g.,reagent
gradeCaC03 vs. standardagriculturallime), the useof wetting and drying cycles
to more closely approximatefield conditions,the most suitabletemperature,and
the appropriatelength of time to conductthe incubation.Additionally, because
microbial activity is often stimulatedwhensoils are incubatedunderwarm, moist
conditions,salts can accumulatein the incubatingsoils, particularly nitratesof
Ca, Mg, and K. Sinceexcessivesolublesaltsare well-known to decreasesoil pH,
it is usually desirableto leach excesssaltsfrom soils prior to determiningpH at
the conclusionof the incubation.
Soil-BaseTitrations
Another methodto estimatethe amountof lime requiredto neutralizesuf-

ficient soil acidity to attaina desiredsoil pH is to titrate (or equilibrate)a soil sus-
pensionwith a basic solution, such as Ca(OH)z or NaOH (Alley & Zelazny,
1987; McLean et aI., 1978). Since soils are similar in many respectsto weak
acids, a properly conductedsoil-basetitration should provide an accuratemea-
sure of both active and potential acidity and thus be well-correlatedwith lime
requirement.Severalprocedureshavebeendevelopedto conductsoil-basetitra-
UME REQUIREMENT 501
tions, but most havethe following featuresin common:(i) the soil is suspended
in a relatively concentratedsalt solution, suchas 1 M KCI, to displacesomeof
the less readily accessibleforms of nonexchangeable acidity associatedwith
clays, oxides and organic matter; (ii) a basic solution is addedeither through
direct titration or by equilibratingseparatealiquotsof the soil-salt mixture with
increasingincrementsof basefor severaldays. Direct titration is often a rather
slow and tediousprocedurebecauseof the time requiredfor nonexchangeable
forms of soil acidity to reactwith the base,hencethe latter methodis often pre-
ferred; (iii) soil pH is measuredafter sufficient time haspassedto allow for equi-
libration of the soil and the basicsolution. Assessinglime requirementby soil-
basetitrations also requiresthat appropriateconversionfactorsto converttitrat-
able acidity into units of practicalvalue (e.g., Mg lime ha-1) be developedand
verified, usually though soil-lime incubationstudies.Recentadvancesin titri-
metric instrumentationhavemadeit possibleto assesslime requirementthrough
rapid (e.g., minutes) soil-basetitrations. Few routine soil testing laboratories
haveadoptedrapid titration methodsat presentbecauseof the practicaldifficul-
ties of integratingtheseproceduresinto the laboratoryand the lack of adequate
calibrationdatabetweentitratableacidity and actualfield lime requirementval-
ues.
Soil-Buffer Equilibrations
All of the proceduresto measurelime requirementdescribedso far have

significant limitations that preventtheir use by routine soil testinglaboratories.
This doesnot in any way diminish the role of field lime responsestudies,soil-
lime incubations,andsoil-basetitrationsin the overall developmentof lime man-
agementprograms.Thesemethodshavecontributedgreatlyto our understanding
of lime reactionsin soils and are the foundationof the more rapid and inexpen-
sive proceduresneededto provideroutineliming recommendations. However,as
clearly seenin Fig. 17-1, the mostwidespreadapproachto assesslime require-
ment in the USA is throughthe useof soil-buffer equilibrations.
Soil-buffer equilibrationsare conductedby adding a buffered chemical
solution to a soil sample,allowing the soil and buffer to equilibratefor a rela-
tively short period of time (e.g., 15-30 min), and then measuringthe pH of the
soil-buffer mixture (McLean, 1978).The buffer solution containsa mixture of a
weakacid anda salt of the sameweakacid andthuscanneutralizeboth acidsand
bases,resulting in a strong tendencyfor the buffer to resist markedchangesin
pH when soil is added.The pH of most buffer solutionsdecreasesin a linear
manneras the acidity in the soil reactswith the chemicalbuffering agentsin the
solution.The decreasein buffer pH is a measureof the amountof soil acidity that
mustbe neutralizedby liming to raisethe soil from its presentpH to the desired
pH. Soil-buffer equilibrationshave the advantageof allowing for a slower neu-
tralization of soil acidity at a lower and more constantpH than can easily be
accomplishedin soil-basetitrations whererapid additionsof basicsolutionscan
causetemporaryincreasesin pH well abovethat encounteredin normal liming
situations.The chemicalpropertiesof buffer solutionsvary by designaccording
to the propertiesof the soils wherethe buffer will be used.For example,the two
S02 SIMS
most commonbuffers now usedin the USA are: (i) the SMP buffer (Shoemaker
et aI., 1961), developedand usedon soils with lime requirements>4.5 Mg ha-1,
soil pH valuesof <5.8 and organic matter contents<10%; and (ii) the Adams-
Evansbuffer (Adams& Evans,1962),designedto measurelime requirementson
low organicmatter,low CEC soils suchas thosecommonto the southeastern and
mid-Atlantic regionsof the USA.
Both single and double buffer methodshave beendevelopedand evaluat-
ed for a wide range of soils. Double buffer approacheshave the advantagesof
providing very soil-specific lime recommendationsbut are not commonly used
by routine soil testinglaboratoriesbecausethey require more time, reagents,and
more complex,albeit not particularly difficult, mathematicalinterpretations.For
this reasonthe emphasisin this chapter is on single buffer lime requirement
methods.Details on double buffer methodsare available in appropriaterefer-
ences(McLean, 1982; McLean et aI., 1977, 1978; Yuan, 1974, 1976).
Shoemaker-McLean-Pratt
Single ButTer Method
Principles.The SMP buffer methodis basedon the relationshipbetween
soil-buffer pH measurementsand the lime requirementoriginally determined
from a soil-lime (CaC03) incubationstudy with 14 acidic Ohio soils (Shoemaker
et aI., 1961).The SMP hasan initial pH of 7.5 and is intendedfor soils with lime
requirements>4.5 Mg ha-l, pH values<5.8 and organic matter contents<10%
(McLean, 1982).If the soil-buffer pH exceeds6.9, indicative of a soil with a low
lime requirement, the predictive accuracy of the SMP buffer decreases.
Similarly, in soils with very high organicmattercontentsthe decreasedreactivi-
ty of H+ with the buffer causeserrors (underestimates)in the lime requirement
predictedby the SMP buffer. Becauseof this a separatecalibrationtable and tar-
get pH (5.2) was developedfor useof the SMP single buffer methodwith organ-
ic soils (McLean, 1982).
McLeanet al. (1977) developedthe SMP doublebuffer methodto improve
the predictive accuracyof the SMP for soils with low lime requirementvalues.
However, van Lierop (1990) noted the inconvenienceand additional cost of
using double buffer pH methodsand proposeda modified calibrationtable for
the SMP single buffer that was designedto improve predictive accuracyat low
lime requirementvaluesand to indicatethe amountof lime requiredto attain tar-
get pH valuesof 5.5, 6.0, 6.5, and 7.0. The modified calibrationtable was based
on a numberof lime requirementstudies(McLean, 1973, 1982; Soon & Bates,
1986; Tran & van Lierop, 1981, 1982) and reflects the curvilinear relationship
betweenSMP soil-buffer pH valuesand incubationlime requirements,particu-
larly importantfor soils with low lime requirementvaluesand/or low target pH
values.
Equipment and Reagents

1. pH meterequippedwith glassand referenceelectrodes.
2. Automatic pipette.
4. Standardbuffers, pH 7.0 and 4.0.
5. Beakersor papercupsfor pH measurements.
Preparationof SMP Buffer

1. Weigh and place in an 18-L bottle: (a) p-nitrophenol (HO • C6H4 •
N02), 32.4 g; (b) potassiumchromate(K2Cr04), 54.0 g; (c) calcium
chloride dihydrate(CaCI2 • 2H20), 955.8 g.
2. Add approximately9 L of deionizedwater. Shakevigorously as the
water is added,and continueshakingfor a few minutesto preventfor-
mation of a crust over the salts.
3. Weigh 36.0 g of calcium acetateCa(OAc)z into a separatecontainer,
and dissolvein approximately5 L of deionizedwater.
4. Add solution from stepNumber3 to that from stepNumber2, shaking
as they are combined.Shakecombinedsolutionsevery 15 to 20 min
for 2 or 3 h.
5. Add 45 mL of triethanolamine(TEA), againshakingas the addition is
made.
6. Shakethe mixture periodically until it is completely dissolved.This
takesabout8 h.
7. Dilute to a final volume of 18 L with deionizedwater.
8. Adjust to pH 7.5 ± 0.02 with 4 M NaOH or 4 M HCI using the stan-
dardizedpH meter.
9. Filter through a fiberglasssheetor cotton mat.
10. Verify buffer capacityby titrating 20 mL of SMP buffer from pH 7.50
to pH 5.00 with standardized0.1 M HCl. This should take 0.28 ±
0.005 cmole HCI/pH unit.
11. Connectan air inlet with a 2.5- by 30-cm cylinder of drierite, a 2.5- by
30-cm cylinder of ascarite,and a 2.5- by 30-cm cylinder of drierite in
seriesto protect againstcontaminationof the SMP buffer with CO2
and water vapor.
Procedure
1. Standardizethe pH meteras describedin Chapter16 (Thomas,1996,
"Standardizationof the Meter"). Note that since the SMP buffer is
adjustedto pH 7.5 and is protectedfrom contaminationas indicated,
its pH changesvery little from day to day. Hence,it canbe usedas sec-
ondary standardfor checkingproperoperationof the pH meter.
2. Weigh or measure5 g of air-dry, screened«2 mm) soil into 28-g (1-
oz) paper cups, add 5 mL of deionizedwater, mix or stir, let stand,
insert electrodes,and read soil pH in water (or 0.01 M CaCI2), as
described in Chapter 16 (Thomas, 1996, "Fundamentalsof pH
Measurement").
3. Add 10 mL of the SMP buffer solution to the soil suspensions after pH
valuesin deionizedwater (or 0.01 M CaCI2) are measured.
4. Placethe tray in a mechanicalshaker.The top of the shakershould be
fitted with a rubbersheetstretchedover rubberfoam, so that when the
top is closed and fastened,the cups are renderedairtight. Close the
cover tightly, and shake at 250 oscillations/min for 10 min. When
shakingis complete,a few swipeswith a cleanspongecleansthe rub-
ber sheet.
S04 SIMS
5. Openthe lid of the shakerand let the suspensions

stand30 min before
determiningthe pH. An alternatechoiceis 15 min of shakingand 15
min of standingtime beforethe pH is read(McLean,1975).The times
of shakingand standingare of prime importance,but the intensity of
the shaking,suchasat 250 to aslow as200 oscillations/min,may have
little effect on the soil-buffer pH reading.
6. Stir the soil suspensionby swirling the electrodesas describedfor
determiningsoil pH. Readthe pH on the standardizedpH meterto ±
0.01 pH units. Recordas soil-buffer pH.
7. Selectand record the amountof lime requiredto bring the soil to the
pH you chooseto lime the soil, basedon the soil-buffer pH of Table
17-2.
Comments. The SMP method was designedfor soils with high lime
requirementsand appreciablequantitiesof exchangeable AI and is lessaccurate
when usedon soils with very low lime requirementvalues. For soils such as
theseit is bestto useeitherthe double-bufferSMP methoddescribedby McLean
Thble 17-2. Calibrationsto detenninelime requirementof the surface20 em of soil using the SMP
single-buffermethod(McLean, 1982).
Quantity of liming material(in Mglha-I ) requiredto reachdesiredpH
Mineral soils Organicsoils

Soil-buffer pH 7.0 7.0 6.5 6.0 5.2
Pure Ag-ground Iimestonet
Caco3
6.8 2.4 3.2 2.7 2.3 1.5
6.7 4.1 5.3 4.7 3.8 2.9
6.6 5.3 7.6 6.5 5.3 4.0
6.5 7.0 10.1 8.5 7.0 5.3
6.4 9.0 12.3 10.5 8.5 6.5
6.3 10.5 14.6 12.3 10.1 7.8
6.2 12.1 16.8 14.3 11.6 9.0
6.1 13.4 19.2 16.1 13.2 10.3
6.0 15.2 21.5 18.1 14.8 11.4
5.9 17.2 23.8 20.1 16.3 12.8
5.8 18.6 26.2 21.9 17.9 13.9
5.7 20.1 28.5 23.9 19.5 15.0
5.6 21.8 30.6 26.0 21.0 16.3
5.5 23.3 33.2 28.0 22.8 17.5
5.4 25.3 35.4 30.0 24.4 18.8
5.3 26.7 37.8 31.8 26.0 19.9
5.2 28.5 40.1 33.8 27.6 21.0
5.1 30.2 42.5 35.8 29.1 22.4
5.0 31.8 44.8 37.8 30.6 23.5
4.9 33.6 47.2 39.9 32.3 24.7
4.8 34.9 49.5 41.6 33.8 26.0
tAg-Ground lime of >90% total neutralizingpower (fNP) or CaC03 equivalent,and finenessof
40% < 0.15mm,50% < 0.25 mm, 70% < 0.85 mm, and 95% < 2.36 mm. Valuesare basedon the
amountof lime requiredfor the top 20 cm of soil. For other depths,increaseor decreasethe val-
ues in table proportionateto the deviation in soil depth from 20 em. The amountsof pure Caco3
requiredto bring the soil to pH 7.0 aregiven aspointsof referencein caseonewishesto useanoth-
er gradeof lime than Ag-Ground lime.
LIME REQUIREMENT 50S
(1982), the modified calibration table of van Lierop (1990) (Table 17-3) or
anotherlime requirementmethod.
Adamsand EvansSingleButTer Method

Principles. The Adamsand Evansbuffer was developedin the southeast-
ern USA andwasdesignedto be a rapid procedureto determinethe lime require-
ment of soils with low CEC, small amountsof 2: 1 type clays, and low organic
mattercontents(Adams& Evans,1962). Soils suchas thesewill thus havelow
lime requirementsand be susceptibleto overliming that may negatively affect
plantgrowth andyields. It is basedon the generalrelationshipbetweensoil water
pH and the degreeof baseunsaturationof Ultisols, the ability of the Adams-
Evansbuffer mixture to indicateexchangeacidity, and a computationof acid to
be neutralized(Le., the lime requirement)from exchangeacidity divided by the
degree of initial base unsaturation,which is then multiplied by the desired
changein basesaturation(Adams& Evans,1962;Soil & PlantAnalysisCouncil,
1992).In this approachsoil pH is usedto estimateacid saturationof the soil (H-
Satt) at the currentpH from the relationship
Soil pH =7.79 - 5.55 (H - sat1) + 2.27 (H - satl)2
Table 17-3. Relationshipsbetweensoil-SMP-bufferpH and lime requirementvaluesto achievepH

5.5,6.0,6.5,and7.0 of mineralsoils. Calibrationdatain this tablerepresentmodified lime require-
ment valuesdevelopedfor the SMP buffer by van Lierop, 1990.
Quantity of liming material(in Mg/ha-t ) requiredto reachdesiredpHt
Soil buffer 5.5 6.0 6.5 7.0

pH
6.9 0.5 0.6 0.7 0.9
6.8 0.6 1.0 1.2 1.5
6.7 0.7 1.4 1.8 2.2
6.6 0.9 1.8 2.5 2.8
6.5 1.2 2.3 3.3 3.6
6.4 1.6 2.9 4.0 4.4
6.3 2.0 3.5 4.9 5.2
6.2 2.5 4.2 5.7 6.0
6.1 3.1 4.9 6.6 7.0
6.0 3.8 5.6 7.5 8.0
5.9 4.5 6.5 8.5 9.0
5.8 5.3 7.3 9.5 10.0
5.7 6.1 8.2 10.5 11.2
5.6 7.0 9.2 11.6 12.4
5.5 8.0 10.2 12.7 13.6
5.4 9.1 11.3 14.0 14.9
5.3 10.2 12.4 15.0 16.2
5.2 11.4 13.6 16.2 17.6
5.1 12.7 14.8 17.5 19.0
5.0 14.0 16.1 18.8 20.4
4.9 15.5 17.4 20.1 22.0
t Lime requirementin megagramsof CaC03 ha-i for a furrow layer of 20-cm depthof soil.
506 SIMS
The samerelationshipis used to calculatethe acid saturationof the soil at the

desiredpH (H-sat2). The Adams-Evansbuffer solution is then usedto estimate
exchangeacidity (soil-H) basedon the fact that each 0.008 mmolc of acidity
decreasesthe pH of 20 mL of a 1:1 buffer/watermixture by 0.01 pH units, a lin-
earchangebetweenpH 7.0 and 8.0
Soil H (cmoIJL) =8 (8.00 - buffer pH).
The amountof acidity to be neutralizedis calculatedfrom the desiredchangein

soil H-saturation
Acid to be neutralized+ Soil-H x (H-satJ - H-sat2).

H-satJ
The original interpretationof the Adams-Evansbuffer includeda correctionfac-

tor of 1.5 basedon the premise that agricultural limestone is not completely
effective in neutralizingsoil acidity. The data in Table 17-4 are basedon target
pH valuesof 6.0 (targetpH most commonlyfound in stateswhereAdams-Evans
is now used)or 6.5 (original interpretation)andwere generatedbasedon the fol-
lowing relationshipwhich includesthis correctionfactor
CaC03 (Mglha-J) =8000(8.00-buffer pH) x (H-satJ- H-sat2) x (1.5)x (2.24)

H-satJ
Table 17-4. Calibrationsto determinelime requirementof the surface20 em of soil usingthe Adams
and Evansbuffer method(Adams& Evans,1962).
Soil pH in Adams-Evansbuffer solution
Soil pH in water 7.8 7.6 7.4 7.2 7.0
Ag-ground limestone
TargetpH in water =6.0
5.9 0.2 0.4 0.5 0.7 0.9
5.7 0.5 1.0 1.4 1.0 2.4
5.5 0.7 1.5 2.2 2.9 3.7
5.3 1.0 1.9 2.9 3.8 4.8
5.1 1.2 2.3 3.4 4.6 5.7
4.9 1.3 2.6 4.0 5.3 6.6
4.7 1.5 3.0 4.5 6.0 7.5
4.5 1.7 3.3 5.0 6.7 8.4
TargetpH in water =6.5
6.3 0.5 1.0 1.5 2.0 2.5
6.1 0.9 1.7 2.6 3.5 4.4
5.9 1.2 2.3 3.5 4.7 5.9
5.7 1.4 2.8 4.3 5.7 7.1
5.5 1.6 3.3 4.9 6.5 8.1
5.3 1.8 3.6 5.4 7.2 9.0
5.1 2.0 3.9 5.9 7.9 9.8
4.9 2.1 4.2 6.3 8.4 10.5
4.7 2.2 4.5 6.7 9.0 11.2
4.5 2.4 4.8 7.2 9.6 12.0

1. pH meterequippedwith glass(indicating) and referenceelectrodes.
2. StandardpH buffers: pH 7.0 and 4.0.
3. Automatic stirrer.
Preparationof Adams-EvansBuffer
1. Dissolve,in a 1-L volumetricflask, 74 g of KCI in approximately500
mL of deionizedwater.
2. Add 10.5 g of KOH to the flask and stir to bring into solution.
3. Add 20 g of p-nitrophenol(HO • CJl4 • N02) and continueto stir.
4. Add 15 g of boric acid (H3B03).
5. Stir the entiremixture, heatingasnecessary,until completedissolution
of all reagentshas occurred.Cool and dilute to a fmal volume of 1L
using deionizedwater.
6. Adjust to a fmal pH of 8.0 ± 0.1 using either KOH or HCI solutions.
Procedure
1. Weigh 10 g of air-dry, sieved«2 mm) soil into a 50-mL beaker(or
other suitablevessel),add 10 mL of deionizedwater and mix briefly
with a glassrod. Let standfor 10 min and measuresoil-waterpH on
standardizedpH meter by the proceduresoutlined in Chapter 16
(Thomas,1996,"Fundamentalsof pH Measurement").
2. Add 10 mL of Adams-Evansbuffer solution to the soil-watermixture
from Step 1. Stir intermittently for 10 min, let standfor 30 min, and
readsoil-bufferpH to the nearest0.01 pH unit. Stir the soil suspension
just prior to measuringpH.
3. Selectthe lime requirementfor a target pH of either 6.0 or 6.5 from
Table 17-4.
Comments. The Adamsand Evansmethoddetectsextremelysmall differ-

encesin lime requirementvalueswhere such differencesmay effect relatively
large changesin soil pH dueto very low CEC (buffer capacity)of the soils. It is
most commonly usedin the Southeasternand Mid-Atlantic regionsof the USA
(Fig. 17-1) and is best suited for soils with a maximum soil H contentof 8.00
cmolJkg (Adams, 1984). Some studies, however, have shown that while the
Adams-Evanslime requirementis well correlatedwith soil-lime incubationval-
ues, it has a tendencyto overestimatelime requirement,perhapsdue to the ini-
tially high pH of the buffer (PH 8.0) which may include pH dependentacidity
that need not be neutralizedto achievea desiredpH (Alabi et aI., 1986; Fox,
1980; Tran & van Lierop, 1981).It also shouldbe notedthat the correctionfac-
tor proposedin the original Adams-Evanscalibration study to adjust for the
effectivenessof agriculturalgradelimestonein neutralizingsoil acidity also may
contribute to the overestimationof lime requirementnoted in these studies.
Several laboratoriesalso have developed state or region-specific regression
equationsthat usetargetpH, soil pH and Adams-Evansbuffer pH to more accu-
rately quantify lime requirementrecommendations (SRIEG-18,1988).
508 SIMS
Mehlich Single ButTer Method

Principles. The Mehlich buffer was originally basedon the amountof lime
requiredto neutralizethe permanent,"neutral-salt"exchangeableacidity found
in acid, mineral soils (Mehlich, 1976). van Lierop(1990) reviewedthe develop-
ment of this buffer and notedseveraldifferencesbetweenthe Mehlich buffer and
the SMP andAdams-Evansbuffers. First,the calibrationof the buffer was based
on achievingoptimum crop yields rather than attaining a specific, desiredpH.
Secondthe predictionof lime requirementvaluesfrom buffer pH valuesis based
on acurvilinear relationship.Finally, somequestionswere raisedabout the cali-
bration accuracywhen "target pH" valuesof 6.0 or 6.5 are desiredmost likely
becausethe basisof the original calibration was neutralizationof exchangeable
acidity, which tendsto predominatein acid soils at pH valuesless than 5.5. van
Lierop (1990) summarizedthe useof the Mehlich buffer as follows: " .. .it is par-
ticularly well suitedfor determiningthe lime requirementfor neutralizingacidi-
ty harmful to crop productivity and will generally recommendsufficient lime-
stoneto achievea pH slightly above5.5. This pH is sufficient to eliminate pos-
sible Al toxicity." Calibration data for the Mehlich buffer developedby van
Lierop (1990) for target pH values of 5.5, 6.0, and 6.5 are provided in Table
17-5.

1. pH meterequippedwith glass(indicating) and referenceelectrodes.
3. Standardbuffers, pH 7.0 and 4.0.
4. Beakersor papercups.
5. Automatic pipette.
Preparationof Mehlich Buffer
1. Two liters of Mehlich buffer are preparedby dissolvingthe following
chemicalsin about 1500 mL of deionizedwater while stirring vigor-
ously: (i) 5 mL of glacial acetic acid (CH3COOH); (ii) 9 mL of tri-
ethanolamine[TEA : N(CH2 • CH20H)3] stock solution, or, for ease
of delivery in pipetting, add18 mL of a 1:1 TENdeionizedwatermix-
ture; (iii) 86 g of NH4CI; and (iv) 40 g of BaCl2 • 2H20.
2. Separatelydissolve 36 g of sodium glycerophosphate,also named
sodium salt of beta-glycerophosphoric acid [(HOCH2)2CHOP03Na2
·5H20], in about400 mL of deionizedwater.
3. Mix solutionsfrom Steps1 and 2 while swirling vigorously. Allow the
mixture to cool to room temperature,then dilute to 2 L with deionized
water and mix thoroughly.
4. Check pH of buffer by mixing equal aliquots of buffer solution and
deionized water (e.g., 10 mL Mehlich buffer + 10 mL deionized
water).The pH ofthe 1:1 mixture shouldbe 6.60± 0.04.Adjust buffer
solution as necessarywith glacial acetic acid or TEA stock solution
until desiredpH is attained.
5. Verify buffering capacityby mixing 10 mL of the Mehlich buffer solu-
tion with 10 mL of deionizedwater and 10 mL of a 0.05 M HCI +
0.017 M AlCl 3 solution.The pH of the resultingmixture shouldbe 4.1

± 0.05. Make the 0.05 M HCI + 0.017M AlCl 3 solution by dissolving
4.024g AlCl 3 • 6H20 in 100 mL of 0.05 M HCl.
Procedure
1. Measure10 mL of dry soil into a beakeror papercup.
2. Add 10 mL of deionizedwater,stir with a glassrod to wet samplethor-
oughly, allow to standfor 30 minutes,and measurepH as describedin
Chapter16 (Thomas,1996, "Fundamentalsof pH Measurement").
3. Add 10 mL of MehIich buffer to soil-water suspensionfrom Step 2.
Stir thoroughlywith a glassrod, allow mixture to sit for 1 h, and then
measuresoil-buffer pH to nearest0.05 pH units.
4. Obtain lime requirementvaluesfor desiredtargetpH from Table 17-5.
Comments. The capacity of the MehIich buffer is 10.4 cmolc aciditylL,
approximatelyequivalentto 10.4 Mg/ha-1 of pure CaC03, henceit is similar to
the Adams-Evansbuffer in that it is bestsuitedfor soils with lower CEC values;
Table 17-5. Relationshipsbetweensoil-buffer pH and lime requirementsfor targetpH valuesof 5.5,

6.0, and 6.5 for mineral soils with the Mehlich buffer (from van Lierop, 1990).
Lime requirement
Soil buffer pH Mehlicht pH 5.5 pH 6.0 pH 6.5

Mglha-1
6.5 0.4 0 0 0
6.4 0.9 0 0.2 0.3
6.3 0.9 0 1.4 1.2
6.2 1.9 0.2 2.6 2.2
6.1 2.4 1.1 3.8 3.2
6.0 3.0 2.0 4.9 4.3
5.9 3.6 2.9 6.1 5.4
5.8 4.2 3.8 7.3 6.6
5.7 4.9 4.6 8.5 7.9
5.6 5.6 5.5 9.6 9.2
5.5 6.3 6.4 10.8 10.6
5.4 7.1 7.3 12.0 12.0
5.3 7.9 8.2 13.2 13.6
5.2 8.7 9.1 14.4 15.1
5.1 9.6 10.0 15.5 16.7
5.0 10.5 10.9 15.5 16.7
4.9 11.4 11.7 17.9 20.2
4.8 12.4 12.6 19.0 22.0
4.7 13.4 13.5 20.2 23.9
4.6 14.4 14.4 21.4 25.8
4.5 15.5 15.3 22.6 27.8
4.4 16.5 16.2 23.7 29.8
4.3 17.7 17.0 24.9 32.0
4.2 18.8 18.0 26.1 34.1
4.1 20.0 18.8 27.3 36.4
4.0 21.2 19.7 28.4 38.7
3.9 22.5 20.6 29.6 41.0
t Lime requirementcalibrationsfor Mehlich lime requirementvalueswere obtainedfrom Mehlich

(1976) basedon optimizing crop yields. Lime requirementvalues to soil pH 5.5 and 6.0 were
developedby Tran and van Lierop (1982), and to soil pH 6.5 by Ssali and Nuwamanya(1981).
510 SIMS
however,it also has beenusedsuccessfullywith Histosolsand soils with Histic

epipedons.If soil-buffer pH values near 6.6 are obtainedwith coarse-textured
soils, cautionshouldbe exercisedwhenselectinga lime rate to avoid overliming
and possible micronutrient deficiencies.The following equations weredevel-
oped to compute lime requirementrecommendationsfor mineral and organic
soils using the Mehlich buffer (Mehlich et aI., 1976):
Acidity (AC), in cmolcfL soil = (6.6 - Mehlich buffer pH)

0.25
Mineral soils (for plantswith slight to moderatetoleranceto soil acidity)
Lime requirement(Mg/ha-1) =[(0.1) x (AC)2] + [AC]

Histosolsor soils with Histic epipedon
Lime requirement(Mg/ha-1) = [-7.4 + 1.6 x (AC)] x 1.3
Other Single ButTer Methods

The Nommik single-bufferis the newestsingle buffer procedurefor mea-
suring lime requirement(Nommik, 1983). It uses imidazole, maleic acid, and
acetateas buffering componentsand was originally calibrated using a lO-wk
Ca(OH)zincubationstudy and a targetpH of 7.0. The principlesand limitations
of this buffer were reviewed by van Lierop (1990) who stated that while the
Nommik buffer was reasonablyprecise,its accuracyis uncertainbecauseof a
lack of calibration data. Further study is neededbefore widespreadadoptionof
this buffer is likely to occur.
The Woodruff buffer (Woodruff, 1948)was the first widely usedbuffer pH
method and consequentlyhas been evaluatedin numerousstudies(Fox, 1980;
Loynachan,1981; McLean et aI., 1960, 1966; Tan & van Lierop, 1981, Webber
et aI., 1987). Most found that the Woodruff buffer was reasonablyprecisewhen
higher target pH valuesare used(e.g., pH 6.5), that it was much more accurate
in soils with lower lime requirements,and that a curvilinear relationshipexisted
betweensoil-buffer pH andlime requirement.Becauseof these concerns, and the
developmentof other buffer solutions,the original Woodruff buffer (Woodruff,
1947) is not used in the USA. However, Brown et ai. (1977) and Brown and
Cisco (1984) have subsequentlymodified and evaluateda new Woodruff buffer
that is believedto be more accuratethan the original; the modified version rec-
ommendsabout 1.6 times as much lime, suggestinga more effective displace-
mentof soil acidity. van Lierop (1990) reviewedthe calibrationstudiesby Brown
and Cisco(1984) andAlabi et ai. (1986) and found that the new Woodruff buffer
lime requirementvalueswere reasonablefor soils with low lime requirements,
but again cautionedthat more calibration studieswere needed,particularly on
soils with higher lime requirements.Threestatesin the USA (ID, MO, MS) cur-
rently use the modified Woodruff buffer to make lime recommendations(Table
17-1).
ExchangeableAluminum
Principles
Researchon crop responsesto lime on highly weathered,acid mineralsoils
hasfrequently shownthat applicationof sufficient lime to neutralizethe amount
of exchangeableAI extractedwith a neutral, unbufferedsalt solution (e.g., 1 M
KCI) is adequateto eliminate soil acidity as a yield-limiting factor for many
crops. This is becausein most soils with thesepropertiesthe major factor asso-
ciatedwith acid soil infertility is toxic levels of exchangeable AI. Generallythis
approachto liming resultsin a lower soil pH thanwould be achievedif otherlime
requirementmethodswere used,typically in the rangeof pH 5.5 to 5.8. The most
widespreadadoptionof this approachto liming hasbeenin areaswherecrop pro-
duction is limited by highly acidic, aluminoussoils and inadequateavailability
of limestone. Liming soils such as these basedon KCI exchangeableAI (or
exchangeableacidity) eliminatesa major limitation to plant growth at minimal
cost.
ExchangeableAI is rarely used by soil testing laboratoriesin the USA to
makelime recommendations for severalreasons.First, from a practicalperspec-
tive, the time and expenserequiredto handleand analyzea separatesoil sample
than is usedfor a routine soil test is usually prohibitive, relative to the use of a
buffer method. Second, other factors can reduce plant growth in acid soils
besidesAI, including excessMn, low levels of exchangeablebases(Ca, Mg),
reducedmicrobial activity, and lowered efficacy of certain pesticides.In these
situationsliming soils merely to neutralizeexchangeable AI may not raise pH to
a level that optimizesplant growth and a correctionfactor to insure thatsoil pH
is adjustedto 6.0 to 6.5 may be necessary.McLean (1982) also noted that plant
toleranceto AI shouldbe consideredandsuggestedthat the lime requiredto neu-
tralize exchangeableAI would be adequatefor AI-tolerant plants, but that this
value should be multiplied by 1.5 and 2.0 for plants with medium and low AI-
tolerance,respectively.

1. Potassiumchloride (KCl), 1 M: dissolve 74.56 g of KCI in 1 L of
deionizedwater.
2. Aluminum complexingsolution (1 M KF). Dissolve58.1 g of KF in 1
L of deionizedwater and titrate to a phenolphthaleinendpoint with
standardized0.1 M NaOH.
3. Sodium hydroxide,0.1 M, standardized.
4. Hydrochloric acid, 0.1 M, standardized.
3. Phenolphthaleinsolution-dissolve1 g of phenolphthaleinin 100 mL
of ethanol.
4. 100 mL polyethylenecentrifugetubesor suitablealternative.
5. Shakerappropriateto extracting vessel(e.g., end-over-endpreferred
for centrifugetubes).
6. Centrifugeor Buchnerfiltration funnels with suction apparatus.
7. Whatmanno. 42 filter paper orequivalent.
512 SIMS
Procedures
1. Weigh 5 g of soil into a 100-mL polyethylenecentrifugetube.
2. Add 50 mL of 1 M KCI and shakefor 30 min.
3. Centrifugefor 10 min at 700 x g or allow sampleto settle and filter
supernatantthrough Buchner funnels using no. 42 Whatman filter
paperand low suction.
4. Analyze for KCI exchangeableacidity and exchangeable AI by direct
titration (seenext steps)or for exchangeableAI by appropriatespec-
troscopicor colorimetric procedures.
5. Titration procedurefor KCI exchangeableacidity: (i) transferKCI fil-
trate into suitable titration vessel; (ii) add four drops of phenolph-
thalein solution; (iii) titrate with standardized0.1 M NaOH until the
first permanentpink endpoint is observed,(Note: A deep pink end-
point indicatesovertitration); and (iv) calculatecentimolesof change
per kilogram of KCI exchangeableacidity as follows
Exchangeableacidity =
(cmolJkg)
(mL NaOH, sample-mLNaOH, blank)
x (molarity of NaOH, mmo1!mL) x (0.1)
sampleweight (kg)
6. Titration procedurefor exchangeable AI: (i) add 10 mL of KF solution

to KCI solution that was titrated in Step5; (ii) titrate with standardized
0.1 M HCl until the pink color disappears,wait 30 s, and then contin-
ue to titrate with 0.1 M HCI until a final clearendpointis reached;and
(iii) calculatecentimolesof chargeper kilogram of exchangeableAI
as follows
ExchangeableAI =
(cmoIJkg)
(mL HCI, sample-mLHCL, blank) x (molarity of HC1!mL) x (0.1)
sampleweight (kg)
7. ExchangeableH also can be determinedfrom this procedureas
ExchangeableH (cmolc kg) =

Exchangeableacidity - ExchangeableAI
8. Lime requirementfor a 20-cm soil depth is calculatedby converting

cmolJkgof exchangeable AI (or acidity) into Mglha of CaC03 as fol-
lows. Multiply this value by 1.5 or 2.0 for plants that are moderately
and highly sensitiveto AI.
Lime Requirement(Mglha) =
ExchangeableAI (or acidity) (cmolJkg)
Comments. Determinationof KCI exchangeableAi (or acidity) by direct

titration is a simple, reliable methodwith no interferences.It can readily detect
as little as 0.05 cmolJkgof exchangeable Ai (or acidity). In addition toits use as
a lime requirementtest, exchangeableacidity by this methodalso can be usedto
determine effective CEC (CEC at the pH of the soil) by summation with
exchangeablebases.Most laboratoriesthat use this approachbaselime require-
ment recommendationson exchangeableAi rather than exchangeableacidity.
However,since there is very little exchangeableH in most soils, the differences
in lime recommendationsbetweenthe two methodsare often minimal and the
use of exchangeableacidity as an index of lime requirementmay be a simpler
and equally satisfactorymethodas is exchangeableAi.
Other Lime Requirement Methods
Most routine soil testing laboratoriesin the USA use buffer methodsto
determinelime requirementbecauseof the accuracy,speed,ease, andrelatively
low cost of this approach.Further,the SMP and Adams-Evansbuffers are by far
the most commonly usedbuffer methods.However, as can be seenin Fig. 17-1
a significant numberof laboratoriesuse other methods(or buffers)to determine
lime requirement.The most commonalternativesto buffers are someestimateof
lime requirementbasedon soil pH and a measuredor recordedfactor that isasso-
ciated with soil buffer capacity. Examplesinclude soil organic matter content,
estimatedCEC, and soil series.Table 17-1 presentsa summaryof the most com-
monly usedalternativesto the SMP and Adams-Evansbuffer methodsfor lime
requirementdetermination, based on a surveyconductedby U.S. soil testinglab-
oratoriesin 1995 and on information from recently publishedregional soil test-
ing publications(NEC-67, 1995; SRIEG-18,1988).
REFERENCES
Adams, F. 1984. Crop responsesto liming in the southernUnited States.p. 211-266.In F. Adams
(ed.) Soil acidity and liming. 2nd ed. Agron. Monogr. 12. ASA, Madison,WI.
Adams, F., and C.E. Evans. 1962. A rapid methodfor measuringlime requirementof Red-Yellow
Podzolicsoils. Soil Sci. Soc. Am. Proc. 26:355--357.
Alabi, K.E., R.C. Sorensen,D. Knudsen,and G.w. Rehm. 1986.Comparisonof severallime require-
ment methods on coarse textured soils of northeasternNebraska. Soil Sci. Soc. Am. 1.
50:937-941.
Alley, M.M., and L.w. Zelazny. 1987. Soil acidity: Soil pH and lime needs.p. 65-72. In 1.R. Brown
(ed.) Soil testing:Sampling,correlation,calibration,and interpretation.SSSASpec.Publ. 21,
SSSA,Madison,WI.
Brown, J.R., J. Garett,andT.R. Fisher. 1977. Soil testingin Missouri. Univ. Missouri-ColumbiaExt.
Div. Ext. Cir. 923.
Brown, J.R., and J.R. Cisco. 1984. An improved Woodruff buffer for estimationof lime require-
ments.Soil Sci. Soc. Am. J. 48:587-592.
Doerge,T.A., and E.H. Gardner.1988. Comparisonof four methodsfor interpretingthe Shoemaker-
Mclean-Pratt(SMP) lime requirementtest. Soil Sci. Soc. Am. 1. 52:1054-1059.
Evans,C.E., and E. J. Kamprath. 1970. Lime responseas relatedto percentAl saturation,solution
AI, and organicmattercontent.Soil Sci. Soc. Am. Proc. 34:893--896.
Foy, C.D., and J.C. Brown. 1964. Toxic factors in acid soils: II. Differential aluminum toleranceof
plant species.Soil Sci. Soc. Am. Proc. 28:27-32.
514 SIMS
Fox, RH. 1980. Comparison of several lime requirement methods for agricultural soils in
Pennsylvania.Commun.Soil Sci. Plant Anal. 11:57-69.
Kamprath,EJ. 1970. Exchangeablealuminum as a criterion for liming leachedmineral soils. Soil
Sci. Soc. Am. Proc. 34:252-254.
Keeney,D.R., and RB. Corey. 1963. Factorsaffectingthe lime requirementsof Wisconsinsoils. Soil
Sci. Soc. Am. Proc. 27:277-280.
Loynachan,T.E. 1981. Lime requirementmethodsfor cold regions.Soil Sci. Soc. Am. J. 45:77-80.
McLean, E.O., H.E. Shoemaker,and W.R Hourigan. 1960. Some effects of aluminum on lime
requirementtestsof soils. p. 142-151.Trans. Int. Congr. Soil Sci., 7th. Vol. 2.
McLean, E.O. 1970. Lime requirementsof soils-inactivetoxic substancesor favorable pH range?
McLean, E.O., 1971. Potentially benenficialeffects from liming: Chemicaland physical. Proc. Soil
Crop Sci. Soc. Fla. 31:189-196.
McLean, E.O. 1973. Testing soilsfor pH and lime requirement.In L.M. Walsh and J.D. Beaton(ed.)
Soil testingand plant analysis.SSSA,Madison,WI.
McLean, E.O. 1975. RecommendedpH and lime requirement tests. p. 6-9. In Recommendedchem-
ical soil test proceduresfor the North Central Region. North Central Region. Publ. 221.
McLean,E.O. 1978. Principlesunderlyingthe practiceof determininglime requirementsof acid soils
by use of buffer methods.Commun.Soil Sci. Plant Anal. 9:699-715.
McLean, E.O. 1982. Soil pH and lime requirement.p. 199-224.In AL. Pageet al. (ed.) Methodsof
soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
McLean, E.O., and J.R Brown. 1984. Crop responsesto lime in the MidwesternUnited States.In E
Adams(ed.) Soil acidity and liming. 2nd ed. ASA, CSSA, and SSSA, Madison,WI.
McLean, E.O., S.w. Dumford, and E Coronel. 1966. A comparisonof severalmethodsof determin-
ing lime requirementsof soils. Soil Sci. Soc. Am. Proc. 30:26-30.
McLean, E.O., D.l. Eckert, G.Y. Reddy, and FJ. Trierweiler. 1978. An improved SMP soil lime
requirementmethodincorporatingdouble-bufferand quick-testfeatures.Soil Sci. Soc. Am.
J.42:311-316.
McLean, E.O., J.E Trierweiler, and OJ. Eckert. 1977. Improved SMP buffer methodfor determin-
ing lime requirementof acid soils. Commun.Soil Sci. Plant Anal. 8:667-675.
Mehlich, A 1953. Rapid determinationof cation and anion exchangepropertiesof soils and pHc of
soils. J. Assoc. Off. Agric. Chern. 36:445-457.
Mehlich, A 1976. New buffer pH method for rapid estimation of exchangeableacidity and lime
requirementsof soils. Commun.Soil Sci. Plant Anal. 7:253-263.
Mehlich, A, S.S. Bowling, and A.L. Hatfield. 1976. Buffer pH acidity in relation to nature of soil
acidity and expressionof lime requirement.Cornmun.Soil Sci. Plant Anal. 7:253-263.
Mohebbi, S., and RL. Mahler. 1988. Evaluationof lime requirementtestsfor northern Idaho soils.
NCR-13. 1988. Recommended chemicalsoil test proceduresfor the North Central region. Bull. 499.
North DakotaAgric. Exp. Stn., Fargo,NO.
NEC-67. 1995. Recommendedsoil testingproceduresfor the NortheasternUnited States.Bull. 493.
2nd ed. Univ. DelawareAgric. Exp. Stn, Newark, DE.
Nommik, H. 1983. A modified procedurefor rapid determinationof titratable acidity and lime
requirementin soils. Acta Agric. Scand.33:337-348.
Peech,M., RL. Cowan,and 1.H. Baker. 1962. A critical study of the BaCl2-triethanolamineand the
ammoniumacetatemethodsfor determiningthe exchangeablehydrogencontentof soils. Soil
Sci. Soc. Am. Proc. 26:37-40.
Reeve,N.G., and M.E. Sumner.1970. Lime requirementsof Natal. Oxisols basedon exchangeable
aluminum. Soil Sci. Soc. Am. Proc. 34:595-598.
Shoemaker,H.E., E.O. McLean, and P.E Pratt. 1961. Buffer methodsfor determinationof lime
requirementsof soils with appreciableamountof exchangeable aluminum.Soil Sci. Soc. Am.
Proc. 25:274-277.
Sims, J.T., and L. Dennis. 1989. Evaluation of lime requirement methods for Delaware soils.
Soil and PlantAnalysisCouncil. 1992.Handbookon referencemethodsfor soil analysis.Counc.Soil
Test. Plant Anal., Athens,GA.
Soon, Y.K., and T.E. Bates. 1986. Determinationof the lime requirementfor acid soils in Ontario
using the SMP buffer methods.Can. J. Soil Sci. 66:373-376.
SRIEG-18. 1988. Proceduresand practicesfollowed by southernstate soil testing laboratoriesfor
making lime recommendations.Bull. 332. Univ. Florida Inst. Food Agric. Sci., Gainesville,
FL.
SRIEG-18, 1992. Referencesoil and media diagnosticproceduresfor the Southernregion of the

United States. Bull. 374. Virginia Agric. Exp. Stn., Virginia Polytech. State Univ.,
Blacksburg,VA.
Ssali,H., andJ.K. Nuwamanya.1981.Buffer pH methodsfor estimationof lime requirementof trop-
ical acid soils. Commun.Soil Sci. PlantAnal. 12:64~59.
Thomas,G.W. 1982. Exchangeablecations. p. 87-110. In A.L. Pageet al. (ed.) Methods of soil
analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Thomas,G.w., and W.H. Hargrove. 1984.The chemistryof soil acidity. p. 3-56. In F. Adams(ed.)
Soil acidity and liming. 2nd ed. Agron. Monogr. 12. ASA, CSSA, and SSSA,Madison,WI.
Tran, T.S., and W. van lierop. 1981. Evaluation and improvementof buffer-pH lime requirement
methods.Soil Sci. 131:178-188.
Tran, T.S., and W. van Lierop. 1982. Lime requirementdeterminationfor attainingpH 5.5 and6.0 of
coarsetexturedsoils using buffer-pH methods.Soil Sci. Soc. Am. J. 46:1008-1014.
van Lierop, W. 1983. Lime requirementdeterminationof acid organicsoils usingbuffer-pH methods.
Can. J. Soil Sci. 63:411-423.
van Lierop, W. 1990.Soil pH and lime requirement.p. 73-126.In R.L. Westerman(ed.) Soil testing
and plant analysis.2nd ed. SSSA,Madison,WI.
Webber,M.D., P.B. Hoyt, and D. Comeau.1982. SolubleAI, exchangeableAI, basesaturationand
pH in relation to barley yield on Canadianacid soils. Can. J. Soil Sci. 62:397-405.
Webber, M.D., P.B. Hoyt, M. Hyborg, and D. Comeau.1977. A comparisonof lime requirement
methodsfor acid Canadiansoils. Can. J. Soil Sci. 57:361-370.
Woodruff, C.M. 1947. Determinationof exchangeablehydrogenand lime requirementof the soil by
meansof the glasselectrodeand a bufferedsolution. Soil Sci. Soc. Am. Proc. 12:141-142.
Woodruff, C.M. 1948.Testingsoils for lime requirementby meansof a buffer solution and the glass
electrode.Soil Sci. 66:5~3.
Yuan, T.L. 1974.A doublebuffer methodfor the determinationof lime requirementof acid soils. Soil
Sci. Soc. Am. J. 38:437-440.
Yuan, T.L. 1976. Anomaly and modification of pH-acidity relationshipsin the doublebuffer method
for lime requirementdetermination.Soil Sci. Soc. Am. J. 40:80(}...802.
Published 1996
Chapter18
Aluminum
PAUL M. BERTSCH, Savannah River Ecology Laboratory,

University of Georgia, Aiken, South Carolina
PAUL R. BLOOM, University of Minnesota, St. Paul, Minnesota
Aluminum is amongthe more importantand commonlyanalyzedconstituentsin

natural waters, soils, sediments,geological materials, and plant tissues, both
becauseit is an ubiquitouselementin soil and geologicalsystemsand because,
when presentin elevatedconcentrations,Al can be a powerful toxicant to plants
and aquaticorganisms.Minerals containingsignificant quantitiesof Al are the
aluminosilicateswhich include the feldspars,micas,kaolins, smectitesand most
other phyllosilicate minerals. Aluminum also is a primary componentof other
nonsilicatemineralsthat occur in soils and geologicalmaterials,including such
minerals as gibbsite [Al(OH)3], variscite [AlP04 • 2H20], and Al sulfateslike
alunite [KAl 3(OHMS04)2] or basaluminite[Al4(OH)lOS04• SH20].
Chemicallyactive or labile soil Al can havea variety of forms that are con-
trolled primarily by thepH and the mineralogical composition of the system.For
example,Al can be bound to negatively chargedclay surfacesby electrostatic
forces and thus can be freely exchangeablewith other cations such as Ca2+,
Mg2+, or K+, or it can be bound to carboxylateand phenolicgroupson organic
matterandbe only partially exchangeable. In addition,Al canbe presentas non-
crystalline hydroxidesand oxides and as complex hydroxopolynUclearcompo-
nentswhich occupy the interstitial spacesof 2:1 clay mineralsand be partially
soluble in neutral salt extracts.In strongly acidic soils (pH < 4.0), hexaaquaAl
ions are the predominantexchangeablespecies.As the pH is increased,the
amount of monomeric hexaaquaAl(H20)~+ decreaseswith the formation of
hydroxo-Al complexes.Thesehydroxo speciesmay be comprisedof the simple
mononuclearions of the generalform Al(H20)6-n(OH)~-n)+, polynuclearspecies
of various sizesand degreesof basicity or, more probably,of a complex admix-
ture of both. If the pH is raisedhigh enough,Al precipitatesas the trihydroxide
mineral phase,either crystallinegibbsite or a poorly orderedanalogue.At pH >
8, the amphotericnatureof Al is expressedwith the formation of the aluminate
ion Al(OH)4".
Book Seriesno. 5.
517
518 BERTSCH& BLOOM
Becauseof its complexchemistry,its ubiquitouspresencein most soil and

geologicalmaterials,and becauseof the dramaticchangesin its solubility with
pH in the rangecommonto many naturalsystems,Al is ratherunique in func-
tioning as both a traceand abundantelement.For example,Al concentrationsin
acidic surfacewatersassociatedwith acid mine drainagecan reachconcentra-
tions of 50 mg L-1 or greater,while Al concentrationsin surfacewater or soil
solutionsin systemsof circumneutralpH can be in the low microgramsper liter
range.
The role of Al in soil acidity is discussedelsewhere(Thomas,1996; Sims,
1996;seeChapters16 and 17). Aluminum concentrationsin acid soils or surface
watersmay be sufficiently high to be toxic to manyplantsandaquaticorganisms.
Aluminum determinationin soil solutions,surfacewaters,and various extracts
of soils, sludges,andgeologicalmaterialsare often madein studiesrelatedto Al
mobility andtoxicity, andmany methodshavebeenused.Aluminum determina-
tion alsois importantin the studyof mineralweatheringandpedogenesis. For all
of theseapplications,the rangeof Al concentrationscan be quite large and the
matricesextremelyvariable. At elevatedconcentrationsand in the absenceof
complexmatricesor interferingsolutes,it is relatively simpleto accuratelydeter-
mine Al by severalmethods.At low Al concentrationor in the presenceof some
interfering ions commonlyfound in naturalsystems,the determinationof Al is
often fraught with difficulties. In theseinstances,samplehandlingis critical and
methodsmust be usedthat separateAl from the interferingsolutesso that more
preciseand accuratemeasurements can be made.Another factor that can com-
plicate the analytical determinationof aqueousAl is the rangeof Al chemical
speciesin solution. Many inorganicand organiccomplexesof AI are not detect-
ed effectively by someanalyticaltechniques.This can be consideredas interfer-
encein the detectionof total solubleAI. On the other hand, many methodsare
designedto provideinformation regardingthe distributionof Al betweenits var-
ious complexedforms and often attemptto capitalizeon differential kinetics or
incompletereactivity.
In this chapter,we discussthe pertinentaspectsof the various methods
which are commonlyusedto exchange,extract,or solubilize a given fraction of
solid phaseAl andof the methodscommonlyusedto determinetotal aqueousAI
or variouschemicalspecies.We haveattemptedto provide the appropriate back-
groundinformationso that investigatorscanchoosethe bestmethodor methods
for their specificapplications.
TOTAL ALUMINUM
Introduction
Total aluminumis often determinedon soils and sedimentsbecauseit pro-

videsuseful informationon the characterizationof soils with respectto origin of
parentmaterialsand weathering,and it servesas a basisfor computingthe min-
eralogicalcompositionof the sample.Traditionally, total dissolution has been
performedby fusion at high temperaturewith Na2C03in platinum crucibles,fol-
ALUMINUM 519
lowed by dehydrationof silica with HCI04 and dissolutionof the otherelements

in HCI. Another commonlyusedmethodhasbeendigestionin HF-HCI04 mix-
tures; however,this procedureis potentially dangerousand requiresa specially
constructedfume hood.
More recent proceduresthat are less laborious and more rapid involve
eitherthe fusion in sodiumtetraboratein graphitecruciblesor digestionin sealed
containersat elevatedtemperaturesusingoxidizing acid mixturescontainingHE
Digestion in HF mixtures requiresthe use of polymer containersstableat high
temperature(e.g.,Teflon) that canbe sealedto allow for heatingto temperatures
in excessof 100°C and to containSF4 gasgeneratedin dissolutionof silicates.
Heatingcan be either in a conventionaloven (Bernas,1968),or in a microwave
oven (Warrenet aI., 1990) if a metal jacket is not usedfor the container.When
the containersareopened,additionof boric acid is necessaryto tie up F- and pre-
vent loss of SiF4• With the boratefusion methodin graphitecruciblesthe fused
materialis in the form of a pellet that is easilydissolvedin dilute acid (Meyer &
Bloom, 1993). Aluminum and other elementscan be determineddirectly using
atomicabsorption(AA) spectrometryor plasmaemissionspectrometry,eitherby
inductively coupledplasma(ICP) or direct currentplasma(DCP), (seeAtomic
Absorption and Inductively Coupled PlasmaOptical Emission Spectrometry)
andwith properadjustmentof pH andremovalof interferingsubstances by spec-
t.rophotometric techniques(see SpectrophotometricMethods fo~ Aluminum
Analysis).For somespectrophotometric methods,interferencemay be a problem
for soils that are high in Fe.
Principles
The proceduredescribedin Lithium MetaborateFusionis for the fusion of

the samplewith LiB0 3 in graphitecruciblesfollowed by dissolution in dilute
HCI. Aluminum and other elementscan be determineddirectly using plasma
emissionspectrometry,eitherby ICP or DCP, or with AA. The acid usedfor dis-
solutionof the fusedmaterialcanbe varied accordingto the type of matrix solu-
tion preferredby the analyst.Meyer and Bloom (1993) used2 M HCL and ICP
analysisfor plant ashsampleswhile Cantillo et al. (1984)used5% nitric acid and
DCP analysisfor sediments.In analyticallaboratorieswhereemissionspectrom-
etersalso are usedfor analysisof B, high boratein the analytical solutionscan
cause·a buildup of B in the sampledelivery system.This may requirea cleanup
procedureafter analysisof boratefusion samples.Spectrophotometricanalysis
using Al complexingagentsalso can be usedfor total Al analysisif the pH is
properly adjusted.For somespectrophotometric methods,interferencemay be a
problemfor soils that are high in Fe.
Poor fusion can result in low results(Cantillo et aI., 1984). Good fusion
requiresthat the soil samplebe in contactonly with the LiB0 3 and not contact
the graphite.This can be accomplishedby placingthe samplein a small well of
boratein the crucible. The cruciblesdo lose about 10% of their weight during
fusion, and flakes of graphiteare typically found after dissolutionof the fused
sample.Thesegraphiteparticlesreadily settleto the bottomanddo not causeany
difficulties during analysis.The cruciblescan be reuseduntil they becometoo
520 BERTSCH& BLOOM
fragile. If samplesare high in silica, the solubility of silica in acid may be

exceededand a precipitatewill form. The precipitationof silica is readily visible
and the ratio of acid volume to samplesizecan be increasedto eliminatethe pre-
cipitation.
Alternative proceduresfor digestion of soil materials involve the use of
Parr bombsor a microwaveoven to heat the samples;the Parr bomb procedure
is presentedin the sectionon Reagents.
Fusionand DissolutionMethodsfor Total Aluminum
Lithium MetaborateFusion
Special Apparatus
1. lO-roL graphitecrucibles(SpexIndustries).
2. Stainlesssteel trays 14 gauge (2.11 rom thickness)with edgesbent
upward to provide for a I-em borderto preventcruciblesfrom sliding
off.
3. 250-mL polyethyleneor polypropylenebottles.
4. Muffle furnace.
5. Long handledtongs(40-50 cm).
6. Leathergloves.
Reagents
1. Lithium metaborate(LiB03)'
2. Hydrochloric acid (2 M).
Procedure.Destroy the organic matter using either H20 2 or NaCI04
(Sawhney,1996, seeChapter36) and grind to pass200 mesh«75 Jlm). Weigh
0.5 g of LiB0 3 in graphitecrucibles. Weigh100 mg of sampleto the nearestmii.-
ligram and place in a small depressionin the borate.The sampleshould not be
in contactwith the graphite.Placecruciblesonto a stainlesssteeltray (a 14- by
28-em tray holds about 20 crucibles). With long-handled tongs and leather
gloves,placethe tray in a 950°C muffle furnacefor 15 min or until the furnace
temperaturereturnsto 950°C (whicheveris longer). Removethe tray and place
on a heat-resistant slab to cool. Pourthe smallsphericalbeadto fusedLiB0 3 into
100-mL polyethyleneor polypropylenebottle containing50.0 mL of 2 M HCI.
Pouringthe molten LiB0 3 as suggestedby Cantillo et al. (1984) is difficult, dan-
gerous,and not necessary.The bottlesshouldbe cappedand placedon a shaker
for 2 h. If silica remains undissolved in the bottom of the bottle, a largervolume
of acid shouldbe used.The following referencescan be consultedfor additional
information: Suhr and Ingamells,1966; Cantillo et aI., 1984; Meyer and Bloom,
1993.
HydrochloricAcid-Nitric Acid-Hydrotlouric Acid

Special Apparatus
1. Parracid digestionbombswith stainlesssteeljackets.
2. Oven capableof heatingto 130°C.
ALUMINUM 521
Reagents
1. Aqua regia (1:1 mixture of concentratedHN03 and HCI).
2. Hydroflouric acid, 48%.
3. Boric acid.
Procedure.Grind to pass150 mesh«100/lm) and weigh 100 mg of sam-
ple to the nearestmilligram and placein a Teflon reactionvessel.Add 1.0 mL of
aquaregia and let standovernight.Add 3.0 mL of HF, insert reactionvesselinto
a stainlesssteel jacket, securetightly and place in an oven at 130°C for 1 h.
Transfersolution to a 250-mL polyethyleneor polypropylenebeakerusing dis-
tilled water. Add 2.8 g of H3B03 and stir with a polyethyleneor polypropylene
stirring rod. Addition of water may be necessaryto completely dissolve the H3
B03. Transferto a 100 mL volumetric flask and dilute to volume. The solutions
shouldnot remain in glassvolumetricflasks for more than 2 h. Transferto poly-
ethyleneor polypropylenebottles for storage.The following referencecan be
consultedfor additional information: Bernas,1968.
Comments
Very small (O.l-g) samplesof clay, soil, mineral, or rock are analyzedfor
total AI by theseprocedures.In dealing with small samples,extremecaution is
neededin selectinga representativesample;one that is finely groundand mixed
thoroughly.
The boratefusion methodcan causeB contaminationproblemsin analyti-
cal laboratorieswhere emissionspectrometersalso are used for analysisof B.
The borate in the analytical solutions can causea buildup of B in the sample
delivery system.This may require a cleanupprocedureafter analysisof borate
fusion samples.Poorfusion can result in low results(Cantillo et aI., 1984).Good
fusion requiresthat the soil samplebe in contact only with the LiB0 3 and not
contactthe graphite.This can be accomplishedby placing the samplein a small
well of boratein the crucible. Acids other than 2 M HCI can be usedfor dissolu-
tion of the fused sample,dependingon the acid matrix preferredfor analysis.
Meyer & Bloom (1993) used 2 M HCI and ICP analysisfor plant ash samples
while Cantillo et al. (1984) used5% HN03 and DCP analysisfor sediments.
The graphitecruciblesdo lose about10% of their weight during fusion and
flakes of graphiteare typically found after dissolutionof the fused sample.These
graphiteparticles readily settle to the bottom and do not causeany difficulties
during analysis. The crucibles can be reuseduntil they becometoo fragile. If
samplesare high in silica, the solubility of silica in acid may be exceededand a
precipitatewill form. The precipitationof silica is readily visible and the ratio of
acid volume to samplesize can be increasedto eliminatethe precipitation.
Microwave digestionin HNOrHF is evenmore rapid than the Parr bomb
procedure.Warren et al. (1990) reportedthat they were able to digest soil sam-
ples in a HCI-HN03 mixture in 7 min. Becausethe vesselsare sealed,Si can be
recoveredby adding H3B03 as soon as the vesselsare opened.Excessiveinter-
nal pressurecan be a problem with some digestion vesselsand Warren et al.
(1990) recommendedthe removal of organic matter before digestion. More
522 BERTSCH& BLOOM
recently,Teflon vesselswith rigid outershellsand safetyreleaseports havebeen

developedwhich allow much greaterinternal pressures.Vesselsmanufactured
from polymer tetrafluoroethylene(PTFE) can tolerate even higher pressures.
With thesevesselsdestructionof organicmatterbeforedigestionof mineralsoils
should not be necessary.Systemswith temperatureand pressuremonitoring and
temperatureprogrammingare available.SeeChapter3 (Hossner,1996)for more
detailson microwavedigestion.
Proper precautionsshould be used when handling concentratedHCI,
HN03 or HE Rubbergloves and eye protectionshould be usedwhen handling
theseacids. Hydrofluoric acid will dissolveglass,hencepolypropylenelabware
must beusedwhen dispensingthis acid.
Alternative Procedurefor Total Aluminum
A HCI04-HF mixture can be used for total dissolution of soil minerals.

Details of HF-HCI04 digestion techniquesare given in Chapter 3 (Hossner,
1996). The analyst should be familiar with the potential explosive dangersof
HCI04 when this acid is in contactwith organic matterand certain inorganics.
This methodrequiresa proper stainlesssteel-linedventilation hood with wash-
down capability in order to avoid a buildup of perchloratesin the hood. As with
methodsdescribedabove,AI can be determinedwith ICP, DCP, AA, or spec-
trophotometricmethods. Becausedigestion is done in open containersin the
presenceof HF, Si is lost as SiF4 gasand accurateanalysisof Si is not possible.
Cautioncannotbe overemphasized when handlingHCI04 or HE Perchloricacid
is a rapid oxidant, and when it is placedwith organics,an explosionmay result.
Rubberglovesand eye protectionshould be usedwhen handlingeither acid.
EXCHANGEABLE AND EXTRACTABLE ALUMINUM
Introduction
The involvementof AI in exchangeand soil acidity reactionshas received

significantattentionfrom researchersinvolved in soil chemistry,mineralogy,and
fertility. The voluminousbody of publicationswill not be reviewed here; how-
ever, many of the "established"principlesmay be found in detailedreview arti-
cles dealing with AI chemistry in soils, solutions, and geological materials
(Jenny, 1961; Coleman& Thomas,1967; Coulter, 1969; Frink, 1973; Thomas,
1977; Hargrove & Thomas, 1984). Traditionally, there have been two primary
usesfor exchangeableAI values. The first is the formulation of lime require-
mentsfor acid soils basedon somemeasureof exchangeableor extractableAI
(Kamprath,1970; Reeve& Sumner,1970; Amedee& Peech,1976; Farinaet aI.,
1980; Juo & Kamprath, 1979; Oates& Kamprath, 1983a,b).For this purpose,
methodsare developedbasedon the efficacy for predicting plant responseto
limestoneadditionsand tend to be less concernedwith AI exchangeequilibria
per se. Second,becauseof its importanceas a predominantcation in acid soils,
exchangeableAI is a critical variable in establishingeffective cation exchange
ALUMINUM S23
capacity(ECEC) values,which are utilized for soil managementand classifica-

tion purposes,and in evaluatingchangesin forestedsoils influencedby acidic
depositionandland-usepractices (e.g.,Juoet aI., 1976;Pavanet aI., 1984;Evans
& Zelazny,1987; Mulder et aI., 1987; Lilieholm & Feagley,1988; Adamset aI.,
1990; Reusset aI., 1990; Rasmussen et aI., 1991).For theseapplications,inves-
tigatorsare interestedin arriving at a reproduciblemeasureof exchangeable Ai3+
that reflectsAI exchangeequilibria as accuratelyas possible.
The characterizationof AI in exchangereactionsis complicatedby the
coexistenceof complexmultiphaseAI componentsin soils, sediments,and min-
eral exchangers.No cleardemarcationexistsbetweenAI that is exchangedfrom
a permanentlychargedsurfacein equilibrium with an unbufferedsalt solution
and that which is concomitantlyreleasedvia solubilization from other nonex-
changeablesourcessuchas mineral structuresor hydroxo-polymericintedayers
(Juo & Kamprath,1979; Lee et aI., 1985; Rasmussenet aI., 1991). It has been
demonstratedthat the quantity of AI displacedfrom soil by inorganiccationscan
be highly dependenton the experimentalconditions used,such as the time of
extraction,the pH and ionic strengthof the extractant,and the natureof the dis-
placing cation (e.g., Skeen & Sumner, 1967; Bloom et aI., 1979; Oates &
Kamprath, 1983a,b;Jarvis, 1986). Another important factor that is sometimes
overlookedis the mineralogicalcompositionof the soil. Variations in soil min-
eral compositionwill greatly influencewhat forms of AI are releasedby given
extractant. Indeed, Kotze et ai. (1984) demonstratedthat 28AI isotopically
exchangeable AI was a variablefraction of salt (KCI and NH4CI) extractableAI.
This studydemonstratedthat most soils releasesomenonisotopicallyexchange-
able AI in 1 M salt extracts,the degreeof which was soil specific. Furthermore,
it is very difficult to estimatethe equivalentsof AI exchangedin a soil system
which may deviatefrom the theoreticalvalue(+3) due to the presenceof reduced
chargedinner-sphereAI complexeswith a numberof organicand inorganiclig-
ands.Becauseof the operationalnatureof the exchangeable AI determination,it
is commonly referredto as extractable AI. Therefore,the selectionof a method
for estimatingexchangephaseAI shouldbe madebasedon the specific applica-
tion and the natureof the soil beinginvestigated.
The most commonly employedextractantis 1 M KCI. This extractantis
usedboth for the determinationof extractableAI, which is utilized in the calcu-
lation of effective CEC and AI saturation,and in somecasesfor the determina-
tion of lime requirement.Liming to 1.5 or 2 times the -extractableAI has been
found to be a reasonablemetric for basinglime requirementsof many acid soils
to minimize acidity inducedyield reductionsfor most crops (Kamprath,1970).
Othershave found that 0.01 M CaCI2, a much less aggressiveextractantof AI,
was superiorfor predicting the pWAI toxicity response(Khalid & Silva, 1979;
Webberet aI., 1982; Wright et aI., 1989; Shuman,1990). Furthermore,0.01 M
CaCl2 extractableAI hasbeenshownto be well correlatedto the free ion activi-
ty of AI {AI3+} in soil solution and it has beenarguedthat this is preciselythe
reasonfor its superior efficacy in predicting AI toxicity as a function of pH
(Wright et aI., 1989). Kotze et ai. (1984) demonstratedthat 0.01 M CaCl2
removed~1O% of the 28AI isotopically exchangeable AI for the sevensoils they
examined.
524 BERTSCH& BLOOM
For soils high in organic matter, 1 M KCI is still employedin the opera-
tionally defined ECEC measurement;however, if the quantity of organically
boundAI is desireda more aggressiveextractantis needed.Both LaCl3 or CuCl2
have been shown to be far more effective than KCI in displacing organically
bound AI (Bloom et aI., 1979; Juo & Kamprath, 1979; Hargrove & Thomas,
1981,1984;Oates& Kamprath,1983a,b).However,Cu2+ also hasbeenfound to
remove some interlayer Al and possibly other structural AI via dissolution of
minerals (Juo & Kamprath, 1979; Hargrove & Thomas, 1981; Oates &
Kamprath, 1983b).The organic bound AI fraction will not necessarilyrelate to
toxicity or soil solutionAI without knowledgeof AI saturationof weakacid sites
(Walker et aI., 1990).
The natureof the Al speciesinvolved in exchangereactionshas beenthe
topic of considerableinterest. Earlier studies suggestedthat exchangeableAI
specieswere predominatelyof zero basicity, i.e., the mononuclearhexaaqua
species(Lin & Coleman, 1960; Rich, 1960; Thomas, 1960; Jackson, 1960;
Coleman & Thomas, 1967). Other studies, however, have indicated that
hydroxo-AI mononuclearand polynuclear species are involved in exchange
processes,and in some instances,preferentialadsorptionof these specieshas
beendemonstrated(Turner, 1967; Hsu, 1968; Brown & Newman, 1973; Bache
& Sharp, 1976; Veith, 1977, 1978; Bloom et aI., 1977; Rengasamy& Oades,
1978; Hodges& Zelazny, 1983; Jardineet aI., 1985; Hsu, 1992). The existence
of polynuclearhydroxo-AI speciesin dilute solutionsand their structural charac-
teristics are now well established(Bertsch, 1987, 1989a; Parker & Bertsch,
1992a; Furrer, 1993). Furthermore,there has been a report of a particular AI
polynuclear,the AI13 species([AI04AI120Hz4Hz012f+), associatedwith an Oa
horizon of an acid spodosol(Hunter & Ross, 1990). This polynuclearalso has
beenshownto form at relatively low solutionAI concentrationsunderconditions
indicative of natural systems(Bertsch,1989a; Parker& Bertsch, 1992b; Furrer
et aI., 1992). Additionally, there is overwhelmingevidenceto suggestthat this
polynuclear is more toxic to some plants and aquatic organisms than the
AI(H 20)g+ (Parkeret aI., 1988,1989;Shann& Bertsch,1993). Thereis still great
uncertainty surrounding the specific forms and stability of polynuclear AI
speciesin soil systemsand their ability to be exchangedfrom mineral or organ-
ic surfaces.
Additional investigationshavedemonstratedstrongevidencethat otherAI
complexes can participate in reversible exchange reactions. Bertsch and
Anderson(1989)demonstratedthat fluoro-, oxalato-,and citrato-aluminumcom-
plexes could be separatedin an ion exchangecolumn during ion chromato-
graphicdeterminationof AI. This methodwas usedby Willett (1989) to provide
evidencethat fluoro-AI complexeswere presentin 0.01 M CaCl2 soil extracts.
Andersonet ai. (1991) alsoprovidedstrongindirect evidencefor the existenceof
fluoro-AI complexeson model exchangers,a referencekaolinite, and a highly
weatheredsoil. More recently, direct evidencefor the existenceof mono- and
difluoro-AI complexeson the surfacesof cationexchangeresinsand mineralsur-
faces has been provided by 27AI and 19F nuclear magnetic resonance(NMR)
spectroscopicinvestigations(Wilkinson et aI., 1993).
ALUMINUM 525
There are important implications surroundingthe existenceof exchange-

able ternary AI surfacec~mplexes. Thesecomplexesare generally not consid-
ered in chemicalspeciationor ion exchangemodels.Furthermore,AI measured
in salt extractsis normally assignedthree mol (+) in ion exchangecalculations.
Thus, the widespreadpresenceof these complexescould result in significant
errorsin determinationof the ECEC and AI saturation.Additional studiesaimed
at defining the importanceof thesecomplexesin acid soils are requiredin light
of theseobservations.
ExtractionFrom Mineral Soils with UnbufferedSalts
Principles
Exchangeable or extractableAl ions are most commonlydisplacedwith an
unbuffered salt solution, such as 1 M KCI, 0.5 M CaCI2, or 0.5 M BaCI2.
Extraction techniquesemploying rather concentrated(0.5-1 M) unbufferedsalt
solutionsareprobablybestsuitedfor estimating"truly" exchangeable AI, at least
to the first approximation.More dilute (0.01-0.1M) neutral salt solutionshave
been demonstratedto be less effective at displacing "truly" exchangeableAI,
albeit they removea fraction that may be relatedto plant responseunder certain
experiment.alconditionswhere they reflect soil solution AI. Other complexi.ng
agentsand acid saltsolutionsmay extractAI from both exchangeable and nonex-
changeablesources,induding structuralhydroxo-polymericinterlayers, organi-
cally bound species,and other noncrystallinemetastableforms that collectively
can be referredto as "labile" AI. It can be arguedthat someof the nonexchange-
able metastableforms representa plant-active"AI pool," which should be con-
sideredin lime recommendations(Arai, 1975; Juo & Kamprath, 1979; Oates&
Kamprath,1983a).Many of theseclaims may be appropriate;however,the meth-
ods employedin the estimationof suchlabile AI pools are soil specific and plant
responseto chemical labile forms will be both speciesand cultivar dependent.
Thus, the utilization of methodsthat attempt to estimatechemically labile AI
require extensivecalibrationbefore they can be effectively implemented.Other
methodsused to estimateorganically bound AI and noncrystallineAI phases,
componentsthat are important in spodosolsand in relation to phosphatereten-
tion are discussedin the sectionon Extractionof AIuminum from OrganicMatter
and OrganicSoils.
Investigationsdealingwith soil classification orsoil acidification process-
es require an accurateassessment of the quantity of exchangephaseAI relative
to exchangeable basecations (Ca 2+, Mg2+, K+, and Na+), and how thesemay dif-
fer over time or as a function of acid inputs(Richardson& Riecken,1977; Pavan
et aI., 1984; Evans& Zelazny, 1987; Mulder et aI., 1987; Lilieholm & Feagley,
1988; Reusset aI., 1990; David et aI., 1991). For theseapplications,it is desir-
able to extractas little AI as possiblefrom sourcesotherthan the exchangephase.
This is particularly important for highly weatheredacid soils which have an
inherently low cation exchangecapacity,thus making errors introducedby min-
eral dissolutionpotentially significant.
526 BERTSCH& BLOOM
"Exchangeable"Aluminum
Method
Reagents
1. Potassiumchloride (KCI), 1 M or ammoniumchloride (NH4CI), 1M.
2. Concentrated nitricacid.
Procedure.Weigh out 5 g of <2 mm soil, sludge,sediment,etc., into a 50-
mL polyethylenecentrifuge tube or some other appropriatecontainer.Add 25
mL of 1 M KCI (or NH4CI), stopper,and shakefor 30 min. If centrifugetubes
are used,centrifuge 10 min at 1000 ~ g, and then filter the supernatantsolution
through either a prewashedWhatmanno. 42 filter (where exchangeableAI val-
ues are high suchthat contaminationfrom filter materialswill be minor; seethe
introductory matter on Sampling Aluminum in Soil Solutions) or a 0.45-Jlm
polycarbonatefilter (whereexchangeable AI or CEC valuesare low and possible
contaminationfrom filter materials could induce errors) to remove floating
organicmatterand soil particles.Acidify the filtrate to pH <3.0 with a few drops
of concentratedHN03. The filtrate should be stored in plastic containers.For
soils low in exchangeableAI where accurateexchangephaseAI values are
required,use Ultrex HN03 or someother appropriatehigh-purity HN03 source.
For very acid soils, AA analysisusing N20 air flame can be usedfor AI analy-
sis. For lessacid soils whereAI concentrationsin the extractswill be lower, ICP-
MS (massspectrometry),graphite furnace AA, or spectrophotometricmethods
are required.
Dilute Salt ExtractableAluminum

Reagents
1. Calcium chloride (CaCI2), 0.01 M.
2. Concentrated nitricacid (HN03).
Procedure.Weigh out 10 g of soil into a 50-mL polyethylenecentrifuge
tube or someother appropriatecontainer.Add 20 mL of 0.01 M CaCI2, stopper,
and shakefor 5 min. Centrifuge,filter, and acidify filtrate as describedabove.
Exceptfor extremelyacid soils, ICP-MS, graphitefurnaceAA or spectrophoto-
metric methodsof AI analysisare necessaryfor theseextracts.The following ref-
erencecan be consultedfor additional information: Hoyt and Webber,1974.
Comments.The use of NH4CI in place of KCI is useful where a single
extractantfor exchangeablebasesand Al is preferredsince NUt is as effective
as K at displacingAI (Lee et aI., 1985). In light of the many problemsinvolved
in determining"exchangeable"AI and the operationalnatureof the determina-
tion, it is not unusual to find various researchersemploying different
sample/solutionratios, shakingperiods,cations,numbersof successiveextrac-
tions, and even concentrationsand pH of displacing unbufferedsalt solutions.
Oatesand Kamprath (1983a) found that two extractionswere required for the
most efficient removalof AI from soils low in organicmatter,while soils high in
organic matter continued to releasesignificant quantities of AI in subsequent
ALUMINUM 527
extractions(Bloom et aI., 1979; Oates & Kamprath, 1983a). Becauseof the

potentialfor Al solubilizationfrom many acid soils during extraction,we prefer
a singleextractionfor routinemeasurements. In mostinstances,choiceof extrac-
tantsand extractionprotocolwill dependon the specificpurposeor natureof the
studybeingconducted,andto a certaindegreeon the individual preferenceof the
researcher.Theseparametersalso are sometimesbasedon mineralogicalconsid-
erations.Thus, no single procedureis best or appropriatefor all situations-
being consistentand statingclearly which methodsare being usedare of prime
importance.
Extraction of Aluminum from Organic Matter and Organic Soils
Principles
OrganicallyboundAl hasthe importantrole of controlling Al in soil solu-
tions in the surfacehorizonsof mineral soils (Bloom et aI., 1979)and in O-hori-
zonsof forest soils (Walker et aI., 1990).This aluminumis boundto carboxylate
groupsin soil organicmatter(Bloom, 1981) and while it is lessreactivethan Al
on the exchangesitesof clays, it is more reactivethan both crystallineand non-
crystallinealuminumhydroxideand Al in aluminosilicates(Bloom et aI., 1979;
Cronan et aI., 1986; Walker et aI., 1990). As describedpreviously, extractants
suchas KCI that havebeentraditionally employedto estimateexchangeable Al,
are ineffective at displacingAl from organicmatter(Bloom et aI., 1979; Juo &
Kamprath,1979; Hargrove& Thomas,1981).Dependingon Alloadings andthe
natureof the organicmatter,KCI typically extractslessthan 50% and as little as
10% or lessof the total organicallybound Al (Bloom et aI., 1979; Hargrove&
Thomas,1981, 1984).
The greaterreactivity of organically boundAl comparedto mineral Al
allows for its estimationby "selective"extractiontechniques. Traditionally, sodi-
um (or potassium)pyrophosphate,0.1 M Na4P207,hasbeenusedto "selective-
ly" extract organic metals, including Al (McKeague, 1967; Bascomb,1968;
McKeagueet aI., 1971).This reagent,however,forms strongcomplexeswith Al,
that, coupledwith its high pH (pH 10), also can removepoorly orderedor non-
crystalline Al phases(Bascomb,1968; McKeague& Schuppli,1982; Page& De
Kimpe, 1989).This noncrystallineAl may be colloidal phaseslargely coassoci-
ated with organic matter that is releasedvia.dispersionof the organic material
during the pyrophosphateextraction(McKeague& Schuppli,1982).The amount
of Al removedby pyrophosphateis generally well correlatedwith organic C,
providing some support for this contention.Furthermore,Jersakand McColl
(1989) found that the pyrophosphateextractableAl fraction representedan
important labile Al pool that correlatedwell with the fraction of Al initially
leachedfrom a surfaceforest soil horizon with dilute citric acid.
It shouldbe mentionedthat there is little consistencyin the literaturecon-
cerningthe useof the Na or K form of pyrophosphateor in the degreeor method
of centrifugation,flocculation,or filtration of the extracts.If dispersionof organ-
ic-associatedcolloidal phasesis an important fraction of the extractableAl in
studiesnot employingfiltration, then Na ratherthan the K salt of pyrophosphate
528 BERTSCH& BLOOM
may be more effective and has been demonstratedto be more reproducible

(Loveland& Digby, 1984). Regardlessof the exactnatureof the solid phaseAI
componentssolubilizedby pyrophosphate,this fractionof extractableAI can be
significant and has beenusedextensivelyas a criterion for the identification of
spodichorizonsand for evaluatingother pedogenicprocesses(Bascomb,1968;
Wada & Higashi, 1976; McKeague& Schuppli, 1982; Jarvis, 1986; Jersak&
McColl, 1989). This extractantis not appropriate,however, for estimatingthe
organicallyboundAI fraction that is usedto estimatethe ECECof an organicsoil
or horizon since it removesAI phasesthat are clearly not exchangeablewith
other cations.
AIternative extractantsusedto removeAI from organic mattervia ligand
exchangereactionsinclude 0.5 M CuCl2 and 0.33 M LaCl3 (Bloom et al., 1979;
Juo & Kamprath, 1979; Hargrove & Thomas,1981, 1984; Oates& Kamprath,
1983a,b;Walker et al., 1990).Copperhasa high affinity for the carboxylatesites
that bind AI andcan readily replaceAI boundto organicmatter.AIthough CuCl2
extractionwas found not to removesignificantAI from aluminosilicateminerals
(Juo & Kamprath, 1979), it has beenshown to extract somepoorly orderedAI
associatedwith soil mineralswith hydroxy interlayeredvermiculiteandsmectite,
and AI associatedwith soil mineral phasesasso,(::iatedwith a subsoil (Juo &
Kamprath, 1979; Oates& Kamprath, 1983b). Lanthanumhas beenfound to be
somewhatless effectivein displacingorganically bound AI, removing between
50 and 100% of the total boundAI dependingon the amountof organicmatter
and AI loadings.Oatesand Kamprath(1983a)found thatfor soils low in organ-
ic matter, both KCl and LaCl3 extractedsimilar amountsof AI, whereasLaCl3
extractedincreasinglymoreAI than KCI as the organicmattercontentincreased.
Copperchloride extractedmore AI evenfor the soils having <1 % organic
matter,suggestingsomesolubilizationof mineralAI phases(Oates& Kamprath,
1983b). Bloom et al. (1979) found thatLa3+ was far more effective than K+ at
displacing AI from organic matter, but only removed 50% of titratable AI.
Hargroveand Thomas(1984) demonstratedthat at low AI loadingson a humi-
fied muck, both Cu2+and La3+ extractedsimilar amountsof AI from organicmat-
ter, with both extractingclose to 100% of the total boundfraction. At higherAI
loadings,however,they found Cu2+ to be more effective than La3+ and speculat-
ed that this was a result of more hydrolyzed and polymerizedAI that, like the
interlayerAI components,were solubilized by Cu2+ but not La3+. Additionally,
Oatesand Kamprath(1983b)found thatLaCl3 extractableAI was the mosteffec-
tive predictor of lime requirementfor acid soils having varying organic matter
comparedto either KCI, which extractedtoo little, and CuCI2, which extracted
too much AI.
Few studieshavedirectly comparedpyrophosphatewith Cu or La chloride
for their efficacy in extractingAI. Jarvis(1986) did comparepyrophosphatewith
LaCl3for AI extractionefficiency andfound thatLaCl3 extractedonly 6 and74%
of the AI removedby pyrophosphate.Pyrophosphatealso was found to remove
significant quantitiesof AI from soils low in organicmatter,providing addition-
al evidencethat this extractantremovessignificant quantitiesof noncrystalline
and possiblycrystallineAI phasesin someacid soils (Jarvis, 1986).
ALUMINUM 529
Methods
Copper Chloride
1. Copperchloride (CuCI2), O.S M.
2. ConcentratedHN03•
Procedure.Weigh out 3 g of <2 mm soil into a SO-mL polyethylenecen-
trifuge tube or some other appropriatecontainer.Add 30 mL of 0.5 M CuCI2,
stopper,and shakefor 2 h. Centrifuge, filter, and acidify filtrate as described
above.Aluminum analysishas to be conductedby ICP or AA due to the inter-
ferenceof high Cu concentrationswith the spectrophotometric methods.The fol-
lowing referencecan be consultedfor additionalinformation: Juo and Kamprath,
1979.
SodiumPyrophosphate.Sodiumpyrophosphate(Na4P207),0.1 M, pH =
10.
Procedure.Weigh out 2 g of <0.5 mm soil into 2S0-mLpolyethylenecen-
trifuge bottlesor someappropriatecontainer.Add 200 mL of 0.1 M Na4P207and
place on an end-over-endshakerfor 16 h at 22 to 2S% C, then centrifuge at
~20 000 g for 10 min. Keep samplesrefrigeratedprior to analysisand analyzeas
soonas possibleafter extraction.The samplesmust be analyzedfor Al with AA
or ICP since the matrix is not suitablefor spectrophotometricanalysis.The fol-
lowing referencecan be consulted for additional information: Loveland and
Digby, 1984.
Comments
The estimationof organically bound Al is highly dependenton th~ soils
beinginvestigatedandthe methodutilized. For soils or soil horizonsthat arevery
high in organic matter, the CuCl2 extraction method should provide the most
accurateassessment of Al associatedwith carboxylateor phenolicgroups.Soils
with increasing mineral content containing interlayered minerals or poorly
ordered Al phases,some solubilization of nonorganically bound Al can be
expectedwith this extractant.
The pyrophosphateextraction clearly removesnoncarboxylatebound Al
and can causethe solubilizationof poorly orderedAl phasesin somesoils. The
relative amountsof Al that are extractedwith this methodfor a given soil are
highly dependenton the centrifugationand filtration methodsemployedsince
colloidal Al phasesare important componentsdispersedwith this extractant
(McKeague& Schuppli, 1982; Loveland & Digby, 1984). Our experiencehas
been that a significant fraction of the colloidal Al dispersedby Na4P207still
passesas O.4S-llm filter, but thatthis fraction is much lower than if no filtration
or a coarserfilter mediais used.Also we havefound that O.l-llm filters are effi-
cient atcapturingmuch of the colloidal Al. For soils or soil horizonscontaining
considerableorganicmatter,the noncarboxylateboundand colloidal Al released
may be coassociatedwith organic polymers,thus providing someuseful infor-
mation for pedogenicassessment purposes.However,examinationof the litera-
ture reveals that the centrifugation, filtration, and flocculation techniques
530 BERTSCH& BLOOM
employedby variousinvestigatorson pyrophosphate extractsarehighly variable.

Therefore,the operationalnatureandsoil specificity of thesemeasurements must
be recognized.
Extractionof Aluminum from Poorly OrderedPhases
Traditionally therehasbeenandcontinuesto be stronginterestin the quan-

tity of poorly ordered"amorphous"or noncrystallineAl phasesin soil. This
interesthasbeendriven from the strong emphasis on researchrelatedto andosol
and spodosolpedogenesis, aswell as by the well-known correlationbetweenthe
noncrystallineAl (and Fe) phasesand importantsoil chemicalproperties,suchas
P fixation or chemically labileAl that may be importantin acid soils (e.g., Chao
& Zhou, 1983;Alvarado & Buol, 1985;Jarvis,1986; Lekwa & Whiteside,1986;
Wang et aI., 1986, 1987; Parfitt et aI., 1988; Lee et aI., 1989; Kodama& Wang,
1989; Bartoli & Philippy, 1990; Mokama, 1993). In most applications,investi-
gatorshavebeeninterestedin differentiatingorganicallyboundAl from Al asso-
ciated with both crystalline and poorly orderedphasessuch as allophane,pro-
toimogolite, or imogolite. Much of the literaturedealingwith methodsdesigned
to provide this differentiation have commonly referredto a seriesof extractants
as comprising "selective" dissolution procedures.Although somerelationships
betweenthe operationallydefinedAl pools and the choiceof a given extractant
do exist, it is clear that the extractantsare not selective for specific phases.
Considerableoverlap betweenthe forms of Al solubilized by a given extractant
exists and the degreeto which an extractantprovides differentiation between
operationallydefined phasesis soil specific. Other investigationshave focused
on defining that fraction of Al presentas high surfaceareaphaseswhich might
be important in anion and cation sorption processesor AI solubilization events.
In theseinstances,empirical relationshipsbetweennoncrystallineAI phasesand
some index of chemical reactivity are established.The literature in this area is
extensiveandthereis little indication that interestin estimatingnoncrystallineAl
componentsis waning. It is not our intent to provide an exhaustivereview of the
various methods,nor to provide recommendedmethods.Rather webriefly dis-
cuss some of the more commonly employed methodsfor extracting noncrys-
talline Al componentsand provide appropriatereferencesfor thoseinterestedin
pursuingthis areafurther.
In addition to the pyrophosphateextractantused to estimateorganically
boundAl (vide supra), sodiumcitrate-dithionite(NaCD) and ammoniumoxalate
in the dark (NH40x-D) have been commonly employedto remove crystalline
andnoncrystallineoxide phases,respectively(McKeagueet al., 1971).Theseex-
tractantswere initially developedand calibratedfor Fe phasesand, thus, cannot
be directly appliedto Al containingphases,especiallysincethe dithionite extrac-
tion involves efficient solubilization of Fe as a result of the reduction from
Fe(III) to Fe(II). The NR.Ox-D extractantis usually assumedto extract non-
crystallineAl phases,whereasit is not totally clearwhat Al phasesare removed
by the NaCD extractant.Poorly orderedAl phases,high-surfaceareacrystalline
Al phases,such as gibbsite, and perhapsAl containedin crystalline and non-
crystalline Fe oxyhydroxidephasescan be extractedwith NaCD. In contrastto
ALUMINUM 531
Fe, whereNaCD extractablevaluesare generallygreateror equalto the NH40x-

D values, the NaCD extractableAI can be greater,equal to, or less than the
NH 40x-D extractableAI dependingon the soil. Iyengeret al. (1981) found that
NH40x-D and NaCD did not significantly remove interlayer AI from the
hydroxy interlayeredminerals,whereasammoniumoxalatereactedunder ultra-
violet radiation removedsignificant quantitiesof AI from hydroxy interlayers.
The soils usedin this study had very well-developedinterlayercomponents,and
resultswith other soils containinghydroxy interlayeredmineralsare lessconsis-
tent with both NH40x-D and NaCD removing some interlayer components
(Barnhisel& Bertsch,1989).
A rapid methodusing 4 M KOH extractionfor AI hasbeenshown to com-
parewell with the NH40x-D extract, and has beensuggestedas a field identifi-
cation aid for spodic and andic horizons(Holmgren & Kimble, 1984; Holmgren
& Yeck, 1984; Kimble et aI., 1984). Many otherextractants,including hydroxyl-
amine,tiron, andsodiumtetraboratealso havebeenusedto extractAI from poor-
ly ordered phases and often compared to both oxalate and pyrophosphate
extracts.Since all of the extractantsare operationallydefined and there exist no
standardsto referencethe various methodsand techniquesagainst,popularity
and analyticalconsistencyis often usedas the basisfor judging appropriateness
of a given extractant.
The differencebetweenthe NH 40x-D and the pyrophosphateextractable
AI is sometimesdefined as inorganic noncrystalline AI, with the implicit as-
sumptionthat the oxalateremovesboth the organicand inorganicpoorly ordered
AI phaseswhile the pyrophosphateremovesonly the organicphases(Wanget aI.,
1986). As discussedpreviously, this distinction is soil and method dependent
and, therefore,is only a grossapproximationthat may be useful for comparing
relative valuesbetweensimilar soils or soil profiles within a soil but not espe-
cially useful for comparisonsbasedon the absolutevalues.Methodsfor many of
the extractantsdescribedaboveare discussedin detail by Jacksonet al. (1986).
SAMPLING ALUMINUM IN SOIL SOLUTIONS
Introduction
Soluble AI in soils is an important parameterfor studying the impact of

acidificationon forest soils andwatersheds,the formation (or dissolution)or sec-
ondary soil minerals, as well as for assessingAI toxicity to plants in acid soils
and aquaticorganismsin acidified watersheds.Thus, determinationof AI in soil
solutionsis often desired.Most of the techniquesusedto obtain soil solutionsfor
chemicalanalysisof the typical predominantsoil cations(Ca2+, Mg2+, Na2+ and
K2+) also can be usedfor AI, but more care is neededsince AI is typically pre-
sent at much lower concentrations.As discussedpreviously,the solubility of Al
is pH dependentand factors that result in changeof pH to a value nearneutrali-
ty can result in loss by precipitation.AIso, many commonly utilized sampling
devicescan result in either the removalof AI throughsorption,or contamination
of AI throughdissolutionreactions.Additionally, collection of sampleswith low
AI concentrationsrequires great care to minimize contaminationfrom back-
532 BERTSCH & BLOOM
groundsources.Carefulacid washingandrinsing of equipmentandsamplehold-

ers as well as the useof plastic laboratoryware is a necessity.
In Situ Samplingwith Lysimeters
Samplesrepresentativeof the water moving through soil during rainfall

eventscan be obtainedusing zero-tensionlysimeterswhich can be made from
porouspolyethylenesheets(pore size =20-30 /lm). Theselysimetersare usual-
ly installed under the soil that is to be sampled,from a pit adjacentto the sam-
pling site. Samplesrepresentativeof more tightly held soil watercanbe obtained
using ceramicporouscup samplerswith submicrometerpore sizes.Teflon cups
are not useful becausethey require imbeddingin silica flour for good hydraulic
contact with soils, since the Teflon itself is relatively hydrophobic.The silica
flour can adsorbsoluble AI. Ceramic materialsare aluminosilicates whichcan
removeor contributeAI to solutions(Litaor, 1988; Raulund-Rasmussen, 1989;
Grossmann& Udluft, 1991). It has been demonstratedthat in acidic soils, soil
solutionscan be neareqUilibrium or supersaturated with respectto AI hydroxide
and hydroxo-sulfatephaseswhich can precipitateduring percolationthrough the
suction cup (Grossman& Udluft, 1991). Additionally, in acid soil systems,i.e.,
pH - 4, AI dissolutionfrom the ceramicmatrix hasbeendemonstrated,with the
quantity of releasedAI being dependenton the pH and the samplingrate (Rau-
lund-Rasmussen,1989; Grossmann& Udluft, 1991). The specific source,i.e.,
manufacturerof the ceramiccup, also seems toinfluencethe amountof AI solu-
bilized (Hughes& Reynolds, 1990; Raulund-Rasmussen, 1991). For soil solu-
tions of pH :2!5 it hasbeenshownthat AI contaminationfrom the ceramiccup can
be minimized by careful acid washing followed by thorough rinsing with dis-
tilled waterandproperequilibrationtime onceinstalledin the field (Litaor, 1987,
1988; Grossmann& Udluft, 1991). Ceramiccupscome in varioussizesthat can
be cementedto plastic pipe for installation in bore holes.
Soil solutionstypically contain CO2 at partial pressuresthat are elevated
aboveatmosphericCC02 (Inskeep& Bloom, 1986a).For soil solutionswith pH
valuesgreaterthanabout4.9, degassingof CO2 during and following samplecol-
lection can result in a pH increase(Reuss& Johnson,1985). The low pressure
inside tensionlysimeterscan accentuatethe problemof degassing.This problem
has been discussedin detail by Suarez(1987). With increasingpH values, AI
adsorptionby containerwalls increasesand precipitationof AI(OH)3 can occur.
With zero-tensionlysimetry, the problemof degassingcan be minimized by im-
mediatesamplingafter a rainfall and storagein bottles that are full and tightly
capped.With tensionlysimetry, the problemof pH changein samplesfrom soils
with soil pH (H20) of 4.9 or greateris difficult to avoid.
Extractionof Soil Samplesin the Laboratory
Laboratory extraction of fresh soil samplesremovedfrom the field is an

alternativeto lysimetry. Miscible displacementof soils in packedcolumnsusing
a solution with an indicator ion that can be used to detect breakthroughof the
eluting water can be used. Sodium thiocyanateis a good indicator solution
ALUMINUM 533
becausethe SCN-ion is mobile and it producesa strongly coloredsolution water

when Fe2+ is addedto a portion of the effluent. Miscible displacement,however,
is slow and laborious.
A more rapid techniqueis centrifugationwith or without a heavy liquid
that is immiscible with water. In a high clay or high organic matter soil, com-
pressionof soil during high-speedcentrifugationwill force water out of the soil
matrix and it can be collectedby decantation(Inskeep& Bloom, 1986b).In most
mineral soils, however,packingof silt and sandparticlescreatesvoids that can
hold water. This water can be forced fromthe voids if the soil is first mixed with
an immiscible liquid that is more densethan water, or if an especiallydesigned
centrifugation assemblyhaving a built-in support and filter disc is employed
(Elkhatib et aI., 1987). The best liquids have a high-specific gravity and low
reactivity. Carbon tetrachloridehas been used but it is both volatile and toxic.
Perfluoro hydrocarbons,which are denseand have a low reactivity, are ideal for
soil solution extraction.
Accuratedeterminationof soil solution AI concentrationsthat representin
situ concentrationsrequires immediate laboratory processingof soil samples.
Degassingof CO2 from all but the most acidic soilscan result in a decreased
HC03 concentrationand in soil solution ionic strength (Inskeep & Bloom,
1986b). This can be minimized by keeping soil samplesin airtight containers.
Another concern,even in airtight containers,is microbial respirationwhich can
increaseCO2 and increasesoil solution ionic strength.This can be minimized by
keepingsampleson ice during collection and refrigeratedduring short-termstor-
age. Mineralization of soil N followed by nitrification producesHN03 which,
even if the pH of the soil is well buffered, causesan increasein ionic strength.
Mineralization and nitrification ratescan be significant evenat 4% C.
Filtration of Soil Solution Samples
The well-establisheddelineationbetween"dissolved" and "total" AI has

beenbasedon filtration througha 0.45-llm filter. It haslong beenrecognizedthat
this operationally defined "dissolved"AI fraction, or even a filtrate passinga
O.l-llm filter, can include significant contributionsfrom finely divided colloidal
solid phasescontaining AI either within the matrix or as an adsorbedspecies
(Joneset aI., 1974; Kennedyet aI., 1974; Barnes,1975).The presenceor absence
of thesefinely divided phasesis highly systemspecific; thus, studies demon-
strating that only a small fraction of AI passinga 0.45-llm filter is colloidal in
naturealso can be readily identified (e.g., Royset& Sullivan, 1986).
Soil solution samplesshouldbe filtered throughnonreactivemembranefil-
ters with pore sizesof 0.45-llm or less. Kennedyet al. (1974) found O.l-llm fil-
ters removedsignificantly more particulateAI than did 0.45-llm filters and this
hasbeenour experiencewith soil solutions,surfacewater and groundwater.Care
must be taken to avoid contaminatingthe sampleswith inorganic or organic
materialsfrom the filters, and to avoid sorptionof AI on the filters. Jardineet aI.
(1986) comparedseveral membranefilters and found polycarbonateto be the
best both with respectto releaseof inorganic contaminantsand sorption of AI
from solution.
534 BERTSCH & BLOOM
The handling of soil solution samplesafter filtration dependson whether

the samplesare to be analyzedfor total "soluble" AI or for determining all
speciesof AI that passthrough microporefilters (vide infra). If total AI is to be
determined,the solutionscan be acidified with Ultrex gradeacid to a pH of two
or lower and storedfor later analysis.Before analysis,fine colloidal AI can be
dissolvedand organic complexesdestroyedby heating to drynessat 130°C to
lS0°C followed by ashingat 4S0°Cfor 3 h (Jameset aI., 1983).The ashcan be
dissolvedin 0.1 M HN02 with NH20H • HCI addedto dissolveoxidesof Mn and
Fe.
CHEMICAL SPECIATION OF SOLUTION ALUMINUM
Introduction
Beginningin the early 1980sand continuingfor the betterpart of a decade,

AI speciationwas a major research areain soil and environmental science. While
ampleevidencehasaccumulatedto suggestthat chemicalspeciationregulatesAI
mobility in the environmentand its bioavailability and toxicity, there remain
many questionsregardingthe behaviorof specific chemicalforms of AI in the
environment.Three generalapproachesto the chemicalspeciationof "soluble"
AI can be identified. The first involves the analytical separationof various AI
fractions basedon differential reactionkinetics with complexationagentsor the
physicochemicalseparationof AI fractions basedon size or charge.The distin-
guishablefractions obtained are then operationally categorizedas distinct AI
species.The secondapproachinvolves the computational differentiationof AI
speciesfrom an analytically determined"total" AI fraction utilizing a thermody-
namically basedgeochemicalspeciationmodel, usually employingmassbalance
constraints(Parkeret aI., 1995).The third generalapproachinvolves somecom-
bination of one or more analytical separationtechniquealong with a geochemi-
cal speciationmodel. In all instances,AI speciationmethodsand techniquespro-
duceoperationally defined parametersthat must be interpretedappropriately.
As with many other operationallydefined methods,techniquesand exper-
imental approachesfor speciatingsolubleAI vary widely. Existing methodsare
continually modified by various investigatorsand new methodsare proposed
regularly. A comprehensivediscussionof AI speciationmethodsis beyondthe
scopeof this chapter.The following discussion provides an introductionto chem-
ical speciationmethodsand appropriatereferencesto guide those interestedin
utilizing thesemethodsin their research.
Timed Spectrophotometric
Assays
The classicalanalytical methodsfor speciatingsolubleAI focusedon the

differentiation of mononuclearand hydroxopolynuclearAI complexesin rela-
tively simple solutions(e.g., Bertsch,1989a,b).The earliestand most common-
ly usedmethodsfor this applicationincluded the 8-hydroxyquinoline(HQ) and
ferron (8-hydroxy-7-iodo-5-quinoline-sulfonicacid) timed spectrophotometric
ALUMINUM 535
assays(e.g.,Okura et aI., 1962;Turner, 1969,1976;Smith, 1971; Smith & Hem,

1972, p. 1-51; Bertsch, 1989a,b;Parker & Bertsch, 1992a,b).Experimentally,
thesemethodsare quite simple: the ferron or 8-hydroxyquinolineis addedto the
solutionto b~ speciatedandthe absorbancemonitoredas a function of time, with
the rapidly reactingAI fraction being differentiatedfrom more slowly reacting
fraction(s) via fitting a first-, second-,or fractional-orderreaction model (e.g.,
Bertsch,1989b).
The differentiationof AI species,evenin theserelatively simple solutions,
hasbeensteepedin controversy(Bertsch,1989b).It is not surprising,therefore,
that the extensionof thesemethodsto much more complexsolutionscontaining
a wide variety of ligandshasbeenboth confusingand inconclusive.Much of the
confusion has resulted from the extensionof methodsbeyond the limits for
which they were calibrated,or the extensionof the interpretationbeyondchem-
ical constraintsimposedby the operationallydefined methodor approach.For
example,timed spectrophotometric methodsdevelopedandcalibratedto provide
differentiationof mono-and polynuclearAI in very simple partially neutralized
solutionshave often beenusedin solutionscontaininga numberof complexing
ligandsthat havebeenshownto interferewith the methods(Bloom et aI., 1978,
1979;Jardine& Zelazny,1987a,b;Noble et aI., 1988;AIva et aI., 1989; Bertsch,
1989b).Thus,manyof the studiesthat haveinferredmolecularlevel information
on AI speciesdistribution from the resultsof one or more speciationmethods
haveto be interpretedwith caution(Bertsch,1989b).
Therehavebeena large numberof spectrophotometric methodsthat have
utilized the kinetic approachto AI speciationin natural waters(Barnes,1975;
Bloom et aI., 1978; Jameset aI., 1983; Seip et aI., 1984; Lalande& Hendershot,
1985; Bartlett et aI., 1987). Jameset ai. (1983) employedthe 15-s reactionwith
8-hydroxyquinoline(HQ) buffered at pH 5.2, followed by extractioninto butyl
acetateas a methodfor speciatingAI in soil solutionsand surfacewaters.The
methodemployeda correctionfor Fe by measuringthe absorbanceat a second
wavelength(600 nm) and subtractingthe correspondingcontribution of the Fe
absorbanceat 395 nm. In samplescontainingonly OH-ligands,the methodwas
demonstratedto predominantlymeasurethe hexaaquaand hydroxo-monomers
after 15 s of reaction,consistentwith previousfindings. The presenceof other
complexing ligands such as F", citrate, and Si, however, producedvariable
results that were dependenton ligand/AI mole ratios and pH, with generally
greaterAI beingrecoveredfollowing the 15-sreactionthanonly the aquatedand
hydroxo-complexedforms. This variability with model ligands led Jameset ai.
(1983) to operationallydefine the 15-sreactiveAI as the "labile" AI fraction and
the slowly reactingAI as "nonlabile." This methodis attractivebecauseof its
simplicity and becauseit placesno molecularsignificanceto the differentiated
fractions. A similar approachhasbeen proposed by Bartlett et ai. (1987) utiliz-
ing pyrochatecholviolet with the operationaldelineationof three AI fractions,
the rapidly reacting(RR AI), moderatelyrapid (MR AI), and total reactive(TR
AI) AI. Othermethodsbasedon the eqUilibrium complexdistribution of AI with
a chromophore,fluorophore, or with fluoride also have been employed(e.g.,
Hodges,1987; Bertsch,1989b; Browne et aI., 1990a,b).AIthough a numberof
thesemethodshave a numberof advantages,they tend to be rather tediousand
536 BERTSCH & BLOOM
rely on assumptionsbasedon achievingthermodynamicequilibrium within the

samplesanalyzedand equilibrium thermodynamiccalculations,both of which
introduceuncertainty(Schecher& Driscoll, 1987; Bertsch,1989b).
Otherapplicationsof the timed spectrophotometric methodshllve involved
extensive calibration and comparisonto 27Al NMR spectroscopicdata (e.g.,
Bertschet aI., 1986a,b;Bertsch,1987; Denney& Hsu, 1986; Parker& Bertsch,
1992a,b).In general,thesestudieshave demonstratedthe efficacy of the ferron
timed spectrophotometricmethodfor differentiating mono- and polynuclearAl
speciesin simple aqueoussolutionsfree of complexingligands. They also have
supportedthe notion that the primary Al polynuclearpresentin dilute solutions
is the [AlO~dOH)24+n(H20)12_n](7-n)+ or the Al13 species.This polynuclear
has beenshownto be more toxic to plants and aquaticorganismsthan mononu-
clear Al and may be an important intermediatein AI hydroxide mineral forma-
tion in certain environments.Although the importance of this polynuclear
speciesin soils has yet to be demonstratedunequivocally,severalstudieshave
demonstratedits formation underconditionsrepresentativeof acid soils and sur-
face waters(Parker& Bertsch,1992b;Furreret at, 1992),and thereis one report
of its possibleexistencein an acid forest soil (Hunter & Ross,1990).
PhysicochemicalSeparationMethods
Perhapsthe most fundamental,yet often trivialized physicochemicalsepa-

ration methodis simple filtration. As discussedpreviously,it has long beenrec-
ognizedthat the operationallydefined"dissolved"Al fraction basedon filtration
through a 0.45- and even O.l-l1m filters can include significant contributions
from finely divided colloidal solid phases(Joneset aI., 1974; Kennedy et at,
1974; Barnes, 1975). It was precisely this observationthat stimulated earlier
attemptsto "speciate"aqueousAI from surfacewatersby differentiation of sol-
uble and solid-phase componentsby utilizing a timed spectrophotometric
method (Barnes,1975; Bertsch, 1989b). The choice of filter materialsand the
properpretreatmentof thesematerialscan be quite importantin termsof sample
contamination,loss of dissolved AI, or dissolved organicC (Campbell et at,
1983; Jardineet aI., 1986; Hodges,1987; Royset& Sullivan, 1986).
Several AI fractionation methods differentiate various complexed Al
speciesbasedon their size and/orcharge.Methodsthat havebeenutilized for this
purposeinclude dialysis, ultrafiltration, size exclusion chromatography(SEC),
ion exclusion, ion chromatography,capillary zone electrophoresis,and C-18
reversephasechromatography(Campbellet aI., 1983; LaZerte, 1984; Driscoll,
1984; Parthasarthy& Buffle, 1985; Gardineret aI., 1987; Bertsch& Anderson,
1989; Willett, 1989; Bertsch,1989b).Thereare advantagesand disadvantages to
all of thesemethodsand approachesas well as various limitations that are criti-
cally discussedin detail elsewhere(Bloom & Erich, 1989; Bertsch,1989b).
Perhapsthe most popular of thesespeciationmethodsutilizes a strongly
acidic sulfonic resin to separate"organically" and "inorganically" complexedAI
forms via noninteractionof neutralor exclusionof negativelychargedAI-organ-
ic complexes.The methodoriginally proposedby Driscoll (1980) utilized a sul-
fonic resin column for species"separation"followed by AI analysisvia the fer-
ALUMINUM 537
ron assay,which was later changedto the 8-hydroxyquinolineassay(Driscoll,

1984). Subsequentstudieshave demonstratedthe utility of pyrochatecholviolet
in an automatedversionof the cationexchangecolumnseparationwhich increas-
es precision and reduces the required sample volume (Seip et aI., 1984;
Rogebereg& Hendricksen,1985; LaZerteet aI., 1988; McAvoy et aI., 1992).
In most applicationsthe method is used to delineatethree operationally
definedAl fractions: (i) a total reactiveAl fraction determinedfollowing acidifi-
cation of the sampleto pH =1 for 1 h (Al), (ii) a "total" monomericAl fraction
(Al m) determinedon a nontreatedsampleby the methodof Barnes(1975), and
(iii) the "nonlabile" monomeric Al fraction determinedon a sample passed
through the sulfonic resin column (No). The No fraction has been ascribedto
organicallycomplexedAl, which is assumedto bypassthe ion exchangecolumn
without interacting. It was further assumedthat all inorganically complexed
mononuclearinorganicAl specieswould completelydissociateand be retained
within the ion exchangecolumn. Thus,the Al m lessthe Alo fraction wasassigned
to a labile monomericfraction assumedto be predominantlymononuclear,inor-
ganically complexedAl, for example,hydroxo-mononuclear Al species,and the
sulfato-, fluoro-, and phosphato-Alcomplexes.Driscoll (1984) pointed out the
importanceof matchingas closely as possiblethe ionic strengthand pH within
the column environmentto that of the original samplesand discussedproper
resin pretreatmentmethodsfor this purpose.Other sourcesof significant varia-
tion in this methodare column size, flow rate, resin beadsize, numberof sam-
ples run through a column, or any factor influencing the sampleresidencetime
within the column. Thesevariablesare particularly critical for sampleshigh in
r, where stable fluoro-Al complexesmay form, those containing weak Al-
organic complexesthat may dissociatewith increasedresidencetime, or those
containingstrongligands,suchas citrate, that can desorbinorganicAl accumu-
latedon the exchanger phase(Hodges,1987;Bertsch,1989b).A numberof stud-
ies have demonstratedthe efficacy of the operational differentiation of the
mononuclearinorganicand organicallyboundAl utilizing this method.Because
of its widespreadacceptance, easeof implementationin field situations,and con-
ceptualsimplicity, it is a very attractivemethodfor Al speciationof naturalsam-
ples.
An ion chromatographic(IC) techniquewas found to separateuncom-
plexedandouter-spherecomplexesfrom inner-sphereinorganicandorganicacid
Al complexesand provide quantitative estimatesof the various inner-sphere
complexescoexistingin solution (Bertsch& Anderson,1989).The methodwas
employedon partially neutralizedAl solutionsand was found to provide values
for mononuclearaqua-hydroxo-Alspecieswithin 2% of the valuesderivedfrom
a quantitative27Al NMR method(Bertschet aI., 1986a,b).Complex redistribu-
tion and coelution of some complex peaksresulting from differencesbetween
sampleand eluent ionic strengthand pH was influencedby the specific ligand
and ligand/Al mole ratios, thus limiting the applicationof this methodfor natur-
al samples.The methodhas, however,beendemonstrateduseful in characteriz-
ing well-defined Al solutions used in kinetic and 'toxicological investigations
(Anderson& Bertsch, 1988). Willett (1989) also employedthe method on salt
extractsof acid soils and demonstratedthe existenceof fluoro-Al complexesin
538 BERTSCH & BLOOM
the extracts,which he inferred as evidencefor their presenceon the exchange

phase.
SPECTROPHOTOMETRICMETHODS
FOR ALUMINUM ANALYSIS
Introduction
Aluminum forms ultravioletor visible absorbingcomplexeswith a number

of organicligandsand the absorbanceis readily usedto determine concentration.
In addition, someAl complexesstrongly fluoresceand measurementof fluores-
cenceintensity can be usedfor quantification of Al at very low concentration.
Both ultraviolet/visible and fluorescencemethodsfor Al determinationrequire
that solutionsnot containstronglycomplexingligandsthat will competewith the
chromophoreor fluorophorefor Al3+. Also, metal ions that cancompetefor com-
plexation with the chromophoreor fluorophore may interfere with Al determi-
nation. Many spectrophotometricmethodsfor the quantitativedeterminationof
Al have beenpublished,all with certain advantagesand disadvantagesfor spe-
cific applications.A numberof the methods,suchas the commonly usedalumi-
non (aurine tricarboxylic acid) assay,involves a complexing organic requiring
colloidal "lake" formation. These methodsshould generally be avoided, espe-
cially for applicationsinvolving speciation,becausethe reaction pathwaysand
stoichiometriesare so poorly defined. A more complete discussionof a wide
rangeof spectrophotometric methodsis given by Bloom and Erich (1989).
8-Hydroxyquinoline-ButylAcetate
SpecialApparatus
1. Extraction tubes,200-mm borosilicateglassculture tubeswith Teflon-
lined screwcaps.
2. Spectrometer.
Reagents
All chemicalsare reagent-gradeexceptwherenotedand are storedin poly-
ethyleneor polypropylenebottles.
1. Aluminum standard,1 x 10--4 M: Dissolve 4.744 g of potassiumalu-
minum sulfate [KAl(S04)2 • 12H20] in water containing10 mL of 1 M
HN03 in a 1-L volumetric flask, and make to volume with deionized
water. This is 1 x 10-2 M, which is then diluted to 1 x 10-4 M with
deionizedwater. Alternately, usea properly dilutedcertified [preferably
NIST (Natl. Inst. Standards& Technol.) traceable] atomic absorption
standardsolution.
2. Sodium acetate (NaOAc), 1 M, containing 0.2% phenanthroline:
Dissolve 2 g of 1,1O-phenanthrolinein 800 mL of deionizedwater by
warming. Add 82 g of NaOAc (anhydroussalt), cool, make to 1-L vol-
ume, and mix thoroughly.
ALUMINUM 539
3. Hydroxylaminehydrochloride(NH20H • HCL): Add 200 g of hydrox-

ylamine hydrochlorideto 1 L of deionizedwater.
4. 8-Hydroxyquinoline, 1%: Dissolve 10 g of 8-hydroxyquinolinein 25
mL of glacial acetic acid. Dilute to 1 L with deionizedwater, and mix
thoroughly.
5. Mixed reagent:Combine four volumes of 1 % 8-hydroxyquinoline
(Reagent4) and one volume of hydroxylaminehydrochloridesolution
(Reagent3). Mix fresh daily.
6. Butyl acetate,spectrograde.
Procedure
Add deionizedwater to the extractiontube so that this volume plus that of
the samplecontainingAI equals25 mL. Then add 5 mL eachof Reagent5 fol-
lowed by Reagent2. Adding Reagent2 first can result in excessivelyhigh pH
with precipitationof AI. Add a samplecontaining1 to 20 Jlg of AI, and briefly
shakethe tube. The final samplemixture shouldhavea pH of 5.0, andit is advis-
able to test a few solutionsfor pH. Allow to stand15 min for complexationof Fe
by phenanthroline,then add 5 mL of butyl acetate.Shakethe tubesvigorously
for 15 s, then let them standfor 15 min for completephaseseparation.Transfer
the butyl acetatephaseto a 1.00-cmborosilicateglasscuvette,and measurethe
absorbanceat 395 nm againsta butyl acetateblank solution. The complex issta-
ble for at least 24 h. The following referencecan be consultedfor additional
information: Bloom et aI., 1978.
Comments
Of the many spectrophotometricmethodsavailable for the quantification
of AI, the 8-hydroxyquinolinemethod offers the best combinationof minimal
interferences,ease,sensitivity, precision,and dynamic linear range.Bloom et al.
(1978) found that butyl acetatewas superiorto CHCl3 in this techniquebecause
of the stability of the complex in butyl acetateeven following prolongedexpo-
sure to ultraviolet light. Additionally, Bloom et al. (1978) presenteda fluoro-
metric determinationof AI with the 8-hydroxyquinolineand butyl acetateextrac-
tion. This techniqueallows the determinationof a very small amountof AI with-
out interferencefrom low levels of severalions. The detectionlimit presented
was 0.3 Jlg L-l of AI, with a practical upperlimit of 50 Jlg L- 1 of AI in solution.
For concentrationsgreaterthan this, the spectrophotometric methodis preferred.
Becauseof the sensitivity, AI contaminationis a problem, and double distilled
water is recommended. Storage of reagents should not be in glass.
Contaminationof reagentsby organiccompoundsthat fluorescealso is a concern
and all reagentsmust be of very high purity.
The 15-min contacttime beforeextractionallows for the 8-hydroxyquino-
line to reactwith AI that is complexedby solubleorganicmatter.This slow reac-
tion rate can be used to separateorganic complexed and colloidal AI from
monomericAI. Fluoride at greaterthan 0.2 mg L-l forms complexesthat are not
detected,but this is not a problemin most soil extracts.Transition metal cations,
notable,Fe2+, Fe3+ and Cu2+ can form complexeswith HQ which interfere, but
540 BERTSCH& BLOOM
this is not a problemin most soils studies(Bloom et aI., 1979). Many other sol-
ventsthanbutyl acetate,notably methyl isobutyl ketone(MIBK), chloroformand
toluenehavebeenusedin the extractionof the HQ complex.Butyl acetateoffers
the advantageof being lessdensethan water, so separatorfunnels are not neces-
sary, and it is less toxic. However, butyl acetatehas a significant solubility in
water so the aqueousvolume must be the samefor all samples.Someauthors
prefer complex formation at pH 8.3 (May et aI., 1979). This version of the pro-
cedurereducesinterferencefrom r but increasesMn2+ interference.This pro-
cedureis more reactivewith organiccomplexesthan the procedureat pH 5.
Alternate SpectrophotometricMethods
Among the many spectrophotometricmethodsavailable for the quantita-

tive determinationof soluble AI (Bloom & Erich, 1996), severalmay provide
advantagesover the HQ methodwhen usedfor specific applications.For exam-
ple, in timed spectrophotometricapproachesof AI speciationof well-defined
partially neutralized solutions, the ferron method has been both extensively
developedand calibrated(e.g., Parker& Bertsch,1992a).The HQ methodis not
convenientfor theseapplicationsbecauseof the requirementfor extractingthe
HQ complexesinto butyl acetateor someother appropriateorganicsolvent. For
automatedAI speciationmethodsusing an ion exchangecolumn, or evenjust for
the automateddeterminationof AI, the pyrocatecholviolet methodworks quite
well and requiresmuch smaller samplevolume than the HQ method (Grigg &
Morrison, 1982; McAvoy et aI., 1992).
Both tiron (4,5-dihydroxy-m-benzenedisulfonic acid) and pyrocatechol
violet have been used as postcolumnreagents forquantitative ion chromato-
graphicdeterminationof both total solubleAI and for applicationsinvolving AI
speciation (Bertsch & Anderson, 1988, 1989; Willett, 1989). Although both
appearedto work well, tiron displayedboth better detectionlimits and a larger
dynamic linear range. An ion chromatographicmethod for AI determination
using fluorometric detectionwith HQ-5-sulphonate was found to haveoutstand-
ing detectionlimits (1 fJ.g L-l) and very large linear range (5-10 000 fJ.g L- 1),
demonstratingsignificant promisefor tracelevel AI determinations(Joneset aI.,
1988).
ATOMIC ABSORPTIONAND INDUCTIVELY COUPLED PLASMA

OPTICAL EMISSION SPECTROMETRY
Atomic Absorptionand Atomic EmissionSpectrometry

FlameAtomic Absorption
Atomic absorptionusing the nitrous oxide/acetyleneflame techniqueis a
commonmethodfor analysisof extractsthat containsufficient concentrationsof
AI (e.g., 1 M KCl). The method,however,is not sensitiveenoughfor soil solu-
tion analysis where concentrationsare less than 1 mg L-l (Bloom & Erich,
1989). Nitrous oxide is required as an oxidant becauseAI forms stable com-
ALUMINUM 541
poundsin coolerflames (e.g.,air/acetylene)which can causeseverereductionin

absorbance.Preconcentrationof AI by extraction of the 8-hydroxyquinoline
complex (vide supra) into a nonaqueousphasecan increasesensitivity by as
much as 100 times (Hsu & Pipes,1972; Barnes,1975).
FlamelessAtomic Absorption
Flamelessatomic absorptionspectrometryis a very sensitivemethod for
AI with a detectionlimit of about 0.15g L-1 (Slavin, 1982).FlamelessAA relies
on the vaporizationof samplesinside a small electrically heatedgraphitetube in
the light path of the spectrometer.Samplesare injected into the graphite tube
which mayor may not havea small platform to hold the injecteddroplet. Precise
analysisrequiresautomaticsampleinjection. After injection, a heatingprogram
is initiated to first dry the sample,then char and atomizethe sample.The chem-
istry and physicsof the charringand atomizationprocessesare complexand pro-
cedural modifications such as use of pyrolytically coatedtubes (Slavin et aI.,
1982) thorium-treatedplatforms(Carnick & Slavin, 1986) and matrix modifiers
(Slavin, 1982) (see Bloom & Erich, 1989, for a more completediscussion)are
usedto modify conditionsto give betterresults.
Matrix modifiers are addedto delay vaporizationof AI and allow for high-
er charring temperatures.For example,an addition of 0.16% Mg(N03)z causes
the AI to becomeimbeddedin a MgO matrix andAI vaporizationis delayeduntil
-MgO is vaporized.Perchloricacid and other chlorine-containingmatricescan
causesevereinterferenceby chlorine binding of AI in the vapor phase.This
interferencecan be reducedby using pyrolytically coatedtubes (Slavin et aI.,
1982).The coatingreducesthe porosity of graphiteand preventsentrainmentof
CI. The quality of the coating is important for minimizing the effects of CI
(Slavin, 1982). Sensitivity can changeas graphitetubesage.Thesechangesmay
be the most pronouncedin the first few firings of a new tube and investigators
should monitor sensitivity.The atomizationof a graphite furnace can be much
more completethan in a flame. Analysis of zeolite suspensionshasshown a 95
to 100% recoveryof AI (Carrondoet aI., 1979)while with kaolinite the recovery
was 85% (Playle et aI., 1982).
PlasmaEmissionand PlasmaMassSpectrometry
The high temperaturesin an Ar-plasma(10 000 K) and the high densityof
free electronsare ideal for atomizingAI. Two typesof plasmageneratorscan be
used: ICP which utilizes microwaveenergy and DCP. The ICP plasmasources
are much more commonand the discussionbelow will refer mostly to ICP. Until
recently,the only meansof detectionhas beenthe optical measurementof emit-
ted light. More recently, MS detectorshave beenused.Atomic emissionICP is
sensitiveenough for detection of AI in most natural systemswith a reported
detectionlimit of 10 Ilg L -1 (Janssenset aI., 1982; Roura et aI., 1982). Because
of the high temperaturein the plasma,most matrix interferencesare eliminated.
The only significant matrix effect is due to changesin viscosity of the sample
(Botto, 1981). Spectral interferences,however, can be significant due to the
overlapof emissionlines.
542 BERTSCH& BLOOM
Reportedinterferencesinclude Ag, B, Mn and Ca. Of these,Mn and Ca

were the most commonin soils studies.Computercorrectionfor spectraloverlap
can eliminate much of the problem. Atomic emission ICP spectrometersare
commonin soil and plant analytical laboratoriesand are usedto analyzea wide
variety of samplesfrom plants and soils (Jones,1977; Soltanpouret aI., 1979;
Lechleret aI., 1980).In addition to injection of solubilizedsamples,someinves-
tigations have analyzedaqueousclay suspensions(Laird et aI., 1991).
ALTERNATIVE METHODS FORALUMINUM ANALYSIS
X-Ray FluorescenceSpectrometry
The use of x-ray spectrometricanalysishas increasedwith the adventof

microprocessorsand the developmentof computersoftwarepackagesthat were
developedin the mid-1970s.The new systemsare fully automatedand allow for
rapid total chemicalanalysisof bulk samples.The generallack of sensitivity lim-
its the use of x-ray fluorescencefor AI analysisto data requirements in the per-
centage range. This lack of sensitivity for AI analysisis largely relatedto its low
atomicweight, but is still useful for AI determinationin bulk soil or mineralsam-
ples,obviating the needfor chemicaldissolution.For complete detailsof sample
preparationand operationalprocedures,see Chapter7 (Karathanasis& Hajek,
1996).
Titrimetric Methods
A number of titrimetric methods have been employed in determining

monomeric AI concentrationsin solutions. The majority of these techniques
involve the titration of soil extractsor suspensionswith dilute alkali. Monitoring
of the various systemsmay be done by severalmeans:pH, as in potentiometric
titration analyses;relative solution conductivity, as is the casefor conductomet-
ric titrations; and the color changein the presenceof an organic indicator, as in
the methodof Yuan (1959). Titrations with complexingagentssuchas F- or eth-
ylenediaminetetraacetic acid (EDTA) also have beenusedto determinesolution
AI, or to estimatechemically labile soil AI (Ares, 1986a,b;Ares & Ziechman,
1988).
The titration of KCI extractsto the endpointof phenolthaleinprovidesan
estimateof "exchangeable"soil acidity. This method is presentedin detail by
McLean (1965) with a summaryof all appropriatechemicalreactions.The end-
point, however, is not as abrupt as with most acid-basetitrations because
AI(OH)3 formation during the titration is sluggish.Potentiometrictitration analy-
sis has beena useful techniquein the speciationof acidic componentsin solu-
tions. This methodhas beengreatly enhancedin recentyearsby the availability
of computercontrolledautomatictitrators which allow for careful control of the
neutralizationreactionand the titration conditions(Bertschet aI., 1989). These
titrations can be conductedon solutions containing dissolvedAI such as KCI
extracts or whole soil systems,generally reasonableendpoints for solution
ALUMINUM 543
monomericAI are obtained.The presenceof hydronium ions as the hydrolytic

product of exchangeableor nonexchangeable AI sources(or both) will titrate
beforethe AI, and this reactionmay produceweak nonidentifiableendpoints.
Solution AI concentrationsalso have been determinedusing complexo-
metric titration analyseswith such substancesas oxine, EDTA, and F (Kolthoff
& Elving, 1966,p. 402-404).Advancesin F specificion electrodeshaveallowed
very convenientand accuratedeterminationsof nanomolaramountsof solution
AI (Radic, 1976). Thesemethodsinclude potentiometrictitrations with P-- solu-
tions (Baumann, 1970; laselskis & Bandermer, 1969) and monitoring the
decreasein P-- activity with time. AIthough thesemethodsare quite easy, they
seemto have applicability only for very "clean" systemsbecauseof the direct
interferenceof both positively and negatively charged ions. The techniques
requiredfor removalof interfering ions seriouslyrestrict the usefulnessor prac-
ticality of thesemethods,especiallywhen other suitable methodsfor AI deter-
mination are available(Baumann,1970).
Fluoride-titratableAI has beenusedto estimate"reactive" soil AI in rela-
tion to plant growth (e.g., Stilwell & Asscott, 1978; Ares, 1986a,b; Ares &
Ziechman,1988).Thesestudiesfound no significant correlationbetweenKCl or
dilute CaCl2 extractableAI and plant growth; however,there was a significant
correlation with plant growth and maximum levels of P-- reactive AI in soils.
They alsoindicatethat the superiorityof P-- titrationsfor estimating"reactiveAI"
resultedfrom the inclusion of noncrystallinemetastableAI forms that they con-
sider a potential significant sourceof phytotoxic hexa-aquaAI 3+ species.Large
variationsin resultsobtainedwith this methodwould be expected based on min-
eralogicalconsiderationsand the natureand stability of noncrystallineAI frac-
tions from soils of varying degreesof weathering,etc.
REFERENCES
Adams, W.A, AY. Ali, and P.I. Lewis. 1990. Releaseof cationic aluminum from acidic soils into
drainagewater and relationshipswith land use.I. Soil Sci. 41:255-268.
Alva, AK., M.D. Sumner,Y.c. Li, and w.P. Miller. 1989. Evaluationof threealuminumassaytech-
niques for excluding aluminum complexedwith fluoride or sulfate. Soil Sci. Soc. Am. J.
53:38-44.
Alvarado,A, and S.W. Buol. 1985. Field estimationof phosphateretentionby andepts.Soil Sci. Soc.
Am. J. 49:911-914.
Amedee,G., and M. Peech.1976. The significanceof KCl extractableAl(III) as an index to lime
requirementof soils of the humid tropics. Soil Sci. 121:227-233.
Anderson,M.A., and P.M. Bertsch. 1988. Dynamicsof aluminum complexationin multiple ligand
systems.Soil Sci. Soc. Am. J. 52:1597-1602.
Anderson,M.A, L.W. Zelazny,andP.M. Bertsch. 1991. Fluoro-aluminumcomplexeson model and
soil exchangers.Soil Sci. Soc. Am. J. 55:71-75.
Arai, S. 1975. Extractionof active aluminum forms from acid soils in Japanwith different reagents.
Geoderma14:63-74.
Ares, J. 1986a.Identification of aluminum speciesin acid forest soil solutionson the basisof Al:F
reactionkinetics: 1. Reactionpathsin pure solutions.Soil Sci. 141:399-407.
Ares, J. 1986b. Identification of aluminum speciesin acid forest soil solutionson the basisof Al:F
reactionkinetics: 2. An exampleat the solling area.Soil Sci. 142:13-19.
Ares, J., and W. Ziecheman.1988. Interactionof organicmatterand aluminumions in acid forest soil
solutions:Metal complexation,flocculation, and precipitation.Soil Sci. 145:437--447.
Bache,B.W., and G.S. Sharp.1976. Soluble polymeric hydroxy-aluminumions in acid soils. J. Soil
Sci. 27:167-174.
S44 BERTSCH& BLOOM
Barnes,R.B. 1975.The detenninationof specificfonns of aluminumin naturalwater. Chem.Geol.

15:177-191.
Barnhisel,R.I., and P.M. Bertsch.1989.Chloritesandhydroxy-interlayeredvenniculiteandsmectite.
p. 729-788.In 1. Dixon and S.B. Weed (ed.) Minerals in soil environments.2nd ed. SSSA
Book Ser. 1. SSSA,Madison,WI.
Bartlett, R.I., D.S. Ross,andF.R. Magdoff. 1987. Simple kinetic fractionationof reactivealuminum
in soil "solutions."Soil Sci. Soc.Am. J. 51:1479-1482.
Bartoli, F., and R. Philippy. 1990.AI-organic matterassociationsas cementingsubstances of ochre-
ousbrown soil aggregates: preliminary examination.Soil Sci. 150:745-751.
Bascomb,C.L. 1968.Distribution of pyrophosphateextractableiron and organiccarbon.1. Soil Sci.
19:251-268.
Baumann,E.W.. 1970. Detenninationof aluminum by potentiometrictitration with fluoride. Anal.
Chem.42:110-111.
Bernas,B. 1968. A new methodfor the decompositionand comprehensiveanalysisof silicatesby
atomicabsorptionspectrometry.Anal. Chem.40:1682-1686.
Bertsch,P.M. 1987. Conditionsfor Al13 polymer fonnation in partially neutralizedaluminumsolu-
tions. Soil. Sci. Soc.Am. 1. 51:825-828.
Bertsch,P.M. 1989a.Aqueouspolynuclearaluminum species.p. 87-115.In G. Sposito(ed.) The
environmentalchemistryof aluminum.CRC Press,Inc., BocaRaton,FL.
Bertsch, P.M. 1989b. Aluminum speciation: Methodology and applications.p. 63-105. In D.C.
AdrianoandW. Salomons(ed.)Advancesin environmentalscience.Acidic precipitation.Vol.
4. Springer-Verlag,New York.
Bertsch,P.M., and M.A. Anderson.1988. Detenninationof aluminum extractedfrom soils by ion
chromatography.Soil Sci. Soc.Am. 1. 52:540-542.
Bertsch,P.M., and M.A. Anderson.1989. Speciationof aluminum in aqueoussolutionsusing ion
chromatography.Anal. Chem.61:535-539.
Bertschi P.M., G.W. Thomas,andR.1. Barnihisel.1986a.Characterization of hydroxy-AI solutionsby
7A1 NMR spectroscopy.Soil Sci. Soc.Am. 1. 50:825-828.
Bertsch,P.M., W.l. Layton, and R.1. Barnhisel. 1986b. Speciationof hydroxy-AI solutionsby wet
chemicalandAI-27 NMR methods.Soil Sci. Soc. Am. 1. 50:1449-1454.
Bertsch,P.M., W.P. Miller, M.A. Anderson,andL. W. Zelazny.1989.Coprecipitationof iron andalu-
minum during titration of mixed A13+, Fe3+ solutions.Clays Clay Miner. 37:12-18.
Bloom, P.R. 1981. Metal-organicmatter interactionsin soil. p. 129-150.In R.H. Dowdy (ed.)
Chemistryin the soil environment.ASA, Madison,WI.
Bloom, P.R., and M.S. Erich. 1996. The quantitationof aqueousaluminum. p. 1-27. In G. Sposito
(ed.) Geochemistryof aluminum.CRC Press,Boca Raton,FL.
Bloom, P.R., M.B. McBride, and B. Chadbourne.1977. Adsorption of aluminum by a smectite:I.
Surfacehydrolysisduring Ca2+_Al3+ exchange.Soil Sci. Soc. Am. 1. 41:1068-1073.
Bloom, P.R.,R.M. Weaver,andM.B. McBride. 1978.The spectrophotometric andfluorometricdeter-
minationof AI with 8-hydroxyquinolineand butyl acetate.Soil Sci. Soc.Am. 1. 42:713-716.
Bloom, P.R., M.B. McBride, and R.M. Weaver. 1979. Aluminum organic matter in acid soils:
Buffering and solution aluminumactivity. Soil Sci. Soc.Am. 1. 43:488-493.
Botto, R.I. 1981.Interferencecorrectionfor simultaneousmultielementdetenninationsby inductive-
ly coupledplasma.p. 141. In R.M. Barnes(ed.) Developmentsin atomic plasmaspectro-
chemicalanalysis.Heyden,London.
Brown, G., and A.C.D. Newman.1973.The reactionsof solublealuminumwith montmorillonite.1.
Soil Sci. 24:339-354.
Browne, B.A., 1.G. McColl, and c.T. Driscoll. 1990a.Aluminum speciationusing morin: I. Morin
and its complexeswith aluminum.1. Environ. Qual. 19:65-72.
Browne, B.A., 1.G. McColl, and C.T. Driscoll. 199Ob. Aluminum speciation using morin: II.
Principlesand procedures.1. Environ. Qual. 19:73-82.
Campbell,P.G.C., M. Bisson, R. Bougie, A. Tessier,and l.-P. Villeneuve. 1983. Speciationof alu-
minum in acidic freshwaters.Anal. Chern.55:224~2252.
Cantillo, A. Y., S.A. Sinex,and G.R. Helz. 1984.Elementalanalysisfor estuarinesedimentsby lithi-
um metaboratefusion and direct current plasma emission spectrometry.Anal. Chern.
56:33-37.
Carnick,G.R., and W. Slavin. 1986.Use ofTh treatedplatfonnsfor the detenninationof AI and Pd.
AI. Spectrosc.7:175.
Carrondo,M.J.T., J.N. Lester,andR. Perry. 1979.Electrothennalatomicabsorptiondetenninationof
total aluminum (including zeolite type A) in waters and waste waters. Anal. Chim. Acta
111:291-295.
ALUMINUM 545
Chao, T.T., and L. Zhou. 1983. Extraction techniquesfor selectivedissolution of amorphousiron

oxidesfrom soils and sediments.Soil Sci. Soc. Am. 1. 47:225-232.
Coleman, N.T., and G.w. Thomas. 1967. The basic chemistry of soil acidity. p. 1-34. In R.W.
Pearsonand F. Adams (ed.) Soil acidity and liming. Agron. Monogr. 12. ASA, CSSA, and
SSSA,Madison,WI.
Coulter, B.S. 1969.The chemistryof hydrogenand aluminum ions in soils, clay mineralsand resins.
Soil Fert. 32:215-223.
Cronan, C.S., W.J. Walker, and P.R Bloom. 1986. Predicting aluminum in natural waters. Nature
(London) 324:140-143.
David, M.B., G.F. Vance,and W.J. Fasth.1991. Forestsoil responseto acid and salt additionsof sul-
fate: II. Aluminum and basecations.Soil Sci. 151:208-219.
Denney,D.Z., and P.H. Hsu. 1986.27AI nuclearmagneticresonanceand ferron kinetic studiesof par-
tially neutralizedAlCI 3 solutions.Clays Clay Miner. 34:604-607.
Driscoll, C.T. 1980. Chemistryand characterizationof somedilute acidified lakes andstreamsin the
Adirondack region of New York State.Ph.D. Thesis,Cornell Univ., Ithaca, NY.
Driscoll, C.T. 1984. A procedurefor the fractionationof aqueousaluminum in dilute acidic waters.
Int. 1. Environ. Anal. Chem. 16:267-284.
Elkhatib, E.A., J.L. Hem, and T.E. Staley. 1987. A rapid centrifugationmethod for obtaining soil
solution. Soil Sci. Soc. Am. J. 51:578-583.
Evans,A., Jr., and L.W. Zelazny. 1987.Effects of sulfateadditionson the statusof exchangeablealu-
minum in a cecil soil. Soil Sci. 143:410-417.
Farina,M.P.W., M.E. Sumner,C.O. Plank, and W.S. Letzsch.1980.Exchangeablealuminumand pH
as indicatorsof lime requirementfor corn. Soil Sci. Soc. Am. J. 44:1036-1041.
Frink, C.R 1973. Aluminum chemistry in acid sulfate soils. p. l31-168.In H. Dost (ed.) Acid sul-
fate soils. Vol. 1. Int. Inst. Land Reclam.Improve. Publ. 18. Wageningen,the Netherlands.
Furrer, G., B. Trusch,and C. Muller. 1992. The formation of polynuclearAll3 under simulatednat-
ural conditions.Geochim.Cosmochim.Acta 56:3831-3838.
Furrer, G. 1993. New aspectson the chemistryof aluminum in soils. Aquat. Sci. 55:281-290.
Gardiner,P.E., R. Schierl, and K. Kreutzer. 1987.Aluminum speciationin soil solution as studiedby
size exclusionchromotography.Plant Soil 103:151-154.
Grigg, J.L., and D. Morrison. 1982. An automaticcolorimetric determinationof aluminum in soil
extractsusing catecholviolet. Commun.Soil Sci. Plant Anal. l3:351-367.
Grossmann,J., and P. Udluft. 1991.The extractionof soil waterby the suction-cupmethod:A review.
J. Soil Sci. 42:83-93.
Hargrove,w.L., and G.W. Thomas.1981. Effect of organic matter on exchangeablealuminum and
plantgrowth in acid soils. In R.H. Dowdy (ed.) Chemistryin the soil environment.ASA Spec.
Publ. 40. ASA and SSSA,Madison,WI.
Hargrove,W.L., and G.W. Thomas.1984. Extractionof aluminum from aluminum-organicmatterin
relation to titratable acidity. Soil Sci. Soc. Am. J. 48:1458-1460.
Hodges,S.C. 1987. Aluminum speciation: A comparisonof five methods. Soil Sci. Soc. Am. J.
51:57-M.
Hodges,S.C., and L.W. Zelazny. 1983. Influencesof OH/AI ratios and loading rates in aluminum-
kaolinite interactions.Soil Sci. Soc. Am. 1. 47:221-225.
Holmgren,G.G.S.,and J.M. Kimble. 1984. Field estimationof amorphousaluminum with 4M potas-
sium hydroxide. Soil Sci. Soc. Am. J. 48:l378-l382.
Holmgren, G.G.S., and RD. Yeck. 1984. Field identification of spodic horizons with potassium
hydroxide extractablealuminum and humic acid color. Soil Sci. Soc. Am. J. 48:l370-l374.
Hossner,L.R. 1996. Dissolution for total elementalanalysis.p. 49~4. In D.L. Sparkse tal. (ed.)
Methods of soil analysis. Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and ASA,
Madison,WI.
Hoyt, P.B.,and M.D. Webber.1974.Rapid measurement of plant-availablealuminumand manganese
in acid Canadiansoils. Can J. Soil Sci. 54:53~1.
Hsu, P.H. 1968. Heterogeneityof montmorillonite surfaceand its effect on the natureof hydroxy-AI
interlayers.Clays Clay Miner. 16:303-311.
Hsu, P.H. 1992. Reaction of OH-AI polymers with smectitesand vermiculite. Clays Clay Miner.
40:300-305.
Hsu, Y., and O. Pipes. 1972. Modification of techniquefor determinationof aluminum in water by
atomic absorptionspectrophotometry.Environ. Sci. Technol. 6:645~47.
Hughes,S., and B. Reynolds.1990.Evaluationof porousceramiccupsfor monitoring soil-wateralu-
minum in acid soils: comment on a paper by Raulund-Rasmussen (1989). J. Soil Sci.
41:325-328.
546 BERTSCH & BLOOM
Hunter, D., and D.S. Ross. 1990. Evidencefor phytotoxic Hydroxy-aluminumpolymer in organic
soil horizons.Science(Washington,DC) 251:1056-1058.
Inskeep,W.P., and P.R. Bloom. 1986a.Calcium carbonatesupersaturationin soil solutionsof calci-
aquolls.Soil Sci. Soc. Am. J. 50:1431-1437.
Inskeep,W.P., and P.R. Bloom. 1986b.Effectsof soil moistureon soil pC02 soil solution bicarbon-
ate and iron chlorosisin soybeans.Soil Sci. Soc.Am. J. 50:946-952.
Iyengar,S.S.,L W. Zelazny,and D.C. Martens.1981. Effect of photolytic oxalatetreatmenton soil
hydroxy-interJayered vermiculites.Clays Clay Miner. 29:429--434.
Jackson,M.L 1960. Structuralrole of hydronium in layer silicatesduring soil genesis.Trans. Int.
Congr.Soil Sci. 7th. 11:445-455.
Jackson,M.L, C.H. Lim, and L W. Zelazny. 1986. Oxides, hydroxides,and aluminosilicates.p.
101-150. In A. Klute (ed.) Methodsof soil analysis.Part 1. 2nd ed. Agron. Monogr. 9. ASA
James,B.R., C.J. Clark, and S.J.Riha. 1983.An 8 hydroxyquinolinemethodfor labile and total alu-
minum in soil extracts.Soil Sci. Soc.Am. J. 47:893-897.
Janssens, E., P. Schutyser,andR. Dams.1982.ICP automated,sequentialanalysisof AI, Be, Cd, Co,
Cr, Cu, Fe, Mn, Ni and Zn in surfacewatersamples.Environ. Technol.Lett. 3:35-42.
Jardine,P.M., L.W. Zelazny, and J.C. Parker. 1985. Mechanismsof aluminum adsorptionon clay
mineralsand peat.Soil Sci. Soc.Am. J. 49:862-867.
Jardine,P.M., L W. Zelazny, and A. Evans,Jr. 1986. Solution aluminum anomaliesresulting from
variousfiltering materials.Soil Sci. Soc. Am. J. 50:891-894.
Jardine,P.M., and L.W. Zelazny. 1987a.Influence of organicanionson the speciationof mononu-
clearand polynuclearaluminumby ferron. Soil Sci. Soc. Am. J. 51:885-889.
Jardine,P.M., and L.W. Zelazny.1987b.Influenceof inorganicanionson the speciationof mononu-
clearand polynuclearaluminumby ferron. Siol Sci. Soc.Am. J. 51:889-892.
Jarvis, S.C. 1986. Forms of aluminum in some acid permanentgrasslandsoils. J. Soil Sci.
37:211-222.
Jaselskis,B., and M.K. Bandermer.1969. Determinationof micro and sernimicroamountsof alu-
minum using fluoride activity electrodes.Anal. Chern.41:855-857.
Jenny,H. 1961.Reflectionson the soil acidity merry-go-round.Soil Sci. Soc.Am. Proc.25:4~32.
Jersak,J.M., andJ.G. McColl. 1989.Aluminum releasefrom solid-phasecomponentsof forest soils
with citric acid. Soil Sci. Soc.Am. 1. 53:550--555.
Jones,B.F., V.C. Kennedy,and G.W. Zellweger. 1974. Comparisonof observedand calculatedcon-
centrationsof dissolvedAI and Fe in streamwater. WaterResour.Res. 10:791-793.
lones,I.B., lr. 1977.Elementalanalysisof soil extractsandplant tissueashby plasmaemissionspec-
troscopy.Commun.Soil Sci. PlantAnal. 8:349-365.
Jones,P.L. Ebdon,andT. Williams. 1988.Determinationof traceamountsof aluminumby ion chro-
matographywith fluorescencedetection.Analyst 113:641-644.
Juo, A.S.R., S.A. Ayanlaja, and J.A. Ogunwale.1976. An evaluationof cation exchangecapacity
measurements for soils in the tropics. Commun.Soil Sci. PlantAnal. 7:751-761.
luo, A.S.R., andE.J. Kamprath. 1979.Copperchloride as an extractantfor estimatingthe potential-
ly reactivealuminumpool in acid soils. Soil Sci. Soc.Am. J. 43:35-38.
Kamprath,E.J. 1970. Exchangeablealuminum as a criterion for liming leachedmineral soils. Soil
Sci. Soc.Am. Proc. 34:252-254.
Karathanasis,A.D., and B.F. Hajek. 1996.Elementalanalysisby x-ray fluorescencespectroscopy.p.
161-223. In D.L Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.
SSSABook Ser. 5. SSSAandASA, Madison,WI.
Kennedy,V.C., G.w. Zellweger,and B.F. Jones.1974. Filter pore size effectson the analysisof AI,
Fe, Mn, and Ti in water.WaterResour.Res. 10:785-785.
Khalid, R.A., and1.A. Silva. 1979.A study of soil aluminumextractionmethodsin relation to plant
aluminumand yield in tropical soils. Trop. Agric. 56:53-63.
Kimble, J.M., C.S. Holzhey, and G.G.S. Holmgren. 1984. An evaluationof potassiumhydroxide-
extractablealuminumin Andepts(Andisols). Soil Sci. Am. 1. 48:1366-1369.
Kodama,H., and C. Wang. 1989. Distribution andcharacterizationof noncrystallineinorganiccom-
ponentsin spodosolsand spodosol-likesoils. Soil Sci. Soc.Am. 1. 53:526-534.
Kolthoff, I.M., and P.l. Elving. 1966.Treatiseon analyticalchemistry.Part 2. Vol. 4. Intersci.,New
York.
Kotze, w.A.G., M. Jouberg,J.F. de VilJiers, M. van der Westhuizen,and D. van der Bank. 1984.
Measurementof exchangeablealuminumin soils and clay mineralsby isotopicexchange.S.
Afr. J. PlantSoil 1(2):57-60.
ALUMINUM 547
Laird, DA, R.H. Dowdy, and R.C. Munter. 1991. Suspensionnebulization analysis of clays by
inductivley coupledplasma-atomicemissionspectroscopy.Soil Sci. Soc. Am. 1. 55:274-278.
Lalande, H., and W.H. Hendershot. 1985. Aluminum speciation in some synthetic systems:
Comparisonof the fast-oxine,pH 5.0 extractionand dialysismethods.Can.1. Fish Aquat. Sci.
43:231-234.
LaZerte, B.D. 1984. Forms of aqueousaluminum in acidified catchmentsof central Ontario: a
methodologicalanalysis.Can. 1. Fish Aquat. Sci. 41:766-776.
LaZerte, B.D., C. Chun, and D. Evans. 1988. Measurementof aqueous aluminum species:
Comparisonof dialysis and ion-exchangetechniques. Environ.Sci. Technol. 22:1106-1108.
Lechler, P.I., W.R. Roy, and R.K. Leininger. 1980. Major and trace elementanalysisof 12 reference
soils by inductivley coupledplasma-atomicemissionspectrometry.Soil Sci. 130:238-24l.
Lee, R., B.W. Bache,MJ. Wilson, and G.S. Sharp.1985. Aluminum releasein relation to the deter-
mination of cation exchangecapacity of some podzolized New Zealandsoils. J. Soil Sci.
36:239-253.
Lee, EY., T.L. Yuan, and V.W. Carlisle. 1989. Nature of cementingmaterialsin orstein horizonsof
selectedFlorida spodosols:II. Soil propertiesand chemical form(s) of aluminum. Soil Sci.
Soc. Am. J. 52:1796-1801.
Lekwa, G., and E.P. Whiteside. 1986. Coastal plain soils of southeasternNigeria: II. Forms of
extractableiron, aluminum,and phosphorus.Soil Sci. Soc. Am. 1. 50:160-166.
Lilieholm, B.c., and S.E. Feagley.1988. Effects of simulatedacid rain on soil and leachateacidifi-
cation of a Lexington silt loam. Soil Sci. 146:44-50.
Lin, c., and N.T. Coleman.1960. The measurementof exchangeableAI in soils and clays. Soil Sci.
Soc. Am. Proc. 24:444-446.
Litaor, M.1. 1987. Aluminum chemistry: Fractionation,speciation,and mineral equilibria of soil
interstitial watersof an alpine watershed,front range,Colorado.Geochim.Cosmochim.Acta
51:1285-1295.
Litaor, M.1. 1988. Review of soil solution samplers.Water Resour.Res. 24:727-733.
Loveland, PJ., and P. Digby. 1984. The extractionof Fe and AI by 0.1 M phrophosphatesolutions:
A comparisonof sometechniques.1. Soil Sci. 35:243-250.
May, H.M., P.A. Helmke, and M.L. Jackson.1979. Determinationof mononucleardissolved alu-
minum in nearneutral waters.Chem.Geol. 24:259-269.
McAvoy, D.C., R.C. Santore,1.D. Shosa,and c.T. Driscoll. 1992.Comparisonbetweenpyrocatechol
violet and 8-hydroxyquinolineproceduresfor determiningaluminumfractions. Soil Sci. Soc.
Am. J. 56:449-455.
McKeague,1.A.1967.An evaluationof 0.1 M pyrophosphateand pyrophosphate-dithionate in com-
parisonwith oxalate as extractantsof the accumulationproductsin Podzolsand someother
soils. Can. J. Soil Sci. 47:95-99.
McKeague,J.A., 1.E. Brydon, and N.M. Miles. 1971.Differentiation of forms of extractableiron and
aluminum in soils. Soil Sci. Soc. Am. Proc. 35:33-38.
McKeague, J.A., and P.A. Schuppli. 1982. Changesin concentrationof iron and aluminum in
pyrophosphateextractsof soil and compositionof sedimentresultingfrom ultracentrifugation
in relation to spodic horizon criteria. Soil Sci. 134:265-270.
Mclean, E.O. 1965. Aluminum p. 978-998.In c.A. Black et al. (ed.) Methodsof soil analysis.Part
2. SSSABook Ser. 5. ASA and SSSA, Madison,WI.
Meyer, M.L., and P.R. Bloom. 1993. Lithium metaboratefusion for silicon calcium magnesiumand
potassiumanalysisof wild rice. Plant Soil 153:281-285.
Mokama,D.L. 1993.Color and amorphousmaterialsin spodosolsfrom Michigan. Soil Sci. Soc. Am.
J.578:125-128.
Mulder, J., 1.1.M. van Grinsven,and N. van Breemen.1987. Impactsof acid atmosphericdeposition
on woodland soils in The Netherlands:III. Aluminum chemistry. Soil Sci. Soc. Am. J.
51:1640-1646.
Noble, A.D., M.E. Sumner,and A.K. Alva. 1988. Comparisonof aluminon and 8-hydroxyquinoline
methodsin the presenceof fluoride for assayingphytotoxic aluminum. Soil Sci. Soc. Am. 1.
52:1059-1063.
Oates,K.M., and E.1. Kamprath. 1983a.Soil acidity and liming: I. Effect of the extractingsolution
cation and pH on the removalof aluminumfrom acid soils. Soil Sci. Soc. Am. J. 47:686-689.
Oates,K.M., and E.1. Kamprath. 1983b. Soil acidity and liming: II. Evaluationof using aluminum
extractedby variouschloride salts for determininglime requirements.Soili Sci. Soc. Am. J.
47:690-692.
Okura, T., K. Goto, and T. Yotsuyanagi.1962. Formsof aluminumdeterminedby an 8-quinolinolate
extractionmethod.Anal. Res. Chern. 34:581-582.
548 BERTSCH& BLOOM
Page,E, and C.R. DeKimpe. 1989.Dissolution descomposesferrugineuxet alumineuxdes horizons

B pozoliquesde sols du Quebecpar Ie dithionite-citrate-bicarbonate, o'oxalate,Ie pyrophos-
phateet Ie tetraborate.Can. I. Soil Sci. 69:451-459.
Parfitt, R.I., C.W. Childs, and D.N. Eden. 1988. Ferrihydrite and allophanein four Andepts from
Hawaii and implicationsfor their classification.Geoderma41:223-241.
Parker,D.R., R.L. Chaney,W.A. Norvel. 1995. Chemicalequilibrium models: Applications to plant
nutrition research.In R.H. Loeppertet al. (ed.) Chemical equilibrium and reaction models.
SSSASpec.Publ. 42. SSSA,Madison,WI.
Parker,D.R., and P.M. Bertsch. 1992a. Identification and quantification of the'Al n ' tridecameric
polycation using ferron. Environ. Sci. Technol. 26:908-914.
Parker,D.R., and P.M. Bertsch.1992b.Formationof the 'Al l3 ' tridecamericpolycationunderdiverse
synthesisconditions.Environ. Sci. Technol. 26:914-921.
Parker,D.R., L.W. Zelazny,and T.B. Kinraide. 1988. Comparisonof three spectrophotometricmeth-
ods for differentiating mono- and polynuclearhydroxy-aluminumcomplexes.Soil Sci. Soc.
Am. 1. 52:67-75.
Parker,D.R., T.B. Kinraide, and L.W. Zelazny. 1989. On the phytotoxicity of polynuclearhydroxy-
aluminum complexes.Soil Sci. Soc. Am. I. 53:789-796.
Parthasarthy,N., and I. Buffle. 1985. Study of polymeric aluminum (III) hydroxide solutions for
application in waste water treatment.Propertiesof the polymer and optimal conditions of
preparation.Water Resour.Res. 19:25.
Pavan,M.A., ET. Bingham, and P.E Pratt. 1984. Redistributionof exchangeablecalcium, magne-
sium, and aluminum following lime or gypsum applicationsto a Brazilian oxisol. Soil Sci.
Soc. Am. I. 48:33-38.
PlayIe, R., I. Gleed, R. Ionasson,and 1.R. Kramer. 1982. Comparisonof atomic absorptionspec-
trophotometric,spectrophotometricand fluorimetric methodsfor the determinationof alu-
minum in water. Anal. Chim. Acta 134:369-373.
Radic, N.J. 1976. Determinationof Nanomoleamountsof aluminum by use of a fluoride ion-sensi-
tive electrode.Analyst (London) 101:657-660.
Rasmussen,P.E., S.L. Schiff, and H.W. Nesbitt. 1991. The determinationof exchangeablecationsin
acid soils: Errors causedby weatheringreactionsduring neutral salt extraction.Can. 1. Soil
Sci. 71:155-163.
Raulund-Rasmussen, K. 1989.Aluminium contaminationand otherchangesof acid soil solution iso-
lated by meansof porcelainsuction-cups.J. Soil Sci. 40:95-1Ol.
Raulund-Rasmussen, K. 1991. Aluminum contaminationof acid soil solution isolated by meansof
porcelainsuction cups: A reply to a paperby Hughes& Reynolds(1990) and an interpreta-
tion of aluminum release.J. Soil Sci. 42:271-276.
Reeve,N.G., and M.E. Sumner. 1970. Lime requirementsof Natal oxisols basedon exchangeable
aluminum. Soil Sci. Soc. Am. Proc. 34:395-398.
Rengasamy,P., andJ.M. Oades.1978. Interactionof monomericand polymeric speciesof metal ions
with clay surfaces.III. Aluminum (III) and Chromium (III). Aust. J. Soil Res. 16:53-66.
Reuss,J.O., and D.W. Johnson.1985. Effect of soil processeson the acidification of water by acidic
deposition.1. Environ. Qual. 14:26-3l.
Reuss,J.O., P.M. Walthall, E.C. Roswall, and R.W.E. Hopper. 1990. Aluminum solubility, calcium-
aluminum exchange,and pH in acid forest soils. Soil Sci. Soc. Am. 1. 54:374-390.
Rich, c.1. 1960.Aluminum in interlayersof vermiculite. Soil Soc. Am. Proc. 24:26-32.
Richardson,J.L., and EE Riecken. 1977. Differencesin exchangeablealuminum and soil acidity in
Loesssoils ofIowa. Soil Sci. Soc. Am. 1. 41:588-593.
Rogebereg,E.J.S.,and A. Henriksen. 1985. An automaticmethodfor fractionation and determina-
tion of aluminum speciesin fresh waters.Vatten 41:48-53.
Roura, M., M. Baucells,G. Lacort, and G. Rauret. 1982. Determinationof aluminium in river water
by the inductively coupledplasma-atomicemissionspectroscopic(ICP) technique,Analytical
techniques.p. 377. In I. Albaiges (ed.) Environmentalchemistry. Vol. 7. PergamonPress,
Elmsford, NY.
Royset, 0., and T.J. Sullivan. 1986. Effect of dissolvedhumic compoundson the determinationof
aqueousaluminum by three spectrophotometricmethods. In!. J. Environ. Anal. Chern.
27:305-314.
Sawhney,B.L. 1996. Extraction of organic chemicals. p. 1071-1084.In D.L. Sparkset al. (ed.)
Madison,WI.
Schecher,W.D., and C.T. Driscoll. 1987.An evaluationof the uncertaintyassociatedwith aluminum
equilibrium calculations.Water Resour.Res. 23:525-534.
ALUMINUM 549
Seip, H.M., L. Muller, and A. Naas. 1984. Aluminum speciation:Comparisonof two spectrophoto-
metric analytical methodsand observedconcentrationsin some acidic aquatic systemsin
southernNorway. Water Soil Air Pollut. 23:81-95.
Shann,J.R.,and P.M. Bertsch.1993.Differential cultivar responseto polynuclearhydroxo-aluminum
complexes.Soil Sci. Soc. Am. J. 57:116--120.
Shuman,L.M. 1990. Comparisonof exchangeableAI, extractableAI, and AI in soil fractions. Can.
J. Soil Sci. 70:263-275.
Sims, J.T. 1996. Lime requirement.p. 491-515.In D.L. Sparkset al. (ed.) Methodsof soil analysis.
Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Skeen,J.B., and M.E. Sumner. 1967. Exchangeablealuminum. I. The efficiency of various elec-
trolytes for extractingaluminum from acid soils. S. Afr. J. Agric. Sci. 10:3-10.
Slavin, W. 1982. Environmentaltrace analyseswith the stabilizedtemperatureplatform furnaceand
Zeemanbackgroundcorrection. p. 397-405. In J. Albaiges (ed.) Analytical techniquesin
environmentalchemistry.Vol. 7. PergamonPress,Elmsford, NY.
Slavin, w., G.R. Camrick, and D.C. Manning. 1982. Graphite-tubeeffects on perchloric acid inter-
ferenceson aluminum and thallium in the stabilized-temperatureplatform furnace, Anal.
Chim. Acta 138:103-110.
Smith, R.W. 1971. Reactionsamong equilibriumand non-equilibriumaqueousspeciesof aluminum
hydroxy complexes.Adv. Chern. Ser. 106:250-279.
Smith, R.w., and J.D. Hem. 1972.Effect of aging on aluminum hydroxidecomplexesin dilute aque-
ous solutions. U.S. Geol. Surv. Water Supply Pap. 1827-D. U.S. Gov. Print. Office,
Washington,DC.
Soltanpour,P.N., S.M. Workman, and AP. Schwab.1979. Use of inductively-coupledplasmaspec-
trometry for simultaneousdeterminationof macro- and micronutrientsin NH4HC03-DTPA
extractsof soils. Soil Sci. Soc. Am. J. 43:75-78.
Stilwell, T.C., and T.G. Asscott. 1978. The relationship betweenfluoride-titratible (reactive) soil
alauminumand plant growth. Soil Sci. 125:28-33.
Suarez,D.L. 1987. Predictionof pH errors in soil-water extractorsdue to degassing.Soil Sci. Soc.
Am. J. 51:64--67.
Suhr, N.H., and e.O. Ingamells. 1966. Solution techniquefor the analysisof silicates.Anal. Chern.
38:730-734.
Thomas, G.W. 1960. Forms of aluminum in cation exchangers.Trans. Int. Congr. Soil Sci. 7th.
11;364-369.
Thomas,G.w. 1977. Historical developmentsin soil chemistry: Ion exchange.Soil Sci. Soc. Am. J.
41:230-237.
Thomas,G.w. 1996. Soil pH and soil acidity. p. 475-490.In D.L. Sparkset al. (ed.) Methodsof soil
Turner, R.e. 1967. Aluminum removed from solution by montmorillonite. Can. J. Soil Sci.
47:217-222.
Turner, R.C. 1969. Three forms of aluminum in aqueous systems determinedby 8-quinolinolate
extractionmethods.Can. J. Chern.47:2521.
Turner, R.e. 1976.A secondspeciesof polynuclearhydroxy aluminumcation, its formation and some
of its properties.Can. J. Chern. 54:1910.
Veith, J.A. 1977. Basicity of exchangeablealuminum,formation of gibbsite,and the compositionof
the exchangeacidity in the presenceof exchangers.Soil Sci. Soc. Am. J. 41:865-870.
Veith, J.A 1978. Selectivity and adsorptioncapacity of smectiteand vermiculite for aluminum of
varying basicity. Clays Clay Miner. 26:45-50.
Wada, K., and T. Higashi. 1976. The categoriesof aluminum- and iron-humuscomplexesin Ando
soils determinedby selectivedissolution.J. Soil Sci. 27:357-368.
Walker, WJ., C.S. Cronan,and P.R. Bloom. 1990.Aluminum solubility in organicsoil horizonsfrom
northernand southernfroestedwatersheds.Soil Sci. Soc. Am. J. 54:369-374.
Wang, e., J.A McKeague,and H. Kodama.1986. Pedogenicimogolite and soil environments:Case
study of Spodosolsin Quebec,Canada.Soil Sci. Soc. Am. 1.50:711-718.
Wang, e., P.A Schuppli, and GJ. Ross. 1987. A comparisonof hydroxylamine and ammonium
oxalatesolutionsas extractantsfor AI, Fe and Si from Spodosolsand Spodosol-likesoils in
Canada.Geoderma40:345-355.
Warren, CJ., B. Xing, and MJ. Dudas. 1990. Simple microwavedigestion techniquefor elemental
analysisof mineral soil samples.Can. J. Soil Sci. 70:617---620.
Webber,M.D., P.B. Hoyt, and D. Coneau.1982. SolubleAI, exchangeable AI, basesaturationand pH
in relation to barley yield on Canadianacid soils. Can. 1. Soil Sci. 62:397-405.
550 BERTSCH & BLOOM
Wilkinson, K.J., P.M. Bertsch,C.H. Iagoe,and P.G.c. Campbell.1993. Surfacecomplexationof AI

on isolatedfish gill cells. Environ. Sci. Techno\.27:1132-1138.
Willett, I.R. 1989. Direct determinationof aluminum and its cationic fluoro-complexesby ion chro-
matography.Soil Sci. Soc. Am. I. 53:1385-1391.
Wright, R.I., V.C. Baligar, and I.L. Ahlrichs. 1989.The influenceof extractableand soil solutionalu-
minum on root growth of wheatseedlings.Soil Sci. 148:293-302.
Yuan, T.L. 1959. Determinationof exchangeablehydrogenin soils by titration method. Soil Sci.
88:164-167.
Published 1996
Chapter19
Lithium, Sodium, Potassium, Rubidium,

and Cesium
P. A. HELMKE, University of Wisconsin, Madison, Wisconsin
D. L. SPARKS,University of Delaware, Newark, Delaware
GENERAL INTRODUCTION
Propertiesof the Alkali Metals
The elementsin Group IA of the periodic table are known as the alkali metals.
Theseelementshavea single electronin their outermostshell. The low ionization
potentialfor the outer electronsand the fact that the resultingcationshave noble
gas configurationsand are thus sphericaland of low polarizability result in the
chemistry of theseelementsbeing essentiallythat of the single chargedcation.
Theseelements aremore electronegative than H and occur solely as monovalent
cations in nature. The chemistry of the elementsis predominantlyionic, but a
slight degreeof covalent bonding occurs in bonds to 0, Nand C in various
chelate and organometalliccompounds.The tendency to covalenceis greatest
with Li and leastwith Cs, which is expectedfrom their charge/radiusratios. Most
compoundsof theseelementsare solublein water exceptsomesilicatesand a few
saltswith large anions.
One isotope of potassium40K (~-, 1.6 x 109 yr, 0.0119% abundance)is
radioactive.Rubidium also is naturally radioactiveowing to 87 Rb (~-, 6 x 1010 yr,
27.2% abundance).Both of theseisotopescan be used in age determinationsof
rocks and minerals.Franciumis not includedin this chapterbecause allof its iso-
topesare radioactivewith short half-lives. Franciumoccursonly as an ultratrace
elementin soils as a decayproduct of 227Ac.
The relative concentrationof the alkali elementsin soils reflect their rela-
tive cosmicconcentrations,which in turn are controlledby the nuclear properties
of the isotopesof the respectiveelements.The concentrationsof Na and K can
rangeup to severalpercentagepoints in soils while thoseof Li, Rb, and Cs rarely
exceedseveralhundredmicrogramsper gram. The adsorptionand ion exchange
behaviorof the alkali elementsin soils and otherexchangemediais a strongfunc-
Book Seriesno. 5.
551
552 HELMKE & SPARKS
tion of their hydratedradii (fable 19-1). This results in K, Rb, and·Cs being
retainedmore strongly in soils than Li and Na. The Cs+ hasthe lowest hydration
energy and is thereforeadsorbedthe most strongly to soils. Scott and Smith
(1987) recently reviewedthe sources,amounts,and forms of alkali elementsin
soils.
Mineralogy of Alkali Elementsin Soils
Albite (NaAlSi30 s), orthoclase(KAlSi 30 s), otherfeldspars,and micasare

the mostcommonsoil mineralsof Na andK. Variouscarbonateandothersaltsof
sodiumare importantin saline-sodicsoils. Lithium, Rb, and Cs are rarely found
in puremineralphasesin soils, andwhensuchmineralsarepresentthey areusu-
ally inheritedfrom the soil parentmaterial.Lithiophorite, a rare Li, Mn, AI min-
eral,is an exceptionasit may form from alterationof birnessite.Lithium, Rb, and
Cs in soils are normally presentas trace impurities in other compounds, espe-
cially the feldsparsand micas, or adsorbedto the soil solid phase,especially
illites.
The alkali elementsin soils are not easily characterizedin termsof differ-
ent forms. The conceptof forms of theseelementsis deeplyingrainedin the soil
scienceliterature,largely asa resultof attemptsto studythe factorsthat affect the
plant availability of soil K. The forms of the alkali elementsin soils aredelineat-
ed by the nature of the elementslocation in mineral or other phases(bonding
mechanism)or by their extractabilitywith specifiedmethods.Thesedistinctions
are interdependentbecausethe extractability of an elementis affected by its
chemicalstate.While thesemethodsof classificationare not precise,they do pro-
vide empiricalinformationthat is usefulin understandingandmanagingthe alka-
li elementsin soils. The variousforms definedin the following sectionon K are
often usedin discussionsof the otheralkali elements.
Potassium
Potassiumis a macronutrientfor plants.Selectionof the properprocedure

from the manyavailablethat assesses the statusof this importantnutrientrequires
a basicknowledgeof the soil chemistryof K anddefinitionsof the variousforms.
The concentrationof K is soil is usually higher than that of any other major or
secondaryplant nutrient(Reiemeier,1951)and it often reflectsthe K contentof
the soil parentmaterial.For igneousrocks,granitesand syenitescontain46 to 54
g kg-t K, basaltscontain7 g kg-t, and peridotitescontain2.0 g kg-to Among the
Table 19-1. Dataon hydrationof aqueousalkali ions.

u+ Na+ K+ Rb+
Crystal radii,t nm 0.06 0.095 0.133 0.0148 0.0169
Hydratedradii (approximate),nm 0.34 0.276 0.232 0.228 0.228
Hydration numbers(approximate):/: 25.3 16.6 10.5 9.9
Hydration energies,kJ mol-! 520.5 405.8 322.2 300.8 255.6
t For six coordination.
:/: From transferencedata.
ALKAU METAL PROPERTIES 553
sedimentaryrocks, clayey shalescontain30 g K kg-I, while limestoneshave an

averageof only 6 g kg-l (Malavolta, 1985; Sparks,1987).
The K contentof mineral soils generally rangesfrom 0.04 to 3% K. The
total K contentof soils rangesfrom 3 000 to 100 000 kg ha-l in the upper0.2 m
of the soil profile. Of this total K content, 98% is in the mineral form while
approximately2% is in the soil solution and exchangeableforms (Schroeder,
1979; Bertsch& Thomas,1985; Sparks,1987).
The propertiesof the hydratedK+ are compared tothoseof the other alka-
li elementsin Table 19-1. Potassium,amongmineral cationsrequiredby plants,
is the largestin the nonhydratedsize (r =0.133 nm) and the numberof oxygen
ions surroundingit in mineral structuresis high (8 or 12), which suggeststhat the
strengthof each K-O bond is not very strong (Sparks& Huang, 1985; Sparks,
1987).Potassiumhasa polarizability of 0.088nm3, which is higherthanfor Ca2+,
Li+, Mi+, and Na+, but lower than for Ba2+, Cs+, NH.t, and Rb+ (Rich, 1968,
1972; Sparks,1980, 1987; Sparks& Huang,1985). Ions with higher polarizabil-
ity are usually preferredin ion exchangereactions.Potassiumhas a hydration
energyof 322.2kJ mol-I, which indicatesit shouldnot causesoil swelling (Helf-
ferich, 1962).
Soil K exists in four forms in soils: solution, exchangeable,ftxed, and
structuralor mineral. There are equilibrium and kinetic reactionsbetweenthese
four forms that affect the level of soil solution K at any particulartime, and thus,
the amountof readily availableK for plants.The forms of soil K in the order of
their availability to plants and microbes are solution > exchangeable> ftxed
(nonexchangeable) > structural(Martin & Sparks,1985; Sparks& Huang,1985;
Sparks,1987).
Soil solution K is the form of K that is directly taken up by plants and
microbesand also is the form most subject to leaching in soils. Levels of soil
solution K are generallylow, unlessrecentamendmentsof K havebeenmadeto
the soil. Levelsof solution K are affectedby the equilibrium andkinetic reactions
that occur betweenthe forms of soil K, the soil moisturecontent,and the con-
centrationsof bivalent cationsin solution and on the exchangerphase(Sparks&
Huang, 1985).
ExchangeableK is the portion of the soil K that is electrostaticallybound
as an outer-spherecomplexto the surfacesof clay mineraland humic substances.
It is readily exchangedwith othercationsand also is readily availableto plants.
Nonexchangeable or "ftxed" K differs from mineral K in that it is not bond-
ed within the crystal structuresof soil mineral particles.It is held betweenadja-
cent tetrahedrallayersof dioctahedraland trioctahedralmicas,vermiculites,and
intergradeclay mineralssuch as chloritized vermiculite (Rich, 1972; Sparks&
Huang, 1985; Sparks,1987). If one equatesnonexchangeable to "ftxed" K, than
also it can occur in randomgapsin the structureof x-ray amorphousclay-sized
minerals(Barber, 1979). Potassiumbecomes"ftxed" becausethe binding forces
betweenK+ and the clay surfacesare greaterthan the hydration forces between
individual K+. This resultsin a partialcollapseof the crystalstructuresandthe K+
are physically trappedto varying degrees,making K releasea slow, diffusion-
controlled process(Sparks, 1987). NonexchangeableK also can be found in
"wedgezones"of weatheredmicasandvermiculites(Rich, 1964).Only ions with
554 HELMKE & SPARKS
a sizesimilar to K+, suchas NUt and H30+, canexchangeK from wedgezones.

Large hydratedcations,suchas Ca2+ and Mg2+ cannotfit into the wedgezones.
Nonexchangeable K is moderatelyto sparinglyavailableto plants,depend-
ing on severalsoil parameters(Goulding & Talibudeen,1979; Sparks& Huang,
1985; Sparks,1987). Releaseof nonexchangeable K to the exchangeableform
occurswhen levels of exchangeableand soil solution K are decreasedby crop
removal and/or leaching and perhapsby large increasesin microbial activity
(Sparks,1980).
Most of the total K in mostsoils is in the structuralforms (Sparks& Huang,
1985), mainly as K-bearing primary minerals such as muscovite,biotite, and
feldsparssuch as microcline and orthoclase.For example,in some Delaware
soils, Parkeret al. (1989a)found that structuralK averagedabout98% of the total
K (Table 19-1). Most of the structuralK was presentas K-feldsparsin the sand
fractions. Sadusky et al. (1987) studied the rate of K releasefrom sandy,
Delawaresoils and the sand fractions of thesesoils usinga H-saturatedresin
method.large quantitiesof K were releasedwithin 30 d, and significantamounts
werereleasedfrom the coarse,medium,andfme fractionsof the soils. In Kenans-
ville, Ap and B2t horizons57 and 54% of the total K release,respectively,could
be ascribedto the three sand fractions. Using scanningelectron microscopy
(SEM) analyses,Saduskyet al. (1987) found that the feldsparsin the sandfrac-
tions were extensivelyweathered,as evidencedby deepetchpits. Thesefmdings
suggestedthat the feldsparK in the sandfraction could be importantin the over-
all K balanceand K-supplyingpowerof sandy,Delawaresoils, especiallyduring
severalcrop growing seasons.The large quantitiesof structuralK in theseand
otherAtlantic CoastalPlain soils could help in explainingthe often observedlack
of crop responseto K amendments(Liebhardt et ai., 1976; Yuan et ai., 1976;
Sparkset al., 1980; Woodruff & Parks,1980; Parkeret ai., 1989b).
Potassiumfixation and releaseare influencedby severalfactors including:
the typesand amountsof soil minerals,levelsof K in the solutionphase,sizeand
degreeof weatheringof the mineral particles,soil pH, rainfall and temperature,
manuring,soil structure,wetting anddrying andfreezingandthawing,biological
activity and concentrationsof complexingorganic acids, redox potential of the
soil, and plant roots (Rich, 1972; Sparks & Huang, 1985; Goulding, 1987;
Sparks,1987). Of thesefactors, the mineralogyof the soil and the level of Kin
the soil solution havethe greatestimpacton fixation and releaseof K.
Sodium
Sodiumis not an essentialelementfor plants.Thereare reportsthat some

cropsbenefitfrom Na (Bear, 1953;Tinker, 1967; Coughtreyet al., 1983)but the
essentialityof Na remainscontroversial(Brownell, 1979). Plantsvary in their
capacityto tolerateNa and thereare only a few reportsof direct negativeeffects
of Na (Pearson,1960).Thereis no standardmethodthat assesses the plant avail-
ability of soil Na. The content of exchangeableand soluble Na are important
parametersin the managementof salineandsodicsoils. Excesssalinity, of which
Na can be partially responsible,reducesplant growth due to the osmoticeffect
and the negativeeffectsof excessNa on soil properties.High concentrationsof
ALKALI METAL PROPERTIES 555
Na tend to dispersesoil colloids and increasesoil swelling. These phenomena

decreasewater infiltration, aeration,and root penetration.The electricalconduc-
tivity of a saturationextractis normally usedas an index of salinization,while the
Na adsorptionratio is usedto estimatethe hazardof sodification(Rhoades,1996,
seeChapter14).
Lithium, Rubidium, and Cesium
Theseelementshave receivedlittle attention in soil science.Rubidium is

more abundantthanLi andCs in soils, at timesexceeding1000mg kg-t and aver-
aging 100 mg kg-to The concentrationof Rb in sites from Illinois were highest
where illitic materials are presentand lowest when loess is abundant(Jones,
1989). Typicalconcentrationsof Li in soils are 20 to 30 mg kg- t and that for Cs
is about5 mg kg-to Soils derived from granitic/syeniticparentmaterial have the
highestconcentrationsof theseelements.Lithium is much more mobile in soils
than are Rb and Cs. The latter elementsare strongly adsorbedby vermiculitic
clays (Franz & Carlson, 1978). The marked preferenceof siloxane sites for Cs
over Li can be exploited to estimatethe structural chargedensity of 2:1 phyl-
losilicates(Anderson& Sposito,1991).The strongadsorptionof 137Cs(30 yr), a
radionuclidefrom atmospherictestingof nucleardevicesin the 1950sand 1960s,
to soils has servedas a useful marker in soil erosion/depositionand pedoturba-
tion studies(Longmoreet aI., 1983; Southard& Graham,1992).
Of the alkali elements,Li is the mosttoxic to plants.Indigenouslevels have
been reportedto be toxic to citrus (Citrus spp; Bradford, 1966). Rubidium can
partially substitutefor K in plants, as their propertiesare similar (Table 19-1).
This has led to the use of the radiotracer86Rb (18.7 d) in K studiesof the soil-
plant-watersystembecausethe only suitable K radiotracerhas too short a half-
life (42K, 12.4 h) to be practical (Diest & Talibudeen,1967). There is consider-
able controversyover the equivalenceof the Rb and K reactionsin the soil-plant-
water systemand this substitutionshould be usedwith caution (Baligar & Bar-
ber, 1978; Eckert & McLean, 1980). Plant uptakeof Cs hasbeenstudiedbecause
of concernabout the fate of radioactiveisotopesof Cs from nucleartests. The
propertiesof Cs are sufficiently different from thoseof K and Rb that 137Cscan-
not be usedas a tracerfor K and Rb (Souty et aI., 1975).
ANALYTICAL METHODS FOR THE ALKALI METALS
The concentrationof the alkali metals are usually determinedby instru-

mental techniquesno matter how the samplesare prepared.Atomic emission
spectroscopyis now the most popular technique.Instrumentaltechniqueshave
the important advantagesof specificity, sensitivity, rapidity of analysis,and ease
of use. They have therefore almost completely displacedwet chemical proce-
duresfor the analysisof the alkali metals.No new wet chemicalprocedureshave
beenpopularizedin recentyears,and the readeris referredto earliercompilations
if such types of analysesare needed(Knudsen et aI., 1982; Hillebrand et aI.,
1953).
556 HELMKE & SPARKS
Selectionof the analytical method is often dictated by the availability of

instruments.The method used to preparethe sample also must be considered
when selectingan analytical technique.Extracts of soils are most conveniently
analyzed by. the optical techniques, inductively coupled plasma-massspec-
troscopy(ICP-MS) or ion chromatographybecausethey require liquid samples.
Solid soil samplesare most easily analyzedby neutronactivationanalysis(NAA)
or x-ray fluorescence(XRF) becausethesetechniquesdo not require the sample
to be dissolved.Although, the accuracyof XRF is improvedif a glassdisk of the
sampleis preparedby fusion with lithium borate.
The most sensitivewavelengthsand the detectionlimits for eachelement
are given in Table 19-2 for the optical methodsof analysis.The detectionlimits
are for optimumconditions.Practicalworking limits are severaltimeshigherthan
the valuesgiven in Table 19-2. The detectionlimits for Rb and Cs with ICP-AES
are so high that they cannotbe measuredin most soil samples.Atomic absorption
spec~rophotometry (AAS) has the disadvantageof being a sequential~echnique
but the instrumentsare lessexpensiveand widely available.Inductively coupled
plasma-atomicemissionspectrophotometryis capableof simultaneousanalysis,
which greatly increasesthe samplethroughput.The compositionof the matrix
affectsthe resultsfrom the optical techniques.Analystsshouldconsultthe respec-
tive chaptersin this monographand the operationmanualof their instrumentsfor
recommendedionization suppressants and other potentialmatrix interferences.
Flame photometrycan be usedfor determiningNa and K in solution. The
most sensitivelines are the sameas thosegiven in Table 19-2 for the otheremis-
sion techniques.The detectionlimit is usually low enoughthat theseelementscan
be measuredin most extractsof soils. The exact detectionlimit varies with type
of instrument and type of flame. Specific analysis techniquessuitable for the
availableinstrumentshouldbe used(Hossner,1996,seeChapter3).
Ion chromatographyalso is suitable for measuringthe alkali elementsin
solution, althoughit seemsto be not commonly usedfor soils (Nieto & Franken-
berger,1985; Basta& Tabatabai,1985). The detectionlimits of modemcolumns
and detectorsfor the alkali ions are estimatedto be 10 to 100 ng mL-l.
The detectionlimits and information on the alkali elementsfor ICP-MS,
NAA, and XRF are given in Table 19-3. The sensitivity of ICP-MS is extremely
low exceptfor K. The relativeinsensitivity for K occursbecausethe dominantK
isotope,39K, falls on the low masstail of the very large peak resultingfrom 40Ar
Table 19-2. Approximatelimits of detectionin single elementsystemsfor flame and graphitefurnace

AAS and ICP-atomicemissionspectroscopy(AES).
Element Wavelength Flame AASt FurnaceAAS* ICP-AES
nm ng mL- 1 pg ngmL-1
Li 670.8 35 2.5 5
Na 589.0 15 0.2 100
K 766.5 40 0.5 125
Rb 780.0 100 1 NO§
Cs 852.1 200 6 NO
t Concentrationto give 1% adsorption.
* Characteristicmassneededto give 1% adsorption.
§ NO =not determinable.
in the carriergas. Inductively coupledplasma-massspectroscopyis intolerantof

high concentrationsof dissolvedsolids. This limits the rangeof suitablesample
preparationtechniquesunless the samplesare diluted. The easiestmethod to
determinethe alkali elementsin solid samplesis NAA becausethe samplesdo
not have to be dissolved.The distribution of researchreactorsunfortunatelylim-
its the availability of this technique,although most academicreactor facilities
will analyzesamplesfor a modestfee. Very few soil researchfacilities haveXRF
equipment,but t.his techniquecan determineNa and K in solid samples.
ANALYSIS FOR TOTAL ALKALI ELEMENTS
Methods to determinetotal K and the other alkali elementsin soils use

acids or a high temperaturefusion to decomposethe soil. The most widely
employeddigestiontechniquesfor total elementsin soils and mineralshave used
combinationsof HF and eitherH2S04 or HCl04 (Klesta& Bartz, 1996,seeChap-
ter 2). The use of two acidsservesseveralpurposes.The HF decomposesthe sil-
icate mineralsby reactionof F with Si to form SiF4, which is volatile when it is
heatedwith strong acids. The secondacid should be an oxidizer to dissolveany
humic substancesin the soil sample.Since the alkali elementsare not a part of
any organicmolecule,the purposeof the oxidant is to preventtheir retentionon
organiccolloids, removeexcessHF from the soil sample,and avoid any interfer-
encesin the subsequentanalytical procedures.
The solution resulting from HF digestionsmust be treatedwith boric acid
to convertexcessF to fluoroboric acid beforethe samplesare transferredto glass
containers.Even boric acid-treated solutionscontainenoughF to etch glasscon-
tainerswith time and all HF-digestedsamplesmust be storedin plastic contain-
ers. Long-term analysisof solutions containing F also damagesthe quartz and
glassnebulizersfound on ICP instruments.Soils that havehigh concentrationsof
Ca may form so much CaF2that a portion of it remainsinsolubleafter the diges-
Table 19-3. Approximate limits of detectionfor ICP-MS, instrumentalneutron activation analysis,

andXRF.
ICP-MS NAA
Stable Radioactiveisotope XRF
Element isotope Detectiontlimit from (n, 1) reaction Detectionlimit:j: detectionlimit§
ngmL·1 mgkg·1
Li 7Li 0.1 ND~ ND ND
Na 23Na 0.06 24Na 0.8 (0.01) 15
K 39K 10 42K 30 (0.5) 30
Rb 85Rb 0.02 86Rb 0.4 1
Cs \33Cs 0.02 134Cs 0.03 1
t Three standarddeviationsabovebackground.
:j: Three standarddeviationsabovebackgroundwith a 2-h neutronirradiation of 1 g of whole soil at
1 x 1013 n cm-2 S-1 and a single radioassayafter 5 d. The values in parentheses
are the detection
limits with a lO-min irradiation and radioassaythe sameday.
§ Three standarddeviationsabovebackgroundwith pressedpellet or glassdisk.
'II ND =not determinable.
558 HELMKE & SPARKS
tion and addition of boric acid. Greaterdilution of the digestedsamplecan dis-

solve the CaFz but this raisesthe limit of detection.
Finely groundsoil samplesalsocan be decomposedby fusion with NaZC03
or NazOz (Klesta & Bartz, 1996, seeChapter2). Obviously Na cannotbe deter-
mined by such procedures. Blanks also must be checkedbecausethe flux often
containsappreciableamountsof Li, Rb, and Cs. Fusionwith NaZC03 in platinum
cruciblescan damagethe expensivecruciblesbecausesomesoil elementsform
alloys with Pt. Fusionwith NazOzis very effective in destroyingsoils but NazOz
with sufficiently low blank concentrationsof Li, Rb, and Cs is rarely available.
Zirconium cruciblesmust be usedwith NazOz fusions. Their price is reasonable
and they last for at least50 fusions. Major disadvantagesof fusion techniquesare
the amountof analyteelementintroducedby the fusion flux, andthe largeamount
of flux requiredresultsin high concentrationsof dissolvedsolids in the samples.
High concentrationsof dissolvedsolidsare not toleratedby ICP techniques,espe-
cially ICP-MS.
The proceduredescribedbelow employs a Teflon bomb method and is
basedon those of Bernas(1968) and Buckley and Cranston(1971). It also is
applicablefor microwave digestionwith the proper bomb or the bottle method
describedby Klesta and Bartz (1996) in Chapter2.
Hydrofluoric Acid Digestion for Total Alkali Elements
Apparatus
1. Agate mortar.
2. Teflon bombsand metal containers.
3. Oven.
4. Volumetric flasks, 100 mL.
5. Plasticstirring rods.
6. Plasticbottles, 100 mL.
7. Plasticpipettes,10 mL.
Reagents
1. Aqua regia. Mix one part of concentratedHN03 with three parts of
concentratedHCl. Include one part of deionizedwater if the aquaregia
is to be stored for any length of time. Without water, objectionable
quantitiesof CI and othergasesare evolved.
2. ConcentratedHE
3. Boric acid.
Procedure
1. Grind soil in an agatemortar to passa 0.14-mm(IOO-mesh)screen.
2. Weigh 0.5 g of soil into a Teflon bomb.
3. Add 1 mL of aquaregia and 10 mL of HF to the samplein the Teflon
bomb using plastic pipettes.
4. Placethe coveredTeflon bomb in its metal container.
5. Heat the samplein an oven at 383 K for 3 h.
6. Add 2.8 g of boric acid to a 100-mL plastic volumetric flask.

7. Pour the liquid out of the Teflon bomb into the volumetric flask. Wash
the remainderof the liquid out of the bomb into the flask with deion-
ized water and thoroughly mix the contentsof the flask.
S. Dilute the solution in the flask to 100 mL with deionizedwater and
store in sealedplastic bottles.
9. Analyze the solution for the alkali elementsusing AAS, ICP, or Ie.
MEASUREMENT OF SOIL POTASSIUM
A numberof methodscan be employedto measurethe quantitiesof K in

the various defined forms of soil K. Only the most commonly usedmethodsare
detailedin this chapter.
ExchangeablePotassium
ExchangeableK is that K which is extractedwith a neutralnormalsalt, usu-

ally 1.0 M NH40Ac minus the water-solubleK (Knudsenet aI., 1982). In soils
that are not saline, levels of water-solubleK are minimal and can be ignored.
However,in salinesoils, the levelsof water-solubleK shouldbe determinedfrom
a saturatedextractor somesimilar extract andsubtractedfrom the amountof K
determinedusing NH 40Ac.
It should be noted that in soils that contain weatheredvermiculitic and
micaceousminerals"wedge zones"can be presentthat contain K. This K is not
accessibleto large index cationssuchas Ca and Mg, but can be extractedby N~,
which is of similar size to K. For example,in soils that contain "wedge zones,"
NH40Ac will extract more K than an extractantlike 1 M CaCI2. It is debatable
whetherthis K is truly "exchangeable."Thus, in soils containing"wedge zones"
exchangeableK could be overestimatedwith NH40Ac (Sparks & Liebhardt,
1981; Sparks& Huang, 1985).
Ammonium AcetateMethcd
The methodoutlined below is that of Thomas(1982).
Apparatus
1. Erlenmeyerflask, 125 mL.
2. Centrifugetubes,50 mL.
3. Buchnerfunnels, 5.5 cm.
4. Volumetric flasks, 50, 10, 250 mL.
5. Centrifuge.
Reagents
1. Ammonium acetate(NH40Ac), 1 M: Add 70 mL of reagent-grade
NH40H, sp gr 0.90, to 57 mL of 99.5% aceticacid per liter of the final
solution desired.Do the mixing in about half the final volume of dis-
tilled water desired, make up to volume with additional water, mix,
560 HELMKE & SPARKS
thoroughly cool, and adjust exactly to pH 7.0 with either NH 40H or

aceticacid. (Only a small volume of acid or basewill be required.)
BuchnerFunnelProcedure.Place10 g of <2-mm, air-dried soil in a 125-
mL Erlenmeyerflask, and addapproximately40 mL of 1 M NH40Ac. Swirl and
let standfor 1 h or more. Transferthe soil and solution to a 5.5-cm Buchnerfun-
nel fitted with a number40 or 42 Whatmanfilter paperand connectedto a 250-
mL suctionflask. Transferthe remainingtracesof soil to the funnel by adding10-
mL portionsof NH40Ac to a final volume in the suctionflask of 90 mL for soils
with a cationexchangecapacity(CEC) of 20 cmol kg-! or lessor 225 mL for soils
with higher CECs. Pour the extract into a 100- or 250-mL volumetric flask, and
make up to volume by rinsing the suction flask with fresh 1 M NH40Ac. Deter-
mine the concentrationsusing AAS, rcp, or Ie.
CentrifugeProcedure.Place5 g of <2-mm, air-dried soil in a 50-mL cen-
trifuge tube. Add 25 mL of 1 M NH 40Ac, stopper,and shakefor 30 min. Place
the tube in a centrifuge,and spin at 2000 rpm for 10 min. Pour the supernatant
into a 50-mL volumetricflask. Repeatwith an additional25 mL, and finally bring
up to a volume of 50 mL with 1 M NH40Ac. Determine cationsaswith the Buch-
ner funnel procedure.For soils with CECs in excessof 20 cmol kg-I, use only 2
g of soil.
Nonexchangeable
Potassium
There are a numberof chemicalmethodsthat can be employedto extract
nonexchangeable K. These include:boiling RN03, hot HCl, electroultrafiltration,
Na-tetraphenylboron with EDTA, and ion exchangeresinssuchas H- and Ca-sat-
uratedresins.
The most commonly used methodfor extractionof nonexchangeable K is
the boiling HN03 technique.Most researchersthat use this methodboil the soil
in 1 M HN03 for 10 min over a flame, transferthe slurry to a filter, leachthe soil
with dilute HN03, and then determinethe concentrationin the filtrate. Oneof the
problemswith boiling only 10 min over a flame is that it is difficult to be precise
about the correctboiling time, the time it takesfor boiling to occur, and the vigor
of boiling (Martin & Sparks,1983). Someworkers have attemptedto diminish
these problemsby using a 386 K oil bath for 25 min, including heating time
(Pratt, 1965).This modification is particularly usefulwhen large numbersof sam-
ples are being analyzed.Of course,one of the major concernswith using a boil-
ing HN03 procedureis the potentialto causedissolutionof mineral forms of K.
Other researchershave used continuousleaching of the soil with dilute
acidssuchas0.01 M HCI or with dilute saltssuchas 0.1 M NaCI, repeatedextrac-
tions with 3, 0.3 and 0.03 M NaCl, Sr salts,hot MgCI 2, and sodiumcobaltnitrite
(Martin & Sparks,1983).
Cation exchangeresinssaturatedwith H or Ca also have beenwidely used
to measurenonexchangeable K. Theseresinshave high CECsand when saturat-
ed with an appropriatecation andmixed with soil and with a dilutesolution, they
will adsorbreleasedK. One of the major advantagesof using cation exchange
resinsto extractK is that they act as a "sink" for the releasedK and thus prevent
an inhibition of further K release.This is a problemwith manybatchmethodsthat
ALKALIMETALPROPERTms 561
employ dilute acidsand electrolytes.One major disadvantageof cation exchange

resinsfor extractingK is that the resinsare expensiveand the procedureis time-
consumingand tedious.
In order for electrolyteand acid solutionsand cation exchangeresinsto be
effective in extractingK, the K concentrationin the solution phasemust be kept
very low, or K releaseis inhibited (Rausell-Colomet aI., 1965; Wells & Norrish,
1968; Feigenbaumet aI., 1981; Martin & Sparks,1983). The critical concentra-
tion abovewhich releaseis inhibited is 4 mg L-l for soils in general,2.3 to 16.8
mg L -1 for trioctahedralmicas in dilute solution, and as low as <0.1 mg L -1 for
muscovite and illite (Smith & Scott, 1966; Martin & Sparks, 1983). A low
enoughconcentrationof solution K can be maintainedby employing a continu-
ous flowing extracting or exchangingsolution, cation exchangeresins, or Na-
tetraphenylboron(Scott et aI., 1960).
Two proceduresfor measurementof nonexchangeable K in soils are pre-
sentedbelow. Theseare: a boiling HN03 method,which is essentiallythat out-
lined by Knudsen et al. (1982) and a H-saturatedresin technique that was
employedby Martin and Sparks(1983) and Saduskyand Sparks(1991).
Some researchers have beencritical of using H-saturatedresins and have
preferredthe the employmentof Ca-saturatedresins (Talibudeenet aI., 1978).
Their argumentis that the H-saturatedresin can breakdown soil minerals.How-
ever, Martin and Sparks (1983) showed that no change in clay mineralogy
occurredafter treatmentwith an H -saturatedresin. Moreover,severalresearchers
havefound Ca- andNa-saturatedresinswere unsatisfactoryfor extractingnonex-
changeableK from soils when usedwith any soil mineralsmore stablethan tri-
octahedralmicas(Arnold, 1958; Haagsma& Miller, 1963).
Boiling Nitric Acid ExtractionMethod

Apparatus
1. Erlenmeyerflasks, 125 mL.
2. Plastic funnels, with 15-cm filter paper(Whatmanno. 50 or equiva-
lent), with supportingracks.
4. Hot plate.
5. Automatic dilutor.
Reagents
1. Nitric acid, 1.0 M. Add 63 mL concentratedHN03 to deionizedwater
and bring to 1 L.
Standards
1. Stock solution: 100 mg VI K in 0.33 M HN03. Preparefrom com-
mercially availablestock solution (1000 mg L-l) or oven-driedKCI.
2. Working standards:0, 5, and 10 mg L- 1 Kin 0.33 N RN03.
Procedure
1. Weigh 2.5 g air-dried, crushedsoil into a 125-mL Erlenmeyerflask.
Add 25 mL 1.0 M RN03 and placeon ahot plate.
562 HELMKE & SPARKS
2. When boiling begins,adjustheatand boil gently for 15 min.

3. Remove flasks from heat, cool for 5 min and filter slurries through
Whatman no. 50 paper, collecting leachatein a 100-mL volumetric
flask.
4. Use four 15-mL aliquotsof 0.1 M HN03 to rinse flask and then leach
soil in funnel. Wait until each aliquot has drained through before
addingthe next. Bring flasks to volume with 0.1 M HN03.
5. DetermineK using AAS, ICP, or IC.
Calculations.The nonexchangeable
K contentis calculatedby
K, mg L-1 x 0.1 Lx (2.5 g soilt1 x 1000 g kg-1 = K, mg kg-1 soil
Divide the aboveresultby 391 to obtainthe nonexchangeableK in centimolesper

kilogram. Comment:An effort should be made to maintain uniform rates and
times of boiling in order to obtain reproducibleresults.
Hydrogen-Saturated
ResinExtraction
The methodoutlined below is that of Martin and Sparks(1983).
Apparatus
1. Buchnerfunnels, 5.5 cm.
2. Temperature-controlled reciprocatingshaker.
3. Plasticcentrifugetubes.
4. Sieve,0.23-mm(60 mesh).
Reagents
1. 1M AgN03•
2. 0.5 M CaCI2•
3. 0.001 M HCI.
4. 1.0MHCl.
5. 1 MNH4Cl.
Hydrogen-saturated resin: Leachthe resin with five, 25-mL aliquotsof 1.0
M HCI solution using a Buchner funnel. Wash the resin with deionizedwater
until the leachategives a negativetest for Cl-1 usingAgN03. In the work of Mar-
tin and Sparks(1983) a Bio-RadAG 50WX resin (Bio Rad Lab, Richmond,CA)
was employed.
Calcium-saturatedsoil: Follow the procedureabove for the saturationof
the resin exceptuse 1.0 M CaCl2 rather than 1.0 M HCI. The purposeof saturat-
ing the soil with Ca is to removeany indigenoussolubleand exchangeableK.
Procedure
1. Place2 g of Ca-saturatedsoil in a 80-mL, plasticcentrifugetube along
with 4 g of moist H-saturatedresin and 50 mL of 0_0001M HCI.
2. Gently shake the suspensionuntil an equilibrium in K releasehas
occurred.The time necessaryto attain an equilibrium will dependon
ALKALI METAL PROPERTIES S63
the mineralogicalcompositionof the soil. Thus, it will be necessaryto

conduct a kinetic study whereby multiple samplesare equilibrated
until the amountof extractedK doesnot changesignificantly with time
usingproceduresgiven in Steps3 to 5 below.
3. After equilibration,separatethe soil from the resin on a 0.23-mm(60-
mesh) sieve. Then, leach the resin with 80 mL of 1 M NH4Cl to
removethe nonexchangeable K and collect the leachatein a 100-mL
volumetric flask.
4. Bring the contentsof the volumetricflask to volume with 1 M NH4Cl.
5. Analyze the solution usingAAS, ICP, or Ie.
Mineral PotassiumAnalyses
One can quantitativelyanalyzefor mineral K (K-feldsparsand micas) by
using a selectivedissolutionmethodemployingNa2S207fusion. The technique
and methodfor calculatingthe quantitiesof K-feldsparsand micas that will be
describedbelow are takenfrom Jackson(1956).A semiquantitativeapproachfor
measuringmineral K is to subtractthe quantity of nonexchangeable K, using the
boiling HN03procedure(given in the sectionon "Nonexchangeable Potassium"),
from the quantity of total K, usingthe HF digestionmethod(given in the section
on "Hydrofluoric Acid Digestionfor Total Alkali Elements").Onealsocanquan-
tify K-feldsparsin the sandfraction of soil using petrographicanalyses(Parkeret
al., 1989a).
Fusion of mineralsin Na2S207hasbeenwidely usedas a way to decom-
posemineralsfor many years(Jackson,1956).The fusion can be perceivedas a
treatmentwith H2S04 at a high temperaturecreatedby the presenceof Na2S04
formed during the fusion,
The S03 is a potentdehydroxylatingagentand forms H2S04with water released

from the heatedminerals. In the method of Jackson(1956), that is presented
below, the octahedralcationsof mica, kaolinite, and chlorite are very soluble in
the 3 M (3 N) HCI washingsafter the fusion, while the remainingSi02 is quick-
ly solublein hot 0.5 M NaOH solutionaddedlater. Kaolinite is resistantto disso-
lution in fuming HCI04. Quartzandfeldspars,which are high in K and Na, resist
the fusion-HCI-NaOHtreatmentsand can be weighedas a residue.Analysis for
K and Na in the residueallows for the determinationof thesefeldsparsvia the use
of correctionfactors that considerthe amountsof dissolutionthat occur in each
size fraction. Mica K and Na can then be determinedby differenceof theseele-
mentsfrom the original content.
Method for Mineral Potassium
The methodoutlined below was that of Jackson(1956).
Apparatus
1. Small agatemortar.
2. Rubbertipped glassrod.
564 HELMKE & SPARKS
3. Silica crucible, 50 mL
4. Platinumcrucible or Teflon beaker,40 mL.
5. Nickel or stainlesssteelbeaker,500 mL.
6. Beakers.
7. Water baths,boiling and cold.
8. Centrifuge.
9. Centrifugetubes,70 mL pointed.
10. Drying oven.
Reagents
1. ReagentgradeNa2S207in powderedform. The salt NaHS04 may be
gently heateduntil the melt is complete,indicating that Na2S207has
formed. The melt shouldthen be cooledand crushedto a powder.
2. HCI, 6 M and 3 M.
3.1MNH4Cl.
4. ConcentratedH2S04•
5. 60% HCl04•
6.48%HF.
7. 0.5 M NaOH.
8. 99% methanol.
9. 50% acetone(diluted with 99% methanol).
10. 100% acetone.
Procedure
1. The sampleshould be fmely ground, it should not be coarseaggre-
gates.Saturatethe samplewith NHt by washingfive times with 1 M
N~CI, and then wash once witha small volume (only twice the cake
volume after centrifugationin NH4CI) of water, once with methanol,
twice with 50% acetonein methanol,and once with acetone.Freeze-
dry the sampleandthen rub it with a rubber-tippedrod to makea pow-
der. Rock, gravel,or sandshouldbe groundin a small agatemortarso
that it passesa 0.23-mm(60-mesh)sieve.
2. Weigh a 0.200-gsample,dried at 373 K, into a 50-mL vitreous silica
crucible containingabout 5 g of Na2S207powder. Mix the sampleof
Na2S207with a glassrod and then add more Na2S207so that a total of
12 or 15 g is added.Fusethe Na2S207undera fume head,using a low
flame at first until vigorousbubblingof the melt ceases.Thereafter,the
full flame of a Meker burner should be applied.Swirl the crucible
while cooling to spreadthe melt on the crucible sides.
3. Transferthe solidified melt as a caketo a 150-mL beakerwith 60 mL
of 3 M HCI to fmish the transfer and wash the sample.Discard the
supernatantliquid eachtime.
4. Transferthe residuefrom the tube to a 500-mL nickel or stainlesssteel
beakerwith 0.5 M NaOH and bring the total volume of NaOH in the
beakerto 100 to 150 mL. The suspensionshouldbe broughtrapidly to
boiling, boiled for 2.5 min, and cooled rapidly in a cold-waterbath.
Transferthe suspensionto 70-mL pointed centrifuge tubes, transfer-
ring the insolubleresiduefrom the beakerquantitatively.Separatethe
residueby centrifugationand washthreetimes with 3 M HCI, careful-

ly washingthe sidesand lip of the tube. Transferthe residueto a tared
platinum crucible (or Teflon beaker),dry at 383 K, and weigh.
5. Analyze the residueand the original sampledried at 383 K for K 20,
Na20 and CaO accordingto the HF-HCI04 methodfor K20 and Na20
and the sodiumcarbonatefusion methodfor CaO (Jackson,1956).
Calculationsof Results.The method outlined below is that of Jackson
(1956).
An approximatevalue of quartz plus feldspar content of a sample is
obtainedas the percentageresiduefrom the fusion. X-ray diffraction analysisof
the residuegives an indication of the proportionsof quartzand feldsparspresent
by the diffraction peak intensities.Correctionfactors for mineral solubility may
be applied. Much more quantitative values are available from the elemental
analysesfor K, Na, and Ca beforeand after the fusion (seebelow).
Feldspars. The K20 present in the residue representsfeldspar in the
residue.Then for the original sample,
% K-feldspar= (% K20 in residue)(% residue/lOO)X [1]
% Na-feldspar=
(% Na20 in residue)(% residue/lOO)Y - Y (% K feldspar) [2]
% Ca-feldspar=(% CaO in residue)(% residue/lOO)Z [3]
in which X, Y, and Z are the conversionfactors for residueoxide to the corre-

spondingfeldspar (Table 19-4) and Y is a correctionfor Na that entersthe K-
Table 19-4. Summaryof factorst for conversionof residue K 20, Na20, and CaO to the respective
end-memberequivalent feldspars (Jackson,1956).
ResidueNa20 ResidueCaO to
ResidueK 20 to to albite, Y anorthite,Z
Microcline Feldspar
Particle Na20/CaO Na20/CaO Na20/CaO Na20/CaO
size X K2O,X' >0.82:j: 0.82-0.37 >0.82 0.82-0.37 Y
Il.
2000-500 6.05 1.026
500-50 6.1 1.036 8.9 8.3 5.2 5.9 0.0
50-20 6.5 1.042 9.1 8.3 5.2 6.0 0.02
20-5 7.0 1.10 9.5 8.8 5.5 6.3 0.04
5-2 8.4 1.23 10.3 10.5 6.0 8.5 0.12
2-0.2 13.2 1.72 13.7 19.6 8.1 17.6 0.20
t Thesefactors differ from the theoretical(X = 5.9, Y = 8.5, Z = 4.95) for 16.9% K 20 in microcline-
orthoclase,11.8% Na20 in albite and 20.2% CaO in anorthite,respectively,becauseof dissolution
of feldspars,surfaceloss of K, Na and Ca and replacementof Na for K and Ca during the Na2S207
fusion. Y is usedin the estimationof albite to correctthe residueNa20 for the Na uptakeby micro-
cline during the fusion. X' is usedin the mica determinationto correct for microcline dissolution
during the fusion.
:j: Ratio of the percentageby weight of the oxidespresentin the original sample.
566 HELMKE & SPARKS
feldsparlattice in the NaZSZ07 fusion. When the NazO/CaOratio in the original

sampleis lessthan 0.37, rarely the casefor soil fractions,the Na-feldsparand Ca-
feldspar are best approximatedfrom the NazO and CaO contentof the original
sample.
Micas. The mica KzO is derivedby differencefrom the contentin the orig-
inal sampleand the amountattributableto feldspar
% feldspar KzO =(% KzO in residue)(% residuell00) X [4]
in which X (Table 19-4), which is X/5.9, correctsfor the dissolution of micro-

cline during the fusion-HCI-NaOHprocedure.A similar equation,with a value of
Y/B.5 (Table 19-4) substitutedfor X, is usedto obtain feldsparNazO.
% mica KzO - % total initial KzO - %feldsparKzO [5]
Similarly, the % mica NazO is derived.The percentageof mica is derivedon the

basisof 10% KzO and 7% NazO.
Quartz. The residuefor soils and sedimentsgenerally containsprimarily
quartz and feldspars.
% quartz=(100/D)[% residue- A(% K feldspar/lOO
- B(% Na feldspar/lOO)- C(% Ca feldspar/lOO)] [6]
in which A, B, C, and D are the recoverypercentages(Table 19-5) for various

size fractions of microcline, albite, anorthite,and quartz, respectively.
SOIL TESTS FOR POTASSIUM
Soil test extractantsfor K were developedto easily and rapidly measureK

in soils and to estimateK availability. Basedon the amountsof extractableK, rec-
ommendations--thatare basedon field test calibrations--canthen be made on
Table 19-5..Factorsfor Eq. [6) to obtain the quartzcontent(Jackson,1956).

% weight recovery
A1bite,B Anorthite, C
Size Microcline, Na20/CaO Na20/CaO Na2O/CaO Na20/CaO Quartz,
fraction A >0.82t 0.82-0.27 >0.82 0.82-0.37 D
fl
500-50 96.5 95.2 93.7 95.2 93.7 99.6
50-20 96.0 94.5 90.9 94.5 90.9 99.4
20-5 93.6 92.7 77.4 92.7 77.4 99.1
5-2 84.4 86.2 41.9 86.2 41.9 98.2
2-0.2 64.0 68.3 16.4 68.3 16.4 96.5
t Ratio of the percentageby weight of the oxidespresentin the original sample.
the amountof K that is neededto maximizeplant yields. Soil testsfor K usually

estimatethe quantity of solution and exchangeableK, and sinceacids are usual-
ly employedas extractants,somenonexchangeable K also is extracted(Wolf &
Beegle,1991).A numberof different soil testsareusedto measureextractableK.
These include: Mehlich 1 and Mehlich 3 proceduresin the northeasternand
southeasternUSA, the Morgan and modified Morgan proceduresin partsof the
northeasternUSA, the 1 M N~OAc at pH 7 procedurein the north centralUSA,
and the ammoniumbicarbonate-DTPAextractionin the westernUSA.
Mehlich 1 Extraction
The methodoutlined below is that of Mehlich (1953).
Apparatus
1. Reciprocatingor rotary shaker,capableof at least 180 opm (oscilla-
tions per min).
2. Standardstainless-steel
scoops,1 and 5 g.
3. Erlenmeyerflasks (150 mL) and filter funnels for extraction.
4. Filter funnels
Reagents
1. Mehlich 1 extracting solution (0.05 M H2S04 + 0.05 M HCl): Also
referredto asdilute doubleacid or the North Carolinaextractant.Using
a graduatedcylinder, add 160 mL of concentratedHCl and 27 mL of
concentratedH2S04 to approximately30 L of deionizedwater in a
large polypropylenecarboy.Make to a final volume of 40 L by adding
deionizedwater. Mix well by bubbling air into the solution for 3 h.
2. Activated e.
Procedure
1. Scoop5 g of sieved,air-dried soil into a 150-mL extractionflask.
2. If it is necessaryto obtain a colorlessfiltrate, add 1 g of activatedC to
eachflask.
3. Add 25 mL of the Mehlich 1 extractingsolution and shakefor 5 min
on a reciprocatingshakerset at a minimum of 180 opm.
4. Filter through a medium-porosityfilter paper (Whatman no. 2 or
equivalent).
5. Analyze for K using AAS, ICP, or Ie.
Mehlich 3 Extraction
The methodoutlined below is that of Mehlich (1984).
Apparatus
1. Stainless-steel
soil scoops,1 and 2.5 g.
568 HELMKE & SPARKS
2. Reciprocatingshaker,Capableof 180 opm.

3. Plastic(PVC), wide-mouth,extractionbottles, 100 mL.
4. Filter funnels.
Reagents
1. Mehlich 3 stock solution (3.75 M NH4F + 0.25 M EDTA): Mix 277.8
g ammoniumfluoride (NH4F) with approximately1200 mL distilled
water in a 2-L volumetric flask. Add 146.1 g of ethyleneidaminete-
traaceticacid {EDTA =[(HOOCCH2)2NCH2N(CH2COOHh]}.Dilute
to volume andmix well. This providesenoughstocksolutionfor about
10 000 samples.
2. Mehlich 3 extractionsolution (0.2M CH3COOH + 0.25 M NH4N03 +
0.015 M NH4F + 0.013 M HN03 + 0.001 M EDTA): Dissolve 1000g
ammoniumnitrate (NH4N03) in approximately40 L of distilled water
in a 50-L calibrated,plastic carboy.
3. Activated carbon.
Procedure
1. Scoop2.5 g of air-dried, sievedsoil into a 100-mL extractionbottle.
eachflask.
3. Add 25 mL of the Mehlich 3 extractionsolution to eachbottle.
4. Shakeat 200 opm for 15 min on a reciprocatingshaker.
5. Filter through a medium-porosity filter paper (Whatman no. 2 or
equivalent).
6. Analyze filtrate for K as given earlier.
Ammonium AcetateExtractablePotassium
The methodoutlined below is that of Brown and Warncke(1988).
Apparatus
1. 2-g scoop.
2. Automatic or semiautomaticextracting solution dispenser,10 or 20
mL.
3. Extractingflasks, 50-mL Erlenmeyeror conical flasks.
4. Funnels(or filter holding devices).
5. Receivingreceptacle,20 to 30 mL beakersor test tubes [note-Most
high volume soil testing laboratorieshave racks of extractingflasks,
funnelsand receivingreceptaclesdesignedto handlemultiple soil sam-
ples at one time.]
6. Rotatingor reciprocatingshakercapableof 200 opm.
Reagents
1. Extractingsolution (1 M NH40Ac at pH 7.0): (i) Placeapproximately
500 mL of distilled water into the mixing vessel.Add 57 mL of glacial
acetic acid (99.5%) then add 69 mL of concentratedammonium

hydroxide(note-Mix in a fume hood). Bring the volume to about900
mL with distilled water. Adjust to pH 7.0 with 3 M NH40H or 3 M
acetic acid. After cooling to room temperature,bring the solution to a
volume of 1 L. (ii) (Alternative) Reagent-gradeammonium acetate
may be used. Add 77.1 g of NH40Ac to 900 mL of distilled water.
After dissolutionof the salt adjust the pH to 7.0 as above. Dilute to a
final volume of 1 L. (Checkthis solutionfor K contaminationfrom the
salt.)
Procedure
1. Scoop1 g of preparedsoil into an extractionflask.
2. Add 10 mL of extractingsolution to the extractionflask.
3. Shakefor 5 min at 200 opm.
4. Filter the suspensionsthrough Whatman no. 2 or equivalent filter
paper.Refilter or repeatif the extractis cloudy.
5. Analyze for K as given earlier.
Morgan Extraction
The methodoutlined below is that of Morgan (1941).
Apparatus
1. Stainless-steelsoil scoops,1 and 10 g.
2. Reciprocatingshaker,capableof 180 opm.
3. Extractionflasks, 125 mL.
Reagents
1. Morgan extractant(0.72 M NaOAc + 0.52 M CH3COOH).Add SOOO g
of sodium acetatetrihydrate (CH3COONa.3HzO) to a SO-L carboy
containingapproximately20 L of distilled water. Add 1450mL glacial
acetic acid and mix until the sodium acetateis dissolved.Dilute to SO
L with distilled water and mix well. The pH of the solution shouldbe
4.8 +/- 0.05. If necessary,adjust to 4.8 with sodium acetateor acetic
acid.
2. Activated C.
Procedure
1. Scoop10 g of air-dried, sievedsoil into 12S-mL Erlenmeyerflasks.
eachflask.
3. Add 50 mL of the Morgan extractantto eachflask.
5. Filter througha mediumporosity filter paper(Whatmanno. 2 or equiv-
alent).
570 HELMKE & SPARKS
Modified Morgan Extraction
The methodoutlined below is that of McIntosh (1969).
Apparatus
1. Stainless-steelsoil scoop,1 and 4 g.
3. Extractionflasks, 50 mL.
4. Filter funnels.
Reagents
1. Modified Morgan extractant(0.62 M N~OH + 1.25 M CH3COOH):
Add 2874 mL glacial aceticacid to a 40-L carboycontainingapproxi-
mately 20 L of distilled water. Add 1825 mL concentratedNH40H.
Dilute to 40 L with distilled water and mix well. The pH of the solu-
tion shouldbe 4.8 ± 0.05. If necessary,adjustto 4.8 with concentrated
NH40H or aceticacid.
2. Activated C.
Procedure
1. Scoop4 g of air-dried, sievedsoil into 50-mL extractionflasks.
eachflask.
3. Add 20 mL of the Morgan extractantto eachflask.
5. Filter througha mediumporosityfilter paper(Whatmanno. 2 or equiv-
alent).
Ammonium Bicarbonate-Diethylenetriamine Pentaacetic Acid

Extraction
The methodoutlined below is that of Soltanpourand Schwab(1977).
Apparatus
2. Extraction flasks,125 mL.
3. Filter funnels.
Reagents
1. Ammonium bicarbonate[diethylenetriaminepentaaceticacid (DTPA)
extractingsolution (0.005M DTPA, 7.6 pH) 1.97 g DTPA to 800 mL
water. Add 2 mL of 1:1 NH40H to enhancedissolutionand to prevent
effervescencewhen the bicarbonateis added.When most of the DTPA

is dissolved,add 1 mol (79.06 g) of NH4C03 and stir gently until the
mixture is dissolved. Adjust the solution pH to 7.6 with ~OH.
Dilute the solution to 1 L with water, and either use immediately or
store undermineral oil to maintainstablepH.
Procedure
1. Weigh 10 g of soil into a 125-mL Erlenmeyerflask.
2. Add 20 mL of ammoniumbicarbonate-DTPAextractingsolution.
3. Shakethe soil mixture on a reciprocatingshakerfor 15 min at 180 opm
with the flasks open.
4. Filter the extractsand analyzefor K as given earlier.
EXCHANGEABLE AND SOLUBLE SODIUM
Introduction
The highestconcentrationof soluble alkali cationsin soils is that of Na in

the salt-affectedsoils that are classifiedas saline-sodie(Bresleret aI., 1982).The
concentrationof exchangeableNa varies from trace amountsto a major portion
of the exchangecapacity,dependingon the soil environment.The concentration
of exchangeableNa is generallysmall comparedto the total concentrationof soil
Na.
Procedure
The concentrationof exchangeableNa in soils is usually determinedby

extraction with neutral 1.0 M ammoniumacetate(see section on "Ammonium
AcetateExtractablePotassium").Other extractants,such as thoseusedfor CEC
measurements, also can be usedfor Na (Sumner& Miller, seeChapter40). Since
the extractantalso dissolvessolubleNa, the resultsmustbe correctedfor the con-
centration of water-solubleNa. Water-solubleNa is measuredin a saturation
extract (Rhoades,1996, see Chapter 14). Appropriate adjustmentsfor dilution
factors and soil weights must be madeso that the valuesmeasuredin the ammo-
nium acetateand saturationextractsare consistent.The concentrationof water-
soluble Na is insignificant comparedto exchangeableNa for most soils of the
humid region but can be importantfor soils of dry regions.
LITHIUM, RUBIDIUM, AND CESIUM
Introduction
The dissolvedforms of theseelements haveattractedlittle attentionexcept
for Li, as discussedearlier. Few measurements
of the exchangeablefraction of
theseelementshavebeenmade.Neutral 1.0M ammoniumacetateis the preferred
S72 HELMKE & SPARKS
extractant.ExtractableLi in 12 soils from California rangedfrom 0.1 to 0.9 mg

kg-1 and averaged0.3 mg kg-1 (Bradford & Pratt, 1961). ExtractableRb in S9
soils from Illinois averaged1.18 mg kg-1 (Jones,1992).
Procedurefor ExtractableLithium or Rubidium

Use the ammonium acetateextraction method given in the section on
"Ammonium AcetateExtractablePotassium."
REFERENCES
Anderson,S.l., and G. Sposito.1991. Cesium-adsorptionmethodfor measuringaccessiblestructural
surfacecharge.Soil Sci. Soc. Am. I. 55:1569-1576.
Arnold, P.W. 1958. Potassium uptake by cation exchangeresins from soils and minerals. Nature
(London) 182:1594-1595.
Baligar, V.C., and S.A. Barber. 1978. Potassiumand rubidium adsroptionand diffusion in soils. Soil
Sci. Soc. Am. I. 42:251-254.
Barber,R.G. 1979. Potassiumfixation in someKenyan soils. I. Soil Sci. 30:785-792.
Basta, N.T., and M.A. Tabatabai.1985. Determinationof exchangeablebasesin soil by ion chro-
matography.Soil Sci. Soc. Am. I. 49:84-89.
Bear, F.E. (ed.). 1953. Sodium symposium.Soil Sci. 76:1-96.
ic spectrophotometry. Anal. Chem.40:1682-1687.
Bertsch,P.M., and G.W. Thomas.1985. Potassiumstatusof temperatureregion soils. p. 131-162.In
R.E. Munson(ed.) Potassiumin agriculture.ASA, CSSA, and SSSA, Madison,WI.
Bradford, G.R. 1966. Lithium. p. 218-224.In H.D. Chapman(ed.) Diagnosticcriteria for plantsand
soils. Div. Agric. Sci., Univ. California, Riverside,CA
Bradford,G.R., and P.F. Pratt. 1961. Separationand determinationof lithium in irrigation water,plant
material,and soil extracts.Soil Sci. 91:189-193.
Bresler,E., B.L. McNeal, and D.L. Carter. 1982. Salineand sodic soils. Springer-Verlag,Berlin.
Brown, I.R., and D. Warncke. 1988. Recommendedcation tests and measuresof cation exchange
capacity.p.15-16.In W.C. Dahnke(ed.) Recommendedchemicalsoil test proceduresfor the
North Central Region. North DakotaAgric. Exp. Stn. Bull. 499.
Brownell, P.F. 1979. Sodium as an essentialmicronutrientelementin plants and its role in metabo-
lism. Adv. Bot. Res.7:117-224.
Buckley, D.E., and R.E. Cranston.1971. Atomic absorptionanalysisof 18 elementsfrom a single
decompositionof aluminosilicate.Chem.Geol. 7:273-284.
Coughtrey,P.I., D. Jackson,and M. Thome. 1983. Sodium.p. 1-41. In Radionuclidedistribution and
transportin terrestrialand aquaticecosystems.A critical review of data. Vol. 3. AA Balke-
ma, Rotterdam.
Diest, J., and O. Talibudeen.1967. Rubidium-86as a tracerfor exchangeablepotassiumin soils. Soil
Sci. 104:119-122.
Eckert, OJ., and E.O. McLean. 1980. Differential bondingof potassiumand rubidium-86 in soils of
differing clay type and degreeof weathering.Soil Sci. Soc. Am. J. 44:425-428.
Feigenbaum,S., R. Edelstein,and I. Shainberg.1981. Releaserate of potassiumand structuralcations
from micasto ion exchangersin dilute solutions.Soil Sci. Soc. Am. 1. 45:501-506.
Franz, G., and R.M. Carlson.1987. Effects of rubidium, cesium,and thallium on interlayer potassi-
um releasefrom Transvaalvermiculite. Soil Sci. Soc. Am. I. 47:552-559.
Goulding, K. W.T. 1987. Potassiumfixation and release.Proc. Colloq. Int. PotashInst. 20:137-154.
Goulding, K.W.T., and O. Talibudeen.1963. Potassiumreservesin a sandy clay soil from the Sax-
mundhumexperiment:Kinetics and equilibrium thermodynamics.1. Soil Sci. 30:291-302.
Haagsma,T., and M.H. Miller. 1963. The releaseof nonexchangeablesoil potassium to cation
exchangeresins as influenced by temperature,moisture and exchangingion. Soil Sci. Soc.
Am. Proc. 27:153-156.
Helfferich, F. 1%2. Ion exchange.McGraw-Hili Book Co., New York.
Hillebrand, W.F., G.E.F. Lundell, H.A. Bright, and J.I. Hoffman. 1953. Applied inorganic analysis
with special referenceto the analysisof metals,mineralsand rocks. 2nd ed. John Wiley &
Hossner,L.R. 1996. Dissolution for total elementalanalysis. p. 4~. In D.L. Sparkset al. (ed.)
Methodsof soil analysis.Part 3. Chemicalmethods.SSSABook Ser. 5. SSA and ASA, Madi-
son, WI.
Jackson,M.L. 1956. Soil chemicalanalysis-advanced course.Publishedby author,Madison,WI.
Jones,R.L. 1989. Rubidium in surfacehorizonsof Illinois soils. Soil Sci. Soc. Am. 1. 53:588-591.
Jones,R.L. 1992. Extractablerubidium in surfacehorizons of Illinois soils. Soil Sci. Soc. Am. J.
56:1453-1454.
Klesta, EJ., and J.K. Bartz. 1996. Quality assuranceand quality control. p. 19-48. In D.L. Sparkset
al. (ed.) Methodsof soil analysis. Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and
ASA, Madison,WI.
Knudsen,D., G.A. Peterson,and P.E Pratt. 1982. Lithium, sodium, and potassium.p. 225-246.In
A.L. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA, Madison,WI.
Liebhardt,w.e., L. Svec, and M.R. Teel. 1976. Yield of com as affectedby potassiumon a Coastal
Plain soil. Commun.Soil Sci. Plant Anal. 7:265-277.
Longmore, M.E., B.M. O'Leary, e.w. Rose, and A.L. Chandica. 1983. Mapping soil erosion and
accumulationwith the fallout isotopecaesium-137.Aus!. 1. Soil Res. 21:373-385.
Malavolta, E. 1985. Potassiumstatusof tropical and subtropical region soils. p. 163-200.In R.E.
Munson (ed.) Potassiumin agriculture.ASA, CSSA, SSSA,Madison,WI.
Martin, H.W., and D.L. Sparks. 1983. Kinetics of nonexchangeablepotassium releasefrom two
CoastalPlain soils. Soil Sci. Soc. Am. J. 47:883--887.
Martin, H.W., and D.L. Sparks.1985. On the behaviorof nonexchangeable potassiumin soils. Com-
mun. Soil Sci. Plant Anal. 16:133-162.
McIntosh, J.L. 1969. Bray and Morgan soil test extractantsmodified for testing acid soils from dif-
ferent parentmaterials.Agron. J. 61:259-265.
Mehlich, A. 1953. Determinationof P, Ca, Mg, K, Na, and NH4• Mimeo 1953. North Carolina Soil
Test Div., Raleigh,Ne.
Mehlich, A. 1984. Mehlich 3 soil test extractant:A modification of the Mehlich 2 extractant.Com-
Morgan, M.E 1941. Chemicalsoil diagnosisby the universalsoil testingsystem.ConnecticutAgric.
Exp. Stn. Bull. 450.
Nieto, K.E, and W.T. Frankenberger.1985. Single column ion chromatography:II. Analysis of
ammonium, alkali metals, and alkaline earth cations in soils. Soil Sci. Soc. Am. J.
49:592-596.
Parker,D.R., D.L. Sparks,GJ. Hendricks,and M.e. Sadusky.1989a.Potassiumin Atlantic Coastal
Plain soils: II. Crop responsesand changesin soil potassiumunder intensive management.
Soil Sci. Soc. Am. J. 53:397-401.
Parker,D.R., GJ. Hendricks,and D.L. Sparks.1989b. Potassiumin Atlantic CoastalPlain soils: II.
Crop responsesand changesin soil potassiumunder intensive management.Soil Sci. Soc.
Am. J. 53:397-401.
Pearson,G.A. 1960.Toleranceof cropsto exchangeable sodium.USDA Inform. Bull. 216. U.S. Gov.
Print. Office, Washington,De.
Pratt, P.E 1965. Potassium.p. 1023-1032.In C.A. Black et al. (ed.) Methodsof soil analysis.Part 2.
Agron. Monogr. 9. ASA, Madison,WI.
Rausell-Colom,J.A., T.R. Sweetman,L.B. Wells, and K. Norrish. 1965. Studies in the artificial
weatheringof micas. p. 40-70. In E.G. Hallsworth and D.V. Crawford (ed.) Experimental
pedology.Butterworths,London.
Reitemeier,R.E 1951. The chemistryof soil potassium.Adv. Agron. 3:113-164.
Sparkset al. (ed.) Methods of soil analysis. Part 3. Chemical methods.SSSA Book Ser. 5.
SSSA and ASA, Madison,WI.
Rich, e.1. 1964. Effect of cation size and pH on potassium exchange in Nason soil. Soil Sci.
98:100-106.
Rich, e.1. 1968. Mineralogy of soil potassium.p. 79-96. In VJ. Kilmer et al. (ed.) The role of potas-
sium in agriculture.ASA, CSSA, and SSSA, Madison,WI.
Rich, e.1. 1972. Potassiumin minerals.Proc. Colloq. Int. PotashInst. 9:15-31.
Sadusky,M.C., and D.L. Sparks. 1991. Anionic effects on potassiumreactionsin variable-charge
Atlantic coastalplain soils. Soil Sci. Soc. Am. J. 55:371-375.
Sadusky,M.e., D.L. Sparks,M.R. Noll, and GJ. Hendricks.1987. Kinetics and mechanismsof potas-
sium releasefrom sandysoils. Soil Sci. Soc. Am. J. 51:1460-1465.
574 HELMKE & SPARKS
Schroeder,D. 1979. Structureand weatheringof potassiumcontainingminerals. Proc. Congr. Int.

PotashInst. 11:43--63.
Scott, A.D., A.P. Edwards,and J.M. Bremner.1960. Removalof fixed ammoniumfrom clay miner-
als by cation exchangeresins.Nature(London) 185:792.
Scott,A.D., and S1. Smith. 1987. Sources,amounts,and forms of alkali elementsin soils. Adv. Soil
Sci. 6:101-147.
Smith, S1., and A.D. Scott. 1966. Extractablepotassiumin Grundite illite: 1. Method of extraction.
Soil Sci. 102:115-122.
Soltanpour,P.N., and A.P. Schwab.1977.A new soil test for simultaneousextractionof macro-and
micro-nutrientsin alkaline soils. Commun.Soil Sci. PlantAnal. 8:195-207.
Southard,R1., andR.c. Graham.1992.Cesium-137distributionin a California pelloxerert:Evidence
of pedoturbation.Soil Sci. Soc.Am. I. 56:202-207.
Souty, N.R., R. Guennelon,and C. Rode. 1975. Someobservationsof potassium,rubidium-87,and
caesium-137absorptionby plantsgrown on nutritive solutions.Ann. Agron. 26:41-58.
Sparks,D.L. 1980. Chemistryof soil potassiumin Atlantic CoastalPlain soils: A review. Commun.
Soil Sci. Plant Anal. 11:435-449.
Sparks,D.L. 1987. Potassiumdynamicsin soils. Adv. Soil Sci. 6:1-63.
Sparks,D.L., and P.M. Huang.1985.Physicalchemistryof soil potassium.p. 201-276.In R.E. Mun-
son (ed.) Potassiumin agriculture.ASA, CSSA, and SSSA,Madison,WI.
Sparks,D.L., andW.C. Liebhardt.1981.Effect of long-termlime andpotassiumapplicationson quan-
tity-intensity (0/1) relationshipsin sandysoil. Soil Sci. Soc.Am. I. 45:786-790.
Sparks,D.L., D.C. Martens,andL.w. Zelazny.1980.Plantuptakeand leachingof appliedandindige-
nouspotassiumin Dothansoils. Agron. J. 72:551-555.
Sumner, M.E., and w.P. Miller. 1996. Cation-exchangecapacity and exchangecoefficients. p.
1201-1229. In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.
SSSABook Ser. 5. SSSAandASA, Madison,WI.
Talibudeen,O. J.D. Beasley,P. Leone,and N. Rajendran.1978.Assessment of soil potassiumreserves
availableto plant roots.I. Soil Sci. 29:207-218.
Thomas,G.W. 1982. Exchangeablecations.p. 159-165.In A.L. Pageet al. (ed.) Methodsof soil
Tinker, P.B. 1967. The relationshipof sodium in the soil to uptakeof sodium by sugarbeetsin the
greenhouseand to yield responsesin the field. p. 223. In G.Y. lacks (ed.) Soil chemistryand
fertility. Trans.It. Meet. Commun.2, 4, Int. Soc.Soil Sci. 1966.Univ. Press,Aberdeen,Scot-
land.
Wells, C.B., and K. Norrish. 1968. Acceleratedratesof releaseof interlayerpotassiumfrom micas.
Trans.9th Int. Congr. Soil Sci. 2:683-694.
Wolf, A., and D. Beegle.1991. Recommended soil testsfor macronutrients:Phosphorus,potassium,
calcium and magnesium.p. 25-34.In J.T. Sims and A. Wolf (ed.) Recommended soil testing
proceduresfor the NortheasternRegion.DelawareAgric. Exp. Stn. Bull. 493.
Woodruff, J.R., and C.L. Parks.1980.Topsoil and subsoilpotassiumcalibrationwith leaf K for fer-
tility rating. Agron. J. 72:392-396.
Yuan, L.L., L.w. Zelazny,and A. Ratanaprasatporn. 1976.Potassiumstatusof selectedpaleudultsin
the Lower CoastalPlain. Soil Sci. Soc.Am. Proc.40:229-233.
Published 1996
Chapter20
Beryllium, Magnesium, Calcium,

Strontium, and Barium
D. L. SUAREZ, USDA -ARS, U.S. Salinity Laboratory, Riverside, California
PROPERTIESOF ALKALINE·EARTH METALS
Beryllium, Mg, Ca, Sr, and Ba are known as the alkaline earth elements,which
are characterizedby two electronsin the outermostshell. Beryllium differs in its
chemical behaviorfrom the other Group II elements,in that covalent bonding
predominates.In contrastto Be, the other elementsin the group readily lose their
two outershell electronsand are mostly presentin ionic form as divalent cations.
To date no monovalentcation compoundshave been observedfor these ele-
ments.(Cotton & Wilkinson, 1980). Table 20-1 gives generalchemicalproper-
ties of the alkaline earthelements.A characteristicof theseelementsis the small
ionic radius and high charge/radiusratio. The chemical propertiesof Mg are
somewhatintermediatebetweenBe and the other Group II elementsin that Mg
also can form covalentbondsand both Mg and Be can precipitateas hydroxides
from aqueoussolutions. Calcium, Sr and Ba have a closer affinity, with a sys-
tematic changein their properties,related to increasingion size. Theseproper-
ties include increasingsolubility of the sulfate,chloride and nitrate salts,increas-
ing hydration of the crystalinesalts,and decreasingionization energy.
Calcium, Sr, and Ba, form relatively insoluble precipitatesof carbonates
and sulfates. In contrast, magnesiumsulfates are soluble and magnesiumcar-
bonatesdo not precipitateuntil Mg solution concentrationsare much greaterthan
Ca.
Magnesiumand Ca are abundantelementsin soil and also are amongthe
most commonly analyzed,becausethey are essentialelementsfor plant growth.
Calcium is essentialfor membranepermeability, solute transport and mainte-
nanceof cell integrity (Marschner,1986). Magnesiumis requiredfor many cel-
lular functions including productionof chlorophyll, proteinsynthesis,regulation
of cellular pH and cation-anionbalance(Marschner,1986). Strontium, Ba and
Be are much less abundantin soils and historically of lesserinterestto soil sci-
entiststhan Mg and Ca. Introduction of radioactiveSr into the atmospheredue
SegoeRd., Madison,WI 53711, USA. Methods of Soil Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
575
576 SUAREZ
Table 20-1. Propertiesof Group II elements.

Ionization enthalpiest
Atomic weight Ionic radiiU Usual coordination§ First Second
nm --klmol-1-
Be, 9.012 3.1 4 899 1757
Mg,24.31 7.8 4-6 737 1150
Ca,40.08 1.06 6 590 1146
Sr,87.62 1.27 6-12 549 1064
Ba,137.33 1.43 6-12 503 965
*
t Cotton and Wilkinson, 1980.
Basedon sixfold coordination.
§ Threkianand Wedepahl,1961.
to nuclearweaponstesting, and subsequentfallout into terrestrialenvironments

increasedinterestin Sr analysisand understandingof the biological and chemi-
cal processesassociatedwith Sr in the plant-soil environment.Plant and soil
materialare not commonlyanalyzedfor Ba content.The major interestin Ba for
soil systemshasbeenits useas an extractingcationfor the analysisof exchange-
able ions. Its use for this purposeis not recommendedfor arid land soils where
calciumcarbonateor gypsumare presentdue to the insolubility of theseBa salts.
Beryllium analyseshave rarely beenperformedfor soils, due in part to general-
ly very low concentrations.Nonetheless,Be is highly toxic to animals and
heightenedenvironmentalconcernsin recentyearshas led to increasedinterest
in Be chemistryand analyses.Although Be is an alkaline earth element(Group
lIa), its chemicalbehavior,with regardsto hydrolysis (Baes& Mesmer, 1976),
and complexation(Sillen & Martell, 1971) is closerto AI than it is to the other
alkaline earth elements.Despitethis chemicalsimilarity, Be occurrencein soils
has not correlatedwith AI concentration(Andersonet al., 1990), likely because
of differencesin mineral source.
MINERALOGY AND DISTRIBUTION OF BERYLLIUM,

MAGNESIUM, CALCIUM, STRONTIUM, AND BARIUM
IN ROCKSAND SOILS
The averagecrustal igneousrock compositionfor Be, Mg, Ca, Sr, and Ba

are 2.19,17.7x 1()3, 67.4 x 1()3, 342, and469 mg kg-I, and for sedimentaryrock
1.98, 19.7 x 1()3, 36.5 x 1()3, 338, and 525 mg kg-I, respectively(Turekian &
Wedephohl,1961).Typical soil concentrationsfor Be, Sr and Ba, are given as 6,
300, and 1000 mg kg-I, respectivelyby Ure (1991).
Beryllium is concentratedin dark minerals and muscovite. In granitic
rocks biotite and hornblendecan contain up to 15 mg kg-I Be while for mus-
covite Be content ranges from 10 to 50 mg kg-I. The mineral beryl
(Be2AI2Si03)6also can occur as a result of metamorphicalteration.Within min-
eral groupsthere is a positive relation betweenMg and Be content(Hormann,
1969).Relatively few analysesof Be in soils havebeenreported.Andersonet al.
(1990) reported that water-solubleconcentrationswere below their detection
ALKALINE EARTH ELEMENTS 577
limit (0.2 f1.g L -1) and total Be rangesfrom 0.3 to 30.5 mg kg-1 for southeastern
U.S. Piedontand CoastalPlan soils. Shackletteet a!. (1971) reportedan average
soil concentrationof 0.6 mg kg-1 for the conterminousUSA.
Magnesiumis abundantin many silicate mineralswhere it exists in solid
solution with Fe2+ (which has a similar octahedralradius). The Mg and Fe con-
tent of rocks are positively correlated,and decreasein the seriesfrom gabbroto
granite. Magnesiumis relatively rare in framework silicates such as feldspars,
but is an importantconstituentof many soil minerals,including olivines, pyrox-
enes,amphiboles,micasand clay minerals,as well as dolomite (Burns & Burns,
1974). Magnesiumalso is presentin limestone(calcite) with a substitutionof 1
to 9% for Ca.
Calcium is the fifth most abundantelementand occurs predominatelyin
silicates and carbonates,although phosphatesand sulfates also are important.
Mineral occurrencesinclude plagioclasefeldspars,pyroxenes,amphiboles,cal-
cite, aragonite(rare in soils), gypsumand dolomite (Faure,1978). Most Ca input
to rivers is estimatedto be derivedfrom weatheringof carbonaterocks (Garrels
& Perry, 1974).
Minor amountsof Sr minerals are found in nature (such as strontianite,
SrC03, and celestite,SrS04),however,most Sr is presentas a trace constituent
of other minerals.Strontiumsubstitutesreadily for Ca as well as K (for example,
in potassiumfeldspar). Most of the Sr input to rivers is from weatheringof car-
bonatesand sulfates(Brass,1976).
Barium is presentas a substitutedion in silicates,mostly feldspars,biotite
and muscovite,and rarely forms its own minerals.The most importantsubstitu-
tions are for K and Ca, with potassiumfeldspar being the most important
(Puchelt,1972). The Ba contentof most soils is in the rangeof 100 to 3000 mg
kg-1 of soil.
TOTAL ELEMENTAL MAGNESIUM, CALCIUM,

STRONTIUM, BARIUM, AND BERYLLIUM
Introduction
Total analysisof theseelementsgenerallyrequiresa soil digestionfor com-

plete destructionof the crystallineand organiccomponentsof the soil. The most
commonmethodis still the sodium carbonatefusion (Jackson,1958), although
the lithium borate fusion (Medlin et a!., 1969), the HCI-HF digestion (Isaac &
Kerber, 1971) and more recently microwave digestion (Gilman & Engelhart,
1989) also are often used.Digestionwith HF shouldbe avoidedif Be analysisis
to be performed,as Be is one of the elementswhich forms volatile fluorides,
resulting in incomplete recovery (McLaren, 1987). Once the elementsare in
solution, they can be readily analyzedby the methodsdescribedbelow, most
commonly be Atomic Absorption Spectroscopy(AAS). More recentalternative
methodsfor total analysisinclude high energy,destructivesampling,ion beams
and Inductively CoupledPlasmaSpectroscopy(ICP) using suspendedsolids.
578 SUAREZ
DigestionMethods
Detailedmethodsfor digestionof soil samplesare presentedin Chapter3

(Rossner,1996).
Analysis by Atomic Absorption Spectroscopy
Seesectionon "Atomic Absorption Spectrophotometry,"Tables20-2 and

20-3, and Chapter4 (Wright & Stuczynski,1996).
Analysis by Inductively CoupledPlasma-

Atomic EmissionSpectrometry
Seesectionon "Beryllium" and Chapter 5(Soltanpouret aI., 1996).
X-Ray Fluorescence
Wavelengthdispersiveand energydispersivex-ray fluorescenceoffer the

possibility of rapid analysiswith a minimum of samplepreparation.The spec-
trometer uses either a solid state Li-drifted silicon detector (wavelength) or
"Bragg diffracting crystal" (dispersive).The methodis widely usedfor measur-
ing many major and minor elementsin rock samples,including Mg, Ca, Sr, and
Ba (Potts, 1987). The method is not suitablefor Be analysis(Be is usedas the
window on the x-ray tube). Samplepreparationtypically consistsof preparation
of powderpelletsor glassbeadsformed from the lithium boratefusion, or sodi-
Table 20-2. Atomic absorptionspectroscopyflame & instrumentsettings.

AA setting Mg Ca Sr Ba Be
Grating Ultraviolet Visible Visible Visible Ultraviolet
Wavelength 285.2nm 422.7nm 460.7nm 553.6nm 234.9nm
Oxidant Air Air Air Nitrous oxide Nitrous oxide
Fuel Acetylene Acetylene Acetylene Acetylene Acetylene
Flame Slightly reducing Slightly reducing Slightly reducing Reducingrich Reducingrich
Table 20-3. Atomic absorptionspectroscopyanalysesof alkali earthelements(detectionlimitt and

working rangein IlgIL).
Flame Flame Graphite
detection working detection
Element Wavelength limits range limit furnace Reference
Be 234.9 1.0 0.02 GiII,1993
0.5(N-Ac) 50-2000 0.2 Clesceriet aI., 1989
Mg 285.2 0.3 0.01 GiII,1993
0.5 20-2000 Clesceriet aI., 1989
Ca 422.7 0.5 0.004 GiII,1993
0.3 200-20000 Clesceriet aI., 1989
Sr 460.7 30 300-5000 Clesceriet aI., 1989
Ba 553.6 10 0.04 GiII,1993
30(N-Ac) 1000-20000 2.0 Clesceriet aI., 1989
t 2 x SD of a blank solution.
urn carbonatefusion (as describedin Chapter7, Karathanasis& Hajek, 1996).

Cooling of the glass bead is an important considerationas it must cool suffi-
ciently slowly to avoid crackingand sufficiently fast to avoid crystallizationand
nonhomogeneity.Recentimprovementsin samplepreparationand instrumenta-
tion methodsare describedby Coutureet al. (1993).
Recently total-reflection x-ray fluorescencespectrometryhas been pro-
posedfor analysisof water and soil sampleswith increaseddetectionlimits over
those of ICP, good precisions and small sample size (10 ,uL) (Mukhtar &
Haswell, 1991).
Procedure
Prepareglass bead (fusion) as describedin Chapter 2 (Klesta & Bartz,
1996).Typically the glassbeadis preparedusing a 5- to lO-g soil sampleground
to at least <150 ,urn and preferably <50,um. After fusion, molten samplesare
rapidly pouredinto a graphiteor duraluminplatenwhich is placedon a" hot plate
at 200°C.A preheatedplungeris loweredto flatten the glassbeadinto a flat disc.
Mter heatingfor 10 min, the platen is removedand cooled at room temperature
(Potts, 1987). Also see Chapter 7 (Karathanasis& Hajek, 1996) "Elemental
Analysis by X-Ray FluorescenceSpectroscopy"for preparationof glassbead.
Powderpelletsare preparedby mixing ground rock or soil with a binder if
necessaryto prevent sample crumbling and compressingthe mixture between
tungstencarbideplatensin a die, typically to 2 to 3 X 107 kg m-2 (2-3 tons cm-2,
Potts, 1987). The binder consistsof a solution of polyvinylpyrrolidone-methyl
celloseaddedas a drop of solution per gram of sample(Harvey & Atkin, 1982).
Operatingconditions and proceduresfor x-ray fluorescenceanalysis are
given in Chapter7 (Karathanasis& Hajek, 1996). Typical detectionlimits for
Mg, Ca, Sr, and Ba are 1,000 12, 3, and 24 mg kg-1 respectively(Potts, 1987),
generallysufficient for most soil samples"
SuspendedSolids by Inductively CoupledPlasma-

Atomic EmissionSpectrometry
Principles
Use of specializednebulizers,such as the Babington,modem"V" groove
and cone entrainmentderivativesallow for direct analysisof slurrieswith parti-
cles as large as 1 mm, althoughit is not recommendedto usesuchlarge-sizepar-
ticles. In order to maintainhomogeneityof the slurry, the particles shouldbe less
than lO,um (Sharp,1987). Ebdonand Collier (1988) recommendedonly the sin-
gle pass(conical)spraychamber,demonstratingthat the doublepassandcyclone
spray chambersalteredthe size distribution of the particlesthat were transport-
ed to the torch. This is particularly undesirablefor analysisof soils, where even
after grinding, different mineralsoccur in the different-sizefractions. The slurry
methodoffers excellentpotential as a rapid and accuratetechnique.Fuller et al.
(1981), however,reportedthat despitethe high temperaturesof the ICP method,
matrix dependencies still existedand thus standardshad to be similar to the sam-
ples in composition.See Chapter 5 (Soltanpour et al., 1996) "Inductively
580 SUAREZ
Table 20-4. Inductively coupledplasmadetectionlimit and working range(l1g1L)t.

Concentric
Element Wavelength nebulizer Working range Ultrasonic Reference
Be 0.07 Gill, 1993
313.04 0.3 Clesceriet aI., 1989
234.86 0.2 0.006 Nham,1995
313.0 5-10000 Fishman& Friedman,1985
Mg 0.08 Gill, 1993
279.08 30 Clesceriet aI., 1989
389.2 5-100000
Ca 0.07 Gill, 1993
317.93 10 Clesceriet aI., 1989
Sr 407.77 0.5 Clesceriet aI., 1989
407.77 0.02 0.002 Nham,1995
421.5 0.5-10000 Fishman& Friedman,1985
Ba 455.4 2 Clesceriet aI., 1989
455.4 0.07 0.0012 Nham,1995
2-10000 Fishman& Friedman,1985
t 2 x SO of a blank solution.
CoupledPlasmaEmissionSpectrometryand Inductively CoupledPlasma-Mass

Spectrometry"for more detailedinformation.
Procedure
Use an ICP instrumentfitted with a single passspraychamberand a 3-mm
inside diameter(i.d.) injector. Grind slightly more than 5 g of soil in agatemor-
tar or motorizedgrinder until all particlesare less than 10 rom in size. Amount
of grinding requiredwill vary dependingon the soil mineralogyand will require
somepreliminary evaluation.Preparea dispersingsolution of 1% Triton! (J.T.
Baker Inc., Philipsburg, NJ) in deionizedwater (01). Next preparea slurry by
weighing out 5.00g of groundsoil in 100 mL of dispersingsolution. Shakesus-
pensionimmediately before analysisto maintain sampleuniformity. Use torch
conditions, wavelength lines and working concentrationrangesdescribedbelow
in the sectionon "Analytical Methods"and in Table 20-4.
Ion Beam
Among otherdirect analyticalmethodsfor alkaline earths,electronmicro-
probe, SIMS (secondaryion mass spectrometers),and ESCA (electron spec-
troscopyfor chemicalanalysis)are increasinglybeing used,most commonlyfor
analysisof mineral surfacesrather than total analysis.SeeChapter13 (Fendorf
& Sparks,1996), "X-Ray Absorption Fine StructureSpectroscopy"for discus-
sion. Laser microanalysisusing atomic emissionspectrometry(LM-AES) also
can be usedfor total analysis;for example,Moenke-Blankenburgand Gunther
(1992) usedthe methodto determinethe Ba contentof ferromanganese nodules.
1 Manufacturersbrand namesare supplied for informational purposesonly and do not imply

endorsementor preferentialtreatmentby the USDA.
PARTITIONING OF ALKALINE· EARTH METALS IN SOILS
Introduction
Various approachescan be taken to partition the alkaline earthelementsin

soils. For example,Mokwunye and Melsted (1972) proposeda fractionation of
Mg into organicallycomplexed,exchangeableand acid soluble(mineral). Other
approacheshave included fractionation basedon the strengthof the extractant
used.This methodis not desirablefor the alkaline earthssince it resultsin oper-
ational definitions ratherthan distinct chemicalpools. Nonethelessit is of some
value to evaluateplant availability of theseelementssinceextractabilitywill dif-
fer within as well as amongthe variousfractions. Although useful for agronom-
ic evaluationsit is less useful for understandingthe underlying chemical reac-
tions and controlson soluble ion composition.The alkaline earthsare presentin
distinct, characterizable,soil fractions classified as soluble, organically bound,
exchangeable, carbonates,distinct silicate phases,and sulfates.While the silicate
fraction is almostalwaysthe largest,and providesa long-termsourcefor soluble
cations,it doesnot affect the seasonaland short-termchangesin the other frac-
tions.
Soluble
Principles
In the past,solublecationswere often includedinto the exchangeable frac-
tion, due to the relatively low concentrationsin solution as comparedto the
exchangeablequantities.More recently there has been increasedinterest in the
soil solution chemistry in addition to the traditional evaluation of nutrient
requirements.Study of solution chemistryrequiresthat the solution composition
be evaluatedindependentlyof the exchangeablecations.In addition, salinesoils
have appreciablequantitiesof soluble Ca and Mg, which need to be character-
ized independentlyof the exchangeablefraction. Analyses of soluble alkaline
earth metalsare generallyperformedusing a saturationextract, as describedin
the sectionon "Extractable/Exchangeable Fraction" and in detail in Chapter14
(Rhoades,1996),"Salinity: Electrical Conductivity and Total DissolvedSolids."
This extractmethodprovidesa convenientreferencestatewith which to compare
different samples,but doesnot representthe solution compositionof the sample
underfield conditions,wherethe water contentis lower and the CO2 partial pres-
sureis much greater.Alternative ways of measuringsolublecationsincludesuse
of zero-tensionlysimeters (Holder et aI., 1991) as well as vacuum extractors
(Litaor, 1988), centrifugationand immiscible displacementtechniques(Elkhatib
et ai., 1987).The selectionamongthesemethodsdependson the researchobjec-
tives, specificallywhetheronewantsto examinethe averagesoil composition,or
whetherone is interestedin the compositionof the macroporesolution, which is
pertinentfor solutetransport.
Procedure
Proceduresfor extractionare given in Chapter14 (Rhoades,1996).
582 SUAREZ
Extractable/Exchangeable Fraction
Principles
The extractableor exchangeablefraction is the fraction most commonly
measuredfor alkaline earth elements.In nonarid zone soils the soluble fraction
of the soil is usually sufficiently small relative to the exchangeablefraction that
no correctionfor solublesaltsis necessary.For thesesoils the term extractableis
generallyutilized. For arid zonesoils it is often necessaryto subtractan estimate
of the soluble cation content from the extractedcation content to obtain the
exchangeablecation quantity (mass). We use the term exchangeablebelow,
assumingthat the soluble quantity either can be neglectedor is subtractedfrom
the extractablefraction. Calcium and Mg are almost always the major compo-
nent of the exchangeableions (with the notable exception being acid soils).
Strontium,Ba, and Be are generallyinsignificant contributorsto the sum of the
exchangeablecations,howeverthis fraction may representan importantcompo-
nent of theseelementsin soil. The adsorptionof exchangeablecationsfrom the
solutionis assumedto be a reversibleprocess.Typically the exchangereactionis
conductedunderbufferedor constantpH conditions.This servesto provide a ref-
erencebasefor comparisonbetweensoils but may result in extractionswhich are
different than those conductedat the pH of the soil in its field condition. Ex-
changeis typically carriedout by multiple reactionswith a concentratedsolution
of an ion which is preferentiallyabsorbedon the exchangesites.This processis
designedto ensurethat the exchangecationsare completelyextractedby the dis-
placing solution. Thesestandardizedexchangemethodsare well suited to com-
paring soils but do not always representthe exchangeablecations as existing
under field conditions. This would require matching the samplespH and ionic
strengthwith an exchanging cationof similar chemicalproperties.In addition to
being experimentallytime consuming,thesedesired criteria cannot be readily
met for the alkaline earths.
Calcium, Mg and Sr can be extractedby concentratedsolutionsof Ba, but
this method is not suitablefor arid land soils. Arid land soils require extraction
with NH4 (or other monovalentsalts)if Ca and Mg analysesare required.Use of
Sr and Ba extractantsin calcareoussoils resultsin precipitationof the insoluble
Sr and Ba carbonatesand extensivedissolutionof calcium carbonate,resulting
in erroneouslyhigh Ca concentrations.Similarly useof NaOAc at pH 8.1 (Bower
et aI., 1952) or other buffers may not be satisfactoryas either dissolutionor pre-
cipitation may occur. At presentthis problem has no easysolution. Ratherthan
attemptingto prevent both dissolution and precipitation, correction for sulfate
and alkalinity in the extracted solution (Amrhein & Suarez, 1990) may be
desiredfor calcareousor gypsiferoussoils.
In someinstancesNHt may extractmore or lesscationsthanjust thoseon
the exchangesites. In calcareous soilsthe extractantdissolvescalcium carbon-
ate. Ammonium acetatemay not extract all cations in the caseof soils high in
mica and vermiculite(Barshad,1954a,b).Also, Fisher(1963)determinedgreater
exchangeableMg concentrationsusing NH40Ac than determinedusing a mag-
nesiumradioisotopemethod.This NHt can exchangeirreversibly with interlay-
er cations which are not generally consideredexchangeableions. Although it
may be conceptuallydesirableto standardizeon a uniform extractantsolution it

is not possible due to differences in soil environmentsand properties.As an
examplefor calcareoussoils an extractantsuch as NaOAc at pH 8.1, may be an
acceptablereferencestatebut this is not a suitablereferencefor acid soils, par-
ticularly when the variable chargecomponentof exchangeis important. Under
these circumstancesa more realistic extraction is necessary.Most commonly
ammoniumacetatebufferedat pH 7 is used.For acid soils, BaCl2 extractantsalso
may be used;thesecan be adjustedwith HCL or NaOH to the desiredpH.
Ammonium AcetateMethod
Apparatus
1. Centrifuge.
2. Centrifugetubes.
3. Vortex mixer.
4. Two-liter volumetric.
5. 100-liter volumetrics.
6. Shaker.
Reagents.Ammonium acetate(NH 40Ac), 0.5 M. Add 70 mL of reagent
gradeNH 40H (sp. gr. 0.90) into a 2-L volumetric containingapproximately1.5
L of deionizedwater. Next slowly add 57 mL of 99.5% acetic acid while inter-
mittently swirling the flask. Adjust to pH 8.1 for arid zone soils or 7.0 for non-
calceroussoils, with either NH 40H or aceticacid. Make up to final volume with
deionizedwater.
Procedure.Add 5 g of <2 mm air-dried soil into 40-mL centrifugetubes.
For soils with CEC values in excessof 200 to 300 mmolc kg-1 soil, reducethe
soil addedproportionately.Oven dry anothersubsample of the same soil and
convert the air-dry weight to an oven-dry basis.Add 33 mL of extractingsolu-
tion to each tube. Cap and mix on a vortex mixer and place in a shakerfor 30
min. Next place the samplesin a centrifugeand spin at 5000 relative centrifugal
force (rcf) for 10 min or until the clay hasbeensettled.
Decantthe supernatantinto a 100-mL volumetric. Add another33 mL of
extractingsolution and repeatthe extraction. Repeatagain. Next bring the vol-
ume of the 100-mL volumetric up to the mark with the addition of a few milli-
liters of the extractingsolution. Determinethe exchangeable cationsusing one of
the analytical methodsdescribedbelow in the sectionon "Analytical Methods."
Preparestandardsusing the 0.5-M extractingsolution.
In the caseof soils high in soluble salts it is necessaryto correct for the
concentrationof solublecations.This correctioncan be madeas follows: analyze
the soluble cationsin a separatesubsample,expressthe massof soluble cations
per kilogram of dry soil. Calculatethe massof eachsolublecation in the sample
usedfor determinationof exchangeable cations.Next subtractthis massfrom the
massof the cation determinedin the extract. Sincethe exchangeablecation con-
centrationsare expressedin millimoles of chargeper kilogram soil, the soluble
concentrationsfrom the saturationextractalso must be expressedon a millimoles
of chargeper kilogram basisbefore subtraction.
584 SUAREZ
In the caseof soils with large amountsof smectitesit may be desirableto

correctfor the effectsof anion exclusion(Bolt et aI., 1976). Among thesemeth-
ods the easiestis to determinethe Cl concentrationin the ammonium acetate
extract and subtractan equivalentconcentrationof cations in the proportion in
which they occurredin the saturationextract[seeChapter14 (Rhoades,1996)for
preparationof a saturationextract].
Calculations
mmole kg-1 = L of extractvolume
x analyzedconcentrationof extract(mmole L- 1) x 1000/gsoil.
High SolubleSalts:
mmole kg-1 = [(L of extractvolume x analyzedconcentrationof extract
(mmole L-l) x 1000/gsoil] - mmole kg-1 soil soluble.
Ammonium Chloride Method for Calcareousand GypsiferousSoils

Procedure.Correction for dissolution of calcite and/or gypsum requires
the following modification of the NH 40Ac method. Substitute NH4Cl for
NH 40Ac as the extractingsolution. Adjust the pH to 7.0 with NaOH (solution
will be poorly buffered). Performthe extractionas indicatedabove.The extract-
ing solution will be bufferedby the soil. In almost all casesthis shouldnot be a
problem.Determinethe alkalinity andsulfateconcentrationon the lOO-mL solu-
tion collectedfrom the extraction.The original buffered NH4Cl should be used
as the blank for the alkalinity titration. The sum of the alkalinity and sulfate,
expressedas millimoles of chargeper liter, is equal to the Ca dissolvedduring
the extraction.Subtractthis quantity, expressedas millimoles of chargeper kilo-
gram from the exchangeableCa concentration.Soils very high in pH (>9) may
require modificationsin procedureto avoid dispersionproblems.
Calculations
mmole kg-1 = [(L of extractvolume x analyzedconcentrationof extract
(mmole L -1) x 1000/gsoil)] - mmole kg-1 soil soluble
- mmole kg-1 dissolved.
Acid Soil Extraction

Use ammoniumacetateextractantadjustedto pH 7.0. Correctionsfor sol-
uble salts,and calcite and gypsumdissolutionare not generallyneeded.In cases
where it is not desirableto exchangewith ammonium,and Ba is not being ana-
lyzed, 0.5 M or 0.25 M barium chloride may be utilized as the extractant.In this
casethe solutionsmay be adjustedto the pH desired.It is important to use the
stock extractingsolution when preparingthe standardsas the BaCl2 may contain
other alkali earthsas impurities.
Availability Indices
Various soil extraction methodsare utilized to obtain availability indices

for plants. Exchangeableand soluble Mg, Ca, Sr, and Ba extracted from
NH40Ac at pH 7 are regardedas available to plants. Other indices include
extraction with 0.0125 M CaCl2 (Schachtschabel,1954) and 0.05 M HCI
(Grahamet aI., 1956). Theseindices range from readily available to potentially
available. Welte et ai. (1960) suggestedthat a better understandingof nutrient
availability would requirethat severalindicesbe measured.The conceptof using
severalindicesis compatiblewith the useof the termsintensity (concentrationin
the solution) and capacity (potentially available such as the exchangeableand
organically bound fraction, Marschner,1986). It seemsunlikely that a general-
ized availability index can be establishedthat would be useful for all soils, due
to their differencesin mineralogy,organic matter content, and chemical condi-
tions. It is not surprisingthat different extractantsare utilized in different envi-
ronments,adoptedafter demonstration that the extractantresultsin adequatecor-
relation of the soil and plant factors. In addition, many availability indices are
modified from the quantitative methodswhen they are adoptedby soil testing
laboratoriesfor routine agricultureproductionevaluation.Modifications are gen-
erally implementedto save time and reduce volumes of extractants.Accurate
characterizationof plant availability may be a problem for Mg and Ba (Lanyon
& Heald, 1982).
Specific plant responseto Mg, Ca, Sr, and Ba dependsnot only on avail-
ability but also on the natureof the soil constituents,ion ratios, where ion com-
petition or imbalancesmay occur,as well as specific plant speciesand evenvari-
etal response.Calcium is usually not deficient in agronomiccropsexceptin very
acid soils with low exchangecapacity(Howard & Adams, 1965). It is generally
acceptedthat the positive plant responseto Ca application to soils (liming) is
rarely due to the increasedCa per se, but ratherdue to an increaseor decreasein
other elements(such as decreasedAl toxicity due to increasedsoil pH and thus
decreasedAl availability).
Shortcomingsof the availability indices are illustrated by the caseof Ca
deficienciesin highly alkaline soils. Despitethe largequantitiesof Ca that can
be extractedwith a NH 40Ac solution bufferedat pH 7, the high pH of 10 or more
in its field condition limits the calcium carbonatesolubility. In this case the
extract masksthe potential Ca deficiency or ionic imbalance.In calcareous,arid
land soils, adverseplant responsemay occur due to ion imbalanceswithin these
elements.Barber(1984, p. 262-277)proposedthat the relation of Ca in solution
to other soluble cationsprovided a betterestimateof Ca statusthan did evalua-
tion of the exchangeablefraction. Many crops suffer yield loss due to high
MglCa ratios in solution. Carter et ai. (1979) found Ca deficiency in barley
(Hordeum vulgare L.) when the Ca/Mg in the soil solution was lessthan one, and
Wolt & Adams(1979) reportedCa deficiencyfor peanuts(Arachis hypogaea L.)
when the Ca/total cation ratio (TC) decreasedbelow 0.1 to 0.25 (dependingon
growth stage). Although Ca/TC activity ratios or ion concentrationratios are
consideredto be suitable for evaluation of Ca deficiency (Carter & Webster,
1990), it seemspreferableto considerspecific ion activity ratios. Adverseplant
586 SUAREZ
responsescan occur for Ca2+/Mg2+ ion activity ratios as high as 1.0, in the case
of wheat(Triticum aestivum L.). In salt-affectedsoils, low Ca/Mg ratios are quite
common,becauseMg salts (particularly carbonates)are more soluble than Ca
salts.
Magnesiumdeficienciesfor optimal plant growth are not uncommon.This
deficiency generally occurs in very acid soils, low in exchangeableMg.
Interferenceof other ions includesNH4-inducedMg deficiency (Mulder, 1956)
which can be alleviatedby substitutionof N03 fertilizer, which alsowill serveto
raisethe soil pH. High levelsof K usually due to high fertilizer applications,also
may reduce Mg uptake (Lanyon & Herald, 1982). ElevatedCa concentrations
also can causeMg deficiency (Hansen& Munns, 1988; Plaut & Grieve, 1988).
As expectedthe point at which Mg may becomedeficient dependson plant
species,environmentalfactors such as temperature(Thill & George, 1975) as
well as ion ratios and varietal differences.
Strontium and barium are consideredas nonessentialelementsfor plant
growth, but are still of interest as they substitute readily for Ca and Mg.
Strontium is of particular concerndue to the radioisotope9OSr, which readily
entersthe food chain. The point at which nonradiogenicSr hasan adverseeffect
on plant responseis speciesdependentand expressedin termsof the Sr/Caratio
(Bowen & Dymond, 1956).
Barium toxicity can occur, but usually only if the Ba levels are greatly in
excessof the Mg and Ca concentrations(Robinsonet aI., 1950). Plant toxicity
due to Ba is thus associatedprimarily with contaminatedsites. Also, due to the
low solubility of barium sulfate, elevatedconcentrationsof Ba may result in S
deficiency.
ANALYTICAL METHODS
Introduction
Analysis of solutionsfor alkaline earthelementscan be performedusing a

wide variety of methods.The most common is AAS, which is relatively rapid,
with good accuracyand generallysatisfactorydetectionlimits. Flame emission
spectrometryis less desirablebut can be utilized in many instances.Inductively
CoupledPlasmaEmissionSpectroscopy(ICPES) also is frequently usedand is
well suited for Ca and Mg, with excellent precision, minimal matrix interfer-
encesand suitable detectionlimits. Inductively coupledplasmaemissionspec-
troscopyis lessuseful for Be, Sr, andBa analysisin that detectionlimits are often
not sufficient for soil andwatersamples(AAS is considerablymoresensitivefor
the alkaline earths).Other commonly used methodsinclude High Performance
Liquid Chromatography(HPLC) and ICP-mass spectrometry (ICP-MS).
Titrimetric methodsusing color indicatorsare very precisebut time consuming,
as a result they are presentlyinfrequently used.
Atomic Absorption Spectrophotometry
Introduction
Atomic absorption spectrophotometry has been the most frequently uti-
lized analytical method for analysisof alkaline earth metals in water extracts.
Absorption spectroscopyis basedon the characteristicenergy levels at which
specific elementsabsorbenergyas they are excitedfrom their groundstate.The
methodsare fast, quite accurate,with low detectionlimits and relatively small
samplepreparationrequirements.Interferencescan be classified into chemical,
ionization, matrix, spectralor self-absorption(Isaac & Kerber, 1971). Details on
the theory and application of AAS are presentedin Chapter 4 (Wright &
Stuczynski,1996).
Principles
The elementsMg, Ca and Sr are typically analyzedusing the air-acetylene
oxidant, fuel combination.Important interferencesare causedby the formation
of stable complexes.For soil watersthe major interferencesare causedby the
formation of sulfateand nitrate complexes.Theseinterferencescan be overcome
by the addition of high concentrationsof La, or lessfrequently recommended,Sr
(for the analysisof Mg or Ca). Thesemetalsform strongcomplexes,thus great-
ly reducing the free ligand concentrationand reducing the formation of com-
plexeswith Mg and Ca. Interferencesalso can be overcomeby the use of chela-
tors, such as ethylenediaminetetraacetic acid (EDTA) (West & Cooke, 1960).
Alternatively the interferencescan be reducedby the use of higher flame tem-
perature,although this in tum causesreducedsensitivity due to increasedion-
ization.
Ionization interferencescan be reducedby the addition of more easily ion-
ized elementssuchas Rb, Cs, or K. Ionization interferenceis more of a problem
for Sr and Ba analysesthan for Ca and Mg. Aluminum and silica interferences
also may occur, for Mg, Ca, and Sr analysis.Aluminum interferencesare elimi-
natedwith the use of La or Sr. Phosphatemay interferewith Ca and Sr analyses,
althoughP is rarely at sufficient concentrationin soil watersto causeinterference
and is readily overcomeby the addition of La. Concentrationsof Mg in excess
of 1000 mg L-l reduceCa values(Fishman& Friedman,1985).
Magnesium, Calcium and Strontium

Lanthanumis addedto solutionsto achievea concentrationof 1% (wt/vol)
in the samplesand standards.A stocksolution of La is preparedby mixing 29 g
of La20 3 with a few milliliters of deionizedwater to form a slurry. Next, slowly,
add while stirring, 250 mL of concentratedreagentgradeHCL (reactionmay be
violent). Dilute the solution to a final volume of 500 mL by addition of deion-
ized water. One milliliter of this solution is addedto every 10 mL of standardor
sample.
588 SUAREZ
Typical working rangesfor Mg are 0.1 to 10 mg L-1 when the burner is

positionedparallelto the light beamand2.5 to 50 mg L-l whenthe burneris per-
pendicularto the beam(Fishman& Friedman,1985).The methoddetectlimit is
dependenton the instrumentused. Clesceriet al. (1989) list the Mg detection
limit as 0.0005mg L-1 Mg, and the optimal concentrationrangeis consideredto
be 0.02 to 2 mg L-l. Typical working rangesfor Ca are 0.01 to 5 mg L-l when
the burneris positionedparallel to the light beamand 1 to 60 mg L-l when the
burner is perpendicularto the beam(Fishman& Friedman,1985). Instrument
detectionlimit for Ca is given as 0.003 mg L-l and the optimal concentration
rangeis consideredto be 0.2 to 20 mg L-l (Clesceriet aI., 1989).Typical work-
ing rangesfor Sr are 0.01 to 5 mg L- 1 when the burneris positionedparallel to
the beam(Fishman& Friedman,1985).Instrumentdetectionlimit for Sr is 0.03
mg L-l and the optimal concentrationrangeis consideredto be 0.3 to 5 mg L-l
(Clesceriet aI., 1989).It is recommendedthat 1000mg L-1 of KCL be addedto
standardsand samplesto minimize Sr ionization. Aluminum, P and Si interfer-
encesarecorrectedby additionof La asdescribedabove.Instrumentsettingand
flame adjustmentare given in Table 20--2. Detectionlimits and working ranges
are given in Table 20--3.
Barium
Barium analysisby AAS is performedwith a nitrous oxide burnerand a
nitrous oxide-acetylenegas mixture. This method reducesinterferencesand
increasessensitivity if NaCI is addedto suppressBa ionization. The recom-
mendedNaCI concentrationis 250 mg L-1 in standardsand samples.1Ypica1
working rangesfor Ba are 0.1 to 5 mg L-l when the burneris positionedparal-
lel to the light beam(Fishman& Friedman,1985). Instrumentdetectionlimits
are 0.03 mg L-l and the optimal concentrationrangeis consideredto be 1 to 20
mg L-1 (Clesceriet aI., 1989).
Beryllium
Principles.Although AAS is a desirablemethodfor mostmetals,analysis
of Be by flame AAS requiresovercomingthe severesuppressinginterferenceof
AI and slight enhancement by Mg, Ca and Na. In addition, detectionlimits are
not sufficient for most samples.The following procedureof Robleset al. (1991)
usesan extractionstepto overcomeinterferencesand improve detectionlimits.
Apparatus
1. Atomic absorptionspectroscopywith a N20 burner.
2. Separatingfunnel.
Reagents
1. Prepareextractant,0.1 mol L- 1 N-Beozoyl-N-phenylhydroxylamine
(HBPA) in isobutyl methyl ketone(IBMK).
Extraction Procedure
1. Adjust the pH of a l00-mL sampleto 6.3.
2. Transfer the sample into the separatingfunnel and add 20 mL of

HBPA-IBMK extractant.
3. Equilibrateby shakingfor 3 min.
4. Allow the phasesto separateand discardthe aqueousphase.
5. Preparestandardsusing the sameextractionmethod.
6. Analyze the sampleusing a nitrous oxide-acetyleneflame.
7. Disposeof IBMK as requiredby the pertainentregulations.
Comments.Detectionlimit is 2).lg L -I and responseis linear up to 1.0 mg
L-I (Robleset aI., 1991). Detectionlevel is suitablefor soil analysesbut not for
most natural waters.Adjustmentof pH is requiredfor satisfactoryextraction.
Determinationin Aqueous Extract. Be analysisby AAS is performed
with a nitroul' oxide burner and a nitrous oxide-acetylenegas mixture. This
methodreducesinterferences,and increasessensitivity if CaCl2 is addedto sup-
pressionization. The recommendedCaCl2 concentrationto suppressionization
is 2.5 g L-I in standardsand samples.A typical working rangefor Be is 0.01 to
0.2 mg L- 1 when the burneris positionedparallel to the light beam(Fishman&
Friedman,1985).Instrumentdetectionlimit is 0.005 mg L -I and the optimal con-
centrationrangeis consideredto be 0.05 to 2 mg L- 1 (Clesceriet aI., 1989).
Interferencesare reportedfor Al concentrationsabove0.5 mg L-I and Na
and Si concentrationsabovelOOO mg L-1.
Inductively CoupledPlasma
Introduction
This instrumentis rapidly becomingthe method of choice for many ele-
mentsincluding the alkaline earths.The plasmais producedby passingionized
Ar gas through a quartz torch. A radio frequency(RF) generator,operatedtypi-
cally at 27.1 MHz, results in oscillating magneticfields, electron acceleration
and collisions generatingtemperaturesof approximately 10000 K. The high
temperaturesand increasedresidencetimes, as comparedto flame spectroscopy,
resultsin generallyincreasedsensitivity and lesschemicalinterference,although
spectralinterferenceshave to be carefully evaluated.
Recent developmentof an ultrasonic nebulizer results in an increasein
aerosol production, narrow droplet size distribution and small droplet size.
Elementscomparedto date including Be, Ba, and Sr suggestlO-fold or better
increasein sensitivity, as comparedto the conventionalpneumaticnebulizer.
Beryllium, Magnesium,Calcium, Strontium,and Barium

The wavelength,working rangeand detectionlimits for analysisare given
in Table 20-4. The ultrasonicnebulizeris particularly useful for analysisof Be,
and Ba which occur at low concentrationsin most soils.
Interferences
Interferenceof Ca and Mg on Be, Sr, and Ba determinationshas been
demonstrated,producingreductionsin sensitivity of 5% for Be at concentrations
590 SUAREZ
as low as 1000mg L- 1 for the interfering element(Thompson& Ramsey,1985).

As a result of this interferencethe higher Ca concentrationrange(100--1000mg
L-1) shouldbe avoided,and if possiblethe samplediluted, if other divalent met-
als are to be analyzedin the samesolution.
ElectrothermalAtomic Absorption
Introduction
Electrothermalatomic absorptionis basedon the sameprinciples as AAS,
exceptthat a heatedatomizeror graphitefurnace is used in place of the burner
assemblyto atomizethe sampleand energizethe atomsto a higher energystate.
Sensitivitiesand detectionlimits are 20 to 1000 times betterthan thoseobtained
with the conventionalflame method.The increasein sensitivity stemsfrom an
increasein the residencetime of the atomsin the optical path and an increasein
the percentageof atomsin the ground stateavailableto be energized.Since the
sampleis injectedinto a tube ratherthan aspiratedcontinuously,only small vol-
umes are needed.Generally the procedureis used only if concentrationsare
below the operatingrangeof AAS, as more interferencesexist and samplepro-
ducibility is generally lower. See Chapter4 (Wright & Stuczynski, 1996) for
generaloperatingprocedures.
Barium
It is recommendedthat pyrolytically coated graphite tubes be used to
increasesensitivity. The optimum analytical range is regardedas 10 to 200 f.lg
L -1 with a detectionlimit of 2 f.lg L -1.
Beryllium
It is recommendedthat pyrolytically coatedtubesbe usedto increasesen-
sitivity. The recommendedmethod (Ellis et aI., 1988) is as follows. Dry at
200°C, rampfor 5 s and holdfor 25 s. Ash at 900°C, ramp for 5 s and hold for
25 s. Atomize at 2700°Cand hold for 4 s. During the atomizestep,the Ar carri-
er gas flow is interrupted.Clean at 2700°C, ramp for 1 s and hold for 1 s. Cool
at 50°C, ramp for 1 s and hold for 30 s before injection of next sample.
FlameEmissionPhotometry
Strontium
Flame emissionis less sensitivethan AAS for Sr analysis(about 0.2 mg
L- 1 detection limit), and below the useful range for analysis of most waters.
Samplesmay requirereconcentrationby acidifying with HN03 ($;0.8 mL per 100
mL of sample)and evaporation(HOff, 1959). Emissionis measuredat 460.7 nm
and backgroundintensity is measuredat 466 nm. Strontiumemissionis taken as
the differenceof thesetwo readings.Emissionis a linear function of concentra-
tion. Use of a fuel-rich, nitrous oxide-acetyleneflame is recommended.The
working concentrationrangeis 1 to 100 mg L -1.
Ion Chromatography
Introduction
Ion chromatographyis becoming increasingly popular as an analytical
methoddue to its versatility, and ability to sequentiallydeterminemultiple ions
on the samesampleby chromatographicseparation.Commonly a conductivity
meter is usedas the detector,which is connecteddirectly to the separationcol-
umn. Usually thesystemconsistsof a guardcolumn,suppressorcolumnandsep-
aration column. Details on the principles and operatingmethodsare given in
Chapter8 (Tabatabai& Frankenberger,1996).
Single Column Method

Apparatus.A high-performanceion chromatographwith a conductivity
detectoris required(Gjerde& Fritz, 1987).
Cation ExchangeColumn
Surface-Sulfonated
Reagents.2.00-mMethylenediammonium
tartrateadjustedto pH 4.5.
Procedure.The column consistsof a surface-sulfonated cation exchange
column of low exchangecapacity. Use of a complexingligand reducesthe ion
chargeand allows for more rapid and better chromatographic separationof diva-
lent cations.Magnesium,Ca and Sr are readily separatedusing 2.00 mM ethyl-
enediammoniumtartrateat pH 4.5 as the eluent. Interferencesby large concen-
trations of other divalent ions, such as Fe2+ can be avoided by addition of an
additionalcomplexingagentwhich is preferentiallycomplexedwith the interfer-
ing ion. In the caseof Fe2+, interferencewith Mg, addition of EDTA (0.01 M)
complexesFe2+ but not Ca, Mg, or Sr at pH 4.5. Elution times of about 12 min
are requiredfor theseions.
Other methodsinclude the use of postcolumnreactorsand use of spec-
trophotometricdetection. Zenki (1981) describeda Mg, Ca, Sr, Ba analytical
methodusing 0.7 M sulfosalicylic acid as the eluentin the presenceof 5 x 10-5
M chlorophosphonazo III as the color-forming reagentall at pH 7.5. The column
was connectedto a flow-through cell having a light path of 8 mm and an inter-
nal volume of 8 ilL. Detectionlimits for Mg, Ca, Sr and Ba are 50, 2, 6, and 20
ng respectively(6.2, 0.25, 0.75,and 2.5 mg L-l).
SuppressedGradientMethod
Apparatus
1. Ion chromatographic system [Dionex 4500i (Dionex Corp.,
Sunnyvale,CA); or equivalent] with gradientpump micromembrane
suppressor(Dionex CMMS-II) conductivity detector, autoregenera-
tion accessary.
2. Ion PacCS10cation exchangecolumn (Dionex).
3. Ion PacCG10 guardcolumn.
4. Cation trapcolumn, CTC-I.
592 SUAREZ
Reagents
1. 40 mM HCl analyticalgrade.
2. Eluent A 0.1 M tetrabutylammoniumhydroxide (TBAOH) analytical
grade.
3. Eluent B 40 mM HCI-20 mM 2,3-diaminoproprionicacid monohy-
drochloride.
4. Ultrapurewater (>20 M a /cm resistivity).
Procedure
1. Continuouslyregeneratesuppressorwith TBAOH solution at a flow
rate of 10 mUmin.
2. Eluent flow rate of 1.0 mL.min.
3. Initial systemusing Eluent A.
4. Inject sample(50-J.lL loop).
5. After 3 min, start gradientramp with introductionof Eluent B.
6. After 13 min, end gradientramp (should be 40% eluent A and 60%
eluentB). Maintain eluentdistribution until 39 min.
7. After 39 min end isocraticelution (return to 100% of EluentA).
8. After 39.5-minend run.
9. Equilibrate systemuntil 55 min, and then load newsample(total run
time 55 min).
Comments.The reported coefficient of variation is less than 4%. The
detectionlimits for Mg, Ca, Sr and Ba are respectively0.050, 0.050, 0.040 and
0.40 mgL-1 (seeDabek-Zlotorzynska& Dlouhy, 1993).
Magnesium,Calcium, StrontiumSuppressed
Ion Chromatography
Apparatus
1. Ion chromatographicsystem (Dionex4000i, or equivalent).
2. Ion PacCS12separatorcolumn (Dionex).
3. Ion PacCS guardcolumn (Dionex).
4. Cation self-regeneratingsuppressor.
5. Conductimetric detector.
Reagents
1. 20 mmol L-1 methanesulphonic acid reagentgrade(prepareddaily).
2. 100 mmol L -1 HCI reagentgrade.
3. Ultrapurewater(>20 M a/cm resistivity).
Procedure
1. NeutralizeHC03 and C03 with HCI as needed.
2. Use column flow rate of 1.0 mUmin.
3. adjustdetectoroutput range0.1 J.lS to 10 J.lS.
4. Inject 25 J.lL of sample.
5. Inject next sampleafter 15 min or after baselineis re-establishedafter
Sr peak.
Comments.New CS12 column offers faster separationthan older CSlO

columns. Method is considerablyfaster than method listed under "Procedure"
above,but Ba separationis not presentedfor the presentmethod.Method is well
suited for Ca and Mg concentrationrange of most waters.Strontium detection
limit <0.02 mg L-l. Method also can determineLi, Na, NH4, and K on the same
sample(seeGros & Gorenc,1994).
Titrimetric
Introduction
Before the developmentof AAS and ICP, titrimetric procedureswere the
methodsof choice for Ca and Mg analyses.The methodsare preciseand usual-
ly do not haveseriousinterferences.Undersomeconditions,suchas the presence
of large amountsof Fe or Al, additional maskingreagentsmay be required. In
some instancesit is necessaryto remove excessFe or Al by precipitationas a
hydroxide. Abnormally large amountsof phosphatemay require a separationor
removal of phosphatebefore the titration. Also increasingor decreasingthe
amountof indicator may aid in determiningthe end-point.
The Ca end-pointis difficult to detectwhen Mg > Ca. This difficulty is due
to the absorptionof the Ca color indicator onto the Mg(OH)z particles.A pro-
tective colloid such as Carbocelmay be addedto prevent the absorptionof the
dye (van Schouwenburg,1960). Also, the color changeat the end-pointcan be
more easily seenif addition of most of the EDTA is madebeforethe pH is raised
(Lewis & Melnick, 1960).
Interferencewith Sr and Ba rarely occurs as thesemetals form relatively
weak complexeswith EDTA. Interferencesare most likely for analysesof total
soil. The proceduresto overcometheseinterferencesare difficult. Barium inter-
ference in total soil analysis may be overcomeby use of the HF and H2S04
digestionrather than the Na2C03fusion, as in the former, Ba remainsinsoluble
as BaS04,while in the latter BaC03 is dissolvedupon addition of acid. Use of
the HF and H2S04 digestion requiresthat care be taken not to precipitategyp-
sum.
Calcium and MagnesiumEthylenediaminetetraacetic

Acid Method
Introduction. Calcium and Mg can be determinedseparately,or in total,

by measuringthe concentrationof the metal-EDTA complex. Interfering ions
include Fe, Mn, Cu, Zn, Ni, and phosphate,howevertheserarely occur at con-
centrations which interfere, except in the samples for total soil analysis.
Hydroxylamine-hydrochlorideis usedto reducethe Fe to Fe2+ and Al and Fe2+
are chelatedwith triethanolamine.The simplest way to compensatefor phos-
phateinterferenceis to add an excessof EDTA, adjust the pH slowly and back-
titrate with a known Ca standard.Alternatively, (i) phosphatecan be precipitat-
ed as zirconium phosphateby addition of zirconium nitrate, or, (ii) phosphate
ions can be absorbedon an anion exchangeresin (Brunisholzet aI., 1953).
Severalindicatorscan be used.The sum of Ca and Mg can be determined
by titration at pH 10 using EriochromeBlack T (EBT) as an indicator. Alterna-
594 SUAREZ
tively Calmagiteproducesthe samecolor changebut hasa sharperend-point.At

pH 12 or higher, if no N14 saltsare present,Mg is precipitatedas Mg(OHh and
Ca can be titrated using Calcon indicator. Magnesiumcan be analyzedusing
EBT indicator after first precipitating Ca by addition of tungstate(Shapiro &
Brannock,1955)The following is adaptedfrom Lanyon and Heald (1982).
Reagents
1. Buffer solution. Dissolve 67.5 g of ammoniumchloride (NH4CI) in
200 mL of deionizedwater. Add 570 mL of concentratedammonium
hydroxide (Nl40H), and dilute the solution to a volume of 1 L with
deionizedwater.
2. Ethylenediaminetetraacetic acid disodiumsalt (EDTA disodium)solu-
tion. Dissolve 2.00 g of EDTA disodium (molecular weight =
372.254)and 0.039 g of magnesiumchloride hexahydrate(MgCI2 •
6H20) in deionizedwater, and dilute the solution to a volume of 1 L.
StoreEDTA reagentin polyethylenebottles.
3. StandardCa solution. Dissolve 0.5004 g of "low Mg" reagent-grade
calcium carbonate(CaC03), previously dried at 150°C, into 5 mL of
approximately6 M hydrochloricacid (HCI), and dilute the solution to
a volume of 1 L. This solution is 500 ~ Ca.
4. StandardMg solution. Dissolve 0.1216 g of reagent-gradeMg metal
in dilute HCL, and dilute the solution to a volume of 1 L. This solu-
tion is 500 ~ Mg.
5. Potassiumcyanide(KCN) solution. Dissolve 1 g of KCN in 100 mL
of deionizedwater.
6. Hydroxylamine-hydrochloride(NH20H • HCI) solution. Dissolve5 g
of NH20H • HCl in 100 mL of deionizedwater.
7. Potassium ferrocyanide [~Fe(CN)6] solution. Dissolve 4 g of
reagent-gradepotassiumferrocyanidetrihydrate [~Fe(CN)6 • 3H20]
in 100 mL of deionizedwater.
8. Triethanolamine(TEA), reagentgrade.
9. Tungstate solution. Dissolve 20 g of sodium tungstate dihydrate
(Na2W04 • 2H20) in 100 mL of deionizedwater.
10. EriochromeBlack (EBl) indicator. Dissolve0.2 g of EBT in 50 mL of
methanol.Preparea fresh solution every 3 wk.
11. Calcon indicator. Dissolve 20 mg of Calcon in 50 mL of methanol.
Preparea fresh solution weekly.
12. Sodium hydroxide (NaOH), 10% solution. Dissolve 10 g of reagent-
gradeNaOH in 90 mL of deionizedwater.
Procedure-Calibration. Standardizationof EDTA. Add 5 mL of standard

0.5 mM Ca solution into several 250-mL beakers,and dilute with deionized
waterto approximately150 mL. Add 15 mL of buffer solution,and 0.5 mL each
of KCN, NH20H-HCl, ~Fe(CN)6' TEA, and EBT (or Calgamite) indicator.
Titrate this solution with the disodiumEDTA solution to a permanentblue color
using a microburetteand undernaturalor fluorescentlighting. Add the last few
drops at 3- to 5-s intervalsto allow for color development.The titration should
be performedwithin 5 min after addition of the buffer. Repeatthe titration with

5-mL aliquots of the standardMg solution. Thesetitrations are to determineif
titratablecontaminantsare presentin the standards.
Standardizethe disodiumEDTA solution again, onlynow usingthe Calcon
indicator instead of the EBT indicator. Repeatthe procedureabove, only this
time also add 1 mL of 10% NaOH, or enoughto raise the pH to 12 or slightly
above.Add 0.25 mL of Calconindicator and titrate from a red to permanentblue
color as above. All burette readings should agree within 0.05 mL.
Standardizationshouldbe madefor every setof new samples.Titration of deion-
ized water blanks servesto recheckthe standardizationof the EDTA and pro-
vides a checkon glasswarecontamination.
Calcium and MagnesiumProcedure.Add an aliquot containing 1 to 2
mg of Ca plus Mg into a 250-mL beakerand dilute to approximately150 mL
with deionizedwater. Add 15 mL of buffer solution and 0.5 mL eachof KCN,
NH 20H-HCI, ~Fe(CN)6 and TEA. Allow a few minutes for the reactionsto
take place. Warming the solution speedsup the process.Add 0.5 mL of EBT or
Calmagiteindicatorand titrate the solution as beforeto the samepermanentblue
color.
An excessof phosphatemay make it difficult to determinethe end-point.
Phosphateinterferenceis overcomeby modifying the procedureas follows. To a
new aliquot, add 0.5 mL eachof KCN, NH 20H-HCI, ~Fe(CN)6 and TEA. Add
a known but excessamountof disodium EDTA. Bring the solution to pH 10 by
adding 15 mL, or more if required,of the buffer solution. Heat the solution to
near boiling for several minutes to speedthe reaction.After the solution has
cooled, add 0.5 mL of EBT indicator and titrate from a blue to a permanentred
color with the standardCa solution.
Calcium Procedure.Place an aliquot containing 1 to 2 mg of Ca into a

250-mL beakerand dilute to approximately150 mL with deionizedwater. Add
0.5 mL eachof KCN, NH 20H-HCl, ~Fe(CN)6' TEA, and sufficient 10% NaOH
(usually about 1 mL) to raise the pH to 12 or slightly above. Add 0.25 mL of
Calcon indicator and titrate from red to permanentblue with the standardized
EDTA solution.
An excessof phosphatemay make it difficult to determinethe end-point.
This interferencecan be overcomeby modifying the procedureas follows. To a
new aliquot, add 0.5 mL eachof KCN, NH 20H-HCI, ~Fe(CN)6 and TEA. Add
a known but excessamountof disodium EDTA. Bring the solution to pH 12 by
adding NaOH. Heat the solution to near boiling for severalminutes. After the
solution has cooled, add 0.25 mL of Calcon indicator and titrate from blue to a
permanentred color with the standardCa solution.
MagnesiumProcedure.Placean aliquot containing2 to 4 mg of Mg into
a 250-mL beakerand dilute to approximately100 mL with deionizedwater. Add
20 mL of tungstatesolution and enoughbuffer solution to obtain a pH of 10, this
usually requiresabout 20 mL. Heat the solution and filter through Whatmanno.
42 filter paper.Wash the paperand precipitatewith a solution containing50 mL
of buffer solution per liter. To the filtrate add 0.5 mL eachof KCN, NH20H-HCI,
596 SUAREZ
~Fe(CN)6 and TEA. Allow a few minutesfor the reactionsto take place. Add
0.5 mL of EBT indicatorand titrate the solutionas beforeto the samepermanent
blue color. The Mg end-pointis slow to develop.Heatingthe solution will speed
the developmentof the end-point.
Permanganate
Method
Calcium is precipitatedquantitativelyas calcium oxalate.The precipitated
calcium oxalateis dissolvedin acid and titrated with permanganate. An excess
of oxalateovercomesthe adverseeffectsof magnesium.InterferencesincludeSr,
Si, AI, Fe, Mn, phosphate,and suspendedmatter. The procedureis not recom-
mendedfor total soil analyses.Interferencesof AI, Fe, and Mn, can be eliminat-
ed with precipitationby ammoniumhydroxide after treatmentwith persulfate.
Phosphateis precipitatedasthe ferric salt andsuspendedmatteris avoidedby fil-
tration. This methodis adoptedfrom Clesceriet al. (1989).
Apparatus
1. Methyl red indicator. Dissolve0.1 g methyl red sodiumsalt and dilute
to 100 mL with deionizedwater.
2. Hydrochloric acid, diluted 1:1 with deionizedwater.
Procedure
Use 200 mL of sample,containinglessthan 50 mg of Ca. removeinterfer-
encesas follows:
1. Silica is removedby the gravimetricmethod.Add 5 mL of 1:1 diluted
HCl to samplein a 200-mL platinum dish and evaporateby placing in
a hot water bath. During evaporationadd an additional 10 mL of the
1:1 HCl in severalsteps.Dry residuein a 110°Coven. Next add 5 mL
of warm 1:1 HCVDI then 50 mL of hot DI water and filter through a
medium texturefilter paper.Washdish and residuewith 1:50 HCVDI
and then with DI water(passthroughfilter paper).Collect the filtered
solution for analysis.{See section4500-Si in [Clesceri et a1. (1989)]
for additionaldetails.}
2. Remove oxide interferencesby concentratingfiltrate to 120 to 150
mL. Add sufficient HCl so that the filtrate containsat least 10 mL of
concentratedHC1. Add 0.1 mL of methyl red indicator. Add 3 M
NH40H until the solution turns yellow. Add 1 g of ammoniumper-
sulfateand heat.When boiling beginscarefully add 3 M NH40H until
the solution becomesslightly alkaline, as determinedby litmus paper
test. Boil for 1 to 2 min and let standfor 10 min until the oxidescoag-
ulate (not longer). Filter the precipitateand wash 3 to 4 times with
~CI solution. Savefiltrate for analysisgiven below.
pH adjustment
Add 0.1 to 0.15 mL of methyl red indicator. Neutralizewith HCl and boil
for 1 min. Add 50 mL of ~C204 • H20 solution and if any precipitateforms,
addjust enoughHCl to redissolve.
Precipitationof Calcium Oxalate

Keeping the solution just below boiling, add 3 M NH40H in drops from a
buret, stirring constantly.Continueadding until solution is turbid (about 5 mL).
Digest for 90 min at 90°C. Filter under suction with a filter crucible. Wash
immediatelywith NH40H diluted to 1% by volume. It is not necessaryto trans-
fer all of the precipitateto the filter crucible, but removeall the excessNH4C20 4
• H20 from the beaker.
Titration
Placefilter crucible on its side in beakerand cover with deionizedwater.
Add H2S04 and while stirring, heat rapidly to 90 to 95°C. Titrate rapidly with
KMn04 to a slightly pink end-pointthat remainsfor at least 1 min. Do not let the
temperaturedrop below 85°C. It may be necessaryto heatthe beakerduring the
titration. Agitate the crucible sufficiently to insure reactionof all the oxalatepre-
cipitate.
Colorimetric
Introduction
Although these were the methodsof choice in the past they have been
replacedby instrumentalmethods.In the absenceof theseinstrumentsthesecol-
orimetric methodsare still suitable.
Beryllium Aluminon Method

The useof aluminonfor Be analysisrequiresthat EDTA be addedto avoid
interferencefrom Al (also Co, Cu, Fe, Mn, Ni, Ti, Zn, and Zr). Samplesshould
be acidified to pH 2 with nitric acid to avoid absorptionon the samplevessel
walls. This procedureis adoptedfrom Clesceriet al. (1989) with a reduction in
sample and reagent volumes. Hazardous chemicals include Be salts and
methanol.
Apparatus.A spectrophotometer with a light path of 5 cm is recommend-
ed, with a wavelengthsettingof 515 nm. Alternatively a filter photometercan be
used if equippedwith a green filter with maximum transmittancenear 515 nm
and a 5-cm light path.
Reagents--Ethylenediaminetetraacetic Acid Reagent. 2.5 g of EDTA into
a 100-mL flask. Add 30 mL of deionizedwater and a drop of methyl red (pre-
paredby adding 5 mg of methyl red to 10 mL of ethanol).Neutralizethe solu-
tion by addingammoniumhydroxide and adddeionizedwater for a total of vol-
ume of 100 mL.
Aluminon Reagent. Add 250 g of ammoniumacetateand 500 mL of deion-
ized water to a l-L beaker.Add 40 mL of concentratedacetic acid and stir until
dissolved.Dissolve0.5 g of aluminon(aurintricarboxylicacid triammoniumsalt)
in 25 mL of deionizedwater and add to the l-L beakersolution. Next dissolve
1.5 g of benzoicacid (C7H60 2) in 10 mL of methanol,add to l-L beakersolu-
tion while stirring and dilute to 1 L with deionizedwater. Add 5 g of gelatin and
598 SUAREZ
125 mL of deionizedwater to a 200-mL beaker.Dissolve gelatinby heatingthe

200-mL beakersolution in a boiling water bath. heat and stir until the gelatin is
dissolved.Next add 250 mL of deionizedwaterto a 500-mLvolumetricflask and
add the warm gelatin solution. After cooling to room temperature,dilute to 500
mL with deionizedwater. Add and mix the aluminon solution and the gelatin
solutionsinto a 2-L dark glassbottle (chemicallyresistantglass).Storein a cool
dark place.The useful life of the solution should be at leastone month.
Procedure.Pipet up to 30 mL of sample(containingless than 100 Ilg of
Be) into a 50-mL volumetric. Add 1 mL of EDTA solutionanddilute with deion-
ized water up to 40 mL. Next add 7.5 mL of aluminon reagent,dilute to 50 mL
total volume, mix, and store in dark for 20 min. Read in spectrophotometerat
515 nm using 5-cm cells.
REFERENCES
Anderson, A, P.M. Bertsch, and w.P. Miller. 1990. Beryllium in selectedsoutheasternsoils. J.
Environ. Qual. 19:347-348.
Amrhein, c., and D.L. Suarez.1990. Procedurefor determiningsodium-calciumselectivity in cal-
careousand gypsiferoussoils. Soil Sci. Soc. Am. J. 54:999-1007.
Baes,C.E, and RE. Mesmer. 1976. The hydrolysisof cations.JohnWiley & Sons,New York.
Barber, S.A. 1984. Soil nutrient bioavailability: A mechanisticapproach.p. 262-277. In Calcium.
John Wiley & Sons,New York.
Barshad,I. 1954a.Cation exchangein micaceousminerals:I. Replaceabilityof the interlayercations
of vermiculite with ammoniumand potassiumions. Soil Sci. 77:463-472.
Barshad,I. 1954b. Cation exchangein micaceousminerals:II. Replaceabilityof ammonium and
potassiumfrom vermiculite, biotite and montmorillonite.Soil Sci. 78:57-76.
Bolt, G.H., M.G.M. Bruggenwertand A Kamphors!.1976. Adsorption of cationsby soil. p. 54-90.
In G.H. Bolt and M.G. M. Bruggenwert(ed.) Soil chemistry A Basic elements.Elsevier
North-Holland, Inc., New York.
Bowen, H.J.M., and J.A Dymond. 1956. Strontium and barium in plants and soils. Proc. R Soc.
London, Bl44:355-368.
Bower, C.A, RE Reitemeier,and M. Fireman. 1952. Exchangeablecation analysisof saline and
alkali soils. Soil Sci. 73:251-261.
Brass,G.W. 1976.The variation of the marine87Sr/86Sr ratio during Phanerozoictime: Interpretation
using a flux model. Geochim.Cosmochim.Acta 40:721-730.
Brunisholz,G., J. Genton,and E. Plattner.1953.Sur Ie dosagecomplexometriquedu calciumen pres-
encede magnesiumet de phosphate.Helv. Chim. Acta 36:782-783.
Burns, R.G., and Y.M. Burns. 1974. Magnesium. p. 12-Al-12-A17. In K.H. Wedepahl (ed.)
Handbookof geochemistryII-I. Springer-Verlag,New York.
Carter, M.R, G.R. Webster,and R.R. Cairns. 1979. Calcium deficiency in some Solonetricsoils of
Alberta. J. Soil Sci. 30:161-174.
Carter, M.R, and G.R. Webster. 1990. Use of the calcium-to-total cation ratio in soil saturation
extractsas an index of plant-availablecalcium. Soil Sci. 149:212-217.
Clesceri,L.S., AE. Greenberg,RR. Trussell (ed.). 1989. Standardmethodsfor the examinationof
water and wastewater.17th ed. Public Health Assoc. Washington,DC.
Cotton, EA, and G. Wilkinson. 1980. Advanced inorganic chemistry.John Wiley & Sons, New
York.
Couture, R.A, M.S. Smith, and R.E Dymek. 1993. X-ray fluorescenceanalysisof silicate rocks
using fused glass discs and a side-window Rh sourcetube: Accuracy, precision and repro-
ducibility. Chern. Geol. 110: 315-328.
Dekeb-Zlotorzynska,E., and J.E Dlouhy. 1993. Simultaneousdeterminationof alkali, alkaline-earth
metal cationsand ammoniumin environmentalsamplesby gradiention chromatography.J.
Chroma!. 638:35-41.
Ebdon,L., and A.R. Collier. 1988. Particlesize effectsof kaolin slurry analysisby inductively cou-
pled plasma-atomicemissionspectrometry.Spectrochim.Acta 43B:355-369.
Elkhatib, E.A., J.L. Hean, and T.E. Staley. 1987. A rapid centrifugationmethod for obtaining soil
Ellis, WO., V.F. Hodge,DA Darby, C.L. Jones,and TA Hinners. 1988. Determinationof berylli-
um in liquid and solid wastesamplesby flame and fumaceAAS. At. Spectrosc.9:181-188.
Faure,O. 1978. Strontium. p. 38-EI-E19.lnKH. Wedepahl(ed.) Handbookof geochemistryII-4,
Springer-Verlag,New York.
Fendorf, S.E., and D.L. Sparks.1996. X-ray absorptionfine structurespectroscopy.p. 377-415.In
D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.SSSABook Ser.
5. SSSAand ASA, Madison,WI.
Fisher,T.R. 1963. Measurementsof adsorbedcationsin soils employing radioactiveisotopeequili-
bration methods.Ph.D. diss. Univ. of Missouri, Columbia(Diss. Abstr. 23:2284).
Fishman,M.1., and L.c. Friedman.1985. Methodsfor the determinationof inorganic substancesin
water and fluvial sediments.p. 1-709.In Techniquesof water-resourcesinvestigations.U.S.
Geol. Surv. Book 5. Open File Rep. 85-495. U.S. Geol. Sur., Denver, CO.
Fuller, C.W., R.C. Hutton, and B. Preston.1981. Comparisonof flame, electrothermal,and induc-
tively coupled plasma atomisation techniquesfor the direct analysis of slurries. Analyst
106:913-920.
Garrels,R.M., and E.A. Perry. 1974. Cycling of carbon,sulfur, and oxygenthroughgeologictime. p.
303-336. In E.D. Goldberg(ed.) The sea.Vol. 5. Wiley-Intersci.,New York.
Oill, R. 1993. AAS or ICP-OES:Are they competingtechniques?Am. Lab. p. 24F-24K
Gilman, L.B., and W.G. Engelhart. 1989. Recent advancesin microwave sample preparation.
Spectrosc.Int. 2:14-21.
Gjerde,D.T., and J.S.Fritz. 1987. Ion chromatography.2nd ed. Dr. Alfred Huthig Verlag, New York.
Graham,E.R., S. Powell, and M. Carter. 1956. Soil magnesiumand the growth and chemicalcom-
position of plants.Missouri Agric. Exp. Stn. Res. Bull. 607.
Oros, N., and B. Gorenc. 1994.Ion chromatographicdeterminationof alkali and alkaline earth met-
als in mineral waters.Chromatographia39:448-452.
Hansen,E.H., and D.N. Munns. 1988. Effect of CaS04and NaCI on mineral contentof Leucaena
leucocephala. Plant Soil 107:101-105.
Harvey, P.K, and B.P. Atkin. 1982. Automaedx-ray fluorescenceanalysis.p. 17-26. In Sampling
and analysisfor the mining industry. Inst. Mining Metall., London.
Holder, M., KW Brown, J.C. Thomas,D. Zabcik, and H.E. Murray. 1991. Capillary-wick unsatu-
ratedzone pore water sampler.Soil Sci. Soc. Am. J. 55:1195-1202.
Hormann,P.K. 1969. Beryllium. p. 4-DI-4-07. In K.H. Wedepahl(ed.) Handbookof geochemistry
II-I. Springer-Verlag,New York.
Horr, c.A. 1959.A surveyof analytical methodsfor the determinationof strontium in naturalwaters.
U.S. Geol. Surv. Water Supply Pap 1496A. U.S. Gov. Print. Office, Washington,DC.
Hossner,L.R. 1996. Dissolution for total elementalanalysis. p. 4~4. In D.L. Sparkset al. (ed.)
Madison,WI.
Howard,D.D., and F. Adams.1965. Calcium requirements for penetration ofsubsoilsby primary cot-
ton roots. Soil Sci. Soc. Am. Proc. 29:558-562.
Isaac,R.A., and J.D. Kerber. 1971. Atomic absorptionand flame photometry:Techniquesand uses
in soil, plant, and water analysis. p. 17-37. In L.M. Walsh (ed.) Instrumentalmethodsfor
analysisof soils and plant tissue.SSSA, Madison,WI.
Jackson,M.L. 1958. Soil chemicalanalysis.Prentice-Hall,Inc. EnglewoodCliffs, NJ.
Karathanasis,A.D., and B.F. Hajek. 1996. Elementalanalysisby x-ray fluorescencespectroscopy.p.
SSSA Book Ser. 5. SSSAand ASA, Madison,WI.
K1esta,E.1., and J.K. Bartz. 1996. Quality assuranceand quality control. p. 19-48.In D.L. Sparkset
al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.SSSA Book Ser. 5. SSSA and
ASA, Madison,WI.
Lanyon, L.E., and WR. Heald. 1982. Magnesium,calcium, strontium, and barium. p. 247-261.In
A.L. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA,Madison,WI.
Lewis, L.L., and L.M. Melnick. 1960. Determinationof calcium and magnesiumwith (ethylenedini-
trito) tetraaceticacid. Anal. Chem. 32:1123-1127.
Litaor, M.I. 1988. Review of soil solution samplers.Water Resour.Res.24:727-723.
600 SUAREZ
Marschner,H. 1986. Mineral nutrition of higher plants.Acad. Press,New York.

McLaren,J.w. 1987.Applications: Environmental.p. 48-64.In P.W.J.M.Boumans(ed.) Inductively
coupledplasmaemissionspectroscopy.Part II. JohnWiley & Sons,New York.
Medlin, J.H., N.H. Suhr, and J.B. Bodkin. 1969. Atomic absorptionanalysisof silicatesemploying
LiB0 2 fusion. At. Absorp. Newslett. 8(2):25-29.
Moenke-Blankenburg,L., and D. Gunther.1992.Lasermicroanalysisof geologicalsamplesby atom-
ic emissionspectrometry(LM-AES) and inductively coupledplasma-atomicemissionspec-
trometry (LM-ICP-AES). Chern. Geol. 95:85-92.
Mokwuyne, A.v., and s.w. Melsted. 1972. Magnesiumforms in selectedtemperateand tropical
soils. Soil Sci. Soc. Am. Proc. 36:762-764.
Mulder, E.G. 1956. Nitrogen-magnesiumrelationshipsin crop plants. Plant Soil 7:341-376.
Mukhtar, S., and S.J. Haswell. 1991. Application of total-reflectionx-ray fluorescencespectrometry
to elementaldeterminationsin water, soil, and sewagesludgesamples.Analyst 116:333-338.
Nham, T.T. 1995. Water analysis using ICP-AES with an ultrasonic nebulizer. Am. Lab. Feb.
48L-48V.
Plaut, Z., and C.M. Grieve. 1988.Photosynthesis of salt-stressed
maizeas influencedby Ca:Naratios
in the nutrient solution. Plant Soil 105:283-286.
Potts,P.J. 1987. A handbookof silicate rock analysis.Chapmanand Hall. New York.
Puchelt,H. 1972.Barium 56-D. p. 56-D-56-D18.ln K.H. Wedepahl(ed.) Handbookof geochemistry
11-4. Springer-Verlag,New York.
Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSA Book Ser. 5.
Robinson,W.O., R.R. Whetstone,and G. Edgington.The occurenceof barium in soils and plants.
USDA Tech.Bull. 1013. u.S. Gov. Print. Office, Washington,DC.
Robles,L.C., e. Garcia-Olalla,M.T. A1emany,and A.S. Aller. 1991. Flameatomic absorptionspec-
trometricmethodfor the determinationof beryllium in naturalwatersafter separationwith N-
Benzoyl-N-phenylhydroxylamine. Analyst 116:735-737.
Schachtschebal, von P. 1954.Dasphanzenverfugbare MagnesiumdesBodenund seineBestimmung.
Z. Phflanzenemaehr. Bodenkd.67:9-23.
Shacklette,H.T., J.e. Hamilton, J.G. Boergen,and 1.M. Bowles. 1971. Elementalcompositionof
superficial materialsin the conterminousUnited States.U.S. Geol. Surv. Prof. Pap. 574-D.
U.S. Gov. Print. Office, Washington,DC.
Shapiro,L., and W.W. Brannock.1955.Automatic photometrictitrations of calcium andmagnesium
in carbonaterocks. Anal. Chern. 27:725-728.
Sharp, B.L. 1987. Applications:Agriculture and food. p. 65-99. In P.W.J.M. Boumans (ed.)
Inductively coupledplasmaemissionspectroscopy.Part II. JohnWiley & Sons,New York.
SiIlen, L.G., and A.E. Martell. 1971. Stability constantsof metal-ion complexes.Suppl. 1. Spec.
Publ. 25. Chern. Soc., Burlington House,London.
Soltanpour,P.N., G.W. Johnson,S.M. Workman,J.B. Jones,Jr., and R.O. Miller. 1996. Inductively
coupledplasmaemissionspectrometryand inductively coupledplasma-mass spectrometry.p.
91-139.In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part3. Chemicalmethods.SSSA
Book Ser. 5. SSSAand ASA, Madison,WI.
Tabatabai,M. and W. Frankenberger.1996. p. 225-245.Liquid chromatography.In D.L. Sparkset
al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and
ASA, Madison,WI.
Thill, J.L., and1.R. George.1975. Cation concentrationsand K to Ca + Mg ratio of nine cool-season
grassesand implicationswith hypomagnesaemia. Agron. 1. 67:89-91.
Thompson,M., and M.H. Ramsey.1985. Matrix effects due to calcium in inductively coupledplas-
ma atomic-emissionspectrometry:Their nature,sourceand remedy.Analyst 110:1413-1422.
Turekian, K.K., and K.H. Wedehohl.1961. Distribution of the elementsin some major units of the
earth'scrust. Geol. Soc. Am. Bull. 72:175-192.
Ure, A.M. 1991.Atomic absorptionand flame emissionspectrometry.p. 1-62. In K. Smith (ed.) Soil
analysis-modeminstrumentaltechniques.2nd ed. Marcel Decker, New York.
van Schouwenburg,J.C. 1960. Micro-EDTA titration of calcium: Magnesiuminterference.Anal.
Chern. 32:709-710.
Welte, E., W. Werner, and E.A. Niederbudde.1960. Zur frage der Magnesium-Dynamiqueim
Boden.p. 246-252.
West,A.C., and W.D. Cooke. 1960. Elimination of anion interferencein flame spectroscopy.Use of
ethylenedinitrilotetraaceticacid. Anal. Chern. 32:1471-1474.
Wolt, J.D., and F. Adams. 1979. Critical levels of soil and nutrient-solutioncalcium for vegetative
growth and fruit developmentof Florunnerpeanuts.Soil Sci. Soc. Am. J. 43:1159-1164.
Wright, R.J.,andT. Stuczynski.1996.Atomic absorptionandflame emissionspectrometry.p. 65-90.
In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.SSSABook
Ser. 5. SSSAand ASA, Madison,Wi.
Zenki, M. 1981. Determinationof alkaline earthmetalsby ion-exchangechromatographywith spec-
trophotometricdetection.Anal. Chern.53:968-971.
Published 1996
Chapter 21
Boron
R. KEREN, Institute of Soils and Water, The Volcani Center,

Agricultural Research Organization, Bet Dagan, Israel
Boron is one of the eight essentialmicronutrientsrequiredfor the normal growth

of most plants. Boron hasa markedeffect on plants,from the standpointof both
plant nutrition-if B is deficient in soil, and toxicity-if it is presentin excessive
amounts.A relatively small soil solution concentrationrangedefine B deficien-
cy and toxicity. Boron deficiencyin plantsis more widespreadthan deficiencyof
other micronutrients.Boron deficiency is found primarily in humid regionsor in
sandysoils, while B toxicity occursmost frequently in soils of arid and semiarid
regions.Boron toxicity occursin theseareaseitherdue to high levelsof B in soils
or due to additionsof B in irrigation water.
Most of the total B in soils is found within the crystalline structuresof the
soil minerals(suchas tourmaline)and it rangesfrom 7 to 80 mg kg-1 soil (Kraus-
kopf, 1972).The total B content insoils, however,has little bearingon the status
of availableB to plants.
The thresholdB concentrationfor irrigation water (the maximumpermissi-
ble concentrationfor a given crop speciesthat does not reduceyield or lead to
injury symptoms)rangedfrom as low as 0.3 mg B L -1 for sensitivecrops to 2.0
mg B L-l for tolerantcrops. Thesecriteria, which were obtainedfrom sandcul-
ture experiments,have provided the basis for B toxicity criteria (U.S. Salinity
Lab. Staff, 1954). In assessingB concentrationin irrigation water, however,the
physical-chemicalcharacteristicsof the soil must be taken into consideration
becauseof the interaction betweenB and soil. Boron sorption and desorption
from soil adsorptionsitesregulatethe B concentrationin soil solution depending
on the changesin solution B concentrationand the affinity of soil for B. Thus,
adsorbedB may buffer fluctuations in solution B concentration,and B concen-
tration in soil solution may changeinsignificantly by changingthe soil watercon-
tent during water uptake by plantst. When irrigation with water high in B is
planned,specialattentionshouldbe paid to this interactionbecauseof the narrow
rangebetweenlevels causingdeficiency and toxicity symptomsin plants.
Book Seriesno. 5.
603
604 KEREN
PRINCIPLESAND CONCEPTSIN BORON CHEMISTRY
Boron Chemistryin AqueousMedia
Boron is consideredas a typical metalloid having propertiesintermediate

betweenthe metalsand the electronegativenonmetals.Boron has a tendencyto
form anionic ratherthan cationic complexes.
Boron chemistryis of covalentB compoundsand not of B3+ becauseof its
very high ionization potentials. For example,the rate of proton transferfrom
B(0H)3 to the excited acridine moleculeis 3.8 x lOS L mol-1 S-1 (Weller, 1957;
Leomahardt& Weller, 1960). All B compoundshave a valency basedon the
excited4p stateofB 1s22s12PxI2py I, and accordinglyare trivalent. The remaining
2p level is, from energyconsiderations,availablefor bonding.
The incompleteoctet in BX3 compoundscausesthem to behaveas accep-
tors (Lewis acids),in which B achievesits maximumcoordinationwith approxi-
mately s~ hybridization, toward many Lewis basis such as aminesand phos-
phines.Boron alsocopmletesits octetby forming both anionic and cationiccom-
plexes. Therefore, tricoordinate B compoundshave strong electron acceptor
propertiesand may form tetracoordinateB structures.The chargein tetracoordi-
natederivativesmay rangefrom negativeto positive, dependingupon the nature
of the ligands.
Boron oxide, Bz03' reactswith waterto form boric acid, B(OH)3' The crys-
tal structureof B(OHh is composedof distinct B(0H)3 moleculeslinked togeth-
er by H bonds to form infmite layers of approximately hexagonalsymmetry
(Zachariasen,1954). When heated,B(OH)3 loseswater stepwise.Above 373K,
the orthoacidgraduallyloseswater,changingto metaboricacid, HB02• Upon fur-
ther heating tetraboric acid (H2B40 7) is formed. At higher temperature,all the
water is lost, and anhydrousboric oxide (B20 3) results.Boric acid is moderately
soluble in water. Its solubility increasesmarkedly with temperaturedue to the
large negativeheatof solution.
For the unshared0 atoms bound to B, they are probably always OH
groups. Thus,Edwardset al. (1955) suggestedthat B(OH)3 forms an ion not by
splitting off W to form H2BOj, but by addition of OH- to form B(OH)4". Thus,
in accordance withthe electronconfigurationof B, B(0H)3 actsas a weak Lewis
acid
[1]
The formation of the hydroxyborateion is spontaneous. The first hydrolysiscon-

stant,Khb is 5.8 X 10-10 at 20°C (Owen, 1934),and the otherKh2 andKh3 values
are 5.0 x 10-13 and 5.0 x 10-14, respectively(Konopik & Leberl, 1949).A disso-
ciation beyondB(OH)4" is not necessaryto explain the experimentaldata,at least
below pH 13 (Ingri, 1963;Mesmeret aI., 1972).Boron speciesotherthanB(OH)3
and B(OH)4", however,can be ignoredin soils for most practicalpurposes.
The first hydrolysis constantsof B(OHh at various temperatures(in the
range of 273-328K) were given by Owen and King (1943), and are shown in
Table 21-1, togetherwith the ionic productsof water, K w , which are useful in
conjunctionwith Khl in certaincalculations.
BORON 60S
Table 21-1. Valuesof hydrolysisconstantof Boric acid, Khb and of ionic productof water,K 2, at sev-
eral temperatures.
Temperature(K) -logtoKhl -logtoKw
278 9.4382 14.7338
283 9.3805 14.5346
288 9.3278 14.3463
293 9.2799 14.1669
298 9.2365 13.9965
303 9.1975 13.8330
308 9.1626 13.6801
313 9.1316 13.5348
318 9.1043 13.3960
323 9.0806 13.2617
328 9.0603 13.1369
Resultsof nuclearmagneticresonance(Good & Ritter, 1962) and Raman

spectroscopy(Servoss& Clark, 1957) all lead to the conclusionthat B(OH)3 has
a trigonal planarstructure,whereasthe B(OH).:! in aqueoussolution has a tetra-
hedralstructure.This differencein structurecan lead to differencesin the affin-
ity of clay for thesetwo B species.As previously discussed,both B(OHh and
B(OH).:! speciesare essentiallymonomericin aqueousmedia at low B concen-
tration (~O.025 mol L-l). However,at high B concentration,polyborateions exist
in appreciableamount(Adams, 1964).
The B(OH).:! is expectedto form a variety of complex salts with suitable
metal acceptorions. Somesamplesof alkali and alkaline earthboratecomplex-
es togetherwith their apparentdissociationconstantsare given in Table 21-2.
Thesecationsare the most likely to show significant ion-pairing with B in sea-
water (Byrne & Kester, 1974).
Potassiumand Sr are not likely to have as strong an effect on B. The sta-
bility constantof the [KB(OH)4]O complex is consideredto be zero (Byrne &
Kester, 1974).
Table 21-2. Apparentdissociationconstantsfor alkali and alkaline earth borateion pairs at 298K.
Ion pair Ionic strength pK Referencet
[NaB (0H)4]0 0.68 -0.24 1

[NaB (OH)4]0 0.16 0.25 3
[NaB (OH)4]0 0.50 0.23 3
[CaB (OH)4j+ 0.68 1.11 1
[CaB (OH)4]+ 0.70 0.73 2
[CaB (OH)4j+ 0.02 1.79 3
[CaB (OH)4j+ 0.16 1.83 3
[MgB (OH)4]+ 0.68 0.90 1
[MgB (OH)4]+ 0.70 0.73 2
[MgB (OH)4]+ 0.02 1.63 3
[MgB (OH)4]+ 0.16 1.66 3
[SrB (OH)41' 0.07 1.54 3
[BaB (OH)41' 0.07 1.54 3
606 KEREN
Boron Adsorption-Desorption by Soil Constituents
Boron can be specifically adsorbedby different clay minerals, hydroxy

oxides of AI, Fe, and Mg, and organic matter. Boron is adsorbedmainly on the
particle edgesof the clay minerals rather than the planar surfaces.The crystal
structureof thosesurfacesgives rise to surfacecationswhich are not fully coor-
dinatedinto the "ideal" ionic lattice and may result in a residualchargeon sur-
face ligands.In an aqueoussolution suchsurfacesites(representedby an S) will
be hydroxylatedand chargedin a mannerindicatedbelow
ow
The B adsorptionprocesscan be explainedby the surfacecomplexation

approach,in which the surfaceis consideredas a ligand. Somepossibleclay sur-
face complexconfigurationsfor B are shownin Fig. 21-1. Suchspecific adsorp-
tion, which occurs irrespectiveof the sign of the net surfacecharge,can occur
theoreticallyfor any species capable of coordinationwith the surfacemetal ions.
However, because0 is the ligand commonly coordinatedto the metal ions in
clay minerals, the B speciesB(OH)3 and B(OH)4" are particularly involved in
such reactions.
Keren and Bingham (1985) review the factors that affect the adsorption
and desorptionof B by soil constituentsand the mechanismsof adsorption.Soil
pH is one of the most importantfactors affecting B uptakeby plants. Increasing
pH enhancesB adsorptionon clay minerals,hydroxy -AI and soils, showing a
maximum in the alkaline pH range.
The responseof B adsorptionon clays to variation in pH can be explained
as follows. Below pH 7, B(OHh predominatesand sincethe affinity of the clay
for this speciesis relatively low, the amountof adsorptionis small. Both B(OH)4"
and OH- concentrationsare low at this pH; thus, their contribution to total B
adsorptionis small despitetheir relatively strongaffinity for the clay. As the pH
is increasedto aboutnine, the B(OH)4" concentrationincreasesrapidly. Sincethe
HO-B-OH
OH
o
I
I
I
HO
""/
0/ ""-0
I
B
OH
I
nn77 777 All 777?/ 7771l AI177l77 AI,l7i','l'J
HO~ /OH
B
I
o
I
////1/1 AlI/77/117
Fig. 21-1. Possiblesurfacecomplexconfigurationsfor B on broken edgesof clay minerals.

BORON 607
Table 21-3. Distribution of B in commonrock types.

Rock class Rock type Boron content
mg kg-1
Igneous Granite 15
Basalt 5
Sedimentary Limestone 20
Sandstone 35
Shale 100
Soils 7-80
OH- concentrationis still low relative to the B concentration,the amount of

adsorbedB increasesrapidly. Furtherincreasesin pH result in an enhancedOH-
concentrationrelative to B(OH)4", and B adsorptiondecreasesrapidly due to the
competitionby OH- at the adsorptionsites.
Wetting anddrying processeshavea significanteffect on B adsorptionand
desorptionby soils and clay minerals(Biggar & Fireman,1960; Keren & Gast,
1981). Biggar and Fireman(1960) have noted that repeatedwetting and drying
increasedB adsorptionin soils. This increasein adsorptionis most evidentdur-
ing the first wetting and drying cycle, with very little adsorptionoccurringafter
the fifth cycle. This adsorptionalso is dependentupon the B concentrationin
solution.As the concentrationof B is increased,the effect of drying becomes
more pronounced.Exchangeableion composition,ionic strengthof soil solution,
and temperaturealso havesignificant effect on B adsorption.
BORON EXTRACTION METHODS

Total Boron in Soil-Fusion Method
Principles
Boron occurs mainly in silicate minerals at a concentrationof approxi-
mately 10 mg kg-1 in the earth'scrust.The distribution of B amongthe common
rock typesis shownin Table 21-3 (Krauskopf,1972).
Soils whosesoluble-Bcontentshavebeenloweredto nontoxic levels for a
given plant by leachingmay regeneratetoxic levelsof solubleB with time in the
absenceof further leaching.The sourcesof this additional Bare B-containing
primary minerals.
Someof the morecommonB-containingmineralsaregiven in Table21-4.
Becauseof the limited solubility of such B-containingmineralsin soils, a
chemicaldeterminationof total B entailssoil decompositionby a Na2C03fusion,
dissolutionof the melt with dilute sulfuric acid (H2S04), and separationof B as
an esterwith alcohol beforedeterminingB. The final reactionin the presenceof
H2S04 takesplacewithin the pH rangeof 5.5 to 6, hastensthe disintegrationof
the melt and leavesmost of the sesquioxidesand silica in insolubleform. Since
mostof the B may be adsorbedby the sesquioxidesand silica surfaces(Keren &
Gast,1981), the determinedB contentcould be lower than what actually exists.
Addition of alcohol up to 60 to 70% by volume servesto precipitatemost
of the large amountof Na2S04which hasbeenformed during the dissolutionof
608 KEREN
Table 21-4. Boron-containingminerals.

Mineral class Mineral subclasses Chemicalformula
Hydrousborates Borax Na2 B4 0 7 • 10 H20
Kemite Na2 B4 0 7 • 4 H20
Colemanite Ca2 B6 0 11 • 5 H20
Ulexite Na Ca Bs 0 9 • 8 H20
Anhydrousborate Ludwigite Mg2Fe BOs
Kotoite Mg3 (B03)2
Complexborosilicates Tourmaline Na (Mg,Fe,Mn,Li,Al)3
Al6Si6 018 (B03)3
(OH, F)4
Axinite (Ca,Mn,Fe)3Al2B03Si4012(OH)
melt. This leavesall of the B and only a small amountof saltsin solution. After
the final evaporation,the samplemust be ignited to volatilize the small amount
of nonvolatileorganicmatterusually introducedwith the alcohol.
Total Soil Boron Content

The following procedureis essentiallythe procedureintroducedby Berger
and Truog (1939) with modificationsby Wear (1965) and Gupta(1966).
SpecialApparatus
1. Platinumcrucible, 30-mL capacity.
2. Glasswarelow in boron.
Reagents
1. Sodiumcarbonate(NazC03), anhydrous(reagentgrade).
2. Sodium carbonate(NazC03), 30%. Dissolve 30 g of NazC03 (reagent
grade) in distilled water and adjust the solution volume to 100 mL
with distilled water. Store the solution in a polyethylenecontainer.
3. Sulfuric acid, 2 M. Add 112 mL of concentratedHzS04 (reagent
grade)to approximately800 mL of distilled water, mix, cool to labo-
ratory temperature,and adjustthe solutionvolume to 1 L with distilled
water.
4. Hydrochloric acid (HCI), 4 M. Add 324 mL of concentratedHCI
(reagentgrade)to approximately600 mL of distilled water, mix, cool
to laboratorytemperature,and adjust the solution volume to 1 L with
distilled water.
5. Hydrochloric acid, 0.1 M. Add 8.1 mL of concentratedHCI (reagent
grade)to approximatley900 mL of distilled water, mix, cool to labo-
ratory temperature,and adjustthe solution volume to 1 L with distilled
water.
6. Ethyl alcohol, 95% (reagentgrade).
7. Phenolphthaleinindicator. Dissolve 0.05 g of phenolphthaleinin 50
mL of 95% ethyl alcoholand 50 mL of distilled water.
8. Bromothymolblue indicator. In a mortar, grind 0.1 g of bromothymol
blue with 16 mL of 0.01 M sodium hydroxide (NaOH). Dilute the
solution to 250 mL with distilled water.
BORON 609
Procedure
Fuse1.0 g of 1oo-mesh(0.149 mm) soil with 6 g of anhydrousNa2C03in
a crucible, as describedby Hossner,1996 (seeChapter3). Include a blank (6 g
of Na2C03)with eachsetof determinations.Cool the crucible,and placeit on its
side in a 2S0-mLbeaker(low-B glass)containingSO mL of distilled water. Place
a cover glasson the beaker,and add 2 M H2S04 in 4- to S-mL incrementsuntil
the melt hasdisintegratedandthe solution is within a pH rangeof 6.0 to 6.8 (test
with an externalindicatorsuchas bromothymolblue or with a pH meter).Trans-
fer the solution to a SOO-mL volumetric flask. Wash the contentsof the beaker
and crucible into a flask with distilled water, keepingthe solution volume <ISO
mL. Add ethyl alcohol to the flask until the volume is nearly SOO mL, and mix
the contentsthoroughly. Add a few drops of phenolphthaleinto the flask, then
adjustthe pH to slightly alkalineby addingthe Na2C03solution. Adjust the vol-
ume to SOO mL with ethyl alcohol, mix, and filter.
Placea 400-mL aliquot of filtrate in a SOO-mL beaker(low-B glassware),
add 100 mL of distilled water, and reducethe solutionvolume to 10 to 20 mL by
heatingon a hot plate or steambath. Transferthe solution to a platinum dish,
evaporatethe contentsto dryness,andgently heatthe dish over an openflame to
destroy any organic matter. Cool the dish, add S mL of 0.1 M HCI, and thor-
oughly triturate the residuewith a rubberpoliceman.Transfera 1-mL aliquot to
a lO-mL test tube, and proceedwith the B determinationby one of the methods
given in the sectionon "Boron Determination."
Gupta(1966) found that the H2S04 extractionof soils interferedwith the
adsorbanceof the B carminecomplexresultingin high valuesof B. No interfer-
enceswere encounteredwith HCI extracts.Thus,underthesecircumstances,4 M
HCI solution shouldbe addedinsteadof 2 M H2S04 solution.
Boron Availability Assessmentfor Plant Uptake
Principles
The suitability of irrigation water hasbeenevaluatedon the basisof crite-
ria which determinethe potential of the water to causeplant injury and yield
reduction.In assessingthe B in irrigation water, however,the physicochemical
characteristicsof the soil must be taken into considerationbecausethe B uptake
by plantsis dependentonly on B activity in soil solution (Keren et aI., 1985a,b).
Boron uptakeby plantsgrown in a soil of low clay contentis significantly greater
than that of plantsgrown in a soil of high clay contentat a given level of added
B (Keren et aI., 1985a,b).
Boron can be specifically adsorbedby different clay mineralswhich vary
in their adsorptioncapabilities. On a weight basis, illite is the most reactive
amongthe commonclay mineralswhereaskaolinite is the least reactive. Simi-
larly, B canbe adsorbedon organicmatterandsesquioxidesby a ligand exchange
mechanism(Kustin & Pizer,1969;Sims& Bingham,1968; Keren & Gast, 1983;
Yermiyahuet aI., 1988).
The availability of soil B dependsalso on pH, ionic strength,adsorbedion
composition,and soil moisturecontent.Soil pH is oneof the mostimportantfac-
610 KEREN
tors affecting B uptakeby plants.Boron adsorptionon clay minerals,hydroxy-AI

and soils, showsa maximumin the alkaline pH range.The effect of exchangeable
ions on B adsorptionby clay minerals is dependentupon the conditions under
which the systemexists.Little effect was observedat pH around7 but significant
effect was found for pH greaterthan 8, with the Ca systemsadsorbingmore B.
There are severalextractionproceduresfor predictingcrop responseto B:
(i) hot-water soluble B, (ii) Mannitol exchangeable B and (iii) Mehlich-III
extractableB. Thesetests have been calibratedfor a numberof field and veg-
etablecrops; hence,realistic standardsare availablefor interpretingsoil analysis.
More recently,a curvilinear relationshipwas found betweenB in com (Zea
mays L.) tissueand hot-watersoluble B, mannitol exchangeableB, NH4-acetate
extractableBand Mehlich-III extractableB (Jin et aI.,1988). Among thesefour
procedures,mannitol exchangeableB correlatedmost closely with B in com tis-
suefrom native B. A linear relationshipbetweenB in leaveplantsand B in satu-
ration extract from soils also was found for Bell pepper(Capsicum annuum L.
var. annuum)andwheat(Triticum aestivum L.) (Keren et aI., 1985a,b).Details of
the hot-watersolubleB, saturationextractof soil pasteand Manitol-CaClz extract
procedurefor availableB for plants are given below.
Hot-WaterSolubleBoron
The most commonly usedmethodfor extractingeasily soluble B from soil
involves refluxing with hot water for a period of 5 min using a soil/waterratio of
1:2 (Berger& Truog, 1939). This procedurehas beenfound to be a useful index
of plant responsesto B fertilization in deficient soils and also of the B contentof
plants (Berger & Truog, 1940; Wear & Patterson,1962; Singh & Sinha, 1976;
Cartwright et aI., 1983). In general,suchrelationshipswere obtainedwith miner-
al soils having concentrationsof extractedB up to about 3 mg kg-I, although
good correlationwas obtainedbetweenextractedB from soil and B contentof
plants in highly organicsoils with B addedto increasesaturationextractedB up
to 50 mg kg-I. Odom (1980) found that the recommendedproceduredid not
achieveequilibrium statefor the B extracted.A longer extractionperiod was rec-
ommended.
The following procedureis essentiallythe procedureintroducedby Berger
and Truog (1939, 1944) with modificationsby Dible et ai. (1954), Baker (1964),
Cartwright et ai. (1983) and Parkerand Gardner(1981).
Special Apparatus
1. Low-boron glassware.
2. Water-cooledreflux condensers.
3. Evaporatingdishes(e.g., porcelaincrucible).
Reagents
1. Calcium hydroxide[Ca(OH)2] suspension.Add 0.4 g of Ca(OHh]
(reagentgrade)to 100 mL of distilled water.
grade)to approximately900 mL of distilled water, mix, cool to room
temperature,and adjustthe solution volume to 1 L with distilled water.
BORON 611
3. Calciumchloride (CaCI2), 0.02M. Dissolve2.22 g of anhydrousCaCl2

in approximately900 mL of distilled water,and adjustthe solutionvol-
ume to 1 L with distilled water.
Procedure.Place a given amount of air-dried soil (equal to 20 g 105°C
oven dried) in a 250 mL low-boron flat-bottom flask, and add40 mL of 0.02 M
CaCl2 solution to obtain clear hot-water extractsfor B determination.Attach a
water-cooledreflux condenserto the flask. Heat the flask until initiation of boil,
reflux the suspensionfor precisely 5 min, and then cool the flask and filter the
suspensionthrough filter paper (Milipore Corp., Bedford, MA) and take an
appropriatealiquot (i.e., 1 mL) for B determination(seesectionon "Boron Deter-
mination"). The presenceof CaCl2 in the extractingsolution does not alter the
amountof B extracted,and give a clear, colorlessextractwell suitableto analy-
sis by inductively coupled plasma (ICP) and by spectrophotometricmethods
using reagentssuchas Azomethine-H.In casenitratesand organicmatterare pre-
sentin solution,transfera 20-mL aliquot of the filtrate to an evaporatingdish, add
2 mL of the Ca(OHhsuspension,and evaporatethe solution to dryness.Heat the
evaporatingdish gently over a flame to destroyorganic matter. After the evapo-
rating dish hascooledto room temperature,add 5 mL of 0.1 M HCl. Triturate the
residue in the evaporatingdish, using a rubber policemanto assuresolution of
residue.
SaturationExtract of Soil Paste

Developmentof injury symptomsand reductionof yield due to excessive
accumulationof B dependson both the crop speciesand the B concentrationin
the soil solution, which can be approximatedby the B level in the soil saturation
extract. Keren et al. (1985a,b) observeda linear correlation betweendry leaf
weight of bell pepperand wheat with the B contentin the leaf tissueand with B
activity in soil solution.
Special Apparatus
1. Buchnerfunnel or Richardsfilter funnel (Richards,1949).
Reagents
1. Calcium hydroxide [Ca(OH)21 suspension.Add 0.4 g of Ca(OHh to
100 mL of distilled water.
grade) to approximately950 mL of distilled water, and mix. When it
hascooledto room temperature,adjustthe volume to 1 L with distilled
water.
Procedure.Weigh approximately 400 g of air-dried soil into a plastic
beaker,and while mixing, add distilled water until the soil pastebecomesplastic
but not saturatedwith water. Cover the beaker,and set the beakerasidefor 2 h to
permit betterdistribution of water throughoutthe soil. Then uncoverthe beaker,
stir the paste,and addwater until the soil appearsto be saturated.After a mini-
mum of 4 h or, preferably,an overnightperiod, stir the pasteand collect a small
sample(10-50 g) for a moisture determination.Transferthe remaining pasteto
612 KEREN
the Buchnerfunnel fitted with a retentivefilter paper,and apply a vacuumto col-

lect the saturationextractin a testtube (Loeppert& Suarez,1996,seeChapterIS)
for additional detailson preparationof saturationpaste).
Filter the solution through filter paper(or milipore filters) and take an ap-
propriatealiquot for B determination(see"References").In caseof organicmat-
ter presencein saturationextract, pipette a lO-mL aliquot of the extract into an
evaporatingdish, add 2 mL of saturatedCa(OH)2suspension,and taketo dryness.
Gently heat the dish over a flame to destroyorganic matter, and then cool it to
room temperature.Add S mL of 0.1 M HCI, and thoroughly triturate the residue
with a rubberpoliceman.Take a 1-mL aliquot and proceedwith one of the deter-
mination methodsgiven in the sectionon "Boron Determination."
Mannitol-CalciumChloride Extract
Principles. Mannitol was included in the extracted solution becauseit
forms a high-affinity, soluble complex with B(OHh and B(OH)4" thus mainti-
naingsolution activities of thesetwo speciesat low levels.The complexationfor-
mation constantis 104 for the 1:1 complexand 8.24 for the 1:2 complex(Nies &
Campbell,1964, p. 92).
R R
I
R-C-OH
I
- R-C-O", /OH]-
I
R-C-OH
]
... I
R-C-O
/B\.
\.OH
I I
R R
Borate-diolcomplex
The ability of mannitol to complexB is dependenton pH, with the concentration

of mannitol-B complexdecreasingwith decreasingpH (Knoeck& Taylor, 1969).
Knoeck and Taylor (1969) concluded that mannitol reacted only with the
B(OH)4", thus an unbufferedmannitol solution would not be expectedto extract
much more B than a water extractfrom acid soils.
. In an examination of B adsorption and desorption by clay minerals,
Hingston(1964)found that 0.01 M mannitol in 0.01 M CaCl2solutionwas unable
to copmletelyremoveB from the clay at low pH values,whereasat pH valuesof
around8.S to 9.0 most of the adsorbedB was desorbed.Thus, this methodcan be
usedas a B extractantin either alkaline or calcareoussoils.
Boron extractionfrom soil with a 0.01 M mannitol-O.01M CaCl2 solution
removessoluble (saturationextract B) plus adsorbedB. Rhoadeset al. (1970)
considerextractableB to be "leachable"B, it is a relevantsoil propertyto include
in assessingB toxicity.
Extraction of B from soil with 0.01 M CaClz-O.OSM mannitol was sug-
gestedby Cartwright et al. (1983). An extractionperiod of 1 h was found to be
appropriatefor routine operationsand minimized timing errors while recovering
BORON 613
most of the solubleB. The methodgives a clear,colorlessextractwell suitableto

analysisby ICP and also by spectrophotometricmethodsusing reagentssuch as
azomethine- H.
Special Apparatus
1. Polyethylenecentrifugetubes,50-mL capacity.
2. Centrifuge
Reagent
1. Mannitol-calcium chloride, 0.01 M. Dissolve 1.82 g of mannitol
(reagentgrade)and 1.1 g of CaCl2 (anhydrous,reagentgrade) in dis-
tilled water. Adjust the solution volume to 1 L with distilled water.
Procedure.Weigh 5 g of air-dried soil into a 50-mL centrifuge tube, add
25 mL of 0.01 M mannitol-O.Q1M CaCl2 solution, and shakethe suspensionfor
16 h. Collect the solution phaseby centrifugingthe suspensionfor approximate-
ly 15 min at 1000g and filtering the supernatantthrough a no. 42 filter paperinto
a plastic 30-mL bottle.
Transfer a I-mL aliquot of the filtered solution to a 50-mL polyethylene
centrifuge tube, and proceedwith determiningB. [The extracting solutiondoes
not interfere with developmentof the colored complex of azomethine-Hand
B(OHh·]
BORON DETERMINATION
Colorimetric Methods
Becauseof the low B concentrationsencounteredin samplesof waters,

soils and plant materials,photocolorimetricproceduresare generally used. Four
methodsare discussedbelow: (i) the carmine procedureintroducedby Hatcher
and Wilcox (1950); (ii) the curcumin procedurerefined by Dible et ai. (1954);
(iii) a modified curcumin procedureusing 2-ethyl-l, 3-hexanediol(Goldman et
aI., 1975); and, (iv) Azomethine-Hmethod.
CarmineMethod
Principals.The carmineprocedurefor determiningB (Hatcher& Wilcox,
1950) is based on the fact that carmine (an anthraquinonedye) complexes
B(OHh in concentratedH2S04• This complexforms within 45 min, retainsmax-
imum absorbanceat the wavelengthof 585 nm for approximately4 h, and is unaf-
fected by the presenceof a wide variety of electrolytes.With this procedure,B
concentrationsranging from 0.5 to 10 Ilg mL-l may be determined.The princi-
pal objectionto the procedureis that developmentof the complex in concentrat-
ed H2S04 makesthe proceduretediousand hazardous.
Special Apparatus
1. Spectrophotometer.
2. Erlenmeyerflask, 125 mL.
614 KEREN
Reagents
1. Sodium hydroxide (NaOH), 0.1 M. Dissolve 4.00 g of NaOH in dis-
tilled water, and adjustsolution volume to 1 L with distilled water.
2. Hydrochloric acid, concentrated(reagentgrade).
3. Hydrochloric acid, approximately0.6 M. Add 48.6 mL of concentrat-
ed HCI (reagentgrade) to approximately900 mL of distilled water,
cool, and adjustvolume with distilled water to 1 L.
4. Sulfuric acid, concentrated(reagentgrade).
5. Carminesolution,0.05% (wt/wt). Add 0.92g of carmineto 1 L of con-
centratedH2S04 (reagent grade), and shake until the carmine dis-
solves.Storesolution in the dark.
6. Standardboric acid B(OH)3 solutions:Dissolve0.5714g of B(OH)3 in
distilled water, and adjust volume to 1 L. One milliliter of this stock
solution contains100 flg of B. Dilute 2, 4, 6, 8, and 10 mL of the stock
solution to 100 mL with distilled water to have B solution concentra-
tions of 2,4,6,8,and 10 flg mL-l, respectively,for preparationof cal-
ibration curve.
Procedure.Transfer a 2-mL aliquot of the diluted standardB solutions,
including distilled water or samplesolution, into an Erlenmeyerflask, and add
two dropsof concentratedHCl. Then slowly add 10 mL of concentratedH2S04,
mix, and cool. Add 10 mL of carminesolution, mix, and let color developfor 45
min. Determineabsorbanceat 585 nm againsta referencesolutionof 2 mL of dis-
tilled water carried through the identical procedure.Preparean absorbance-B
concentration curve from absorbancereadingsof standardsolutionscontainingB
up to 10 flg mL-l.
If the B concentrationof the sampleis too high for a reliable absorbance
reading,dilute the samplewith distilled water to a known volume, mix, pipette2
mL of diluted sampleinto a flask, and proceedas prescribedabove.For samples
too low in B for a reliable absorbancereading,pipette a suitablealiquot into an
evaporatingdish, make alkaline with 0.1 M Ca(OH)z, and take to drynesson a
steambath or hot plate at low temperature.Cool, add 5 mL of 0.6 M Het, tritu-
rate with a rubber policeman,and filter into a lO-mL test tube. Pipette2 mL of
filtrate into a flask, and proceedwith the determinationas describedabove.
CurcuminMethod
Principles.Curcumin, 1,7-bis (4-hydroxy-3-methoxyphenyl)-1,6-heptadi-
ene-3,5-dione,is a natural dyestuff. Upon evaporationto drynessof an acidified
solution containingB(OHh and curcumin,a red reactionproductsolublein alco-
hol is formed in amountproportionalto the amountof B(OH)3 present.This prod-
uct has been identified as an isomeric form of curcumin and was namedroso-
cyanin for the rose color of the acid form and the blue color of its metallic salt.
The rosocyamineis possibly formed by the reactionof B(OH)4" with one of the
hydroxyl groupsof the curcumin molecule.The formation of a complex by the
reactionof B(OH)3 with curcumin is a very sensitivetest for B.
Most of the methodsdescribedfor forming the B-curcumin complexhave,
as their basis,Naftel's(1939) procedures.The procedureis basedon formation of
BORON 615
a coloredproductsolublein alcohol when solution B(OHh, oxalic acid, and cur-

cumin are evaporatedto dryness.At 540 nm, absorbanceof an alcoholic solution
of the product(rosocyanin)is proportionalto that of the B concentration.Subse-
quently, Dible et al. (1954) modified the proceduredescribedby Naftel (1939) for
rapid and routine determinationsof B in extractsof soil and plant samples.The
techniquedescribedbelow is basedon their procedure,with modificationssug-
gestedby Johnsonand Ulrich (1959). The procedureis appropriatefor B con-
centrationsrangingfrom 0.1 to 2.0 Ilg mL- 1. The determinationofB by curcum-
in methodis sensitiveand accurate,but it also is complex and time consuming.
Special Apparatus
2. Porcelainevaporatingdishes.
3. Water bath with temperaturecontrol for 55 ± 3°C.
Reagents
1. Curcumin-oxalicacid solution. Dissolve 0.04 g of finely ground cur-
cumin and 5 g of oxalic acidin 100 mL of 95% ethyl alcohol. If stored
in a dark bottle in a refrigerator,this reagentcan be usedsatisfactorily
for a week.
2. Ethyl alcohol, 95% (reagentgrade).
3. StandardB solutions.Dissolve 0.2859g of B(OHh (reagentgrade)in
distilled water, and adjust volume of solution to 1 L with distilled
water. One milliliter of this solution contains50 Ilg of B. Dilute 0.2,
0.5, 1.0, 2.0, and 4.0 mL of the 50-llg B per milliliter solution to 100
mL with distilled water to have a seriesof standardsolutionscontain-
ing 0.1,0.25,0.50,1.0, and 2.0 Ilg B mL-l, respectively.Include a dis-
tilled water samplefor the 0 Ilg of B per milliliter standardsolution.
Thesesolutionsare used to preparea calibration curve.
Procedure.Pipettea 1-mL aliquot of either a samplecontaining0.1 to 2.0
Ilg of B per milliliter or of the standardB solutionsinto a porcelainevaporating
dish. Add 4 mL of the curcumin-oxalicacid solution, and mix. Placeon a water
bath maintainedat 55 ± 3°C, and take to dryness.Maintain disheson a steambath
at this temperaturefor at least15 min after the solutionsevaporate.Cool the dish-
es to room temperature,and then add 25 mL of 95% ethyl alcohol. Rub the
residue with a rubber policeman to ensurecomplete extraction of the colored
compoundby the alcohol. Using a Whatman no. 2 filter paper, filter into an
appropriatecontainer,and read absorbanceof solutionsat the wavelengthof 540
nm.
Preparean absorbance-Bconcentrationcurve from absorbancereadingsof
the standardsolutionscontainingup to 2.0 Ilg of B per milliliter.
Modified Curcumin Method: 2-Ethyl-l,3-Hexanediol

in Chloroform Extraction
Principles. Goldman et al. (1975) modified a spectrophotometricproce-
dure for the determinationof submicrogramconcentrationsof B in water sam-
ples. The procedureentails complexingB(OH)3 with 2-ethyl-1,3-hexanediol(in
616 KEREN
CHCI3), convertingthe complexto the roscyanincomplexusinga solutionof cur-

cumin in glacial aceticacid, followed by the addition of concentratedH2S04• The
concentrateis diluted with ethanol,and absorbanceof the diluted solution is read
at the wavelengthof 550 nm. The procedureis rapid, precise,and in particular,
convenientfor determiningB in water samples.
Their procedureis essentiallythe same as that of Mair and Day (1972)
spectrophotometricdeterminationof B extractedfrom ashedanimal tissues.
Special Apparatus
2. Polypropylenetest tubes,17 mm in diameterand 100 mm long.
3. V:>rtex mixer.
Reagents
1. Hydrochloric acid, 1 M. Add 81 mL of concentratedHCL (reagent
grade)to approximately900 mL of distilled water, and mix. When the
solution is cool, adjustvolume to 1000 mL with distilled water.
2. Sulfuric acid, concentrated(reagentgrade).
3. Ethanol(C2H50H), 95% (reagentgrade).
4. 2-Ethyl-1,3-Hexanediolin chloroform (CHCI3). Dilute 10 mL of 2-
ethyl-1,3-hexanediolto 100 mL with CHCl3 (reagentgrade).
5. Curcuminsolu!ion. Dissolve0.375g of finely ground curcuminin 100
mL of glacial acetic acid (reagent grade). The curcumin solution
shouldbe fresh, preparedat the sameday of determination.
6. StandardB solution. Dissolve 0.57175g of boric acid B(OHh in dis-
tilled water. This solution contains100 Ilg of B per milliliter. Prepare
standardsolutionscontaining0.1, 0.25, 0.50,0.75, and 1.0 mg of B per
milliliter by diluting to 100 mL, respectivley,0.1, 0.25, 0.50, 0.75.and
1.0 mL of the 100 Ilg mL-l B solution.
Procedure.Pipette 1.0 mL of a samplecontaining0.1 to 1.0 Ilg of B or of

the standardB solutionsinto a polypropylenetesttube, add 2 mL of 1 M HCI, and
mix with a Vortex mixer (Scientific Industries,Inc., New York). Add 3 mL of the
2-ethyl-1,3-hexanediolsolution (in CHCl3), and mix for approximately30 s with
the Vortex mixer. Transfera 0.5-mL aliquot of the organicphaseto an empty test
tube, add 1 mL of the curcumin solution, followed by 0.3 mL of concentrated
H2S04, and mix thoroughly. Allow the reactionto proceedfor 15 min, dilute the
solution to 25 mL with ethanol,and read absorbanceof the solution at the wave-
length of 550 nm within 20 min.
If the aliquot of a samplecontainsB concentration> 1.0 Ilg, dilute the sam-
ple with enough distilled water so that a 1-mL aliquot contains <1.0 Ilg B. In
caseswhere a 1-mL aliquot of the sampledoesnot contain sufficient B to make
a reliable determination,a larger aliquot of the organic phasecan be taken and
then concentratedto 0.5 mL by evaporatingthe volume using a slight vacuum.
Preparean absorbance-Bconcentrationcurve from absorbancereadingsof
the B standardsolution «(}"'1.0 Ilg mL-l).
BORON 617
Azomethine-Hydrogen Method
Principles. Sincethe introductionof the azomethine-Hreagent(Shaninaet
aI., 1967)it hasrapidly gainedfavor as a methodfor determiningB in plant mate-
rials, soil extracts,andwater.The methodis sensitive,simple,fast, and subjectto
little interference.
The method employed azomethine-Has the reagent to form a colored
copmlex of B(OHh in aqueousmedia. Solutionsof azomethine-Hwith B(OHh
were observedto form a complex very rapidly independentof the presenceof a
wide variety of salts.
Basson et ai. (1969) modified the azomethine-Hprocedurefor routine
analysisof the B contentof citrus (Citrus spp.) leaves.Their modification con-
sistedof a refinementin the synthesisof the reagent,improvementof buffer con-
trol and through use of an ethylenediaminetetraacetic acid (EDTA) solution, a
reduction of interferencesfrom metals presentin citrus leaves.They observed
close agreementbetweendeterminationsof B in citrus leavesby both the im-
proved automatedazomethine-Hmethodand the carmineprocedure(Hatcher&
Wilcox, 1950).Subsequently,Wolf (1971,1974)modified the azomethine-Hpro-
cedurefurther for high-volumeroutine B analysisof soil, plant, fertilizer, sludge,
and water samples.Commercially manufacturedazomethine-H,acetatebuffers,
and EDTA solutionswere employedin the determinationof B.
This methoddoes not require the use of concentratedacids and is subject
to few interferences.Over the B concentrationrange of 0.5 to 1.0 Ilg mL-1,
azomethine-Hsolution reactswith B(OHh to form a stablecomplex at pH 5.1.
Maximum absorbanceoccursat the wavelengthof 420 nm. The major drawback
to the azomethine-Hmethod for soils is error resulting from suspendedor dis-
solved material which imparts a yellow color to the extract. Wolf (1974) and
Gupta (1979) have proposedthe use of charcoal to decolorize soil extracts.
Adding excesscharcoal,however,can lead to a loss of B from solution. The pro-
ceduredescribedin the following sectionis basedon the proceduredescribedby
Guptaand Stewart(1975) with somemodifications.
Special Apparatus
Reagents
1. Azomethine-Hsolution. Dissolve 0.9 g azomethine-Hand add2 g of
L-ascorbicacid in about200 mL of lukewarmdistilled water, cool and
then dilute to 250 mL. If the resulting solution is not clear, heat the
flask gently. The solution can be kept for 7 d under refrigeration.
2. Ethylenediaminetetraacetic acid solution, 0.025M. Dissolve4.563 g of
disodiumsalt of EDTA in 200 mL of distilled waterand adjustvolume
with distilled water to 500 mL.
3. Buffer solution. Dissolve 250 g of ammoniumacetatein 500 mL dis-
tilled water and addacetic acid (approximately300 mL) to bring the
pH of the final solution to 4.8.
4. StandardB solution. Dissolve0.57174g of B(OH)3 (reagentgrade)in
distilled water, and adjust volume of solution to 1 L with distilled
618 KEREN
water. One milliliter of this solution contains 100 Ilg B. Dilute this
solutionl0-foldto havethe 10 Ilg mL-l standardB solution.This solu-
tion is usedto preparea calibrationcurve.
5. Washedactivatedcharcoal.Add 1 L of extractingsolution to 500 g of
activatedcharcoalthrough no. 42 Whatman filter paper on buchner
funnel. Wash the charcoalfour or five times with 200-mL portionsof
respectiveextractingsolutions.Leachwith distilled water and dry the
charcoalin an oven at 60°C.
Procedure
Calibration Curve. Transfer0, 1,2,3,4,and 5 mL of the 10 Ilg mL-l B
stock solution to 25-mL volumetric flasks and add distilled water to bring the
solutionvolume to about10 mL. Add 1 mL of EDTA solutionand 2 mL of buffer
solution and mix well. Then add 5 mL of Azomethine-Hsolution and adjust the
volume to 25 mL with distilled water. Mix the contentsof the flasks and keep
them for 2 h for color development.Readthe absorbanceon spectrophotometer
at a wave length of 420 nm. The final B concentrationsin the volumetric flasks
are 0,0.4,0.8,1.2, 1.6, and 2.0 Ilg mL-l, respectively.
Sample Solution. The solution should be colorless.If not, add activated
charcoal(the minimum necessaryamount), shake for 5 min andfilter throughno.
42 Whatmanfilter paper.Transfera known suitablealiquot (1-17 mL in which
the final B concentrationis in the rangeof the calibrationcurve) from the color-
less solution in a 25-mL volumetric flask. If the aliquot is less than 15 mL add
distilled waterup to a total solutionvolumeof 15 mL, beforeaddingthe reagents.
Add 1 mL of EDTA solution and 2 mL of buffer solution and mix well. Thenadd
5 mL of Azomethine-Hsolution and adjust the volume to 25 mL with distilled
water. Mix the contentof the flask and after 2 h read the absorbanceon spec-
trophotometerat a wavelengthof 420 om.
PotentiometricMethods
Mannitol PotentiometricMethod
Principles. Foote (1932) and Wilcox (1932) developed an analytical
methodfor the dire~t determinationof B in dilute aqueoussolutions.The method
is basedon the fact that B(OH)3 in the presenceof mannitol titrateslike a strong
monobasicacid. This procedurehas beenadoptedby the American Society for
Testingand Materials(ASTM) as a standardtest methodfor B in water. The fol-
lowing descriptionof the chemical procedureis basedon the ASTM (1978, p.
261-265)outline.
Special)lpparatus
1. Burette,10-mL microburettewith a Teflon stopcock.
2. Erlenmeyerflask, widemouth,250 mL.
3. Magneticstirrer and Teflon-coveredstirring bar.
4. pH meterwith single combinedelectrode.
BORON 619
Reagents
1. Sodium hydroxide, 1 M. Dissolve 40.0 g of NaOH pellets in C0z-free
distilled water, cool to ambienttemperature,adjustvolume to 1 L with
C0z-freewater, and store in a plastic bottle with a CO2 trap.
2. Sodiumhydroxide,0.1 M. Dissolve4.0 g of NaOH pelletsin C0z-free
distilled water, cool to ambienttemperature,adjustvolume to 1 L with
C0z-freewater, and store in a plastic bottle with a CO2 trap.
3. Sodium hydroxide,0.02 M. Preparean approximately0.02 M solution
by diluting 20 mL of the 1.0 M NaOH to 1 L with CO2-free distilled
water. Storein a plasticbottle with a CO2 trap. Standardizethe diluted
solution against100 mL of a to mg L-1 B solution.
4. Potassium permanganate (KMn04) solution, 3.3 g L. Dissolve3.3 g of
KMn04 in distilled water, and dilute to 1 L.
5. Ethylenediaminetetraacetic acid disodium salt (EDTA disodium) solu-
tion, 0.1 M. Dissolve37.21 g of EDTA disodiumin distilled water, and
dilute to 1 L.
6. Hydrochloric acid, 1% (vol/vol). Mix 1 volume of concentratedHCI
(reagentgrade)with 99 volumesof distilled water.
7. Mannitol, B free (reagentgrade).
8. B standardsolution, to Ilg mL-1. Dissolve0.57175g of B(OHh in dis-
tilled water, and dilute to 1 L. Dilute this solution to-fold to have the
standardto Ilg mL- 1 B solution.
Procedure
1. Pipettean aliquot containingbetween0.1 to 1.0 mg of B into a 250-mL
Erlenmayerflask, adjustvolume in the flask to approximately100 mL,
and place over a magneticstirrer. Insert the combinedelectrode,and
adjustthe pH of the solution to 2.5 ± 0.2 using the 1 % HCI solution.
2. Removethe electrode,placea watch glassover the mouth of the flask,
and heatthe solution to boiling. Boil for 3 min to expel CO2, then add
1 mL of KMn04 solution, and continueto boil solution with the watch
glass in place. Add to mL of EDTA disodium solution, and boil the
solution with the watch glassin place for 3 min. The solution should
be colorlessat this point.
3. Removethe flask from the heat,stopper,and immerseit in a waterbath
to lower the temperatureto within 3°C of the ambienttemperature.
4. Removethe stopperand insertthe pH electrode.Adjust the solutionpH
to approximately8.0 with the 0.1 M NaOH solution while stirring the
solution gently with the magneticstirrer. After adjusting the solution
pH to just below 7.60 with 1 % HCI solution, carefully back-titratethe
solution to pH 7.60 ± 0.01 with the standardized0.02M NaOH. Add 5
g ± 0.5 of mannitol and stir gently, the solution should clear and
becomeacid. Back-titrateto pH 7.60 ± 0.01 with the standard0.02 M
NaOH. Record the volume of NaOH required to back-titrate to pH
7.60.
Each milliliter of the 0.02 M NaOH requiredto back-titratethe
solution to 7.60 is theoretically equivalent to 0.216 mg of B. The
620 KEREN
equivalencyshould be verified by standardizingthe 0.02 M NaOH

solution against100 mL of a 10 mg L -1 B solution following the pro-
cedureoutlinedabovefor the analysisof a samplecontaining0.1 to 1.0
mgofB.
TetratluoroborateSelectiveElectrodeMethod
Principles. The principles of the method involve two primary steps:(i)
transformationof B in an aqueoussampleto the tetrafluoroborateform (BF;!),
and (ii) subsequentpotentiometricmeasuringof BF4 with a specific liquid ion
exchangemembraneelectrodein conjunctionwith an appropriatejunction refer-
enceelectrode.Boron concentrationin most samplesare usually low. Thus, B in
the samplesolutionis concentratedon a B-specificion exchangeresinbeforeBF4
is formed and subsequentlyeluted form the column. This is done beforemaking
a potentiometricreadingof the solution. The electrodecan be usedfor tetrafluo-
roborateconcentrationsdown to 3 x 10--6 M. The widecalibrationrangefor B (3
x 10-6 -2 X 10-3 M) is a distinct advantageof this method.interferencefrom sev-
eral anionswas estimatedand the B-specific resin also servesto remove these
interfering anionsat a certainlevel. 'The applicability of this methodmay be lim-
ited to certain types of samplesbecauselarge concentrationsof anions such as
nitrate, sulfate, and iodine can interfere with the responseof the fluoroborate
electrodes.The following procedurefor B determinationin water samplesis
basedon the techniquedescribedby Carlsonand Paul (1968, 1969).
Special Apparatus
1. pH meterwith appropriatemillivolt scale.
2. Tetrafluoroborate(BF4) specific ion electrode(Orion Research,Inc.,
model no. 93-05).
3. Plasticsleevereferenceelectrode(Orion Research,Inc., Boston, MA,
model no. 90-04) filled with KCI solution saturatedwith Ag ions.
4. Ion exchangecolumns. Use polyethylenebottles and tubing to con-
structcolumns.Figure 21-2 indicatesdimensionsfor the columncom-
ponentparts. Polyethylenebottleswith the bottomsremovedare used
as reservoirsfor the Amberlite lRA-743 (SigmaChemicals,St. Louis,
MO) and weak acid resin columns.Cotton wool is usedto supportthe
resin. The weak and strongacid resin columnsare allowed to settleby
gravity while the Amberlite is tapped-lightly. To excludeair, a cotton
wool plug is addedabovethe resin. Maintain water in the columnsdur-
ing assembly.If necessary,largecolumnsconstructedin a similar man-
ner can be usedto purify the reagents.
5. Polyethylenelabware.Use this type throughout.Polyethylenebeakers
(50 mL) are convenientto use to collect final eluent solution and to
measurepotentials.
Reagents
1. Ammonium hydroxide (NH40H), 3 M. Add 200 mL of 58% ~OH
(reagentgrade)to distilled water,cool to room temperature,
and adjust
the solution volume to 1 L with distilled water.
BORON 621
A 8 c
so ...1 so"'.
80TTU BOr-I.!:
T
10:"",
T
30"""
l
T
,0......
1
Fig. 21-2. Ion exchangecolumn: (A) Amberlite IRA-743 resin column; (B) Dowex 50 W-X8 resin
column; and (C) Biorex 70 resin column.
2. Hydrofluoric acid (HF), 10%. Add 200 mL of 50% HF (reagentgrade)

to distilled water in a polyethylenecontainer,cool to room tempera-
ture, and adjust the solution volume to 1 L with distilled water.
3. Hydrochloric acid, 3 M. Add 243 mL of concentratedHCL (reagent
grade)to distilled water,cool to room temperature,and adjustthe solu-
tion volume to 1 L with distilled water.
4. Sodiumhydroxide,0.3 M. Dissolve 12.0 g of NaOH pellets in distilled
water, cool to room temperature,and adjustthe solution volume to 1 L
with distilled water.
5. Calcium chloride, 0.15 M. Dissolve 16.7 g of anhydrous CaCl2
(reagentgrade) in distilled water, and adjust the solution volume to 1
L with distilled water.
6. Boric acid [B(OHh], 50 Ilg B mL- i . Dissolve 0.28587g of B(OH)3
(reagentgrade) in distilled water, and adjust the volume to 1 L with
distilled water. Each milliliter of this stock solution contains50 Ilg of
B. Preparestandardsolutionscontaining0.10, 0.25,1.00, and 10.0 mg
of B per milliliter by diluting, respectively,0.20, 0.50, 2.00, 20.0 mL
of the 50 Ilg mL- i B solution to 100 mL with distilled water.
7. Potassiumtetrafluoroborate(KBF4). Preparereferencesolution con-
taining 10.8 Ilg of B per milliliter by placing0.126g of KBF4 (reagent
grade)in a tared 1-L plastic bottle, adding 10 mL of concentratedHF
(reagentgrade) and bringing to a net weight of 1 kg with distilled
water.
8. BioRex 70 (Sigma Chemicals,st. Louis, MO), weak acid resin, 50 to
100 mesh(0-149-O.297-mm).Convertthe resin to an ammoniumform
622 KEREN
before packing the column by washing resin with 3 M NH40H three

times and then twice with distilled water.
9. Dowex 50W-X8 (SigmaChemicals,St. Louis, MO), strongacid resin,
50 to 100 mesh(0.149-O.297-mm).
10. Amberlite lRA-743 Resin,50 to 100 mesh(0.149-O.297-mm).
Procedure
A. Use of resin column to convert B to tetrafluoroborate
1. Neutral and alkaline solutions.Add an aliquot, containing1 to 500
~g of B of the samplesolution, directly into the Amberlite lRA-743
column. After the solution has drainedfrom the column, add 2 mL
of distilled water to the column, followed by 2 mL of NH40H solu-
tion, and 2 mL of water. This stepremovesany anionsthat might be
held by the amine groupsin the Amberlite lRA-743. Add 2 mL of
10% HF solution. After a lO-min wait, add 5 mL of distilled water.
The column now containsBF4" and r held at the aminegroups.
Placethe Amberlite lRA-743 column directly abovea Dowex
50W-X8 column, add exactly 10 mL of 0.3 M NaOH solution. Col-
lect the eluent from the lower column in a 50-mL polyethylene
beaker.Sodiumhydroxidereadily elutesthe BF4" from the weakbase
exchangegroupsof the Amberlite lRA-743 columns.As the solution
passesthrough the strong-acidcolumn, excesssodium hydroxide is
removed,excessfluoride is decreasedby precipitationas CaFz, and
the remainingrand BF4" are convertedto the acids. Finally, deter-
mine the solution'sconcentrationof BF4" potentiometrically.
2. The capacityof the Amberlite lRA-743 to adsorbB from acid solu-
tion is considerablylessthan its capacityto adsorbB from neutralor
alkaline solutions. Therefore,for acid solutions:Place the BioRex
70 resin column immediatelyabovethe Amberlite lRA-743column,
and addan aliquot containing1 to 500 Ilg of B of the acid solution
to the upper column. After the solution has drained through both
columns,add three5-mL aliquotsof distilled waterto the uppercol-
umn. After drainageceasesfrom the columns, remove the BioRex
70 column, and wash the Amberlite lRA-743 column consecutively
with 2 mL of 3 M Na40H solution and 3 mL of distilled water. The
B containedin the Amberlite lRA-743 column is convertedto BF4"
on addition of 2 mL of 10% HF solution to the column. Ten minutes
after the HF treatment,pass5 mL of distilled water through the col-
umn. At this stage,the Amberlite IRA-743 column retainsboth BF4"
and r.
Elute BF4" from the Amberlite lRA-743column by placing the
column immediatelyabovethe Dowex SOW-X8 column and passing
10.0 mL of 0.3 M NaOH solution through both as describedabove
for neutral and alkaline solutions.Collect the eluentfrom the lower
column in a 50-mL polyethylenebeaker,and determinethe concen-
tration of BF4" potentiometrically.
BORON 623
B. Regenerationof columns
The resin columnsneedto be regeneratedafter eachrun. Regeneratethe
Amberlite IRA-743 column by washingthe column sequentiallywith 3
mL of 3 M HCl solution, distilled water, 3 M NH40H solution, and dis-
tilled water. Wash the BioRex 70 column with 3 mL of 3 M NH 40H
solution and twice with 5 mL of distilled water. Regeneratethe Dowex
50W-X8 column by passingthrough the column 10 mL of 3 M HCI
solution, 2 mL of distilled water, 10 mL of 0.3 M CaCl2 solution, and
two 10 mL portionsof distilled water.
C. Potentiometricprocedure
1. Calibration curve. Pass20-mL aliquots of the B standardsolutions
(0.10, 0.25, 0.50, 1.0, 10.0, and 50.0 mg mL-l) through the resin
columns, and collect in 50-mL beakersas outlined for neutral and
alkaline solutions in the previous "Procedure"section.Determine
the potential of the individual eluentswith the BFi -specific elec-
trode-referenceelectrode assembly using an expanding scale pH
meter to measuremillivolts. More consistentreadingsare obtained
if solutionsare stirred at a constantrate with a magneticstirrer and
a Teflon-coatedstirring bar. Immediately adjust the potentiometer
assemblyto read +150 m V when testing it againstthe KEF4 refer-
ence solution. The slope of the standardcurve is approximately58
mV for each10-fold changein B concentration.
2. Samplesolutions(eluents).Measurethe potentialof eluentsolutions
from the Dowex 50-W -8X column with the specific electrode
assemblyusing a pH meter equippedfor electromotiveforce (emf)
readingsin millivolts. As describedabove,adjust the potentiometer
to +150 mV when testingit againstthe referenceKEF4 solution.
As temperaturehas an effect on the slope of the calibration
curve, it is desirableto measureunder conditions of constanttem-
perature.If laboratory temperaturesvary more than 2°C, recalibra-
tion will be necessary.
Inductively CoupledPlasmaSpectrometry
Principles
The inductively coupledplasma-atomicemissionspectrometry(ICP-AES)
comprisesthree basic units: the sourceunit [including radiofrequency(RF) gen-
erator,the plasmatorch and gasflow control system,and the sampleintroduction
system],a spectrometerand a computerfor control and data analysis.
An ICP is formed by coupling the energyfrom a RF field to free electrons
in a suitablegas.The gas, usually Ar, is containedin a plasmatorch constructed
from a high-temperatureresistantmaterial(e.g., quartz)that is transparentto the
RF radiation,and is containedin a coppercoil connectedto the RF generator.The
magneticfield is producedaroundthe coppercoil in the upper part of the torch.
The electronsare acceleratedby the magneticfield oscillation and eventually,
some electronsattain energiesequivalentto the ionization potential of the gas.
624 KEREN
Rapidly, an equilibrium is reachedin which the rateof electronproductionis bal-

ancedby lossesdueto recombinationanddiffusion anda stableplasmais formed.
The plasmais effectivley a conductorand is heatedby the flow of currentinduced
by the radiofrequencyfield.
For atomic emissionspectrometry,energy is required from the flame to
excite the ions and atomsin orderto produceatomic emissionspectra(character-
istic of the element)whoseintensity is not only a function of the atomic concen-
tration in the flame but also of the temperatureof the flame. The energyrequired
for the production of atomic emission spectra increasesas the wavelength
decreases.The extremelyhigh temperatureattainedby the radio-frequencyplas-
ma sources,as distinct from chemicalflames, are sufficient to produceemission
spectrawith excellent sensitivityevenfor elementswhosespectrallines lie well
below 200 nm.
The useof ICP-AES for soil analysisis graduallyreplacingatomic absorp-
tion as the preferredtechniquefor the multielementanalysisof solutionsamples.
This methodis accurateand precisebut requiresexpensiveinstrumentation.Soil
extractsin water,aceticacid, or EDTA may be analyzeddirectly. However,when
strongsalt solutionsareusedfor the extraction,(e.g., 1 M NH4Cl or CaClz) prob-
lems with sampleintroductionmay be encountered,necessitatingthe dilution of
the sample,addition of surfactant(e.g., Triton X-100) or use of high salt nebu-
lizers to improve the flow characteristicsof the material (Helmke, 1996, see
chapter 6). Sample preparationproceduresdevelopedfor Atomic Absorption
Spectrometerare readily adaptedfor ICP-AES (Helmke, 1996, seechapter6).
Application for Boron Determination

The ICP-AES is suitablefor B determination.The detectionlimit by ICP-
AES (27 MHz plasma)for B is 6 ng mL-1 (5.5 x 10-7 M) at wavelengthof
249.773nm. Boron can be determineddirectly in the aqueousextract(Aitken et
aI., 1987), and since the coextractedlevels of Fe are low (<10 mg mL-1), there
areno significantinterferences.However,only free colloidal extractsare suitable
for B analysisby ICP-AES technique.
REFERENCES
Adams, R.M. 1964. Boron, metallo-boroncompoundsand boranes.JohnWiley & Sons, Inc., New
York.
Aitken, R.L., A.J. Jeffrey, and B.L. Compton. 1987. Evaluationof selectedextractants forboron in
someQueenslandsoils. Aust. J. Soil Res. 25:263-273.
American Society for Testing Materials. 1978. Annual book of ASTM standards.Part 31. ASTM,
Philadelphia.
Baker, A.S. 1964. Modifications in the curcumin procedurefor the determinationof boron in soil
extracts.J. Agric. Food Chern. 12:367-370.
Basson,W.O., R.G. Bohmer,and D.A. Stanton.1969.An automatedprocedurefor the determination
of boron in plant tissue.Analyst (London) 94:1135-114l.
Berger,K.C., and E. Truog. 1939.Boron determinationin soils and plants.Ind. Eng. Chern.Anal. Ed.
11:540-545.
Berger,K.C., and E. Truog. 1940. Boron Deficienciesas revealedby plant and soil test. J. Am. Soc.
Agron. 32:297-30l.
Berger, K.c., and E. Truog. 1944. Boron tests and determinationfor soils and plants. Soil Sci.
57:25-36.
BORON 625
Biggar, I.W., and M. Fireman.1960.Boron adsorptionand releaseby soils. Soil Sci. Soc. Am. Proc.
24:115-120.
Byrne, R.I., Ir., and D.R. Kester. 1974. Inorganic speciationof boron in seawater.1. Mar. Res.
32:119--127.
Carlson,R.M., andI.L. Paul. 1968.Potentiometricdeterminationof boron as tetrafluoroborate.Anal.
Chern.40:1292-1295.
Carlson,R.M., and I.L. Paul. 1%9. Potentiometricdeterminationof boron in agricultural samples.
Soil Sci. 108:266-272.
Cartwright, B., KG. Tiller, BA Zarcinas,and L.R. Spouncer.1983.The chemicalassessment of the
boron statusof soils. Aust.I. Soil Res.21:321-332.
Dible, W.T., E. Truog, and K.C. Berger. 1954. Boron determinationin soils and plants.Anal. Chern.
26:418-421.
Dyrssen,D., andI. Hansson.1973.Ionic mediumeffectsin seawater.Comparisonof acidity constants
of carbonicacid in sodiumchloride and syntheticseawater.Mar. Chern. 1:137-149.
Edwards,1.0., G.C. Morrison, V.F. Ross,andI.W. Schultz.1955.The structureof the aqueousborate
ion. 1. Am. Chern.Soc. 77:266-268.
Foote,F.1. 1932.Determinationof boron in waters.Ind. Eng. Chern.Anal. Ed. 4:39--42.
Goldman,E., S. Taormina,and M. Castillo. 1975.A modified curcuminmethodfor determiningtrace
amountsof boron.1. Am. WaterWorks Assoc.67:14.
Good,C.D., andD.M. Rilter. 1962.A1kenylboranes:I. Improvedpreparativemethodsandnew obser-
vationson methylvinylboranes.I.Am. Chern.Soc.84:1162-1166.
Gupta,V.C. 1%6. A modified procedurefor the determinationof total boron from soil fused with
sodiumcarbonate.Soil Sci. Soc.Am. 1. 30:655-656.
Gupta,V.C. 1979.Somefactorsaffecting the determinationof hot-water-solubleboron from podzol
soils usingazomethine-H.Can.1. Soil Sci. 59:241-247.
Gupta,S.K, andI.W.B. Stewart.1975.The extractionand determinationof plant-availableboron in
soils. Schweiz.Landwirtsch.Forsch.14:153-169.
Hatcher, I.T., and L.Y. Wilcox. 1950. Colorimetric determinationof boron using carmine. Anal.
Chern.22:567-569.
Helmke, PA 1996. Neutron activation analysis.p. 141-159.In D.L. Sparkset al. (ed.) Methodsof
soil analysis.Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Hingston,F.1. 1964.Reactionbetweenboron and clays. Aust. 1. Soil Res.2:83-95.
Hossner,L.R. 1996. Dissolution for total elementalanalysis.p. 49--64. In D.L. Sparkset al. (ed.)
Methodsof soil analysis.Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and ASA,
Madison,WI.
Ingri, N. 1963.Equilibrium studiesof the polyanionscontainingBill, SilV, GeIV andVV. Svensk,Kern.
TIdskr.75:199--230.
lin, 1.-y., D.C. Martens,and L.W. Zelazny. 1988. Plant availability of applied and native boron in
soils with diverseproperties.Plant Soil 105:127-132.
Iohnson,C.M., and A. Ulrich. 1959. Analytical methodsfor use in plant analysis.California Agric.
Exp. StD. Bull. 766.
Keren,R., and F.T. Bingham.1985.Boron in water, soils and plants.Adv. Soil Sci. 1:229--276.
Keren, R., F.T. Bingham,andI.D. Rhoades.1985a.Plant uptakeof boron asaffectedby boron distri-
bution betweenliquid and solid phasesin soil. Soil Sci. Soc. Am. 1. 49:297-302.
Keren, R., R.T. Bingham,andI.D. Rhoades.1985b.Effect of clay contentin soil on boronuptakeand
yield of wheat.Soil Sci. Soc. Am. 1. 49:1466-1470.
Keren,R., andR.G. Gast.1981.Effectsof wetting anddrying, andof exchangeable cations,on boron
adsorptionand releaseby montmorillonite.Soil Sci. Soc. Am. 1. 45:478-482.
Keren, R., and R.G. Gast. 1983. pH-dependentboron adsorptionby montmorillonite hydroxy-alu-
minum complexes.Soil Sci. Soc. Am. J. 47:1116-1121.
Knoeck, J., and I.K Taylor. 1969. Aqueous boric acid-borate-mannitolequilibria. Anal. Chern.
41:1730-1734.
Konopik, N., and O. Leberl. 1949. Colorimetric determinationof pH in the range of 10 to 15. 1.
Monatsh.Chern.80:420-429.
Krauskopf,KB. 1972.Geochemistryof micronutrients.p. 7-40.In 1.1. Mortvedt et al. (ed.) Micronu-
trients in agriculture.SSSA,Madison,WI.
Kustin, K, andR. Pizer. 1969.Temperature-jump study of the rate and mechanismof the boric acid-
tartaric acid complexation.1. Am. Chern.Soc. 91:317-321.
Leomahardt,H., and A. Weller. 1960. Kinetische protondonatorwirkunghydratisierter kationer.
Naturwissenschaflen 47:58-59.
626 KEREN
Loeppert,R.H., and D.L. Suarez.1996.Carbonateandgypsum.p. 437-475.In D.L. Sparkset al. (ed.)

Madison,WI.
Mair, 1.W., and H.G. Day. 1972. Curcumin method for spectrophotometricdeterminationof boron
extracted from radiofrequency ashed animal tissues using 2-ethyl-l,3-hexanediol.Anal.
Chern.44:2015-2017.
Mesmer,R.E., C.F. Baes,Ir., and F.H. Sweeton.1972.Acidity measurements at elevatedtemperature.
VI. Boric acid equilibria. Inorg. Chern. 11:537-543.
Naftel, 1. 1939. Colorimetric microdeterminationof boron. Ind. Eng. chern.Anal. Ed. 11:407-409.
Nies, N.P., and G.W. campbell.1964. Boron, metalloboroncompounds,and boranes.R.M. Adamson
(ed.) Intersci. Publ., New York.
Odom,I.W. 1980. Kinetics of the hot water soluble boron soil test. Commun.Soil Sci. Plant Anal.
11:759-765.
Onak, T.P., H. Landesman,R.E. Williams, and I. Shapiro.1959.The Bl1 nuclearmagneticresonance
chemical shifts and spin coupling values for various compounds. 1. Phys. Chern.
63:1533-1535.
Owen, B.B. 1934. The dissociation constantof boric acid from 10 to 50°. I. Am. Chern. Soc.
56:1695-1697.
Owen, B.B., and E.I. King. 1943. The effect of sodium chloride upon the ionization of boric acid at
varioustemperatures.1. Am. Chern. Soc. 65:1612-1620.
Parker,D.R., and E.H. Gardner. 1981. The determinationof hot-water-solubleboron in some acid
Oregon soils using a modified azomethine-Hprocedure.Commun. Soil Sci. Plant Anal.
12:1311-1322.
Reardon,E.I. 1976. Dissociationconstantsfor alkali earth and sodium borate ion pairs from 10 to
50°C. Chern. Geol. 18:309-325.
Rhoades,J.D., R.D. Ingvalson, and 1.T. Hatcher. 1970. Laboratory determinationof leachablesoil
boron. Soil Sci. Soc. Am. Proc. 34:871-875.
Richards,L.A. 1949. Filter funnels for soil extracts.Agron. I. 41:446.
Servoss,R.R., and H.M. Clark. 1957. Vibrational spectraof normal and isotopically labeledboric
acid. 1. Chern. Phys. 26:1175-1178.
Shanina,T.M., N.E. Gelman,and V.S. Mikhailovskaya.1967. Quantitativeanalysisof heteroorganic
compounds. Spectrophotometricmicrodetermination of boron. J. Anal. Chern. USSR
22:663-667.
Sims,I.R., and F.T. Bingham.1968. Retentionof boron by layer silicates,sesquioxidesand soil mate-
rials: II. Sesquioxides.Soil Sci. Soc. Am. Proc. 32:364-369.
Singh, K.P., and H. Sinha. 1976. Availability of boron in relation to certain soil properties.J. Indian
Soil Sci. 24:403-408.
U.S. Salinity Laboratory Staff. 1954. Diagnosisand improvementof saline and alkali soils. USDA
Handb.60. USDA, Washington,DC.
Wear, 1.1. 1965. Boron. p. 1059-1063.In C.A. Black et al. (ed.) Methods of soil analysis.Part 2.
Agronomy Monogr 9. ASA, Madison,WI.
Wear, 1.1., and R.M. Patterson.1962. Effect of soil pH and texture on the availability of water-solu-
ble boron in the soil. Soil Sci. Soc. Am. Proc. 26:344-346.
Weller, A. 1957. Protolytischereaktionendesangeregtenacridins. Z. Elektrochem.61:956-961.
Wilcox, L.V. 1932. Electrometrictitration of boric acid. Ind. Eng. Chern.Anal. Ed. 4:38-39.
Wolf, B. 1971.The determinationof boron in soil extracts,plant materials,composts,manures,water
and nutrient solutions.Commun.Soil Sci. Plant Anal. 2:363-374.
Wolf, B. 1974. Improvementsin the azomethine-Hmethodfor the determinationof boron. Commun.
Yermiyahu,U., R. Keren, and Y. Chen. 1988. Boron sorption on compostedorganic matter. Soil Sci.
Soc. Am. 1. 52:1309-1313.
Zachariasen,W.H. 1954. The precisestructureof orthoboricacid. Acta Cryst.7:305-310.
Published 1996
Chapter22
Silicon
R. LEWIS JONES,University of Illinois, Urbana, Illinois
GARY B. DREHER,Illinois State Geological Survey, Champaign, Illinois
SILICON IN SOILS
Metallic silicon (Si) doesnot occur naturally becauseof the oxidation potentials
°
that prevail in the earth'scrust. Exceptfor forming somecompoundswith C, Si
is invariably covalentlyboundwith assilica (SiOz) which comprisesabout600
g kg-1 (280 g kg-1 as Si) of the crust (Condie, 1974). Bowen (1966) gave the
meanSi contentof soils as 330 g kg-I. Shackletteand Boerngen(1984) reported
the arithmeticmeanSi contentof 406 soil and surficial materialsamplescollect-
ed at about 0.2-m depth at sites throughoutthe conterminousUSA was 310 g
kg-I (standarddeviation, 64.8 g kg-I, range 16-450g kg-I). The abundanceof
Si in natureinsuredthat its determinationwas carried outin the earliestcompre-
hensivechemicalstudiesof soils and the weatheredzone.
Silicon occursin a diversearray of primary mineralsamongthe groupsof
neo-, ino-, soro-, cyclo-, tekto-, and phyllosilicates.Weatheringof theseminer-
als producessecondarysilicatesamongwhich the clay mineralsand poorly crys-
talline and amorphoussiliceous compoundsare especially important in soils.
Special extractionschemeshavebeendevisedto analyzefor silicatesfor varying
crystallinity and knowledgeof the concentrationsof thesemineralsor phaseshas
aidedin understandingsoil genesisand the technologyof soils.
Silicic acid is taken up by vascular plants---especiallygrasses-andis
depositedas opal in roots, stems,and leaves(Piperno,1988; Dreeset aI., 1989).
The silt-size fraction of some soils may contain severaltens of gramsof these
opal phytoliths per kilogram of silt, consequentlythis cycling of Si by plants is
very importantto the Si budgetof most terrestrialecosystems.
PRINCIPLESOF ANALYSES
Although very satisfactoryresults for total Si can be obtainedby nonde-

structive methodslike x-ray fluorescencespectroscopy(Karathanasis& Hajek,
Book Seriesno. 5.
627
628 JONES& DREHER
1996, seeChapter7) most analysesare conductedon solutionspreparedby dis-

solving theSi-bearingsolid phases.
Classical Si analyseslike those carried out early in this century by the
Bureauof Chemistryand Soils of the U.S. Departmentof Agriculture were done
gravimetricallyon soil samplesthat were fusedwith NaZC03. The SiOzwas sub-
sequentlybrought into solution and then precipitated,dehydrated,and weighed
(Robinson,1930). At present,NaOH, anotheralkaline flux, is sometimesused.
Both LiBO z (lithium metaborate)and Li zB40 7 (lithium tetraborate)are acidic
compoundsthat easilyfuse silicates.In either case,the finely groundsoil is inti-
mately mixed with the flux and the mixture is fused over a burneror in a muffle
furnace. Clear glassesare formed with the Li fluxes and opaquelight-colored
fusion cakesare formed with the Na fluxes. In eachcase,the fused material is
dissolvedto form a clear solution for subsequentanalysis.
Various extractants,which are used in assessingsoil forming processes,
selectivelydissolve certain siliceoussoil componentsand are often assayedfor
Si. For example,dithionite-citrate-carbonate (DCB) (Weaver et ai., 1968) and
ammoniumoxalateextracts(Wang & Schuppli, 1986) preparedfor Fe determi-
nationsalso are often analyzedfor Si. AIlophane can be estimatedby extraction
of soil with 0.5 M NaOH and subsequentanalysisof Si and AI (Jackson,1974).
To estimatekaolin, anothersplit of the sampleheatedto 823 K, to dehydroxylate
kaolin, is similarly extractedwith NaOH and analyzedfor Si and AI; the differ-
encein Si and AI between383 and 823 K extractsrepresentskaolin. In this pro-
cedure,if the SiOz to AIZ03 ratio is not close to one, adjustmentsfor smectitic
componentsare made.Recently(Kodama& Ross,1991), Tiron(4,5-dihydroxy-
1,3-benzene-disulfonic acid, disodium salt) hasbeenusedto selectivelydissolve
allophaneand opal, which can then be quantitativelyestimatedfrom Si and AI
determinationsof Tiron extracts.
The analysis of the solutions obtained by these procedurescan be con-
ductedby absorptiometricor instrumentalmethods.Absorptiometricprocedures
rely on the fact that silicic acid forms a complexcompoundwith molybdic acid,
most efficiently in the pH range of 1 to 2. This silicomolybdic acid [H4 (Si
M0 120 40)] occurs asa and ~ structuralisomers,which can be reducedto form a
and ~ silicomolybdousacid. The oxidized form is yellow and the reducedform
is blue. The reducedform is more sensitivefor chemicalanalysis.
Iron and P interfere with the formation of the silicomolybdic acid. Iron
may precipitatemolybdatein the procedurefor the preparationof silicomolybdic
acid (a yellow precipitate)or oxidize reductantin the procedurefor the prepara-
tion of silicomolybdousacid (a blue precipitate).Phosphorus formsa complex
with molybdateto form phosphomolybate.The phosphomolybdate complex ab-
sorbslight in the samewavelengthrange as silicomolybdic acid. Both interfer-
ants may be supressedby the addition of tartaric acid.
A wavelengthof 850 nm is normally usedfor the determinationof Si in the
silicomolybdic acid procedure,however,Fe may interfere by absorbinglight at
this wavelength.This interferencemay be avoidedby determiningSi at 650 nm,
althoughat this wavelengththe determinationis lesssensitive(Jeffery & Wilson,
1960).
SILICON 629
Absorptiometricdeterminationof Si in DCB extractsrequirestreatmentsto

destroydithionite and citrate.Weaveret al. (1968) suggestedbubbling air for 4
h throughthe extractto destroythe dithionite and addingammoniummolybdate
at a concentrationof 60 mM L-1 to complex citrate. Weaveret al. (1968) also
found that adding80 mM L-1 of tartaric acid would effectively complex2.5 mM
L-1 of Fe. Yuan and Breland(1969) evaluatedatomic absorptionspectrometry
(AAS) of Si in DCB extractsby adding20 and40 mg L-1 Si to the extract.They
obtainedan averageSi recoveryof 98%.
A numberof standardreferencematerialsis availablefor use as Si stan-
dardsand for laboratoryquality assurance.GladneyandBums(1983) assembled
a large numberof analysesof U.S. GeologicalSurvey rock standardsuseful in
making choicesamongmatricesof materials.Severalstandardreferencemateri-
als (SRM) available from the National Institute of Standardsand Technology
also can serveas standards;theseinclude potashfeldspar(SRM 70a, 314 g Si
kg-l), sodafeldspar(SRM 99a, 305 g Si kg-l), glasssand(SRM 14513,386.9g
Si kg-I), and obsidianrock (SRM 688, 341.5 g Si kg-l). Threesoils certified as
referencematerials,with Si ranging from 158.6 to 257.2 g kg-I, are available
from the CanadaCentrefor Mineral and EnergyTechnology.All of thesestan-
dardsare suitablefor use in Si determinationsdescribedbelow and the analyst
should preferablychooseseveralreferencestandardsin the concentrationrange
anticipatedfor the unknowns.
The analytical chemistry methodsdiscussedin this chapterare lithium
tetraboratefusion followed by AAS or inductively coupled plasma emission
spectrometry(ICPES),acid digestionfollowed by AAS or ICPES,silicomolyb-
dic acid procedurefollowed by light absorptionspectrometry,and silicomolyb-
dousacid procedurefollowed by light absorptionspectrometry.
SamplePreparationfor Total Silicon: Lithium TetraborateFusion
Reagents
All reagentsare reagentgradeor higher quality.
1. Lithium tetraborate(Li2B407)'
2. 5% hydrochloricacid (HCI) solution.
3. Silicon standard,200 mg L-1. Prepareby fusion of spectrographicgrade
silicon dioxide with lithium tetraborate.Silicon standardsprepared
from aqueoussolutionsof sodiumsilicate are unacceptable.
Procedure
The lithium tetraboratefusion procedureis basedon a standardtestmethod
of the American Society for Testing and Materials(1991). Oven-driedsamples
for analysisshouldbe ashedat 823 K before fusion, to removeorganic matter.
Approximately 1 g of finely ground soil is weighedinto a tared porcelaincru-
cible. The crucible plus sampleis placedin a cool muffle furnaceand gradually
heatedto 823 K, then held at 823 K for 2 h or until organicmatteris no longer
630 JONES & DREHER
visible, whicheveris longer. The sampleand crucible are cooled ina desiccator,
then weighed. Grind the sampleto passa 200-meshsieve (less than 74 f.lm) by
using a mullite or agatemortar and pestle.The sampleand crucible are reignited
at 823 K for 1 h, cooledandweighedagain.If desired,the loss on ignition at 823
K may be calculatedby subtractingthe reignitedweight from the original weight.
Thoroughly mix the ignited samplewith a small platinum spatula.
Platinumcruciblesusedin the fusion of samplesshouldbe cleanedby boil-
ing in 5% HCl, then rinsed with deionizedwater. The cleanedcruciblesshould
be protectedfrom subsequentcontaminationby not placing them directly on
benchtops or in other areaswheresilicon may be picked up.
Immediatelyweigh 0.200 g (to the nearest0.0001 g) of the finely ground
(ashed)soil sampleinto a clean35-mL platinumcrucible.Add 0.5 g (to the near-
est 0.01 g) of lithium tetraborateand carefully mix the flux and soil sampleby
using a platinum spatula.Cover the mixture with a second0.5-g portion of lithi-
um tetraborate.Placethe crucible in a muffle furnacethat has beenpreheatedto
1273 K. Allow the crucible to remain in the muffle furnace for 15 min. This
length of time is usually sufficient for completefusion of the sample.However,
if a clear melt is not obtained,further heatingmay be required.
Removethe cruciblefrom the furnace,swirl the melt in the crucible to mix
it, and allow it to cool in air on a clean surfaceto room temperature.Rinse the
outside surfaceof the crucible with deionizedwater to remove any debris that
may be clinging to it, then place the crucible in a clean250-mL beaker.Placea
small, clean tetrafluoroethylene(TFE)-fluorocarbon-coatedstirring bar in the
bottomof the crucible. Add 150 mL of 5% HCI to the beakercontainingthe cru-
cible and place the beakeron a stirring hot plate. This volume of acid solution
shouldbe enoughto flood the crucible. Cover the beaker,heatto just below boil-
ing temperature,and stir constantly.Heat the beakeruntil all the fusion cakehas
completelydissolvedfrom the crucible(usually lessthan about30 min). Remove
the beakerfrom the hot plate and allow it to cool to room temperature.
Remove the stirring bar from the crucible with a magneticretriever and
rinse the bar with a small quantity of 5% HCI. Catch the rinsate in the beaker.
Remove the crucible from the beaker by means of platinum-tipped crucible
tongs. Invert the crucible over the end of a TFE-fluorocarbonstirring rod above
the beaker;thoroughly rinse the crucible, inside and outside,with small amounts
of 5% HCI. Catch the rinsate in the beaker.Quantitativelytransferthe solution
from the beakertoa 250-mL volumetric flask. Dilute to the mark with 5% HCI.
This is the stock samplesolution. Transfer10 mL of the stock samplesolution
into a 100-mL volumetric flask using a volumetric pipet and dilute to the mark
with 5% HCI.
Preparea primary Si stock standardsolution by following the samefusion
procedureusing 0.1070g of reignitedspectroscopicgradeSi02• Dilute the stock
standardsolution to 250 mL, this solutioncontains200 mg Si L-l. Prepareanaly-
sis standardsolutionsfor AAS or ICPES by diluting aliquots of the stock stan-
dard solution to provide a seriesof calibration standardswith concentrationsin
the rangeof 4 to 50 mg L-l. Preparea blank solution using the aboveprocedure,
but with neithersamplenor silica.
SILICON 631
Atomic Absorption Spectrometry
Use a silicon hollow-cathodelamp and set the atomic absorptionspec-

trometerat 251.6nm. Use a fuel-rich nitrous oxide/acetyleneflame. Adjust other
instrumentparametersaccordingto the instrumentmanufacturer'sinstructions.
Follow the instrumentmanufacturer'sinstructionsfor the safeuseof the nitrous
oxide/acetyleneflame. Calculate the concentrationof Si02 and report on the
basisof the unashed,oven-driedsoil.
Inductively CoupledPlasmaEmissionSpectrometry
Silicon is readily determinedin plasmaemissionspectrometryat a wave-
length of 288.158nm. Spectralinterferencesfrom other elementsshouldbe neg-
ligible becauseof the high concentrationof Si in soils. Follow the instrument
manufacturer'sinstructions for operating the instrument and for any recom-
mendedsafety practices.Calculate the concentrationof Si02 in the unashed,
oven-driedsoil.
Comments
The lithium tetraboratefusion procedureis well suitedfor many soils and
rocks. However, if erratic results are obtained, lithiummetaborateor a mixture
of one part lithium metaborateto two parts lithium tetraboratemay be used
(Shapiro,1975). Silicon in solutionspreparedafter lithium tetraboratefusion is
more stablethan when preparedafter lithium metaborate fusion,and may be sta-
ble for up to severalmonths(Brown et aI., 1969).The solutionobtainedfrom this
proceduremay be usedto determineother major and minor elementsin the soil
sampleby either AAS or ICPES. However, before acceptingresultsof analysis
for trace elementsbasedon this procedure,the analyst must check the lithium
tetraborate and/or lithium metaboratefor the presenceof tracecontaminants.As
part of laboratory quality assurance,samplesof known compositionshould be
preparedand analyzedin the samemanneras unknowns.
Advantages of Lithium Tetraborate Fusion

1. Most soil mineralsare readily dissolvedby lithium tetraborate.
2. Fusion solutions canbe usedfor the determinationof other major and
minor elementsin soil-forming minerals.
Disadvantages of Lithium Tetraborate Fusion

1. Possibleformation of Si02 gel would result in a negativeerror in solu-
tions containinghigh concentrationsof Si. Govett (1961) indicatedthat
Si02 may form in aqueoussolutions containing Si at concentration
greaterthat 110 to 140 mg L-I. The pH of the solution shouldbe main-
tainedundereithervery acidic (pH 1-3) or very alkaline conditions(pH
> 9) to preventthe formation of Si02 gel (Govett, 1961).
632 JONES& DREHER
2. Possibleintroduction of trace contaminantsby the flux. This could be

importantif the analystintendsto use the fusion solution for trace ele-
ment determinations.
SamplePreparationfor Total Silicon: Acid Digestion
Reagents
1. Aqua regia. Mix one part concentratednitric acid (HN03) with three
partsconcentratedHCI. Preparefresh before eachacid decomposition.
2. Hydrofluoric acid (HF), 48%.
3. Boric acid (H3B03) solution. Dissolve 50 g crystalline H3B03 in 1 L
deionizedwater.
Procedure
The acid digestion procedureis basedon methodspublishedby Bernas
(1968), Ruch et al. (1974), and Harvey et al. (1983). Refer to the previoussec-
tion on "Procedure"for the removalof organicmatterfrom the soil. Weigh 0.100
g (to the nearest0.0001 g) of finely ground,ashedsoil sampleinto the TFE-flu-
orocarbonliner of a sampledigestion bomb (Parr InstrumentCo., Moline, IL
61265)1.Use plastic pipetsto add 1.5 mL of aquaregia and 2.5 mL of HF to the
sample.Be careful to wet the entire sample.Closethe bomb and heat for 2 h at
373 K to 383 K. Allow the bomb to cool, then carefully open it and add 25 mL
of H3B03. Heat the sampledigest(not abovethe boiling point), if necessary,for
10 to 15 min after addition of the H3B03, to aid in the dissolutionof any result-
ing precipitate.Somedark-colored,insolubleorganicresiduethat escapedashing
may remain. Transferthe contentsof the TFE-fluorocarbonliner quantitatively
to a 100-mL plastic volumetric flask, dilute to the mark with deionizedwater,
then transferthe solution to an acid-washed125-mL high-densitypolyethylene
bottle. Dilute 10 mL of this solution with deionizedwater to 100 mL. Store the
diluted samplein an acid-washedhigh-densitypolyethylenebottle.
Preparea blank solution using the samemethodas for samples.Ignite 0.2
g of spectroscopicgradeSi02 at 1273 K for 30 min. Preparea stock standardSi
solution by dissolving0.1070g of ignited Si02 in an acid decompositionbomb
as describedabove. Dilute the stock standardsolution to 100 mL, as described
above.This solution contains500 mg Si L-1. Dilute aliquots of the stock stan-
dard solution to provide a seriesof calibration standardswith concentrationsin
the rangeof 4 to 50 mg L- 1.
Atomic Absorption Spectrometry

DetermineSi by AAS using a fuel-rich nitrous oxide/acetyleneflame and
a wavelengthof 251.65nm. Adjust the otherinstrumentparametersaccordingto
I The use of trade namesin this report is for descriptivepurposesonly and does not constitute
endorsementby either the University of Illinois or the Illinois StateGeologicalSurvey.
SILICON 633
the manufacturer'sinstructions.Follow appropriatesafetyprecautionsin using a

nitrous oxide/acetyleneflame. Calculate the concentrationof Si02 in the un-
ashed,oven-driedsoil.
Inductively CoupledPlasmaEmissionSpectrometry
Silicon is determinedby ICPESat 288.158nm. Spectralinterferencesfrom
other elementsshould be negligible, becauseof the usually high concentrations
of Si found in soils. Follow the instrumentmanufacturer'sinstructionsfor oper-
ating the instrumentand follow any recommendedsafety precautions.Calculate
the concentrationof Si02 in the unashed,oven-driedsoil.
Comments
Hydrofluoric acid causesextremelypainful, slowly healingburnsto tissue.
Exerciseextremecaution in working with HF and all work should be done in a
fume hood. Always wearglovesthat are imperviousto HF, and gogglesor a face
shield when working with this or any other acid. Checka chemicalcompatibili-
ty chart for properglove selection.
Boric acid solution is addedto the samplesolution to dissolveprecipitated
fluorides and to complexexcessfluoride as BF4. If free HF is allowed to remain
in the samplesolution, it will be volatilized during analysisby either AAS or
ICPES and may attack the burner or torch parts, the flame or torch ventilation
system,and/or be lost to the laboratory air, where it may attack surfacesin the
laboratory or harm individuals in the lab either by inhalation or by skin or eye
contact.
Silicon forms volatile silicon tetrafluoride(SiF4) with HF. The loss of SiF4
from solution presentsa possiblesourceof negativeerror if the bomb is opened
before its contentshavecooledto room temperatureor if the soil sampleis dis-
solved in an open container.
Tightly capped60-mL high-density polyethylenebottles have been suc-
cessfully substitutedfor the TFE-fluorocarbon-linedacid decompositionvessels
at temperaturesbelow 403 K. If high-densitypolyethylenebottlesare used,they
should be heatedin an ordinary water bath (Langmyhr & Paus,1968) or on a
steambath (Harvey et aI., 1983). Follow the sameprocedureas abovefor addi-
tion of H3B03 solution and dilution. Decomposition methods using vessels
designed for microwave heating and suitable for soil materials have been
describedby Kingston and lassie(1988).
One type of soil that might requirespecialtreatmentin the acid dissolution
methodis calcareoussoil. A weighedsampleof this soil shouldbe initially treat-
ed with dilute HCI to destroy CaC03 and CaMg(C03h- The sampleshould be
takento drynesswithout baking the sample,then it may be treatedin the manner
describedfor the HF-aquaregia dissolution.As part of laboratoryquality assur-
ance, samplesof known composition should be preparedand analyzedin the
samemanneras unknowns.
Advantages of Acid Digestion

1. The dissolvedsolids concentrationafter acid digestionof soil is low.
634 JONES& DREHER
2. Contaminationof the samplesolution by reagentsis low.

3. Solutionspreparedby acid digestionmay be usedfor determinationsof
traceelementsin soils.
Disadvantages ofAcid Digestion
1. Silicon may be volatilized as SiF4 (would result in a negativeerror).
2. Somerock-forming minerals,notably zircon, are not readily solublein
HF (would result in a negativeerror).
3. If all excessHF is not complexedby H3B03, the remainingfree HF may
attack a glass or quartz nebulizeror torch (would result in a positive
error).
SILICON DETERMINATION BY LIGHT ABSORPTION

SPECTROMETRY
Yellow SilicomolybdicAcid Procedure
Reagents
1. Ammonium molybdatetetrahydrate[(NH4)6 M070 24 • 4H20]. Dissolve
54 g in 750 mL of deionizedwater then adjust pH to ~7 with NaOH,
dilute to 1 L and store in a high-densitypolyethylenebottle. The solu-
tion is 0.3 M with respectto MoO]-.
2. Tartaric acid (C4H60 6) solution, 20%. Dissolve 100 g tartaric acid in
waterand dilute to 500 mL. Storein a high-densitypolyethylenebottle.
3. Reducingsolution. Dissolve25 g of sodiumbisulfite (NaHS03) in 200
mL water. Dissolve 2 g of sodium sulfite (Na2S03) and 0.4 g of 1-
amino-2-naphthol-4-sulfonic acid [NH2ClOHs(OH)S03H]in 25 mL of
water. Mix solutionsand dilute to 250 L; store in a high-densitypoly-
ethylenebottle, and refrigerate.
4. Silicon standard,50 mg L-l. Fuse 0.1070 g of spectrographicgrade
Si02with NaOH. Dissolvethe fusion chargein 700 to 800 mL of deion-
ized water (the exactamountof water is not important) and then add
sufficient 0.5 M sulfuric acid (H2S04) to give a pH of 1.5. Dilute to 1 L
in a volumetricflask. Storein a high-densitypolyethylenebottle.
Procedure
The yellow silicomolybdic acid procedure is based on the method of
Govett (1961). Ash the sampleto removeorganicmatter(seethe first sectionon
"Procedure").Weigh 0.100g (to the nearest0.0001 g) into a 50-mL nickel cru-
cible. Add 3 g NaOH, mix well. Placethe crucibleover a bunsenburnerandheat
to a dull rednessfor at least 10 min. After cooling, transferthe crucible and its
contentsto a polyethylenebeakerand addabout 800 mL of deionizedwater.
Allow to standfor at least1 h. Stir the contentsof eachbeakerwith a plasticstir-
ring rod or a TFE-fluorocarbon-coated stirring bar until the fusion chargeis dis-
SILICON 635
solved.Removethe stirring bar andcrucibleand rinse themwith deionizedwater

into the beaker.Add sufficient 0.5 M H2S04 to adjust the pH of the solution to
1.5. Quantitatively transferthe solution to a l-L volumetric flask and dilute to
volume with deionizedwater. Transfera volume(~20 mL) of the preparedsolu-
tion to a 50-mL volumetric flask, so that the volume transferredcontainsless
than 0.4 mg Si. Add 10 mL 0.5 M H2S04 to the volumetricflask, followed by 10
mL ammoniummolybdatesolution, and then 10 mL of tartaric acid solution.
Mix thoroughly and dilute to the mark without delay. Determineabsorbanceof
the samplesolution at 400 nm between2 and 10 min after additionof the ammo-
nium molybdatesolution. Preparea standardcurve to 8 ~g Si mL-l.
Comments
The gram-equivalentsof W should be kept betweenthree and five per
gram-ionof MoOl- and the total ionic strengthshouldbe lessthan 0.5 to prevent
formation of the a.-form of silicomolybdic acid (Strickland, 1952). The a.-form
is lesssensitiveand is more stablethan the ~-form. It forms readily at a pH high-
er than about 2.5 or in excesselectrolyte(Strickland, 1952). Samplesprepared
by sodiumhydroxide(NaOH) fusion shouldbe analyzedwithin severaldays.For
purposesof quality assurance,samplesof known compositionshould be pre-
paredand analyzedin the samemanneras unknowns.For analysesof extractsit
is advisableto spike severalsampleswith Si to evaluaterecovery.
Blue SilicomolybdousAcid Procedure
Procedure
This procedureis basedon methodspublishedby Shapiroand Brannock
(1962) and Weaveret al. (1968).
Use the sameprocedureas in the yellow silicomolybdic acid procedure
(seethe sectionon "Yellow SilicomolybdicAcid Procedure")for fusion and dis-
solution of the sample.Transfera portion of the preparedsolution to a 50-mL
volumetricflask so that the volumecontainslessthan 0.02 mg Si. Add 10 mL of
0.5 M H2S04, and 10 mL of ammonium molybdate solution,and mix by swirling
the contentsof the volumetric flask. After 2 min add 5 mL of tartaric acid and 5
mL of reducingsolution, mix without delay, and then dilute to the 50 mL mark.
Becauseof the instability of the samplematrix, a matrix blank should be ana-
lyzed, however, standardsand samplesshould be determinedvs. a deionized
water blank (Jeffery & Wilson, 1960). Correction for the absorbanceof the
reagentblank should be made in the final calculations. Measure absorbance
againstthe deionizedwater blank at 820 nm at least30 min after addition of the
ammoniummolybdatesolution. Preparea standardcurve to 0.4 ~g Si mL-l.
Comments
As in the molybdic acid procedure,it is importantto analyzesamplespre-
paredby NaOH fusion within 2 d and it also is advisableto spike randomsam-
ples with known quantitiesof Si to evaluaterecovery.A numberof reductants
636 JONES & DREHER
have beenusedincluding stannouschloride, hydroxylamine,and ascorbicacid;

however, the mixed reagent with l-amino-2-napthol-4-sulfonicacid, listed
amongthe reagents,producesthe moststablecolor. Interlaboratorystandardsfor
Si shouldbe carriedthroughthe procedurefor quality assurance.
REFERENCES
American Society for Testing and Materials. 1991. Standardtest methodfor major and minor ele-
mentsin coal and coke ash by atomic absorption,Method D 3682-87.p. 369-374.In RA
Storer(ed.) Annual Book of ASTM Standards,GaseousFuels,Coal and Coke. Version 5.05.
ASTM, Philadelphia,PA.
Bowen, HJ.M. 1966. Traceelementsin biochemistry. Acad.Press,New York.
Brown, D.F.G., AM. MacKay, and A. Turek. 1969. Preparationof stablesilica standardsolutionsin
rock analysesusing lithium tetraborate.Anal. Chern.41:2091.
Condie, K.C. 1974. Abundancein igneous rocks and in the crust. p. 14-E-1 to 14-E-4. In K.H.
Wedepohl(ed.) Handbookgeochemistry.Vol. 2. Springer-Verlag,Berlin.
Drees,R, L.P. Wilding, N.E. Smeck,and AL. Senkayi.1989.Silica in soils: Quartzand disordered
polymorphs.p. 913-976.In I.B. Dixon and S.B. Weed (ed.) Minerals in soil environments.
SSSABook Ser. 1. SSSA,Madison,WI.
Gladney, E.S., and C.E. Bums. 1983. Compilation of elementalconcentrationsin eleven United
StatesGeologicalSurvey rock standards.Geostand.Newsl. 7:3-226.
Govett, GJ.S. 1961. Critical factors in the determinationof silica. Anal. Chim. Acta 25:69-80.
Harvey, RD., RA Cahill, C.-L. Chou, and I.D. Steele.1983. Mineral matterand trace elementsin
the Herrin and Springfield coals, Illinois Basin coal field. Illinois State Geol.Surv.
Contract/GrantRep. 1983-4.
Iackson, M.L. 1974. Soil chemical analysis-advancedcourse. 2nd ed. Dept. Soil Sci., Univ.
Wisconsin,Madison.
Jeffery, P.G., and AD. Wilson. 1960.A combinedgravimetricand photometricprocedurefor deter-
mining silica in silicate rocks and minerals.Analyst (London) 85:478-486.
Karathanasis,AD., and B.F. Hajek. 1996. Elementalanalysisby x-ray fluorescencespectroscopy.p.
Kingston, H.M., and L.B. lassie(ed.). 1988. Introduction to microwavesamplepreparation:Theory
and Practice.Am. Chern.Soc. Prof. ReferenceBook. ACS, Washington,DC.
Kodama, H., and GJ. Ross. 1991. Tiron dissolutionmethodusedto removeand characterizeinor-
ganic componentsin soils. Soil Sci. Soc. Am. I. 55:1180-1187.
Langmyhr,FJ., and P.E. Paus.1968. The analysisof inorganicsiliceousmaterialsby atomic absorp-
tion spectrophotometry andthe hydrofluoric acid decompositiontechnique.Anal. Chim. Acta
43:397-408.
Piperno,D.M. 1988. Phytolith analysis.Acad. Press,San Francisco,CA
Robinson,W.O. 1930. Method and procedureof soil analysisusedin the Division of Soil Chemistry
and Physics.USDA Circ. 139. USDA, Washington,DC.
Ruch, RR, HJ. Gluskoter, andN.F. Shimp. 1974.Occurrenceand distribution of potentially volatile
traceelementsin coal: A final report. Illinois StateGeol. Surv. Environ. Geol. Note No. 72.
Shacklette,H.T., and I.G. Boerngen.1984. Elementconcentrationsin soils and other surficialmate-
rials of the conterminousUnited States.U.S. Geol. Surv. Prof. Pap. 1270. U.S. Gov. Print.
Shapiro, L. 1975. Rapid analysisof silicate, carbonate,and phosphaterocks. Rev. ed. U.S. Geol.
Surv. Bull. 1401. U.S. Gov. Print. Office, Washington,DC.
Shapiro,L., and W.W. Brannock. 1962. Rapid analysisof silicate, carbonateand phosphaterocks.
U.S. Geol. Surv. Bull. 1144-A U.S. Gov. Print. Office, Washington,DC.
Strickland,I.D.H.1952.The preparationand propertiesof silicomolybdicacid. III. The combination
of silicate and molybdate.I. Am. Chern.Soc. 74:872-876.
Yuan, T.L., and H.L. Breland. 1969. Evaluationof atomic absorptionmethodsfor determinationsof
aluminum,iron, and silicon in clay and soil extracts.Soil Sci. Soc. Am. Proc. 33:868-872.
SIUCON 637
Wang,C., andP.A. Schuppli.1986. Determiningammoniumoxalate-extractable Si in soils. Can.1.

Soil Sci. 66:751-755.
Weaver,R.M., J.K. Syers,and M.L. Jackson.1968. Determinationof silica in citrate-bicarbonate-
dithionite extractsof soils. Soil Sci. Soc.Am. Proc. 32:497-501.
Published 1996
Chapter23
Iron
RICHARD H. LOEPPERT,Texas A&M University, College Station, Texas
WILLIAM P. INSKEEP,Montana State University, Bozeman, Montana
Total soil Fe concentrationsvary widely and rangefrom <1 % to >20%; the medi-
an concentrationis approximately3% (Murad & Fischer,1988). Under earthsur-
face conditions,Fe may exist in either the Fe2+ (ferrous)or Fe3+ (ferric) oxidation
state.Although Fe is an abundantelementin primary and secondarymineralsin
soils, it's low availability frequently limits plant growth, especiallyin alkaline
and calcareoussoils, becauseof the low solubilities of the Fe-containingsec-
ondaryminerals(Lindsay & Schwab,1982; Lindsay, 1984). In low pH soils and/
or reducingconditions,soluble forms of Fe can be presentin sufficient concen-
trationsto be toxic to plants.
The objective of this chapteris to summarizemethodologiesfor the deter-
mination of total soil Fe (fusion and digestion procedures),forms of soil Fe
(selectiveextraction procedures)and Fe bioavailability, and for the analysisof
total dissolvedFe, Fe2+ and Fe3+ in soil digestsand extracts.Thesediscussions
will be precededby a summaryof the principal forms of soil Fe and their prop-
erties,especiallywith regardto thosefactorswhich influencebioavailability. The
researcheralso is referredto previousreviews by Olson (1965), Olson and Ellis
(1982), and Loveland(1988).
PRINCIPAL FORMS OF SOIL IRON
The predominantiron oxides in the soil are hematite,goethite, lepidocro-

cite, magnetite,maghemiteand ferrihydrite (Schwertmann,1988; Schwertmann
& Taylor, 1989). Each of theseoxides is an Fe3+-containingmineral, exceptfor
magnetitewhich containsboth Fe2+ and Fe3+, and eachexists predominantlyin
the clay-sizefraction of the soil (exceptfor magnetitewhich can occur predomi-
nantly in the silt- and sand-sizefractions).The soil iron oxide mineralscanexhib-
it considerablesubstitutionof other ions, e.g., Al or Mn, for Fe in the structure.
Goethiteand hematiteare the dominantmineralsin well-oxidizedor arid or semi-
arid environments,and lepidocrocite is found predominantlyin hydromorphic
Book Seriesno. 5.
639
640 LOEPPERT& INSKEEP
environments.Magnetiteis most commonin reducedor slightly-weatheredsoils.

Ferrihydriteis likely to exist in most soils, thoughusually in small quantities,but
is frequently a major componentin soils exhibiting fluctuating redox potentials.
Each of the soil iron oxide mineralsexists in relatively well-orderedcrystalline
forms in the soil, exceptfor ferrihydrite which is more highly hydrated,with only
short-rangecrystallineorderand with particle sizesof 10 nm or less.Ferrihydrite
often exists in associationwith layer silicates, amorphoussilicates and organic
matter. Positive identification of the predominantFe oxides is accomplishedby
powder x-ray diffraction, infrared spectroscopyand mossbauerspectroscopy
(Schwertmann& Taylor, 1989).Identification is facilitated by isolation of the
clay-sizecomponentsby particle-sizefractionationprocedures,followed by mag-
netic separationof the iron-containingminerals(Schulze& Dixon, 1979). Posi-
tive identification of ferrihydrite in the soil is especiallydifficult due to its usual-
ly low concentrationand poor crystallinity, though selective-extractionproce-
duresindicatethat ferrihydrite or a ferrihydritelike phasemay be presentin most
soils. The relative stabilities of the soil iron oxide mineralsdecreasein the fol-
lowing order (Schwertmann,1991)
hematite= goethite:>lepidocrocite= magnetite:>ferrihydrite.
The major factor which influencesthe bioavailability of a soil iron oxide is its rel-
ative easeof dissolution,which is influencedby the particlesizeand surfacereac-
tivity of the mineral phase.Kinetic studieshaveindicatedthat ferrihydrite, due to
its small particle size, high surfaceareaand high surfacereactivity, is consider-
ably more reactivethan equivalentquantitiesof goethiteor hematite.Ferrihydrite
and similar poorly crystallineiron oxidesplaya dominantrole in influencing the
availability of Fe to plants in oxidized calcareoussoils (Loeppert & Hallmark,
1985; Morris et aI., 1990).
Iron is presentas a structuralcomponentof layer silicatesin the soil (2~50
g kg-1 is common),but this Fe is largely unavailablefor plant growth. Iron also
may exist as an exchangeion on layer-silicatesurfaces,but in appreciablequan-
tities only in low-pH «4.5) or low-redox conditions.
Iron is a componentof other primary or secondaryminerals, e.g., pyrox-
enes,amphiboles,pyrite and siderite.The occurrenceof eachof thesemineralsis
usually restrictedto reducedor relatively less-weathered soils. Pyrite (FeS2)is an
importantsourceof Fe in acid mine drainage,wherethe oxidation of Fe2+ and S-
resultsin high concentrationsof dissolvedFe3+ and sOi-. The Fe3+ releaseddur-
ing FeS2oxidationin theseenvironmentsoften forms secondaryminerals,includ-
ingjarosite[KFe3(S04h(OH)6]andferrihydrite (Nordstrom,1982; van Breeman,
1988). Siderite (FeC03) is sometimesimportant in reducedenvironments(e.g.,
submergedsedimentsor flooded soils) where Fe2+ is stablerelative to Fe3+ and
concentrationsof CO2(g) are high (Lindsay, 1979).
Complexationof Fe by organic matter, e.g., humic substances,in soils is
highly influencedby pH and redox potential.At low pH «5.0), stablecomplex-
esof organicmatterwith Fe, especiallyFe3+, are likely to exist (Goodman,1987).
At pH valuesgreaterthan 6.0, Fe3+-humic complexesbecomelessimportantdue
to hydrolysisof Fe and precipitationof iron oxides.In calcareoussoils, wherethe
IRON 641
pH of the bulk soil solution is usually within the rangeof 7.5 to 8.5, Fe3+ is not
readily retainedby soil humatesagainsthydrolysis; however,in microenviron-
mentsof reducedpH and redox potential,e.g.,in zonesof active microbial activ-
ity, complexationof Fe by organic mattercan be appreciable.Also, in the rhizo-
sphere,complexationof Fe by soluble organicscan be appreciabledue to the
action of plant-andmicrobial-siderophores (chelators),someof which will retain
Fe against hydrolysis at pH values >7.5. In addition, high concentrationsof
organic acidssuchas oxalateand citrate in the rhizosphereof someplant species
can be importantin the complexationof Fe3+ (Inskeep& Comfort, 1986). These
observationsall point to the fact that Fe concentrationscanbe considerablyhigh-
er in the rhizospherethan in the bulk soil solution.
The concentrationof solution-phaseFe is controlled by pH, redox poten-
tial, the concentrationof water-solubleiron-col11plexingagents,the solubility of
soil iron-oxide phases,and the kinetics of dissolutionand precipitationof these
phases(Lindsay, 1979). In the pH rangeof most calcareoussoils (approximate-
ly 7.5-8.5),the concentrationof solution-phaseFe3+ is at an approximatemini-
mum (Lindsay, 1979). In the absenceof organic complexing ligands, the total
dissolvedFe3+ concentrationis approximately10-10 M, which is considerably
less than the approximately 10-7.7 M concentrationthat is required for plant
growth in nutrient culture (Lindsay & Schwab,1982; Lindsay, 1984). The equi-
librium total Fe concentrationof the soil solution increaseswith decreasingre-
dox potential, due to the increasedconcentrationof Fe2+ speciesin solution.
Lindsay and Schwab(1982) calculatedthat for Fe in solution to exceedthe crit-
icallevel for plantsthe redox potentialmust drop below a pe + pH level of 9.75.
Most calcareoussoils havebulk soil solutionpe + pH valuesconsiderablygreater
than 9.75; therefore,except under conditions of flooding or saturationof soil
poresfor extendedperiods,it is likely that the total dissolvedFe concentrationof
a bulk soil is insufficient to meet the immediatenutritional needsof most plant
species.Plantsand microorganismshave evolved mechanismsof increasingthe
solubility and availability of Fe in the rhizosphere,by exudationof H+, organic
acids, reducingagentsand phytosiderophores.
TOTAL IRON
Introduction
Sincesoil Fe is presentpredominantlyin the form of iron oxidesor a com-

ponentof solid silicate, phosphate,carbonateor sulfide phases,the quantitative
determinationof total soil Fe usually involves decompositionof the soil matrix
followed by dissolutionof the Fe, prior to chemicalanalysis.The most common-
ly usedproceduresfor decompositionof the soil matrix are Na2C03or Li meta-
borate(LiB0 2) fusion or acid digestionwith either HF or an HFIHCIOJH2S04
mixture (Loveland, 1988). When only Fe is to be determined,the acid digestion
proceduresare usually preferredsince they are simpler and errors due to loss of
sampleare lesslikely to occur. The disadvantageof acid dissolutionprocedures
is that concentratedacidsare used,for which the researchershouldbe thorough-
ly familiar with hazardsand necessaryprecautions(Hossner,1996, seeChapter

3). Iron solubilizedby either methodcan then be determinedby atomic absorp-
tion spectroscopy(AAS) or inductively coupledplasmaatomic emissionspec-
troscopy (ICPAES), or colorimetrically (most commonly by formation of the
Fez+-phenanthroline complex).The final choiceof procedurefor decomposition
of the soil matrix and Fe analysismay dependprimarily on availability of equip-
ment. Soils for total elementalanalysisshouldbe finely ground, to passa 0.15-
mm pore size (approximately100 mesh in.-1) sieve, to ensureadequaterate of
reactionduring the digestionor fusion process.
Iron may exist in either the 2+ or 3+ oxidation states;therefore,a modifi-
cation of the procedurefor total analysiscan involve determinationof Fe2+ and
Fe3+. This modification usually involves the determinationsof Fe2+ and total Fe;
concentrationof Fe3+ is then determinedby difference.Specialprecautionsare
usually required,becausesome samplepretreatment(e.g., storage,drying) and
fusion or digestion proceduresmay precludean accuratedeterminationof the
original concentrationsof Fez+ and Fe3+ (seeLoveland, 1988).
Decompositionof Sample(SodiumCarbonateFusion)
Principles
Fusion with NazC03 is useful when the concentrationsof other elements
(e.g., Al, Ca, Mg, Mn and Si) in addition to Fe are being determined.The
NazC03 fusion reaction will result in the transformationof Fe to ionic forms
which are totally solublein hydrochloricacid (HCI). The experimentershouldbe
thoroughlyfamiliar with the careand maintenanceof platinum crucibles,which
are utilized in this procedure(Jackson,1958; Lim & Jackson,1982). Graphite
cruciblesare often preferredsincethe melts do not adhereto the crucible walls.
Although they are cheaperthan platinum crucibles,graphitecruciblesmay last
for only 6 to 10 digestions.
SpecialApparatus
1. Platinumcrucible, 30-mL capacity,with lid.
Reagents
1. Sodiumcarbonate,anhydrous.
2. Hydrochloric acid, 6 M.
3. Hydrochloric acid, concentrated,12 M.
Procedure
Place1 g of soil, which has beengroundto passthrough a 100 meshin.-1
(0.15-mmnominal pore size) sieve, into a 15- to 30-mL capacityplatinum cru-
cible (Jackson,1958). Cover the crucibleloosely, with the platinum coverslight-
ly ajar. Heatthe crucibleslowly, to preventa suddenignition of soil organicmat-
ter, with a Mekker burner to about 900°C, and maintain the heat for approxi-
mately 30 min. Cover the crucible, and allow it to cool in a desiccator.
IRON 643
Add approximately5 g of anhydrousNazC03 to the previouslyignited soil,

and mix the contentsthoroughly. Cover the crucible loosely and heat it slowly,
to preventunnecessarysplattering,for 10 min; then increasethe intensity of the
heat until the crucible bottom is red, and maintain this temperaturefor 15 to 20
min. Remove the cover and continue heating for a few additional minutes to
completethe fusion. Discontinueheating, and while the sampleis still molten,
swirl the crucible to distribute the cooling material along the sides of the con-
tainer. Allow the sampleto cool completely,and then placeit in a Pyrex beaker.
The formation of bubbles or small cracks during the initial cooling suggests
incompletefusion. If this occurs,remelt the sampleand continueheatingat a suf-
ficient temperatureto achieve a red crucible bottom for an additional 15 min.
Allow the crucible to cool to room temperature.
Removalfrom the crucible of a samplecontaininga high amountof silica
can be facilitated by addinga sufficient quantity of hot deionizedwater to loosen
the cooled sampleprior to placing it in the beaker.Add water to the crucible to
facilitate the completetransferof sampleto the beaker.Add approximately5 mL
of 6 M HCI to the crucible, and heatslowly to disintegratethe remainingsample.
Samplesthat are difficult to remove may be transferredby placing the cooled
crucibleon its side in the beakerand addingsufficient water to cover the sample;
the crucible is then heatedslowly to loosenthe sampleand rinsed with water to
remove the remaining sample.The whole sample is dissolvedby adding con-
centratedHCI slowly to the beaker,while using care to preventexcessiveeffer-
vescence.The dissolvedFe is then brought to a known volume and analyzedby
AAS, ICPAES or a colorimetric procedure.
Decompositionof Sample(Hydrofluoric AcidlPerchloricAcid/

Sulfuric Acid Digestion)
Principles
Digestion involves reaction of the soil with a mixture of concentrated
acids, usually hydrofluoric acid (HF), perchloric acid (HCI04) and sulfuric acid
(H ZS04). Since concentratedacids are involved in the procedure,the researcher
mustbe cognizantof the hazardsand precautionsin their use(Hossner,1996,see
Chapter3). Eachacid in the digestionmixture playsa specific role in the process.
Hydrofluoric acid promotesdecompositionof the silicate matrix by formation of
gaseousSiF4, HCI04 oxidizesorganicmatter,HZS0 4 moderatesthe violent reac-
tion betweenHF and the sample(Loveland,1988)and promotes volatilizationof
HF and HCI04. Soils that are high in organic matter should be pretreatedwith
nitric acid (HN03) to minimize the possibility of explosionduring the reaction
of organicmatterwith HCl04. The mixed acid systemdescribedbelow (Jackson,
1958; Lim & Jackson,1982) cannot be used to differentiate betweenFe z+ and
Fe3+, since Fe z+ is readily oxidized in the presenceof HCI04 or HN03. Jeffery
and Hutchison(1981) and Kiss (1984) describemixed acid systems,involving
the useof H ZS04 and HF, which havebeenusedto differentiateFe z+ and Fe3+. A
procedureinvolving the digestion of soil with HF/HzS04 in the presenceof
excess1,1O-phenanthrolinein the dark, to prevent the photoreductionof Fe3+,
hasbeenusedto determineFez+spectrophotometrically asthe Fe2+-1,1O-phenan-
throline complex (Stucki, 1981b).Total Fe in the digest is then determinedfol-

lowing the photoreductionof Fe3+ upon exposureto a mercury vapor lamp
(Komadel & Stucki, 1988).
SpecialApparatus
1. Platinumcrucible, 30-mL capacity,with lid, or a 50-mL PTFE (polyte-
trafluoroethylene)beaker.
Reagents
1. Hydrofluoric acid, concentrated,48 to 50%.
2. Perchloricacid, concentrated,69 to 72%.
3. Nitric acid (HN03), concentrated,69 to 71%.
4. Sulfuric acid, concentrated,95 to 98%.
5. Sulfuric acid, 3 M.
DigestionProcedure
This digestion procedure(Jackson,1958; Lim & Jackson,1982) must be
conductedin a HCI04 hood. The proceduredescribedbelow will result in the
loss of Si; therefore,modificationsmust be employedif Si also is to be deter-
mined (Lim & Jackson,1982).
Place 0.5 g of soil, ground to 0.15-mmparticle size (to passa 100 mesh
in.-1 sieve) with a nonmetalgrinder, into a 15 to 30-mL capacityplatinum cru-
cible. The PTFE beakeris heat resistantto 260°C; therefore,if the PTFE beaker
is usedin place of the platinum crucible, specialprecautionsmust be taken that
the beakeris not exposedto temperaturesover 240°C. Glassmust not be used
sinceit will reactwith HE Carry a reagentblank throughthe entire digestionpro-
cedure.With organic soils or soils high in organic matter, pretreatthe sample
overnightat room temperaturewith 3 mL concentratedHN03.
Add a few drops of H2S04, 5 mL of HF, and 0.5 mL of HCI04• Acids
should always be added to room temperaturesamples;they should never be
addedto hot samples,to minimize the dangerof explosion.The digestioncan be
convenientlycarriedout on a sandbath. During digestion,the sampleshouldbe
loosely covered,with the cover slightly ajar. Heat the crucible slowly to prevent
a suddenignition of organicmatter, and then maintain the temperatureat 200 to
225°C until the sampleis dry. Cool the crucible, and add3 mL of water and a
few dropsof HCI04• Heat on the sandbath until dry. Removeheat,and allow the
crucible to cool. Add 5 mL of 3 M H2S04 and approximately5 mL of water, and
heat the crucible to a gentleboil. Repeatthe procedureuntil the residueis com-
pletely dissolved.Dilute the samplewith deionizedwater for the determination
of Fe by AAS, ICPAES or colorimetry.
Decompositionof Sample(Hydrofluoric Acid Digestion)
Digestionwith HF aloneis sufficient to dissolvemost soil minerals,espe-

cially at high temperatureand pressure(Bernas,1968). Hydrofluoric acid diges-
IRON 645
tions are usually performed in sealedcontainers(usually PTFE or polypropy-

lene), comparedto mixed acid digestionswhich are usually performedin open
systems.The HF digestion procedureis summarizedin this book by Hossner
(1996, see Chapter3). This procedureis not suitable for the differentiation of
Fe2+ and Fe3+ due to the presenceof aqua regia. For a discussionof possible
digestionproceduresfor the determinationof Fe2+ and Fe3+, the reader isreferred
to Loveland(1988), Stucki (1981a,b)and Komadel and Stucki (1988).
SELECTIVE EXTRACTION PROCEDURES
Introduction
A large numberof selectivedissolutionproceduresdesignedto targetspe-

cific fractions of soil Fe havebeendevelopedover the last 30 yr. Sincetotal soil
Fe is of limited use in understandingbioavailability or pedogenicprocesses,it is
often helpful to estimatethe amountof Fe presentin different solid-phaseforms.
The principal fractions of soil Fe which are targetedfor selectivedissolution
(extraction) include total or "free" iron oxide, "amorphous"or "active" iron
oxide, organically bound Fe, and exchangeableor solution-phaseFe. The most
commonlyusedselectivedissolutionproceduresare summarizedin Table 23-1,
and the detailed proceduresare presentedin the sectionswhich follow. It is
appropriateto considerthese selective-dissolutionproceduresas operationally
defined basedon the specific extractionprocedure,rather than to think of each
extractionas an accuratemeasureof a specific fraction of soil Fe, since none of
the proceduresis absolutelyselectivefor the specific phasesfor which they are
intended.
Total "Free" Iron Oxide

Introduction
The largestproportion of the total Fe of most soils is in the form of oxide
minerals such as hematite,goethite, lepidocrociteand ferrihydrite. The proce-
Table 23-1. A summaryof commonly usedselective-dissolutionproceduresfor specific fractions of

soil Fe.
Targetphases Procedure/reagent References
Total "free" iron oxides Citrate-dithionite(unbuffered) Holmgren, 1967

Citrate-dithionite-bicarbonate Mehra & Jackson,1960
(buffered at pH 7)
Poorly crystalline ("active") iron NH4-oxalate-oxalicacid (pH 3) Schwertmann,1964
oxides (in the dark) McKeague& Day, 1966
Jacksonet aI., 1986
Organically boundFe Pyrophosphate,pH 10t McKeague,1967
Bascomb,1968
ExchangeableFe MgCl z
Availability indices DTPA, buffered at pH 7.3 Lindsay & Norvell, 1978
Ammonium bicarbonate-DTPA Soltanpour& Schwab,1977
pH 7.6
t This procedureis not specific for organicallybound Fe.
dure for extractionand determinationof free iron oxide involves reductivedisso-

lution, e.g., with zinc powder and ammonium tartrate (Haldane, 1956) or Na
dithionite (Deb, 1950; Kilmer, 1960). During the latter reaction,sodiumdithion-
ite decomposes to hydrogensulfite, resultingin an acid solution (pH 2.6-3.5)and
the possible precipitationof FeS and elementalS. To prevent this precipitation
reaction, Mehra and Jackson(1960) used citrate to chelatedissolved Fe2+ and
NaHC03 to buffer the solutionnearpH 7. Holmgren (1967)concludedthat either
NaHC03 or citrate in adequatequantitieswill result in sufficient pH buffering to
prevent reagentdecompositionand precipitation of reaction products and that
bicarbonateis not necessaryif the citrate/dithioniteratio is greaterthan 20:1. The
bicarbonatemethodrequirestwo sampletreatmentsto ensurethoroughextraction
of Fe and careful control of temperature(75-80°C) to: (i) ensureadequatereac-
tion rate and (ii) minimize the decompositionof dithionite that occursat higher
temperatures(Loveland, 1988).The citrate buffered system requires a single
overnight extraction at room temperature;consequently,it is simpler than the
HC03"-bufferedmethodand is more suitablefor large numbersof samples.Nei-
ther of the dithionite proceduresis suitable for determinationof the iron oxide
Fe2+/Fe3+ mole ratio, since dithionite resultsin the reductionof Fe3+.
Dithionite is a strong reductant and theoretically should be capable of
reducing allFe3+ in iron-containingoxidesto Fe2+ at pH valuesbelow 10 (Borg-
gaard,1988);however, thedissolutionefficiency is affectedby particlesize. With
insufficient grinding, large crystalsof magnetite,goethiteand hematitemay not
be totally dissolved during the dithionite procedure(McKeague & Day, 1966;
McKeagueet aI., 1971; Walker, 1983). Although the dithionite proceduresare
targetedfor "free" iron oxides,the extractswill include small contributionsfrom
water-soluble,exchangeableand organically bound Fe. In addition, dithionite
procedureshave been shown to attack a fraction of the Fe containedin layer-
silicate minerals, especially nontronite (Dudas & Harward, 1971; Ryan &
Gschwend,1991) and montmorillonite and vermiculite containinghydroxy-iron
interlayers (Carsteaet aI., 1970). Nevertheless,the amountsof pedogenically
formed iron oxidesextractedusingdithionite procedureshavebeenshownto cor-
relatewell with the amountsof iron oxidesdeterminedby x-ray diffraction. A rel-
atively new procedure, outlined by Ryan and Gschwend(1991), utilizing a
ternary complex of TI(III), citrate and ethylenediaminetetraacetic acid (EDTA)
showssomepromisefor extractinglessstructuralFe from the layer silicates, than
the dithionite procedures.
Citrate-dithioniteExtractableIron
Reagents
1. Sodiumdithionite (Na2S204)'
2. Sodiumcitrate (Na3C6Hs07• 2 H20).
3. Superfloc16 polyacrylamideflocculating agent(Cytec Industries,Inc.,
West Paterson,NJ), 2 g L-l in water.
Procedure.The procedurebelow is a modification of the Holmgren(1967)
procedure.Place0.5 g of soil previously ground to passa 100 mesh in- l (0.15-
mm nominal pore size) sieve into a 50-mL polypropylenecentrifugetube. Add
IRON 647
0.5 g of Na2S204and 6 g of Na3CJfs07• 2 H20. Add 30 mL of deionizedwater,

cover the tube, and shakeovernight(16 h) on a reciprocatingshaker.Transferthe
suspensionto a 50-mL volumetric flask, and add one to two drops of Superfloc
polyacrylamideflocculating agent. Shakevigorously for 15 s, dilute to volume,
and shakeagain.A black or dark-graycolor of the suspensionis an indication of
precipitationof FeS.In this case,increasethe quantity of Na3C6Hs07in the orig-
inal suspensionand begin the procedureover again.Allow the suspensionto set-
tle for 1 h, and centrifugeif necessary.Dilute the supernatewith deionizedwater
as appropriatefor Fe analysis,and measureFe by AAS or ICPAES. During AAS
analysis,the citrate dithionite reagenttendsto clog the burnerhead;therefore,it
is helpful to aspiratedeionizedwater for at least 20 s betweensamples.
Citrate-bicarbonate-dithionite
Method
The procedurebelow is a modification of that describedby Mehra & Jack-
son (1960) and Jacksonet al. (1986).
Special Apparatus
1. Heatingwater bath capableof attaining80°C.
Reagents
1. Sodium citrate (Na3C6Hs07• 2 H20), 0.3 M. Add 88.2 g of solid
Na3C6Hs07• 2 H20 to approximately200 mL deionizedwater in a l-L
volumetric flask, swirl until dissolved,and bring to volume.
2. Sodiumbicarbonate(NaHC03), 1 M. Add 84 g of NaHC03 to a l-L vol-
umetric flask, and dilute to volume.
3. Sodiumdithionite (NaZS204)'
4. Sodiumchloride (NaCl), saturatedsolution.
5. Acetone,reagentgrade.
6. Superfloc16 polyacrylamideflocculating agent(Cytec Industries,Inc.,
West Paterson,NJ), 2 g L-I in water.
Procedure.Transfer5 g of soil, containing less than 0.5 g of Fez03 and
groundto passa 250-meshin.-l (0.06-mmnominal pore size) sieve,to a 100-mL
polypropylenecentrifuge tube. The samplesize can be adjustedin accordance
with the expectedamountof extractableFe. Add 40 mL of 0.3 M sodium citrate
and 5 mL of 1 M NaHC03. Shakethe tube to mix the contents,and heat the tube
in a water bath at 75 to 80°C for severalminuteswhile stirring the suspension
with a glassrod. Care must be taken to avoid temperatureshigher than 80°C to
preventdecompositionof dithionite and the possibleformation of FeS.When the
temperatureof the soil suspensionhas risen to 75 to 80°C, add about 1 g of
NaZS204powderwith a calibratedspoon;immediatelystir for 1 min, then inter-
mittently for 5 min. Add a secondI-g portion of Na2SZ04,and continuestirring
intermittently for an additional 10 min. After the digestion,add 10 mL of satu-
rated NaCI to promote flocculation, and centrifuge. If the suspensionfails to
flocculate with the addition of NaCI, add 10 mL of acetone(C3H60), mix the
contents,warm in a water bath, and centrifuge for 5 min at 1600 to 2200 rota-
tions per minute (rpm). Decantthe supernateinto a 500-mL volumetric flask. An
alternativeflocculation procedureinvolves the addition of two to four drops of

Superflocsolution, thorough mixing and centrifugationfor 5 min at :WOO rpm
(Holmgren, 1967).
Repeatthe extractionprocedurefor samplesin which a brown or red col-
oration persistsor thosecontainingmore than 5% Fez03'Washthe sampleswith
sodiumcitratesolution(andwith NaCl if necessaryfor flocculation), combinethe
washingswith the previousdecantate,dilute with deionizedwaterto volume, and
mix.
Iron can be determinedby AAS by direct aspirationof the appropriately
diluted citrate-bicarbonate-dithionitesolution (Jenneet aI., 1974). During AAS
analysis,the citrate dithionite reagenttendsto clog the burnerhead;therefore,it
is helpful to aspiratedeionizedwater for at least20 s betweensamples.Alterna-
tively, Fe may be determinedby the Fez+-1,1O-phenanthroline procedure.A rea-
gent blank should be utilized throughoutthe procedureand subtractedfrom the
Fe determinedin the sample.
"Active" or "Amorphous"Iron Oxide
Introduction
The "active" iron oxides(often called noncrystalline,poorly ordered,poor-
1y crystalline,short-rangeorderedor amorphousiron oxides) are the most reac-
tive iron oxides in the soil due to their small size and consequentlyhigh surface
area.This classof oxidesincludesferrihydrite and the ferrihydritelike minerals.
The most commonlyusedprocedurefor obtainingquantitativeestimatesof
the "active" iron oxide componentis pH 3.0, 0.2 M ammoniumoxalateextraction
in the dark for either 2 h (Schwertmann,1964) or 4 h (McKeague& Day, 1966).
Although eachof the iron oxide mineralswill react with ammoniumoxalate to
someextent (Schwertmann,1991), the reactionsproceedat considerablydiffer-
ent rateswhich are dependenton particle size and surfacereactivity. The prefer-
ential dissolutionof poorly crystallineiron oxide (including ferrihydrite) hasbeen
confirmedby differential x-ray diffraction (Schwertmannet aI., 1982); however,
considerabledissolutionof lepidocrocite andmagnetitealso may occur(Schwert-
mann, 1973). Acid ammoniumoxalate has little effect on kaolinite, montmoril-
lonite, vermiculite or illite (McKeague& Day, 1966; Hodges& Zelazny, 1980);
however,Arshad et aI. (1972) observedconsiderabledissolutionof trioctahedral
layer silicates,e.g., biotite and chlorite. The ammoniumoxalate extraction is a
kinetically controlledprocedure;therefore,the quantity of Fe extractedis strong-
ly influencedby reactiontime and temperature,as well as shakingintensity. The
proceduremust be performedin the dark to preventphotoreductionand retard
rate of dissolutionof the crystalline iron oxides.Ammonium oxalateextractable
Fe will include water-solubleFe, exchangeableFe and a fraction of the organi-
cally boundFe.
Most of the variationsof the ammoniumoxalateprocedureinvolve prepa-
ration of pH 3 ammoniumoxalatefrom a mixture of ammoniumoxalateandoxal-
ic acid (Tamm, 1922; Schwertmann,1964; McKeague & Day, 1966; Chao &
Zhou, 1983).Jacksonet ai. (1986) preparedthe reagentby acidification of 0.2 M
IRON 649
ammoniumoxalate to pH 3.0 with HCl. Calcareoussoils must be pretreatedto

removeCaC03, sinceoxalic acid will reactwith CaC03 to changethe pH of the
oxalate/oxalicacid buffer; also, oxalatewill be precipitatedas the Ca salt. Calci-
um carbonatecan be removedby reactionwith ammoniumacetateat pH 5.5.
In a comparisonof a rangeof soils and sediments,Chao and Zhou (1983)
observedthat extraction with a combined solution of 0.2 M hydroxylamine
hydrochlorideand 0.2 M HCI at 50°C for 30 min gave results similar to those
obtainedwith the ammoniumoxalateextractionin the dark. Alkaline EDTA also
has beenused to dissolve noncrystallineiron oxides (Borggaard,1988); howev-
er, reactiontimes of 90 d or more are requiredfor quantitativeextraction.Results
by the acid ammonium oxalate and EDTA proceduresare highly correlated
(Borggaard,1988).
Acid Ammonium Oxalatein Darkness-Thmm's

Reagent
The procedure described below is a modification of the procedureof
Schwertmann(1964) and McKeagueand Day (1966).
Reagents
1. Acidified ammoniumoxalate,pH 3.0. Preparea solutionof 0.175M am-

monium oxalate[(NH4)2C204] + 0.1 M oxalic acid (H 2C20 4). Add 24.87
g of (NH4)2C204• H20 and 12.61 g of H2C20 4 • 2H20 to approximate-
ly 800 mL deionizedwater, adjust to pH 3.0 by addition of NH40H or
HCI, and dilute to 1 L final volume.
2. Ammonium acetate,1.0 M, pH 5.5. Add 60 g of glacial acetic acid to
600 mL of deionizedwater, adjustto pH 5.5 with NH40H, and dilute to
1 L final volume.
Procedure.Place500 mg of soil, which hasbeenpreviouslygroundto pass
a 100 meshin.- I (0.15-mmnominal poresize) sieve,into a 50-mL polypropylene
centrifugetube. Calcareoussoils must be pretreatedto remove CaC03. Add 30
mL of pH 5.5 1.0 M ammoniumacetate,allow to react for 1 h with intermittent
stirring of the ventedcontainer,measurethe pH with a combinationpH electrode,
and readjustthe pH to 5.5 by the dropwise addition of acetic acid. Repeatthe
addition of aceticacid hourly until the pH remainsapproximately constant. Cen-
trifuge, decant, wash with deionizedwater twice to remove dissolved Ca and
acetate,and allow the sampleto air dry. Before the ammoniumoxalate is added
in the stepbelow, care must be taken that the sampleis crushedto a suitablepar-
ticle size.
Add 30 mL of pH 3.0 ammoniumoxalatesolution, stopperthe tube, and
immediately place the tube in a light-proof container.Immediately begin agita-
tion of the sample,and continuefor exactly 2 h on a reciprocatingshaker.Cen-
trifuge the sample, decant, dilute the supernateas appropriatewith deionized
water, and analyzefor Fe by AAS or ICPAES. If the samplecannotbe analyzed
immediatelyit should be storedin the dark to preventphotoinduceddecomposi-
tion of oxalate,which could result in precipitationof Fe (Borggaard,1988). High
concentrationsof oxalatecan result in cloggingof the burnerheadduring the neb-
ulization process;therefore,samplesshould be diluted with deionizedwater to
the extent possibleprior to analysis.Also, to minimize clogging, it is helpful to

aspiratedeionizedwater for at least 20 s betweensamples.High concentrations
of oxalatewill interfere with AAS analysis;therefore,it might be necessaryto
decomposethe oxalateprior to analysis.This stepis accomplishedby evaporat-
ing a known volume of the ammonium oxalate extract to dryness,ashing at
500°C in a porcelaincrucible for 1 h, and redissolvingthe sedimentin a known
volume of 1 M HCl, prior to AAS analysis.Samplescan alternativelybe treated
by digesting1 mL of the ammoniumoxalate extractwith lO mL of concentrated
RN03 to drynessand dissolvingthe sedimentin a knownvolume of 1 M HCI. As
an alternativeto AAS analysis,the samplecan be analyzedspectrophotometri-
cally by the Fe2+-l,lO-phenanthrolinemethod.
Organically Bound Iron

Introduction
Potassium-or Na-pyrophosphate extractions,usually at pH lO, have been
utilized for the estimationof organicallyboundFe (McKeague,1967; Bascomb,
1968).Thereareseveralvery severeproblemswith this procedure,especiallythe
peptization and dispersion of microcrystalline iron oxide by pyrophosphate
(Jeanroy& Guillet, 1981).Becauseof this dispersionphenomenon,results are
highly dependenton the centrifugationand ftltration procedure(Schuppli et aI.,
1983; Loveland & Digby, 1984). The quantity of Fe extractedwith pyrophos-
phate decreaseswith increasingcentrifugation(McKeague& Schuppli, 1982);
therefore,uniform high-speedcentrifugationor micropore filtration treatments
are required (Schuppli et aI., 1983; Loveland & Digby, 1984).Even following
high-speedcentrifugation, microcrystalline clays and oxides may remain sus-
pended(Borggaard,1988); however,high-speedcentrifugationmay be preferred
to ftltration becauseof the problemof cloggedfilters during microporeftltration
procedures(Loveland, 1988). Clariftcation of the extractalso hasbeenachieved
by treatmentwith Superflocor Na2S04(Schuppli et aI., 1983). The dispersion
phenomenon and the quantity of pyrophosphate-extractableFe also is influenced
by pH and whetherthe Na or K salt is utilized (Loveland& Digby, 1984). Love-
land and Digby (1984)concludedthat a techniqueusing 0.1 M Na pyrophosphate
at pH 10 andcentrifugationat 20 000 x G providedthe most reproducibleresults.
Studieshave indicatedthat the pyrophosphateextract obtainedfollowing
high-speedcentrifugation includesmicrocrystalline iron oxide particles which
are mixed or possibly coatedwith organicmatter,though it is not clear whether
theseiron-oxide/organic-mattercomplexeswere originally presentin the soil or
formed during the extraction procedure(McKeague & Schuppli, 1982). Pyro-
phosphate-extractable Fe cannotbe designatedas organically bound Fe, due to
uncertaintyabout the actual sourceof the extractedFe (Schuppli et aI., 1983).
Despite these problems, the procedureis presentedbelow due to its frequent
occurrencein the literature.The researchershouldpreciselystatethe centrifuga-
tion and clariftcation procedureand must take care not to overextendthe inter-
pretationof results.
Acetylacetone (Bascomb & Thanigasalam,1978) and 0.05 M Na2B407at
pH 9.7 have beenusedas alternativesto the pyrophosphateextraction.Sodium
IRON 651
tetraboratewas found to be a poor extractantfor organically bound Fe (Mc-

Keague& Sheldrick, 1977). Bascomband Thanigasalam(1978) concludedthat
acetylacetoneis lesssuitablethan pyrophosphatefor extractingorganicallybound
Fe, sinceacetylacetonedissolvesmore of the mineral forms of Fe than pyrophos-
phate.
SodiumPyrophosphateExtractableIron
Reagents
1. Sodiumpyrophosphate(Na4P207),0.1 M, pH 10.0. Dissolve 44.61 g of
Na4P207.10 H20 in approximately800 mL of deionizedwater. Adjust
to pH 10 with NaOH, and dilute to volume (1 L) with deionizedwater.
Storethe pyrophosphatesolution in the refrigeratorand in a sealedcon-
tainer with a CO2 trap to preventcontactwith atmosphericCO2,
Procedure. Place 250 mg of soil «2-mm particle size) into a 50-mL
polypropylenecentrifugetube. Add 25 mL of 0.1 M Na4P207(pH 10), and shake
16 h at 23°C on a reciprocatingshaker. Centrifuge at 20 000 x G for 30 min.
Decantthe supernate,and analyzefor Fe using AAS or ICPAES.
Water-Solubleand ExchangeableIron
Introduction
Water-solubleFe in the soil is usually presentin such low concentrations
that it is detectableonly in soils of very low pH «4.5; e.g., exposedlignite beds
or soils exposedto acid mine drainage),or very low redox potential (e.g., tidal
marshsedimentsor flooded soils). It is possibleto directly extractsuitablequan-
tities of soil pore waterfor subsequentelementalanalysisonly from wet or water-
saturatedsoils. Immiscible displacementproceduresare required for the dis-
placementof pore water from drier soils (Kinniburgh & Miles, 1983; Kittrick,
1983).Otherproceduresfor obtainingsoil waterextractsincludeextractionof the
soil with deionizedwater (as is discussedin the section below)or preparationof
a water-saturatedsoil paste.
Extractionof soil with neutral(e.g.,CaCl2, MgCl 2or KCl) or buffered(e.g.,
NH4C2H30 2) salt solutionswill result in the displacementof both dissolvedFe
and exchangeable Fe. Acid-bufferedextractantsmay not be suitablefor neutralor
calcareoussoils, sincethey could result in considerabledissolutionof solid-phase
carbonatesand oxideswith the possiblereleaseof Fe from the mineral structures.
Becauseof the usually low concentrationsin the soil, detectionof exchangeable
Fe extractedwith neutral or buffered salts only will be possiblefor soils of low
pH and/or redox potential.
Samplesfor the determinationof water-solubleor salt-extractableFe must
not be air or oven dried or evenexposedto air, sincetheseprocedurescould result
in changesin the forms and concentrationof extractableFe. Samplesextracted
with either deionizedwater or neutral salt solutionsmust be protectedfrom the
atmosphereafter sampling to prevent changesin redox potential, oxidation of
Fe2+ to Fe3+ and precipitationof dissolvedFe as iron oxides.
Microcrystalline iron oxides are readily dispersedduring soil extraction,

especially during extraction with deionized water and the alkali metal salts.
Therefore,extractsshouldbe passedthrough filters with pore sizes<0.20 Ilm to
minimize errors due to the presenceof suspendedFe. Even with this treatment,
precautionsare advisedin the interpretationof results,sinceiron-containingmin-
erals in the soil can have particle sizesas small as 0.005 Ilm.
Iron in the soil solution or in aqueousor salt extracts can be measured
directly by AAS, ICPAES or colorimetric procedures.With very small amounts
of Fe, concentrationof Fe in the soil solution or extract may be requried. Con-
centrationmay be accomplishedby complexationwith a chelatein an immisci-
ble organic solvent, complexationwith a chelatingresin, or by evaporationand
resolubilizationin acid.
Water-ExtractableIron
Reagents
1. Deionizedwater.
2. Cylinder of prepurifiednitrogen(N2) gas.
Sampling Procedure.As the soil is sampled in the field, immediately
place it in a preweighed(tube plus cap) 50-mL centrifugetube, so that approxi-
mately one-thirdof the total tube volume is filled with soil. Then purgethe sam-
ple with N2 gas, and immediatelyseal the tube tightly to preventcontactof the
soil with the atmosphere.This procedureis especiallyimportantfor reducedsoils
in which changesin redox potential could result in changesin the concentration
of dissolvedFe. An alternativesamplingprocedureis to placethe field moist soil
in a Ziploc heavy duty(2.7 mil; 1 mil = 0.001 in. = 0.025 mm) polyethylene
freezerbag, squeezethe bag to excludeair, and closesecurely.The polyethylene
bagsare easy to use, but have relatively high O2 (approximately1.9 x 102 cm3
mil- 1 m-2 atm-1 h-1) and CO2 diffusion rates. For this reason,the polyethylene
bags should be placed in a larger container,e.g., an insulatedcooler, which is
purgedwith N2 gas.Other plastic materialscan be usedwhich haveconsiderably
lower O2 diffusion ratesthan polyethylene,e.g., Mylar, polyvinylidene chloride
(PVDC; 0.62 cm3 mil- 1 m-2 atm-1 h-1), or Saranex(0.31 cm3 mil- 1 m-2 atm-1
h-1). The field moist samplesobtainedby this procedureare transferredto a cen-
trifuge tube as soonas possiblein the laboratoryso that approximatelyone-third
of the total tube volume is filled with soil. The tube is immediatelypurgedwith
N2 and closedtightly to avoid contactwith the atmosphere.
Extraction Procedure.Samples obtained by either procedure above
should be extractedas soon as feasible. The centrifugetube (with cap) plus wet
soil is weighed; then deionizedwater (previously bubbled vigorously with N2
gas)is addedto give an approximate1:1 (v/v) wet-soil/deionized-waterratio, and
the tube is purgedagain with N2 and weighedagain to obtain the tube plus soil
plus waterweight. The sampleis shakenfor 30 min on a reciprocatingshakerand
centrifuged. Decant the supernate,and filter immediately through a 0.20-llm
pore-sizemembranefilter. Acidify the filtrate to pH <2.0 by the dropwiseaddi-
IRON 653
tion of 2 M HCI to minimize oxidationof Fe2+ and precipitationof iron oxide, and
analyzeby AAS, ICPAES or a colorimetric procedure.The soil in the centrifuge
tube is oven dried (110°C), and the tube (with cap) plus dry soil is weighed.Iron
concentrationmay eitherbe reportedon an oven-driedsoil basisor on a soil-solu-
tion basis. The sampling, extraction and analysis steps must be carried out as
rapidly as possibleto minimize the possibility of Fe transformationsduring the
procedure.
Neutral Salt Extractable Iron

Reagents
1. Magnesiumchloride (MgCI 2), 1 M.
Procedure. Use the same procedure as is described above for water-
extractableFe, exceptsubstitute1 M MgCl 2 for deionizedwater in the extraction
step.Other neutral salts (e.g., KCI, CaCI2) can be used in place of MgCI 2; how-
ever, careshouldbe takento avoid saltswhich could result in the precipitationof
Fe-containingphases.For example,KCI should not be used in acid sulfate soils
to prevent the precipitation of jarosite. Neutral or alkaline buffered salts (e.g.,
sodium acetate)should be avoidedto preventthe precipitationof ferric hydrox-
ide.
AVAILABILITY INDICES
Introduction
Water soluble or neutral-salt extractable Fe are not generally useful as

indicesof plant-availableFe due to the low (often nondetectable)quantitiesof Fe
extracted.The extractabilityof Fe can be increasedby the use of reducitlg agents
(reductive dissolution), e.g., 0.5% oxalic acid (Thorne, 1941) or 0.2% hydro-
quinone in ammonium acetate(Thorne & Wallace, 1944), or by the use of an
acidic extractant(H+-mediateddissolution),e.g., pH 3.0,1 M ammoniumacetate
(Jackson,1958); however,noneof theseextractantshasbeenextensivelyutilized.
The two most widely usedproceduresfor assessment of Fe availability to plants
have been EDTA and diethylenetriaminepentaacetic acid (DTPA) extraction
(Lindsay & Norvell, 1978). Ethylenediaminedi-(o-hydroxyphenylaceticacid)
(EDDHA) also has been usedsuccessfully.Each of thesecompoundswill com-
plex Fe3+. The stability of the Fe3+ complex is strongly influenced by pH
(Norvell, 1972; Lindsay, 1979). At a given pH, the stability of the Fe3+ chelate
decreasesin the following order: EDDHA > DTPA> EDTA. The quantity of Fe
extractedgenerallydecreasesin the sameorder. Lindsay and Norvell (1978) con-
cluded that DTPA was the most useful extractantfor the simultaneousextraction
of Fe, Zn, Mn and Cu. A modification of the DTPA procedure,in which the
extractingsolution is bufferedby 1 M NH4HC03 at pH 7.6, hasbeenproposedby
Soltanpourand Schwab(1977) for the simultaneousextractionof NO), K+ and
phosphate,as weltas Fe, Mn, Zn, and Cu.
Diethylenetriaminepentaacetic
Acid (DTPA) Soil Test
Theory
The DTPA solution (0.005 M) is buffered at pH 7.3 with 0.1 M tri-
ethanolamine(TEA) in the presenceof 0.01 M CaCl2 (Lindsay & Norvell, 1978).
The rate of releaseof Fe by DTPA is highly dependenton pH. A pH of 7.3 was
adoptedto preventthe dissolutionof CaC03 and the possiblereleaseof occluded
Fe which could occurat lower pH values,and to avoid the reductionin Fe release
that is observedat higher pH values.Triethanolaminewas selectedas the buffer
becauseof its pKa (7.8) and becauseit burnscleanly with little interferencedur-
ing AAS analysis.At pH 7.3 and 0.01 M Ca2+, the DTPA is fully complexedwith
Ca; in the presenceof Fe3+, the Fe competeseffectively with Ca for complexa-
tion by DTPA. Approximately 50% of the added Ca is complexedby DTPA,
while the remaining50% is in solution as Ca2+. At pH 7.3, approximately75% of
the total TEA is presentin the protonatedtriethanolammonium(HTEA+) form.
The HTEA+ can competewith Ca2+ for cation exchangesites, thereby further
increasingthe Ca2+ concentrationin solution and retarding the dissolution of
CaC03·
Diethylenetriaminepentaacetic acid will form complexeswith soluble and
exchangeableFe, as well as with Fe which is mobilized from the Fe containing
solid phases,predominantlythe iron oxides. The releaseof Fe from iron oxide
occursas a two-stepreaction:(1) a rapid chemisorptionof ligand at the mineral
surfaceand a concomitantweakeningof internal O-Fe-O bonds,and (2) a slow
(rate-limiting) abstractionand chelation of Fe by the surface adsorbedligand
(Stumm & Furrer, 1987; Schwertmann,1991). Steps(1) and (2) are each influ-
encedby the propertiesof the ligand and by the mineralogyand surfaceareaof
the oxide. The rate of reactionof DTPA with soil iron oxidesgenerallydecreas-
es in the following order: ferrihydrite :> lepidocrocite= magnetite:> goethite >
hematite.
The quantity of DTPA-extractableFe is highly dependenton reactiontime
(Lindsay & Norvell, 1978; Geiger & Loeppert,1988).The rate of dissolutionof
Fe from soil decreasescurvilinearly, due to the initial more rapid reaction with
high-energysurfacesitesand a gradualreductionin reactionrate asthe active sur-
face sitesare consumed.In the normal2-hextractionof soil with DTPA, the actu-
al final concentrationof the Fe-DTPA complex (usually within the range of
approximately0.25-20mg L- 1) is significantly less than the theoreticalequilib-
rium concentration(approximately 279 mg L- 1) (Lindsay & Norvell, 1978).
Therefore, the quantity of DTPA-extractableFe is highly dependenton any
experimentalfactor that influencesreaction rate, e.g., grinding intensity, extrac-
tion temperature,shaking time and shaking intensity (Soltanpouret aI., 1976).
Standardizationof the procedureis very important to obtain uniform and repro-
ducible results in the DTPA soil test. Since DTPA is presentin excessof the
micronutrientmetal cationsthat are normally solubilizedduring the extractionof
most agriculturalsoils, the extractionof one micronutrientwill probably not sig-
nificantly affect the amounts of other metals extracted (Lindsay & Norvell,
1978).
The DTPA soil test developedby Lindsay and Norvell (1978) and the
NH4HC03 DTPA test proposedby Soltanpourand Schwab(1977) were primari-
IRON 655
ly intendedfor determiningFe bioavailability in neutraland calcareoussoils. The

useof theseextractantshasbeenquestionedfor acid soils below pH 6, for which
it may not be appropriateto usea bufferedextractantat pH 7.3 or 7.6 (O'Connor,
1988). Furthermore,highly acid soils may exhaustthe buffering capacity of the
TEA presentin the DTPA. extractingsolution. If the final pH of the extracting
solution varies for different soils, one can expect significant variability in the
amountof metal extracteddue to changesin reactionkinetics as a function of pH.
Haynesand Swift (1983) evaluatedthe use of DTPA extraction in moderately
acid soils and found significant variability in extractableFe as a function of the
pH of the extracting solutions. Generally, the amounts of extractablemetals,
including Fe, increasewith decreasingpH of the extractingsolution. At very low
pH values,as may be encounteredin somemine spoil soils, the concentrationof
DTPA may not be sufficient at 0.005 M to avoid secondaryinteractionsamong
metal ions. In summary,althoughit may be more appropriateto usean unbuffered
DTPA for soils with pH valuesbelow six, the concentrationof DTPA may be too
low, resulting in competitionamongmetalsfor the DTPA. O'Connor(1988) has
summarizedseveral limitations and inappropriateuses of the DTPA soil test,
especiallywith regard to soils with high metal loadings. Users should be cog-
nizant that the DTPA soil test in its current form was designedto extract labile
forms of Fe for the purposeof distinguishingbetweenFe deficient and nondefi-
cient neutraland calcareoussoils.
Influenceof Soil Factorson the DTPA Extraction Procedure

The amountof Fe extractedby DTPA (Loeppert& Hallmark, 1985; Geiger
& Loeppert, 1988; Morris et aI., 1990), as well as by EDTA (Borggaard,1976;
Borggaard,1982) and EDDHA (Loeppert & Hallmark, 1985), is positively cor-
relatedwith the "active" iron-oxidecontentof the soil as assessed
by pH 3 ammo-
nium oxalateextractionin the dark, though lower quantitiesof Fe are extracted
in a given time by the chelateextractantsthan by ammoniumoxalate.Dissolution
of soil iron oxide by DTPA occursas a surfacereactionwhich is strongly influ-
encedby the quantity, particle-sizedistribution, surfaceareaand surfacereactiv-
ity of the soil iron oxides. The evidencestrongly suggeststhat the predominant
sourceof DTPA-extractableFe in oxidized calcareoussoils is the highly reactive
poorly crystalline iron-oxide component,which often exists in associationwith
soil organic matter and layer silicates. A much smaller proportion is usually
attributable directly to organically bound Fe2+ and Fe3+ (Geiger & Loeppert,
1988). In acid soils, a significantly higher proportion of DTPA-extractableFe
may be attributableto organicallybound Fe, due to the strongpH dependencyof
the stability of the Fe3+-humic complex.
Influenceof SamplePreparationVariableson the DTPA Extraction

Procedure
The air drying of oxidized field-moist samplesusually resultsin increases
in the level of DTPA-extractableFe (Shuman,1980; Leggett & Argyle, 1983;
Cihacek,1988; Geiger& Loeppert,1994); however,air drying of highly reduced
or flooded soils may result in decreasesin DTPA-extractableFe due to oxidation
656 LOEPPERI'& INSKEEP
of Fe2+ andprecipitationas iron oxides.LeggettandArgyle (1983) observedthat

the increasedextraction of Fe due to air drying was only partially reversedby
reincubationwith water. Ovendrying of the soil prior to extractionusually results
in substantial increasesin DTPA-extractable Fe (Khan & Soltanpour, 1978;
Leggett & Argyle, 1983; Geiger & Loeppert, 1994). This latter phenomenonis
attributable to the formation of surface defects on the poorly crystalline iron
oxidesand increasesin surfacefree energyand reactivity.
Correb.tionof DTPA·ExtractableIron with Iron Chlorosis

Correlationstudiesrelating DTPA-extractableFe with crop growth are nec-
essaryto give meaningful interpretationsto the soil test. Lindsay and Norvell
(1978) observedthat DTPA-extractableFe was highly correlatedwith the inci-
denceof Fe chlorosisof sorghum(Sorghum x alum L. Parodi) and reportedthe
following Fe soil-testcorrelationvaluesfor sorghum:<2.5 mg DTPA- extractable
Fe kg-1 soil, deficient; 2.5 to 4.5 mg kg-I, intermediate;>4.5 mg kg-1, sufficient.
Correlationof DTPA-extractableFe with the incidenceof Fe chlorosisgenerally
has beenmost successfulfor grasses(StrategyII plants; Marschneret aI., 1986;
Romheld& Marschner,1986) in well-drainedcalcareoussoils (Loeppert& Hall-
mark, 1985). For Strategy II plants the primary mechanismsof Fe-deficiency
stressresponseare phytosiderophore(chelator)releaseand subsequentFe mobi-
lization anduptake.The strengthof the DTPA soil test lies in the ability of DTPA
to extractthe samelabile forms of soil Fe as the plant (Geiger& Loeppert,1988).
The DTPA soil test has beenconsiderablyless successfulwith StrategyI
plantsin which the primary Fe-deficiencystress-response mechanismsare acidi-
fication of the rhizosphereand increasedroot plasma-membrane Fe3+-reductase
activity (Marschneret al., 1986; Romheld & Marschner,1986). Diethylenetri-
aminepentaacetic acid-extractableFe predictedonly 14 to 20% of the observed
variation in chlorophyll concentrationof soybean(Glycine max L. Merr.) grown
in 23 calcareoussoils in a growth room (Morris et al., 1990). For the StrategyI
plants, solution HCO)" concentration(Fleming et aI., 1984; Inskeep & Bloom,
1986; Marschneret al., 1986) and soil CaC03 reactivity (Morris et aI., 1990) are
importantsoil factorswhich influence the effectivenessof the plant Fe-deficien-
cy stress-response mechanism.Also, in wet or poorly drained soils, the DTPA
soil test is usually of limited usefulnessin the assessment of Fe availability to
plantsbecauseof the importantinfluenceof soil aerationand root respirationon
Fe mobilization and uptake.Other indigenoussoil factors (suchas soil carbonate
reactivity) and soil environmental factors(such as soil water, aerationand com-
paction), as well as cultivar Fe-deficiencystresstolerance,also must be consid-
eredto adequatelypredict Fe chlorosisof soybean(Morris et aI., 1990). Multipa-
rametermodels(e.g., DTPA-extractableFe, soil-waterstatus,and CaC03 activi-
ty) may be much more generallyuseful, but havenot beenextensivelyutilized.
Procedurefor DTPA·ExtractableIron
Reagents
1. Triethanolamine.
acid.
IRON 657
3. Calcium chloride (CaCI2 • 2 H20).

Preparationof DTPA ExtractingSolution

Dissolve 149.1 g reagentgradeTEA, 19.67 g DTPA, and 14.7 g CaCl2 • 2
H 20 in approximately200 mL of deionizedwater. After the DTPA hasdissolved,
dilute to 9 L with deionizedwater. Adjust the pH to 7.3 ± 0.05 with 6 M HCI
(approximately83 mL), and dilute to 10 L final volume.
Procedure
The samplepretreatmentprocedureshould be precisely defined, since it
will influence the quantityof DTPA-extractableFe. Most researchersprefer air
drying at room temperature,since the exact conditionsof pretreatmentare easier
to reproduce;results also are usually reproducible.More intensedrying proce-
dures, e.g., oven drying, should be avoided. The proceduresummarizedbelow
was originally proposedby Lindsay and Norvell (1978).
Add 20 mL of extracting solution to 10 g of air-dried soil (previously
ground to pass a 2-mm pore-sizesieve) in a SO-mL polypropylenecentrifuge
tube. Shakeon a reciprocatingshakerfor exactly 2 h at room temperature(230 C).
Since this is a kinetically controlled procedure,temperature,shaking time and
shaking intensity are critical experimentalvariables.Centrifuge immediately at
3000 x G, decantthe supernate,and filter through a O.4S-~m pore-sizemem-
branefilter. Following appropriatedilution, the extractshouldbe analyzedfor Fe
by AAS or ICPAES.
Procedurefor Ammonium Bicarbonate-DTPA Extractable

Iron
Theory
The ammonium bicarbonate-DTPA(AB-DTPA) soil test was originally
developedby Soltanpourand Schwab(1977) as a modification of the DTPA soil
test. The primary goal was to develop a soil test which would simultaneously
extractN03, P, K, Zn, Fe, Mn and Cu from neutraland calcareoussoils. This test
also has beenusedto extractother elementsincluding Pb, Cd, Ni, Se, As, B, Mo
and S from mine spoils and soils treatedwith sewagesludge. The relationship
betweenbioavailability and AB-DTPA extractableamountsof severalof these
elementswas reviewedby Soltanpour(1991).
The AB-DTPA extracting solution is composedof 1 M NH4HC03 and
0.005 M DTPA and is adjustedto an initial pH of 7.6. The extractingsolutionwill
releaseCO2(g) when exposedto the atmosphereor during soil extraction,with the
subsequentincreasein pH to valuesas high as 8.5. At 1 M NH 4HC03, the AB-
DTPA extracting solution contains similar concentrationsof NUt and HC03
used in traditional extraction methodologiesfor cations including K+ and for
anionsincluding pol-. The presenceof DTPA allows for extractionof tracemet-
als which complex with DTPA (Lindsay & Norvell, 1978). The solutionmay be
slightly colored due to the solubilization of organic matter; however, carbon
black shouldnot be usedas a clarifying agent,sinceit can result in adsorptionof
DTPA and Fe-DTPA from the extract(Soltanpour& Workman, 1979).
The AB-DTPA soil test hasbeenshownto successfullydistinguishFe defi-

cient from nondeficientsoils for sorghum (Havlin& Soltanpour, 1981, 1982,
1984). Soils with AB-DTPA extractable-Fevalues<4.8 mg kg-1 soil were con-
sideredto be Fe deficient. The AB-DTPA procedurewill have similar problems
as the DTPA soil test for StrategyI plantssuch as soybeanor on wet or flooded
soils (Loeppertet aI., 1994),whereother soil factorsbesidessoil Fe influenceFe
acquisitionand uptake.
Reagents
acid.
3. Ammonium hydroxide(NH40H).
Preparationof Ammonium Bicarbonate-DTPAExtractingSolution

Dissolve 1.97 g of DTPA in 0.8 L of deionizedwater, using 2 mL of 1:1
(v:v) NH40H to facilitate the dissolution.After the DTPA is dissolved,add 79.1
g NH4HC03 and stir until dissolved.Adjust the pH to 7.6 using NH40H or HCI,
and bring the final volume to 1 L. Sincethis solution will releaseCO2 over time,
the solution pH is unstable,and extractingsolutionsshouldbe preparedimmedi-
ately prior to use.
Procedure
As with the DTPA soil test,samplepretreatmentmay influencethe amount
of Fe extracted.Consequently,all samplesshould be treatedconsistently(oven
drying shouldbe avoided).Add 20 mL of extractingsolution to 10 g of soil in a
125-mL erlenmeyerflask, and shakeon a rotary shakerat 180 cycles min-1 for
15 min. Flasks are kept open to allow for CO2 release.Filter or centrifuge the
extract to remove suspendedparticles,and filter the clarified extract through a
0.45-flm pore-sizefilter. The extract can be analyzedfor Fe directly by AAS or
ICPAES; however, a high salt nebulizermay be required. Also,it is helpful to
aspiratedeionizedwater for at least 20 s betweensamples.In the absenceof a
high salt nebulizer,the extractcan be acidified using a known volume of HN03
and swirled for 15 min in an openflask to eliminatecarbonate species. A reagent
blank.shouldbe carriedthrough the entire procedure.
ATOMIC ABSORPfION SPECTROSCOPY
Principles
Filtered soil digestsor extractscan be analyzeddirectly, after appropriate
dilution, by AAS, usingthe air/acetyleneflame. With the graphitefurnace,Fe can
be determinedin the partsper billion range.There are no major interferencesin
the AAS analysisof Fe, thoughsamplesmust be acidified adequatelyto prevent
precipitationof Fe as iron oxides.
SpecialApparatus
1. Atomic absorptionspectrophotometer.
IRON 659
Reagents
1. StandardFe solution, 1000mg kg-I. StandardFe solutionsare commer-

cially available. As an alternative, a standardFe solution can be pre-
paredby dissolving 1 g of analytical reagentgradeiron wire in 100 mL
of 1.75 M H2S04. Warm if necessaryto obtain complete dissolution.
Dilute to 1 L with deionizedwater.
Procedure
The instrumentshould be preparedfor analysisaccordingto the instruc-

tions of the manufacturer.Iron is usually determinedat a wavelengthof 248.3 nm
using an oxidizing (lean) air/acetyleneflame.
Dilute the soil digestor extractto a final concentrationof 0.5 to 5 mg L- 1•
Analyze a few samplesand comparethe absorbanceto that of a 5 mg L- 1 stan-
dard to determinewhetherfurther dilution will be required. Samplesshould be
analyzedaccordingto the instructionsof the manufacturer;however, the usual
linear working rangeis approximately0 to 6 mg Fe L-l. Concentrationsof sam-
ples, within this working range,are determinedby comparisonwith a calibration
curve of standardswhich have beenpreparedfrom dilution of the 1000 mg L-I
stock solution. The standardsolutions should be preparedin the same matrix
solutionasthe unknownsample.The concentrationof Fe in a blank solution(trip-
licate samples),which hasbeencarriedthroughthe entire analyticalprocess,also
is determined.Iron in the digest or extracton a soil weight basisis calculatedas
follows
Fe, mg kg-1 = [(Fetestsolnmg L-I - Feblankmg L- 1)
(extractingsolution volume, L)(dilution)]/(weight of sample,kg)
Onepossiblesourceof error is the precipitationof Fe from samplesand standards

as Fe3+ hydroxideor phosphates.Careshouldbe takenthat samplesand standards
are adequatelyacidified to prevent precipitation, especially samplesprepared
from aqueousor neutral-saltextractsof soil.
COLORIMETRIC DETERMINATION OF FERROUSIRON

AND FERRIC IRON BY THE 1,lO-PHENANTHROLINEMETHOD
Principles
Total dissolvedFe can be determined,at a detectionlimit of approximate-

ly 0.1 mg L-I, by reductionof Fe with hydroxylaminehydrochlorideand reaction
with 1,1O-phenanthrolineto form the tri-(l,lO-phenanthroline)ferrous complex
[Fe(C12HgN2W], which has a red color (Olson, 1965; Olson & Ellis, 1982).
Absorption,determinedat 510 nm, is linear in the concentrationrangeof approx-
imately 0.1 to 5 mg VI. The color is developedunderacid conditions,preferably
pH 3 to 5, to preventthe precipitationof salts,e.g., Ca3(P04)2.The color reaction
is specific for Fe2+; therefore,Fe2+ concentrationin the original solution can be
LOEPPERT& INSKEEP
determinedby the addition of 1,1O-phenanthrolinein the absenceof a reducing

agent.The Fe3+ concentrationis calculatedfrom the total Fe concentrationof the
reducedaliquot minus the Fe2+ concentrationof the unreducedaliquot (Olson &
Ellis, 1982).Hydrofluoric acid can interfere;thereforethis reagentshouldbe kept
to a minimumduring the color developmentstep(Stucki, 1981a,b).The method-
ology mustbe uniform, including the orderof addition of reagents,pH and length
of time of color development.Stucki (1981a,b)recommendedthat the order of
addition of reagentsshouldbe reducingagent,phenanthrolineand buffer. If only
Fe2+ is to be determined,the reactionmust be performedin the dark (or undera
red photographiclight in the dark) to preventthe photoreductionof Fe3+. Also,
pH control is essentialto minimize the spontaneoustransformationof Fe2+ to
Fe3+.
Severalpotentialinterferencesmay be problematicdependingon soil type
and/orthe solution matrix. Strongoxidizing agentssuchas CN- and nitrite, poly-
phosphates(orthophosphateto a lesserdegree),Cr and Zn may interfere when
their concentrationsexceed10 times the amount of Fe present(APHA, 1981).
Cobalt, Cu, Ni and AI may interfere at concentrationsof 5,5,2and 20 mg L-t,
respectively.Organic matter also may interfere with Fe analysis; if noticeable
color is present,it may be necessaryto either ashor digest the sampleto obtain
an extractsuitablefor colorimetric analysis.Details of the recommendedashing
or digestionpretreatmentcan be found in StandardMethodsfor the Examination
of Water and Wastewater(APHA, 1981).
Othercomplexingagentswhich can be useful for the determinationof low
Fe concentrationsin water or neutral salt extractsare 2,4,6-tripyridyl-s-triazine
(TPTZ) (Collins et aI., 1959) and 4,7-diphenyl-1,10-phenanthroline (batho-
phenanthroline)(Smith et aI., 1952; Crawley & Aspinall, 1955).
SpecialApparatus
1. Visible or ultraviolet (UV)/visible spectrophotometer.
Reagents
1. Ammonium acetate(NH4C2H30 2), 5 M.
2. Hydroxylamine hydrochloride(NH20H • HCI), 10% (10 g NH20H •
HCI diluted to 100 mL total volume with deionizedwater).
3. 1,1O-phenanthrolinereagent. Dissolve 0.30 g of 1,1O-phenanthroline
monohydratein waterby heatingthe mixture to 80c C. Cool the solution
and add water to a final volume of 100 mL.
5. StandardFe solution, 100 mg Fe L-t. StandardFe solutions are com-
mercially available,but also can be preparedby dissolving0.1 g of ana-
lytical reagentgradeiron wire in 100 mL of 1.8 M H2S04• Warm if nec-
essary to obtain complete dissolution. Dilute to 1 L with deionized
water.
6. StandardFe solutions,5 mg L-t. Add 10 mL of 18 M H2S04 to 50.0 mL
of standardFe solution having 100 mg L-t of Fe, and dilute the solution
to 1 L with deionizedwater.
IRON 661
Procedure
To determinetotal Fe (Fez+plus Fe3+) concentrationof the extractor digest,

transfera suitablequantity of the sampleto a 50-mL volumetric flask, suchthat
the final concentrationof Fe in the flask will be from 0.25 to 2.25 mg L -1. Add 1
mL of 10% NHzOH • Hel and 1 mL of 1,10-phenanthrolinereagent.Mix the
flask, and add2 mL of 5 M N14CzH30 z. Mix the flask again. Measurethe pH of
the sampleto determineif it is betweenpH 3 and 5. If the pH is not within this
range,add 6 M Hel slowly until pH 4 is obtained,and bring to volume with dis-
tilled water. Record the quantity of Hel required, and addthis amount to each
samplein this sampleset. The color developsalmost immediately and is stable
for at least 15 d. Transferthe samplesto spectrophotometer tubes (or utilize a
flow-through cell) for the determinationof absorbanceat 510 nm. A 1-cm path
length may be acceptablefor total Fe concentrationsranging from 0.25 to 2.25
mg L-l; however,the detectionlimit can be improvedby using photocellswith a
5- or lO-cm light path. The concentrationof Fe in a blank solution (triplicate
samples),which has been carried through the entire analytical process,also is
determined.Iron in the digestor extracton a soil weight basisis calculatedas fol-
lows
Fe, mg kg-1 = [(Fetestsolnmg L-1 - Feblankmg L-l)
(extractingsolution volume, L)(dilution)]/(weight of sample,kg)
To determineFez+ concentration,the sampleis treatedas describedabove,

exceptthat NHzOH • Hel is not addedduring the color developmentstep.Sam-
ples for determinationof Fez+ shouldbe preparedand storedin the dark or under
a red photographicdarkroom lamp to prevent photoreductionof Fe3+ to Fez+.
Also, pH control is essentialto retardrate of reduction.Ferric iron concentration
can then be determinedby subtractingFe z+ concentrationfrom total Fe concen-
tration of the extract.
PrepareFe standardsof 0.00,0.25,0.75,1.25,1.75, and 2.25 mg Fe L-l by
adding0-, 5-, 15-, 25-,35-, and 45-mL portionsof a 5 mg Fe L-l stock solution
to 100-mLvolumetricflasks. Add reagentsso that the samplesand standardswill
havethe identical matrix composition,bring to volume, and developthe color of
the standardsaspreviouslydescribed.Preparea calibrationcurveof the standards
at 51O-nm wavelengthusing a visible or UV/visible spectrophotometer.Deter-
mine concentrationsof Fe in the samplesby comparisonwith the standardcurve.
REFERENCES
American Public Health Association. 1981. Standardmethodsfor the examinationof water and
wastewater.APHA, Washington,DC.
Arshad,M.A., R.I. St. Arnaud, and P.M. Huang. 1972. Dissolutionof trioctahedrallayersilicatesby
ammonium oxalate, sodium dithionite-citrate-bicarbonate,and potassium pyrophosphate.
Can. J. Soil Sci. 52:19--26.
Bascomb,C.L. 1968. Distribution of pyrophosphate-extractable iron and organic carbon in soils of
variousgroups.J. Soil Sci. 19:251-258.
662 LOEPPERT & INSKEEP
Bascomb,C.L., and K. Thanigasalam.1978. Comparisonof aqueousacetylacetoneand potassium

pyrophosphatesolutionsfor selectiveextractionof organic-boundFe from soils. 1. Soil Sci.
29:382-387.
Bernas,B. 1968. A new methodfor decompositionand comprehensiveanalysisof silicatesby atom-
ic absorptionspectrometry.Anal. Chern. 40:1682-1686.
Borggaard,O.K. 1976. Selectiveextraction of amorphousiron oxide by EDTA from a mixture of
amorphousiron oxide, goethite,and hematite.J. Soil Sci. 27:478-486.
Borggaard,O.K. 1982. Selectiveextractionof amorphousiron oxides by EDTA from selectedsili-
catesand mixturesof amorphousand crystalline iron oxides. Clay Miner. 17:365-368.
Borggaard,O.K.1988. Phaseidentification by phasedissolutiontechniques.p. 83-98.In J.w. Stucki
et al. (ed.) Iron in soils and clay minerals.Reidel, Dordrecht,the Netherlands.
Carstea,D.D., M.E. Harward,and E.G. Knox. 1970.Comparisonof iron and aluminumhydroxy inter-
layers in montmorillonite and vermiculite: II. Dissolution. Soil Sci. Soc. Am. Proc.
34:522-526.
Chao,T.T., and L. Zhou. 1983. Extraction techniquesfor dissolutionof amorphousiron oxides from
soils and sediments.Soil Sci. Soc. Am. 1. 47:225-232.
Cihacek,L.I. 1988. DTPA extractableiron soil test correlationwith hybrid grain sorghumproduction
on gypsumaffectedsoils. J. Plant Nutr. 11:1533-1544.
Collins, P.F., H. Diehl, and G.F. Smith. 1959. 2,4,6-tripyridyl-s-triazineas a reagentfor iron. Anal.
Chern.31:1862-1867.
Crawley, R.H.A., and M.L. Aspinall. 1955. Determinationof impurities in tungstenmetal. II. Deter-
mination of iron. Anal. Chim. Acta 13:376-378.
Deb, B.C. 1950. The estimationof free iron oxides in soils and clays and their removal. J. Soil Sci.
1:212-220.
Dudas,M.I., and M.E. Harward. 1971.Effect of dissolutiontreatmenton standardand soil clays. Soil
Sci. Soc. Am. J. 35:134-140.
Fleming, A.L., RL. Chaney,and B.A. Coulombe.1984. Bicarbonateinhibits Fe-stressresponseand
Fe uptake-translocation of chlorosis-susceptiblesoybeancultivars. I. Plant Nutr. 7:699--714.
Geiger, S.c., and R.H. Loeppert. 1988.The effect of soil gas phaseC02 concentrationon Fe defi-
ciency chlorosis of sorghum and soybean grown in calcareous soils. 1. Plant Nutr.
11:1493-1502.
Geiger, S.C., and RH. Loeppert. 1994. Influence of air drying and soil gas phasecarbondioxide on
DTPA-extractableiron in calcareoussoils. Commun.Soil Sci. Plant Anal. 25:341-349.
Goodman,B.A 1987. The characterizationof Fe complexeswith soil organicmatter.p. 677--fJ87.ln
J.W. Stucki et al. (ed.) Iron in soils and clay minerals.Reidel, Dordrecht,the Netherlands.
Haldane,AD. 1956. Determinationof free iron oxide in soil. Soil Sci. 82:483-489.
Havlin, J.L., and P.N. Soltanpour.1981.Evaluationof the N~HC03-DTPA soil test for iron andzinc.
Soil Sci. Soc. Am. J. 45:70--75.
Havlin, J.L., and P.N. Soltanpour.1982.Greenhouseand field evaluationof the NH4HC04-DTPA soil
test for Fe. J. Plant Nutr. 5:769--783.
Havlin, 1.L.,and P.N. Soltanpour.1984. Changesin NH4HC03-DTPA extractablezinc and iron as
affectedby varioussoil properties.Soil Sci. 137:188--193.
Haynes,R.I., and RS. Swift. 1983. An evaluationof the use of DTPA and EDTA as extractantsfor
micronutrientsin moderatelyacid soils. 74:111-122.
Hodges,S.C., and L.W. Zelazny. 1980.Determinationof noncrystallinesoil componentsby weight
differenceafter selectivedissolution.Clays Clay Miner. 28:35-42.
Holmgren, G.G.S. 1967. A rapid citrate-dithionite extractableiron procedure.Soil Sci. Soc. Am.
Proc. 31:210--211.
Hossner,L.R. 1996. Dissolution for total elementalanalysis. p. 49--64. In D.L. Sparkset al. (ed.)
Methods of soil analysis. Part 3. Chemical methods.SSSABook Ser. 5. SSSA and ASA,
Madison,WI.
Inskeep,W.P., and P.R Bloom. 1986.Effects of soil moistureon soil pC02, soil solution bicarbonate,
and iron chlorosisin soybeans.Soil Sci. Soc. Am. J. 50:946-952.
Inskeep,W.P., and S.D. Comfort. 1986. Thermodynamicpredictionsfor the effectsof root exudates
on metal speciationin the rhizosphere.J. Plant Nutr. 9:567-576.
Jackson,M.L. 1958. Soil chemicalanalysis.PrenticeHall, EnglewoodCliffs, NJ.
Jackson,M.L., C.H. Lim and L.W. Zelazny. 1986. Oxides, hydroxides, and aluminosilicates.p.
101-150.In A Klute (ed.) Methodsof soil analysis.Part 1. 2nd ed. Agron. Monogr. 9. ASA
Jeanroy,E., and B. Guillet. 1981.The occurrenceof suspended ferruginousparticlesin pyrophosphate
extractsof somesoil horizons.Geoderma26:95-105.
IRON 663
Jeffery, P.G., and D. Hutchison.1981. Chemicalmethodsof rock analysis.Pergamon,Oxford.

Jenne,E.A., J.w. Ball, and C. Simpson.1974. Determinationof trace metalsin sodiumdithionite-cit-
rate extractsof soils and sedimentsby atomic absorption.J. Environ. Qual. 3:281-287.
Khan, A., and P.N. Soltanpour.1978. Effect of wetting and drying on DTPA-extractableFe, Zn, Mn,
and Cu in soils. Commun.Soil Sci. Plant Anal. 9:193-202.
Kilmer, FJ. 1960. The estimationof free iron oxidesin soils. Soil Sci. Soc. Am. Proc. 24:420-421.
soils and rocks. Environ. Sci. Tech. 17:362-368.
Kiss, E. 1984. Investigationof some asymmetrictriazines as reagentsfor the spectrophotometric
micro-determinationof the iron oxidation statein silicates.Anal. Chim. Acta 161:231-244.
Kittrick, J.A. 1983. Accuracy of several immiscible displacementliquids. Soil Sci. Soc. Am. J.
47:1045-1047.
Komadel, P., and 1.w. Stucki. 1988. Quantitativeassayof mineralsfor Fe2+ and Fe3+ using1,10-
phenanthroline:III. A rapid photochemicalmethod.Clays Clay Miner. 36:379-381.
Leggett,G.E., and D.P. Argyle. 1983.The DTPA extractableiron, manganese,copper,and zinc from
neutral and calcareoussoils underdifferent conditions.Soil Sci. Soc. Am. J. 45:518-522.
Lim, C.H., and 1.L. Jackson.1982. Dissolution for total elementalanalysis.p. 1-12. In A.L. Pageet
al. (ed.) Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Lindsay, w.L. 1979. Chemicalequilibria of soils. JohnWiley & Sons,New York.
Lindsay, w.L. 1984. Soil and plant relationshipsassociatedwith iron deficiency with emphasison
nutrient interactions.1. Plant Nutr. 7:489-500.
Lindsay, W.L., and w.A. Norvell. 1978. Developmentof a DTPA soil test for zinc, iron, manganese,
and copper.Soil Sci. Soc. Am. J. 42:421-428.
Lindsay, w.L., and A.P. Schwab.1982. The chemistryof iron in soils and its availability to plants. J.
Plant Nutr. 5:821--840.
Loeppert,R.H., and C.T. Hallmark. 1985. Indigenoussoil propertiesinfluencing the availability of
iron in calcareoussoils. Soil Sci. Soc. Am. 1. 49:597-603.
Loeppert,R.H., L.c. Wei, and w.R. Ocumpaugh.1994. Soil factors influencing mobilization of trace
metals in calcareoussoils. p. 345-363.In 1.A. Manthey et al. (ed.) Biochemistry of metal
micronutrientsin the rhizospohere.Lewis, Boca Raton.
Loveland,P.J. 1988.The assayfor iron in soil and clay minerals.p. 99-140.In J.W. Stucki et al. (ed.)
Iron in soils and clay minerals.Reidel, Dordrecht,the Netherlands.
Loveland,PJ.,and P. Digby. 1984. The extractionof Fe and Al by 0.1 M pyrophosphatesolutions:A
comparisonof sometechniques.1. Soil Sci. 35:243-250.
Marschner,H., V. Romheld,and M. Kissel. 1986. Different strategiesin higher plantsin mobilization
and uptakeof iron. J. Plant Nutr. 9:695-713.
McKeague,J.A. 1967. An evaluationof 0.1 M pyrophosphateand pyrophosphate-dithionite in com-
parison with oxalate as extractantsof the accumulationproductsin podzolsand some other
soils. Can. I. Soil Sci. 47:95-99.
McKeague,1.A., 1.A. Brydon, and N.M. Miles. 1971. Differentiation of forms of extractableiron and
aluminum in soils. Soil Sci. Soc. Am. Proc. 35:33-38.
McKeague,I.A., and J.H. Day. 1966. Dithionite- and oxalate-extractable Fe and Al as aids in differ-
entiatingvariousclassesof soils. Can. I. Soil Sci. 46:13-22.
McKeague, 1.A., and P.A. Schuppli. 1982. Changesin concentrationof iron and aluminum in
pyrophosphateextractsof soil and compositionof sedimentresultingfrom ultracentrifugation
in relation to spodichorizon criteria. Soil Sci. 134:265-270.
McKeague,1.A., and B.H. Sheldrick. 1977. Sodium hydroxide-tetraboratein comparisonwith sodi-
um pyrophosphateas an extractantof complexescharacteristicof spodichorizons.Geoderma
19:97-104.
Mehra, D.P., and M.L. Jackson.1960. Iron oxide removal from soils and clays by a dithionite-citrate
systembuffered with sodium bicarbonate.In Clays and clay minerals. Proc. 7th Natl. Congr.
Pergamon,London.
Morris, D.R., R.H. Loeppert,and TJ. Moore. 1990. Indigenoussoil factors influencing Fe chlorosis
of soybeanin calcareoussoils. Soil Sci. Soc. Am. 1. 15:1329-1336.
Murad, E., and W.R. Fischer. 1988.The geobiochemicalcycle of iron. p. 1-18. In 1.w. Stucki et al.
(ed.) Iron in soils and clay minerals.Reidel, Dordrecht,the Netherlands.
Nordstrom, D.K. 1982. Aqueous pyrite oxidation and the subsequentformation of secondaryiron
minerals.p. 37-56. In lA. Kittrick et al. (ed.) Acid sulfateweathering.SSSASpec.Publ. 10.
SSSA, Madison,WI.
Norvell, w.A. 1972. Equilibria of metal chelatesin soil solution. p. 115-138.In 1.1. Mortvedt (ed.)
Micronutrientsin agriculture.SSSA,Madison,WI.
LOEPPERT& INSKEEP
O'Connor,G.A. 1988. Use and misuseof the DTPA soil test.J. Environ. Qual. 17:715-718.
Olson, R.Y. 1965. Iron. p. 963-973.In c.A. Black (ed.) Methodsof soil analysis.Part 2. Agron.
Olson,R.V., and R. Ellis. 1982.Iron. p. 301-312.In A.L. Page(ed.) Methodsof soil analysis.Part2.
Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Rtimheld, Y., and H. Marschner.1986. Mobilization of iron in the rhizosphereof different plant
species.Adv. PlantNutr. 2:155-204.
Ryan,J.N., and P.M. Gschwend.1991.Extractionof iron oxidesfrom sedimentsusing reductivedis-
solutionby titanium (III). ClaysClay Miner. 39:509-518.
Schulze,D.G., andJ.B. Dixon. 1979.High gradientmagneticseparationof iron oxidesandothermag-
netic materialsfrom soil clays. Soil Sci. Soc.Am. J. 43:793-799.
Schuppli,P.A., GJ. Ross,and J.A. McKeague.1983.The effective removalof suspendedmaterials
from pyrophosphateextractsof soils from tropical and temperateregions.Soil Sci. Soc. Am.
1. 47:1026-1032.
Schwertmann,U. 1964.The differentiationof iron oxide in soils by a photochemicalextractionwith
acid ammoniumoxalate.Z. PfIanzenemaehr. Dueng.Bodenkund.105:194-201.
Schwertmann,U. 1973. Use of oxalatefor Fe extractionfrom soils. Can. J. Soil Sci. 53:244-246.
Schwertmann,U. 1988. Somepropertiesof soil and syntheticiron oxides.p. 203-251.In J.W. Stuc-
ki et al. (ed.) Iron in soils andclay minerals.Reidel,Dordrecht,the Netherlands.
Schwertmann,U. 1991. Solubility and dissolutionof iron oxides. p. 3-27. In Y. Chen and Y. Hadar
(ed.) Iron nutrition and interactionsin plants.K1uwer, Dordrecht,the Netherlands.
Schwertmann,U., D.G. Schulze,and E. Murad. 1982. Identification of ferrihydrite in soils by disso-
lution kinetics, differential x-ray diffraction and mtissbauerspectroscopy.Soil Sci. Soc.Am.
J. 46:869-875.
Schwertmann,U., andR.M. Taylor. 1989.Iron oxides.p. 379-438.In J.B. Dixon andS.B. Weed(ed.)
Minerals in soil environments.SSSA,Madison,WI.
Shuman,L.M. 1980. Effectsof soil temperature,moisture,and air-drying on extractablemanganese,
iron, copper,and zinc. Soil Sci. 130:336-343.
Smith, F.G., W.H. McCurdy, and H. Diehl. 1952.The colorimetric determinationof iron in raw and
treatedmunicipal water suppliesby use of 4,7-diphenyl-l,1O-phenanthroline. Analyst (Lon-
don) 77:418-422.
Soltanpour,P.N. 1991.Determinationof nutrientavailability andelementaltoxicity by AB-DTPA soil
testand ICPS.p. 165-190.In B.A Stewart(ed.) Advancesin soil science.Vol. 16. Springer-
Verlag, New York.
Soltanpour,P.N., A. Khan, and W.L. Lindsay. 1976.Factorsaffecting DTPA-extractableZn, Fe, Mn,
and Co from soils. Commun.Soil Sci. PlantAnal. 7:797-821.
SoItanpour,P.N., and AP. Schwab.1977.A new soil test for simultaneousextractionof macro-and
micro-nutrientsin alkaline soils. Commun.Soil Sci. PlantAnal. 8:195-207.
Soltanpour,P.N., and S. Workman. 1979. Modification of the ~HC03-DTPA test to omit carbon
black. Commun.Soil Sci. PlantAnal. 10:1411-1420.
Stucki, J.W. 1981a.The quantitativeassayof mineralsfor Fe2+ and Fe3+ using 1,100phenanthroline:
II. Sourcesof variability. Soil Sci. Soc.Am. J. 45:633-637.
Stucki, J.W. 1981b.The quantitativeassayof mineralsfor Fe2+ and Fe3+ using 1,IO-phenanthroline:
II. A photochemicalmethod.Soil Sci. Soc.Am. J. 45:638-641.
Stumm,w., and G. Furrer. 1987.The dissolutionof oxidesandaluminumsilicates:Examplesof sur-
face coordinationcontrolled kinetics. p. 197-219.In W. Stumm(ed.) Aquatic surfacechem-
istry. JohnWiley & Sons,New York.
Tamm,O. 1922.Eine methodezur bestimunungder anorganischen kamponentedesgelkomplexesin
boden.Medd. Stat.Skogsforsoks.(Stockholm)19:387-404.
Thome,D.W. 1941. Factorsinfluencing the solubility of iron and phosphorusin chlorotic and non-
chlorotic areasof Hyrum clay loam. Iowa StateColi. J. Sci. 15:433-445.
Thome,D.W., and A Wallace. 1944. Somefactorsaffecting chlorosison high-lime soils. I. Ferrous
and ferric iron. Soil Sci. 57:299-312.
van Breeman,N. 1988. Redox processesin iron and sulfur involved in the formation of acid sulfate
soils. p. 825-841.In J.W. Stucki et a1. (ed.) Iron in soil andclay minerals. Dordrecht,Reidel,
the Netherlands.
Walker, AL. 1983.The effectsof magnetiteon oxalate-anddithionite-extractableiron. Soil Sci. Soc.
Am. J. 47:1022-1026.
Published 1996
Chapter 24
Manganese
ROBERT P. GAMBRELL, Wetland Biogeochemistry Institute, Louisiana

State University, Baton Rouge, Louisiana
INTRODUCTION
Manganese(Mn) is an essentialtrace metal for plant nutrition. Most soil testing

researchhasfocusedon deficiency problems,but Mn also is known to causetox-
icity problemsto plants when soil levels and physicochemicalconditionsfavor
excessplant availability (Duncan et aI., 1983; Fales & Ohki, 1982; Foy et aI.,
1981; Nelson, 1983; and Ritchie, 1989). Thus soil analysisfor Mn is of interest
from both deficiency and toxicity perspectives.
Manganeseis similar to Fe in its chemicalbehaviorin soils and geological
materials.This similarity is due in part to the different valencestatesin which
both commonly can be found in soils, and, the influence of soil pH and redox
conditionson their valencestatesand various chemical forms. Manganesemay
exist in many valencestates,but underearth surfaceconditions,only the 2+, 3+,
and 4+ valencesare found. However,the trivalent form is reportedto be unstable
in solution.
Total Mn is found over a wide concentrationrangein soils: from <20 Ilglg
to concentrations>3000 /lg/g (Krauskopf, 1972) with typical total levels around
600 /lg/g (Fuller & Warrick, 1985).Somevolcanicashsoils in Hawaii are notable
exceptionswith Mn contentsexceeding10% (Stevenson,1986). Levelsof chem-
ically mobile and plant-availableforms tend to be less, but also exhibit a wide
range. Soil pH and redox potential are the primary factors affecting Mn avail-
ability. Waterloggedconditionsand/orpH levels below six favor reductionto the
more mobile and plant availabledivalent Mn form (Mn2+), while higher pH and
oxidized soil conditionsfavor Mn 2+ oxidation to the Mn 4+ form which is gener-
ally found in insolubleoxides in soils.
Manganeseis found in a numberof generalchemical forms in soils. Ste-
venson(1986) lists thesegeneralforms or pools for trace metalsas:
1. Soluble. This includes free cations or cations complexedwith organic
and inorganicligands.
2. Exchangeable.Exchangeablerefersto cationsweakly held on the cation
exchangesites of clay mineralssuch that other cations may causedis-
Book Seriesno. 5.
665
666 GAMBRELL
placementto solubleor otherforms. Soil organic matteralso contributes

to this form.
3. Specifically adsorbedor retained.This includesspecific adsorptionre-
actionscontributingto more tight bondingthan exchangeprocesses,and
adsorptionto oxidesof Fe. Also, Mn may form its own oxidesthat can
adsorbor occludeother trace metalsin soils.
4. Adsorbed or complexedby soil organic matter. This can include ex-
changeablecations, but a considerableportion of potentially available
metalscan be held much more strongly by organic chelationprocesses.
Most researchon the stability of metal-humicmaterial complexessup-
ports the Irving-Williams seriesin which Mn is found to be much less
strongly associatedwith humic materialthan many other traceand toxic
metalsof interest in soils (McBride, 1978; Schnitzer& Hansen,1970;
Verloo & Cottenie,1972).
5. Insolubleprecipitates.This includesprecipitatedcompoundsof Mn and
large molecularweight oxide forms.
6. A constituentof minerals. This would include Mn minerals as well as
Mn that may be presentby isomorphoussubstitutionfor Al and/orFe in
crystallineclay minerals.
SolubleMn is the most mobile and plant availableform. ExchangeableMn
also is consideredreadily availabledue to the easewith which it can be displaced
and become soluble.Manganesepresentwithin the crystallinelattice structureof
mineralseither as a primary constituentor as an impurity resultingfrom isomor-
phoussubstitutionis unavailablefor leachingor plant uptake and may become
availableonly as a consequence of the very slow processof mineral weathering.
Only a small proportionof the total Mn in soils is found in the readily avail-
able solubleand exchangeable forms. Betweenthe readily availableand unavail-
able forms are the potentially availableforms that may be releasedto more avail-
able forms dependingon soil chemicaland microbial conditions,and especially
as changesin theseconditionsoccur.
Adams(1965) statedthat plant-availableMn is more dependenton pH than
any otherfactor, althoughlevelsof organic matterandsoil aerationdo exertsome
influence on availability. A changein soil pH or oxidation-reductionconditions
(or both) may have markedinfluence on transformationsbetweenreadily avail-
able and potentially availableforms. Acid and reducingsoil conditionsfavor di-
valent Mn forms that result in increasedlevels of dissolved and exchangeable
Mn. Where Mn toxicity is a problem, the soils are probably acid, reducing, or
both. Increasingsoil pH and oxidation conditionsfavor the formation and stabil-
ity of sparinglysolublehigheroxide Mn minerals,thoughconsiderableadsorbed
Mn may persist.
Manganesedeficienciesmost likely occur under oxidizing conditions in
high pH soils, especiallyin leachedsoils that havebeenlimed. Organicsoils, de-
pletedof Mn underreducingconditions,are often deficient in Mn after drainage
(Levesque& Mathur, 1988).
Kubota and Allaway (1972) reviewed the literature on the distribution of
traceelementproblemsand reportedMn deficiencytendsto be more prevalentin
humic regionsof the East than in the West. Liming acid Southeasternsoils often
MANGANESE 667
resultsin Mn deficiencies(Gettier et aI., 1985; Martini & Mutters, 1985). Where

problemsexist in the West, it was reported to be mostly in Mn-sensitivefruit
trees. Also, Mn toxicity has beennoted in many parts of the country in moder-
ately to strongly acid soils.
Much of the potentially available Mn in typical aerobic agricultural soils
may be in the form of higher Mn oxide minerals,or adsorbedor coprecipitated
with hydrousoxidesof Fe. The chemistryof Mn in oxidizedsoils is complexdue
to the existenceof many nonstoichiometricoxides with mixed valencies.The
associationof many other trace and toxic metals with Mn oxides makes these
materialsof interestfrom geochemicaland environmentalperspectivesin soils
and sedimentsas well.
ANALYSIS
InstrumentalApproaches-Introduction
Improvementsin analytical instrumentationand procedurescontinueto be

madewith time. Whereas15 yr ago most Mn analysesfor agricultural purposes
in industrializedcountrieswould have been done by flame atomic absorption
spectroscopy(AA), today, inductively coupled plasma emission spectroscopy
(ICP) is becomingthe most common instrumentfor researchas well as routine
soil and plant analyses.However,a textsuchas this alsoshouldbe useful to agri-
cultural scientistsand chemistsin less-developedcountrieswhere more recent
instrumentationfor chemicalanalysesmay not be available.Therefore,this chap-
ter will include the colorimetric analysesfor Mn reportedby Adams (1965) and
Gambrell and Patrick (1982) in earlier editionsof this publication.
Other instrumentscapableof Mn analysisinclude x-ray emission,neutron
activation, and ion chromatographymethods. While these instrumental tech-
niquescan be appliedto Mn in soil samplesor soil sampleextracts,they will not
be coveredhereas their usefor tracemetal analysesin agriculturalsamples isnot
common,and for most, not as convenientas atomic absorptionor plasmaemis-
sion spectroscopymethods.
Inductively CoupledArgon PlasmaEmissionSpectroscopy
Samplepreparationmethodsare generallythe samefor both AA and ICP

spectroscopy.
Watsonand Isaac(1990) reviewedanalyticalinstrumentationapplicableto
agricultural samplesand reported the work of others who showed ICP results
comparedfavorably with AA, sparkemission,x-ray fluorescence,energydisper-
sive x-ray fluorescence,neutron activation analysis, and, rotating disk spark
emissionspectrometry.Studieswere cited showing ICP gave excellentcompar-
isons with Mn results from atomic absorptionanalysesof various soil extracts
including the Mehlich I and DTPA extracts,two important soil testing proce-
duresfor Mn to be discussedelsewherein this chapter.One of the studiesthey
668 GAMBRELL
cited concluded systematic and random errors associatedwith ICP were either
equivalentor smallerthan thoseof other instruments.Reportscomparingresults
of the samesamplesby many different laboratoriesshowedMn to be amongthe
elementsgiving the best precision.
Preparationof water, soil, and tissuesamplesfor Mn analysisby ICP spec-
troscopy is essentiallythe same as for AA spectroscopy,but there are a few
specific precautionsthat will be discussedbelow. General considerationsand
operatingparametersfor an ICP, such as detectionlimits, linear range, rf power
levels, argon flow rates,signal readingposition in the plasma,and type of nebu-
lizer, will not be discussedhere.
One important precautionwhen preparingcombinedelementalcalibration
standardsfrom stocksolutionsis that the stocksolutionsmust not containsignif-
icant contaminationwith other elementsmeasured by the ICP. This is not critical
with atomicabsorptionasexplainedbelow. As a simpleexample,considera com-
bined Fe and Mn ICP calibration standard(dependingon the instrumentand its
setup,most ICP calibrationstandardscontainmore than two elements).If a com-
bined Fe-Mn standardwere madewith an Fe compoundthat unknowingly con-
tained 10% as much Mn as Fe, the calibrationand ICP analysiswould give good
resultsfor Fe but would be in error for Mn becausethe unknown extra Mn from
the Fe sourcewould makethe Mn concentrationhigherthan intended.Thus a cal-
ibration solution preparedto contain 10 mgIL might contain 11 mgIL which
would causea problem when the computercalibration file is programmedto
processthis solution'semissionsignal as 10 mgIL. This is not a problem with a
one-element-at-a-time methodsuch as AA spectroscopy,but it can contributeto
errorswherevermultielementstandardsare usedwith simultaneousmultielement
analysessuch as occurswith an ICP.
Another problem with ICP over AA is interelementinterferences.Some-
times the emissionline of an elementotherthan the oneof interestcan be so close
to the analytical line of interestbeing measuredthat a positive interferencewill
occur. This is a potentialproblem thatcan be managedby one or a combination
of methods.Sincemost elementshaveseveraluseableemissionlines, experience
with particulartypesof samples(soil, ore, or tissue,for example)enablesone to
select analytical lines to eliminate or minimize interelementinterference.The
type of samplesto be analyzedand anticipatedinterelementinterferencesare usu-
ally a considerationwhen selectinganalytical lines to use when a simultaneous
ICP is ordered,or when setting up a sequentialICP. However, it is not usually
possibleto select emissionlines to read that eliminate all interelementemission
interference.
Another methodof reducing interelementinterferenceis backgroundcor-
rection that is available in most instruments.Backgroundcorrectionmay some-
times solve interelementinterferenceproblems or only somewhatreduce the
interference,dependingon the instrumentand other factors.
A third methodusedby the authoris to establishfor eachelementmeasured
what the interfering elementsare (if any), quantify the interferenceand calculate
a correction factor, and then run all of the generatedelementaldata through a
spreadsheet program that automaticallymakescorrections,if necessary,for inter-
fering elements.The correction factors are determinedusing high-purity single
MANGANESE 669
elementsolutionsat a concentrationjust below the linear rangefor that element

looking for apparentmeasurableamountsof otherelements.The correctionfac-
tors are redeterminedperiodically, but usually no significant changesare neces-
sary unlessinstrumentaloperatingconditionshave beenaltered.This approach
hasbeendescribedelsewhere(USEPA, 1986).The successfulapplicationof the
spreadsheetinterferencecorrection technique requires the instrument to be
equippedto measurethose elementsthat may be presentin a samplecausing
interference.For tracemetalsin soils work, the instrumentshouldhavethe abil-
ity to measureall the major metalsfound in soils to enablecheckingfor interele-
ment interferenceand making a correction.For example,on the ICP with which
the authoris mostfamiliar, the analyticalline for Mn is 257.6nm and it hasbeen
determinedthat the major soil ions do not measureablyinterferewith Mn, and no
correctionon Mn is needed.Manganese,however,doesexhibit some interele-
ment interferencefor a coupleof other traceelements,and a correctionis made
for Mn interferenceon these.Thesecorrectionsare usuallysignificantonly where
a large concentrationof the interfering elementis present,suchas occurswith a
strong acid extractof a soil or sediment,or an acid digest of someplant tissue
samples.
Reagents
Generally,ICP is a multielementinstrumentalmethodthat usescalibration
standardscontaining known concentrationsof several elements of interest,
whereascalibration standardsfor AA normally contain single elements.Induc-
tively coupledplasmaemissionspectroscopycalibrationstandardsare available
commercially,either as custom-madesolutionscontainingcombinationsof the
desiredelementsat requiredconcentrations,or as individual stocksolutionscon-
taining single elementswhich one can use to makecombinedelementalcalibra-
tion standards.
Furtherdetailsare not given herebecausethe elementsto be includedin a
multielementstandardand the concentrationsof theseelementsdependson the
configurationof a particular ICP and operatorpreferences.Should one wish to
make a single elementstandardfor Mn for ICP analysis,a commerciallyavail-
able stock atomic absorptionMn standardwould be suitablefor making appro-
priate dilutions. If the stock solution describedfor atomic absorptionelsewhere
in this chapteris usedfor calibratingan ICP, it is not necessaryto addthe Ca solu-
tion to eliminateSi interferencefor Mn as describedin the proceduresectionfor
atomic absorption.
Procedure
The procedurefor preparing the appropriateICP calibration standards
dependson the configuration of a particular instrument.Generally, a reagent
blank is usedto establishthe zero concentrationfor the instrument'scalibration
curve, and, dependingon the instrumentcapabilities,a secondsolution contain-
ing Mn (and other elements)at a specifiedconcentrationwithin the linear range
is usedto get the upper point on a two-point standardcalibration curve. Some
instrumentsand associatedcomputersoftware permit multiple point standards
670 GAMBRELL
that may even extendbeyondthe linear rangefor the metal of interest.Thus the
manufacturer'sinstructionsand instrumentconfigurationwill determinehow to
make up calibrationstandards.
Samplesare analyzedaccordingto operatinginstructionsfor a particular
instrument.Precautionsare takento be sureproblemshave notcompromisedper-
formance.These includemakingfrequentmanualor automaticcalibrationchecks
for instrumentdrift, for nebulizerclogging problemsthat contribute to reduced
sampleuptakerate.
Atomic Absorption
For more informationon the Atomic AbsorptionMethod,seeGambrelland

Patrick (1982).
The two commonatomizationmethodsfor atomic absorptionare flame and
electrothermal(for example,graphite furnace). For purposesof Mn analysisof
soil and plant samples,the readily availableflame atomizationinstrumentsusu-
ally have adequatesensitivity.
Reagents
1. Manganesestock solution, 1000 mgIL. Dissolve 3.076 g of manganous
sulfatemonohydrate(MnS04 • H20) in approximately200 mL of high-
purity water in a 1000-mLvolumetric flask. Add 1.5 mL of concentrat-
ed nitric acid (HN03), and make to volume with additionalwater. Each
milliliter of this solution contains1.00 mg of Mn.
2. Calcium solution. Dissolve630 mg of calcium carbonate(CaC03) in 10
mL of concentratedhydrochloricacid (HCI). Add 200 mL of high-puri-
ty water, and heatgently to obtain completesolution. Cool and dilute to
1000 mL with additional water. (This amendmentis not necessaryfor
ICP analysis.)
3. Nitric acid (HN03), concentrated.
Procedure
Flame atomic absorptionwith direct aspirationinto an air-acetyleneflame
shouldprovide sufficient sensitivity for most Mn determinations(APHA, 1980).
Silica can sometimesinterfere with the determinationof Mn by AA (APHA,
1980). Where this may be a problem, interferencecan be eliminatedby the addi-
tion of Ca. Twenty-five milliliters of Ca solution should be mixed with 100 mL
of sample and standards(or an equivalent dilution) before analysis (APHA,
1980). Calibration curves should be preparedbasedon the original concentra-
tions of the standards before dilution with the Ca solution. The working concen-
tration rangewill vary dependingon the instrumentusedand instrumentoperat-
ing conditions. Working standardsare preparedby the appropriatedilution of
stocksolutionsand the addition of 1.5 mL of concentratedHN03 per 1000 mL of
calibrationstandard.Nitric acid is addedfor pH control to ensurestability of dis-
solved Mn forms. For example,10 mL of Mn stocksolution plus 1.5 mL of con-
centratedHN03 diluted to 1000 mL with high-purity water will give a working
standardof 10 mgIL.
MANGANESE 671
Colorimetric (periodate)Method
For further information on the Colorimetric (Periodate)Method seeAdams

(1965) and Gambrell and Patrick (1982).
The periodatemethoddevelopedby Willard and Greathouse(1917) is rapid
and reliable over a wide rangeof Mn concentrations.The color developedis sta-
ble for long periodsof time and is little affectedby variation in acid or periodate
concentration,but color developmentmay be affectedby the presenceof reduc-
ing substances.In this colorimetric method,Mn 2+ is oxidized to Mn04 by either
potassiummetaperiodateor sodiumparaperiodatein a strongly acid solution
2Mn2+ + 5104 + 3H20 ~ 2Mn04 + 5103" + 6W
The following discussionof the periodatemethod is taken primarily from

Adams(1965).
Chloride and organic matter are the commonly encounteredreductantsin
soils that may causeinterference,and thesecan be effectively removed.Interfer-
encein the Mn determinationby Cl- is due to the following reactionof Cl- with
permanganate
1OCl- + 2Mn04 + 16W ~ 5Cl2 + 2Mn2+ + 8H20
Additional periodateoxidizesthe Mn 2+ producedby Cl-. Small amountsof

Cl- will not causeinterferenceas long as excessperiodateis added.Most soils do
not contain sufficient Cl- to causeinterference,but of course,reagentscontain-
ing Cl- should be avoided. In soils containing excessCl-, the Cl- may be
removed by repeating boiling with sulfuric acid (H 2S04), carefully washing
down the sidesof the beakerwith water betweenboilings. A statementrecom-
mendingICP or AA as the preferredmethodsof analysisis included with each
extractionprocedurein this chapterthat includesCl--containingreagents.
Interferencefrom organic mattercan be eliminatedby heatingthe sample
with HN03 and H20 2 (describedelsewherein this section), or, heating with
H2S04 and RN03 to remove both organic matter and chloride (also described
elsewherein this section).
The concentrationand kinds of acids that have beensuccessfullyusedfor
the developmentof the Mn04 color vary, but HCl shouldnot be usedbecauseof
Cl- interference.Where there is sufficient Ca in the samplefor the precipitation
of calcium sulfate (CaS04),H2S04 should be used with care becauseturbidity
may develop. Otherwise, repeatedevaporationswith H2S04 is effective in re-
moving excessCl-. Nitric acid does not interfere with the developmentof the
Mn04 color, and evaporationof a samplewith HN03 usually eliminatesreduc-
ing substancesthat may interferewith color development.Ferric periodateis in-
soluble in HN03 but is soluble in H2S04 and phosphoricacid (H 3P04).
Interferencealso may occur from metalssuch as Cr, Ni, and Fe that impart
color to the solution. Ferric iron is the only one likely to be of importancein soil
analyses,and this interferenceis removedby the useof R3P04 which forms a col-
orlesscomplexwith Fe. For soils high in Ti, H3P04 alone is not suitablebecause
672 GAMBRELL
of the precipitation of titanium phosphate.This turbidity can be removed by

addingH2S04.
The procedurefor the periodatecolorimetricmethodis given below (adapt-
ed from Shermanet aI., 1942; Adams, 1965).
Transferan aliquot of a water sampleor water extract of a soil containing
from 0.01 to 0.3 mg of Mn to a 50-mL beaker.Add 5 mL of HN03 and 2 mL of
30% hydrogenperoxide(H20 2), cover the beakerwith a watch glass,and digest
the contentson a steambath or hot plate for 30 min to destroy organic matter.
Removethe cover, and evaporatethe contentsto dryness.
An alternativefor removing organicmatterand Cl- is to add 5 mL of con-
centratedH2S04 and 5 mL of concentratedHN03 to the filtrate, mixing between
additions.Evaporateuntil S03 fumesevolve, then cool. Add 20 mL of high-puri-
ty water and cool again.Add 2 mL of concentratedHN03, 5 mL of 85% H3P04,
and 0.3 g of potassiummetaperiodate(KI0 4) or 0.5 g of sodium paraperiodate
(Na3HzI06).If the amountof Mn is 10 ~g or less,20 mg of AgN03 can be added
to improve sensitivity (APHA, 1980). Coverthe beakerwith a watch glass,bring
the solution to a gentle boil on a hot plate, and continueto boil the solution gent-
ly for at least 5 min after the color develops.For very small amountsof Mn, a
longerboiling periodmay be required.Cool, dilute the solution to 50 mL in a vol-
umetric flask, mix contents thoroughly, and measureabsorbanceat 540 nm.
Determinethe Mn contentof the solution by comparingsampleabsorbancewith
a plot of the absorbanceof variousMn standardsencompassing the concentration
rangeof the samples.
Background
A large numberof soil extractionstudieshave beenconductedto evaluate
the plant availability of soil Mn, and yet there is still needfor improvedsoil test-
ing proceduresfor both deficiencyand toxicity levels.
Ritchie (1989) suggestedthe predictiveability of a numberof soil Mn tests
might be improved by a multiple regressionevaluationincluding soil properties
suchas pH, organic mattercontent,baseratios, manganese oxide levels,and per-
hapsselectedplant properties.
Depending on soil conditions and plant factors, a number of chemical
forms of Mn may contributeto plant availableMn. However,successfulcomplex
or sequentialfractionationschemesmay be necessaryto fully characterizelevels
and forms of Mn. Ritchie (1989) has pointedout that becauseof a lack of speci-
ficity for someextractants,the resultsof some sequentialextractionprocedures
are questionable.Recently,Wardenand Reisenauer(1991) reporteda sequential
chemical fractionation procedurefor Mn that they indicate avoids the use of
extractantsthat reduce Mn oxides until the last step, and includes carbonate-
bound Mn. The procedureis believed to be especially applicable to soil-plant
studiesof Mn accordingto Wardenand Reisenauer(1991).
Total concentrationsof trace elementsin soils are usually not as important
as soil propertiesin determiningplant availability. Thoughmethodswill be given
in this chapterfor determiningtotal soil Mn for soil characterizationpurposes,
MANGANESE 673
soil test proceduresfor Mn as well as other trace metalsare generallydesignedto

predict plant availability. Soluble and exchangeableMn levels are obviously
importantto plant availability, and soil testsfor Mn focusedon theseforms orig-
inally (Ritchie, 1989). Adams(1965) suggestedsoluble Mn in soil is a good indi-
cator of Mn in low-pH soils while exchangeableMn would probablybe a good
estimateof available Mn in high-pH soils. Methodsfor theseforms are present-
ed in this chapter. However, the more successfulextraction proceduresin use
today will extract more than dissolvedand exchangeableforms of Mn indicating
someof the potentially availableforms contributeto plant availability as well.
Martensand Lindsay (1990) recently reviewedsoil testing proceduresfor
trace metalsincluding Mn. In addition to discussingextractionprocedures,they
reviewed the effects of soil samplepreparationmethodsand extractionparame-
ters. In this review, variablesincluded crops, soil types, and the reportedcritical
levels for identifying possibledeficiency conditions. They reportedthe diethyl-
enetriaminepentaacetic acid-triethanolamine[DTPA-TEA, or 0.005 M DTPA;
0.01 M calcium chloride (CaCI2), 0.1 M TEA, buffered at pH 7.3] procedureis
one of the more widely used for identifying deficiency problemswith Mn (and
also for Cu, Fe, and Zn).
This test procedure has found widespread applicability since initially
reported (Lindsay & Norvell, 1978). In a review, Ritchie (1989) reported this
extractant has been shown, in different studies, to include water-soluble,
exchangeable,reducible,and manganeseoxide forms. Norvell (1984) compared
chelatingagentsfor metalsincluding Mn in diversesoil materials.He statedthat
the commonly used DTPA soil test was designedfor weakly acid and alkaline
soils. Though it has apparentlybeensuccessfullyusedwith someacid, reduced,
and contaminatedsoils, he discussedreasonsthe original test might not always
be adequatefor acid, or reducedsoils (for example,sediments)or contaminated
soils, andpointedout thesematerialsmay requireincreasingthe chelatingreagent
concentration,multiple extractions,or higher extractant/soilratios.
For soybean[Glycine max (L.) Merr.] in the southeasternUSA in particu-
lar, the Mehlich I or the Mehlich II procedureand variationsof theseare impor-
tant soil tests(Gettier et at., 1985; Mascagni& Cox, 1988). A procedureusing
0.033 M H3P04 is reportedto be currently usedin somenorth central statesas a
soil test for Mn.
Many of the proceduresin usehavebeenfound to be effective on a region-
al basis. Levesqueand Mathur (1988) evaluatedeight extractantsfor active and
reserveforms of Mn and other elementsin 55 cultivated Histosols. For these
soils, for example,extractantsother than 1 M ammoniumacetate(NH 40Ac) and
the frequently usedDTPA-TEA gave better results.Neilsen et at. (1990) worked
with Mn concentrationin apple (Malus domestica Borkh.) leaf tissue, soil pH,
and severalsoil extractionmethodsin orchardsoils. They concludedsoil testsfor
Mn availability to apple treesmay not perform equally well in deficient and tox-
icity situations.Sharpeand Parks(1982) worked with 71 soil samplesrepresent-
ing sevensoil seriesin the Ultisol and Alfisol orderscomparingthree soil extrac-
tion proceduresfor determiningplant-availableMn. Extractantsstudiedwere the
Mehlich-l (double acid), DTPA, and NH 40Ac. Plants included corn (Zea mays
L.), tobacco(Nicotiana tabacum L.), cotton (Gossypium hirsutum L.), and soy-
674 GAMBRELL
bean.It was reportedthat the Mehlich-1 extractantwas a poor predictor for Mn

uptakeby com and cotton while DTPA-extractableMn was consistentlyreliable
for cotton and soybean.It also was reportedthat any of the soil test procedures
would predict plant Mn levels if a multiple regressionequationwere used that
includedsoil pH, Fe, and Ca. Otherreviews(Ritchie, 1989)also point out that the
most suitableMn extractantmay dependon soil pH.
In this chapter,only a few of the more commonlyusedmethodscan be pre-
sented.Omissionof others doesnot suggesttheir unsuitability, but one shouldbe
awarefrom other sourcesof the appropriateextractantsfor soil typesof research
and agronomicinterest.
Method for Water-solubleManganese
The methodfor water-solublemanganesewas adaptedfrom Shermanet al.

(1942) and Adams(1965).
Comments
Water-solubleMn generallywill not be suitablefor determiningthe labile
Mn pool in Mn-deficient soils. Water-solubleMn may be a valuableparameter
whereMn toxicity is suspected,especiallyin acid soils or poorly aeratedor flood-
ed soils. For flooded, reducedsoils, opportunity for sampleaerationshould be
minimized. Preferably,the determinationshould be made as soon as possible
after obtaining samples.The usual soil samplepreparationstepsof drying and
grinding should be avoided [Bartlett & James,1980; (also Bartlett & James,
1996, seeChapter 25)].The wet or moist soil shouldbe extractedand a separate
aliquot usedfor determiningmoisturecontent(Jackson,1958).
Reagents
1. Nitric acid, concentrated.
2. Hydrogenperoxide,30%.
3. Phosphoricacid, 85%.
4. Potassiumperiodate,solid.
Procedure
Place10 g of soil in a 250-mL Erlenmeyerflask, and add 100 mL of high-
purity water. Shakethe flask for 30 min on amechanicalshaker,then centrifuge
and/orfilter to recoverthe solution phase.If Mn is to be determinedby AA, add
the filtrate to a 100-mL volumetric flask, add 0.15 mL of concentratedHN03 as
a preservative,and bring the sample to volume with high-purity water. Induc-
tively coupledplasmaemissionspectroscopyor AA is the preferred methodof
analysis.If the colorimetric methodof analysisis to be used,seethe sectionon
"Colorimetric (Periodate)Method."
ExchangeableManganese
For more information on exchangeablemanganeseseeAdams (1965).

MANGANESE 675
Comments
This procedureis normally consideredto include both exchangeableand
water-solubleMn. This proceduremay not quantitatively recoverwater-soluble
Mn unlessprecautionsare taken to minimize sampledrying, and perhapsoxida-
tion in the caseof flooded soils, as mentionedfor water soluble Mn (seesection
on "Method for Water-SolubleManganese."
ExchangeableMn is probably a good estimateof readily available Mn in
most soils and is frequently reported in the literature. In anaerobicor low pH
soils, water-solubleMn may be a better estimateof plant-availableMn. An un-
buffered, 1 N solution of NH40Ac is usually the strongsalt solution selectedfor
extractingexchangeablesoil Mn. However, 1 N NH40Ac buffered at variouspH
levels as well as other salt solutionssuchas Ca(N03hor Mg(N03h and varying
concentrationsof CaCl2 are often employed.
Browman et al. (1969) in a greenhousestudy, evaluatedeight soil testsfor
predicting uptakeof native Mn by corn in 63 Wisconsinsoils. In thesestudies,a
syntheticorganicchelateand a H3P04 proceduregave the best resultswhen used
alone, but, when combinedwith a measureof soil pH, a 1 N NH40Ac solution
buffered to pH 7.0 gave a betterprediction equation.
In a review, Cox and Kamprath (1972) reported studies that showed
exchangeableMn (including 1 N NH 40Ac) has been found to be significantly
correlatedwith plant uptake.A neutral 1 N NH40Ac extractof soil also hasbeen
useful for predicting uptake of Mn for wheat (Triticum aestivum L.) in India
(Gajbhiye et aI., 1984).
Reagents
5. Ammonium acetate,1 N, pH 7.
Procedure
Transferto a 100-mLvolumetric flask an aliquot of a neutral,1 N NH 40Ac
extract [obtainedas describedin Chapter40 (Sumner& Miller, 1996) for cation
exchangecapacity]. This extract may be made to volume in the l00-mL volu-
metric flask and analyzeddirectly, or after dilution, by ICP or AA spectroscopy.
Unless the analysis is to be completedsoon after extraction,HN03 should be
addedto ensurestability of the Mn in a soluble form. Becauseof the buffering
capacityof the 1 N NH40Ac, considerablymore HN03 must be addedto lower
the pH to less than two comparedto that amount necessaryfor preserving a
water-solubleMn sample.
Alternatively, if Mn is the only exchangeablecation to be determined,add
100 mL of neutral 1 N NH40Ac to 10 g of soil. Shakethe mixture continuously
for 30 min on a mechanicalshakerand then intermittently for at least 6 h. Cen-
trifuge the suspensionand/orpassa known volume of the solution phasethrough
676 GAMBRELL
a filter. Transferto a 100-mL beakerand analyzeby AA or ICP with appropriate

dilutions if necessary.
If the colorimetric determinationis to be used, proceedas follows. An
aliquot containing from 0.01 to 0.3 mg of Mn should be added to a 100-mL
beaker.Evaporatethe solution to drynesson ahot plate, and continueheatingthe
beakeruntil the NH40Ac ceasesto fume. Completethe procedureas described
for the periodatecolorimetricmethodafter the stepto destroyorganicmatter[see
sectionon "Colorimetric (Periodate)Method"].
Double Acid (Hydrochloric Acid + Sulfuric Acid)-

ExtractableManganese
For more information on the Double Acid-ExtractableManganeseMethod

seeCox (1968).
Comments
This also is known as the Mehlich-1 extractionmethod.The colorimetric
methodmay not be suitablefor this extractionbecauseof substantialCl- addition
with the extractingsolution. A greaterquantity of the double-acidextractantcan
be used as long as the soil sampleis increasedto maintain a 1:4 soil/extractant
ratio. The presenceof charcoalincreasesthe level of Mn extractedby this proce-
dure for soils with low extractableMn.
This extractanthas found widespreadapplication,especiallyin the south-
easternUSA. Cox and Kamprath(1972) suggestthat the typical critical concen-
tration rangeusing this procedureis 5 to 9 mgIL.
Reagents
1. Acid extractantmixture. hydrochloricacid 0.05 N; and H2S04 0.025N.
2. Charcoal.
Procedure
Weigh 5 g of soil and approximately0.2 g of powderedcharcoalinto a 100-
mL flask. Then add 20 mL of a mixture of 0.05 N HCI and 0.025N H2S04 (Cox,
1968). Shakefor 15 min. Centrifuge and/or filter the suspensionto obtain the
solutionphase,and analyze.The preferredmethodof analysisis ICP or AA spec-
troscopy.
acid-ExtractableManganese
Reagents
1. Diethylenetriaminepentaaceticacid extraction solution. Diethylenetri-
aminepentaaceticacid, 0.005 M; CaCI2, 0.01 M; TEA, 0.1 M; adjusted
to pH 7.30.
MANGANESE 677
Procedure
A 1:2 soil/extractantratio is used.Typically, a lO-g aliquot of air-dried soil
is shakenwith 20 mL of the extractingsolutionfor 2 h and centrifugedand/orfil-
tered to obtain the solution phase(Follett & Lindsay, 1971). Inductively coupled
plasma emission spectroscopyor AA are the preferred methods of analysis.
Under most conditions,the Mn concentrationof the extract will be suitable for
direct analysis.
A samplepreservativeis not necessaryif the analysiscan be completed
within a day or two. Otherwise,refrigerationmay be requiredto retard microbial
growth.
Easily ReducibleManganese
For more information on the Easily Reducible ManganeseMethod see

Adams(1965), Shermanet al. (1942).
Comments
ReducibleMn is important in soils becausea large proportion of the total
soil Mn may be present~s oxidesand hydroxidesin typically aerobicagricultur-
al soils. This procedureis still commonlyusedwhereseveralsoil extractionsare
beingtestedto predictplant-availableMn, and Cox and Kamprath(1972)suggest
that 25 to 65 Jlg of Mn per gram of s0il as the critical range for this extraction
procedure.However, exchangeableor certain other of the more recently devel-
opedextractantsare usually found to be betterfor predictingplant availability of
soil Mn.
Reagents
5. Neutral ammoniumacetate,1 N, containing0.2% hydroquinone.
Procedure
Add 100 mL of NH40Ac-hydroquinonesolution to 10 g of soil. Shakethe
mixture on a mechanicalshakerfor 30 min and then intermittently for an addi-
tional 6 h. Centrifuge and/or filter the suspension.This filtrate can be analyzed
directly or after appropriatedilution by ICP or AA spectroscopy.
If the colorimetric methodis to be used,proceedas follows. Transferto a
100-mL beakeran aliquot containingfrom 0.01 to 0.3 mg of Mn and evaporate
the solution to dryness. Continue heating until NH40Ac fumes are no longer
emitted.Cool the beaker,add 5 mL of HN03 and 2 mL of 30% H20 2 (10 mL for
reducedsoils). Cover the beakerwith a watch glass. Digest the contentsfor 30
min on a steambathor hot plate to destroyorganicmatter.Removethe coverand
678 GAMBRELL
evaporatethe contentsto dryness.Cool the beaker,add 30 mL of high-purity

water,2 mL of 85% H3P04, andabout0.3 g of KI0 4, andcompletethe procedure
as describedin the sectionon "Colorimetric (Periodate)Method" for the perio-
date colorimetric method.Subtractthe amountof exchangeableand water solu-
ble Mn determinedseparatelyto obtaineasily reducibleMn.
SelectiveDissolutionof ManganeseOxidesfrom Soils and Sediments
For more information on the SelectiveDissolution of ManganeseOxides

for Soils and Sediments,seeChao(1972).
Comments
This proceduremay be of greaterinterestfor geochemicalstudiesthan for
agronomic purposes. For example,ZasoskiandBurau(1988) examinedthe sorp-
tion of Cd and Zn and interactioneffects of thesetrace and toxic metalson 0-
Mn02. This is just one of many papersdemonstratingthe importanceof man-
ganeseoxideson the retentionof traceandtoxic metals.The procedureis report-
ed to selectivelydissolvehydrousMn oxidesin soils andsedimentswith minimal
attack on coexisting Fe oxides. Chao (1972) discussesthe effect of varying
NH20H • HCI concentrationas well as extractionpH and time on the efficiency
of Mn oxide dissolutionin relation to Fe oxide dissolution.
This procedureis especiallyuseful for determiningheavy metal ions that
may be coprecipitatedwith hydrousMn oxidesas distinguishedfrom Fe oxides.
For work involving traceandpotentiallytoxic metals,careshouldbe usedin min-
imizing metalscontaminationof the samplefrom sieving through metal sieves,
contaminatedwater or reagents,and contaminatedglasswareor stoppersused
during the samplepreparationsteps.
Reagents
1. Hydroxylamine hydrochloride (NH20H • HCI) extractant, 0.1 M
NH20H • HCI in 0.01 M HN03•
Procedure
Add 0.5 g of a finely ground,air-driedsoil or sedimentsampleto 25 mL of
NH20H • HCI solution (or equivalentratios) in a flask, and shakefor 30 min on
a mechanicalshaker.Centrifugeand/or filter the suspensionand determinethe
Mn content of the solution phase.Inductively coupled plasmaemissionspec-
troscopyor AA spectroscopyare the preferredmethodsof analysis.
Total Manganeseby Atomic Absorptionor Inductively Coupled

PlasmaEmissionSpectroscopy
For more information on the total manganeseby atomic absorptionor

inductively coupledplasmaemissionspectroscopymethod,seeFiskell (1965).
MANGANESE 679
Reagents
3. Hydrofluoric acid, 48%.
5. Perchloricacid, 70%.
Procedure
A O.5-g finely ground, oven-dry soil sampleis placedin a 100-mL Teflon
or polypropylenebeaker.Add 5 mL of H20 2 and evaporatethe solution to near
drynessat 90°C, repeatingthis treatmentuntil the sampleis no longer efferves-
cent upon further additionsof H20 2• Add two drops of concentratedH2S04 and
5 mL of HF and placethe beakerson a sandbath. Slowly raisethe temperatureto
about200°Cand evaporateto dryness.Repeatthe addition of H2S04, HF, and the
evaporationstep. Then add10 mL of concentratedHN03, 1 mL of concentrated
H2S04, 3 mL of HCI04, and continueheatinguntil strong,white fumes of S03
are produced.Perchloric acidis a very strongoxidizing agent.It is important to
destroymost of the organicmatterby the H20 2 treatmentbefore the addition of
HCI04• Also, a fume hood designedfor HCI04 digestionshould be used. Cool
the beakerand add 25 mL of high-purity water. Inductively coupledplasmaemis-
sion spectroscopyor AA spectroscopyare the preferredmethodsof analysis.
Total Manganese,Direct Soil Analysesby Inductively Coupled

PlasmaEmissionSpectroscopy
For more informationon the total manganeseby direct aspirationof dilute

soil suspensioninto an ICP, seeDick et al. (1983).
Comments
Dick et al. (1985) reporteda rapid and efficient methodfor the simultane-
ous determinationof total concentrationsof multiple elementsin soils by direct
analysisof soil suspensionsamplesby ICP. This was reportedto eliminate the
time-consumingacid or fusion solubilization steps necessaryfor other total
analysismethods.It was statedGilbert (1962) was the first to report using the
slurry introductiontechniquewith atomic absorption.However,the much higher
temperatureof ICP spectroscopyshouldgreatly increaseatomizationefficiency.
This direct ICP analysis method for soil suspensionsshould be useful
where the total concentrationof severalelementsmust be determinedin large
numbersof soil samples.
Procedure
The soil should be ground to a fine consistencyfor 25 min in a microniz-
ing mill. Suspenda 2-g aliquot in high-purity waterwith continuousstirring. The
slurry is to be analyzeddirectly by ICP to determinetotal concentrationsof Mn
680 GAMBRELL
(aswell asAI, Ba, Ca, Fe, Mg, and Sr as reportedby Dick et al. (1985).The aver-
ageconcentrationsobtainedfor the elementsstudiedin 10 soils agreedto within
2% of resultsobtainedwhere the samesampleswere first solubilizedby sodium
carbonate(Na2C03) fusion. Grinding the soils reducedthe variation causedby
the particle-sizedifferences.The importanceof utilizing a fine particle size soil
material also has beendocumentedby Laird et al. (1991).
The ICP is operatedthe sameas for liquid samples.They reportedit is nec-
essaryto determinethe atomizationefficiency (E) for eachelementand to divide
the ICP resultsfor the indicatedconcentrationfor eachelementby E to get the
correct elementalconcentration.Dick et al. (1985) reported determiningthe E
factor on three standardsoils where the "true value" was determinedby atomic
absorptionanalysisfor samplespreparedby the fusion method, then the direct
analysisvalue was divided by the true value to calculateE for eachelement.
The atomization efficiency may be different for different ICP types and
would especiallybe a function of certainoperatingparameters.
Of the sevenelementsstudied,Mn gavethe highestatomizationefficiency,
or E valuesfor three standardsoils ranging from 0.715 to 0.789. Though some
elementsgavedifferent resultsdependingon the concentrationof soil in the sus-
pension,the differencefor Mn was slight whether1 or 10 g of soil was suspend-
ed per liter. The procedurewas reportedto be reproduciblefor soils with sandlev-
els between80 and 10%. For Mn, the analyticalline usedwas 257.6 nm.
Laird et al. (1991) also provideduseful information on this procedure.
Total Manganeseby the Colorimetric Method

Reagents
Procedure
Place1 g of finely groundsoil into a 100-or 150-mLplatinum or porcelain
dish. Add 10 mL ofHN03 (addcarefully to soils containingcarbonates).Heat the
beakerand contentsslowly in a sandbath on a hot plate and evaporatethe con-
tents to dryness.Placethe residuein a muffle furnaceat 500 aC for 30 min. Cool
the dish. If a porcelaindish was used,transferthe contentsto a 100-mL Teflon or
polypropylenebeaker,rinsing with high-purity water. Placethe beakerin a sand
bath on a hot plate and evaporatethe water, then cool. Add 5 mL of concentrat-
ed H2S04 and 10 mL of HF, and stir the mixture with a platinum or Teflon rod.
Heat slowly on a sandbath undera hood until the H2S04 hasfumed strongly for
30 min. Avoid overheatingwhich may causebumping.Cool the dish, then add 20
mL of deionizedor distilled water. Filter the solution into a 100-mL volumetric
flask and continue as describedfor the periodatecolorimetric method. [see the
sectionon "Colorimetric (Periodate)Method"].
If insoluble mineral matter remainsafter the acid treatment,this material
should be separated,washed,ignited, and fused with Na2C03.The melt should
be dissolvedand addedto the original solution (Adams, 1965).
MANGANESE 681
REFERENCES
Adams, F. 1965. Manganese.p. 1011-1018.In C.A. Black et al. (ed.) Methodsof soil analysis.Part
2. Agron. Monogr. 9. ASA, Madison,WI.
AmericanPublic Health Association.1980. Standardmethodsfor the examinationof waterand waste
water. 15th ed. APHA, Washington,DC.
Bartlett, R, and B. James.1980. Studying dried, stored soil samples-somepitfalls. Soil Sci. Soc.
Am. 1. 44:721-724.
Bartlett, R., and B. James.1966. Chromium. p. 683-701.In D.L. Sparkset al. (ed.) Methodsof soil
Browman,M.G., G. Chester,and H.B. Pionke. 1969. Evaluationof testsfor predictingthe availabil-
ity of soil manganeseto plants.1. Agric. Sci. 72:335-340.
Chao,T.T. 1972. Selectivedissolutionof manganeseoxides from soils and sedimentswith acidified
hydroxylaminehydrochloride.Soil Sci. Soc. Am. Proc. 36:764-768.
Cox, F.R. 1968.Developmentof a yield responsepredictionand manganesesoil test interpretationfor
soybeans.Agron. 1. 60:521-524.
Cox, F.R, and EJ. Kamprath. 1972. Micronutrient soil tests.p. 289-318.In J.J. Mortvedt et al. (ed.)
Micronutrientsin agriculture.SSSA, Madison,WI.
Dick, W.A, 1.R. Page,and KE. Jewell. 1985. Direct analysisof soil suspensions by inductively cou-
pled plasma-atomicemission spectrometry for determination of total metals. Soil Sci.
139:211-218.
Duncan,RR., RB. Clark, and P.R. Furlani. 1983. Laboratory and field evaluationsof sorghumfor
responseto aluminum and acid soil. Agron. 1. 75:1023-1026.
Fales,S.L., and K Ohki. 1982. Manganesedeficiencyand toxicity in wheat: Influenceon growth and
forage quality of herbage.Agron. 1. 74:1070-1073.
Fiskell, J.G.A. 1965. Copper.p. 1078-1089.In C.A Black et al. (ed.) Methodsof soil analysis.Part
2. Agron. Monogr. 9. ASA, Madison,WI.
Follett, RH., and W.L. Lindsay. 1971. Changesin DTPA-extractablezinc, iron, manganese,and cop-
per in soils following fertilization. Soil Sci. Soc. Am. Proc. 35:600-602.
Foy, C.D., H.W. Webb, and J.E. Jones.1981. Adaptationof cotton genotypesto an acid, manganese
toxic soil. Agron. 1. 73:107-111.
Fuller, W.H., and AW. Warrick. 1985. Soils in wastetreatmentand utilization. Volume I. Land treat-
ment. CRC PressInc., Boca Raton, FL.
Gambrell,RP., and W.H. Patrick,Jr. 1982. Manganese.p. 313-322.In AL. Pageet al. (ed.) Methods
of soil analysis.2nd ed. ASA and SSSA,Madison,WI.
Gajbhiye, K.S., N.N. Goswami, N.K. Banerjee,R. De, and R.K. Singh. 1984. Evaluationof a com-
mon extractantfor estimatingavailable Fe, Mn, Zn, and Cu in soil. J. Indian Soc. Soil Sci.
32:309-312.
Gettier, S.w., D.C. Martens,and SJ.Donohue.1985. Soybeanyield responsepredictionfrom soil test
and tissuemanganeselevels. Agron. 1. 77:63-67.
Gilbert, P.T. 1962. Direct flame-photometric analysis of powdered materials. Anal. Chem.
34:1025-1026.
Krauskopf,KB. 1972.Geochemistryof micronutrients.p. 7-40. In J.J. Mortvedt et al. (ed.) Micronu-
trients in agriculture.SSSA, Madison,WI.
Kubota,1., and W.H. Allaway. 1972. Geographicdistribution of traceelementproblems.p. 525-554.
In J.J. Mortvedt et al. (ed.) Micronutrientsin agriculture.SSSA, Madison,WI.
Laird, D.A, RH. Dowdy, and RC. Munter. 1991. Suspensionnebulizationanalysisof clays by induc-
tively coupledplasma-atomicemissionspectroscopy.Soil Sci. Soc. Am. J. 55:274-278.
Levesque,M.P., and S.P. Mathur. 1988. Soil testsfor copper,iron, manganese,and zinc in Histosols:
3. A comparisonof eight extractantsfor measuringactive and reserveforms of the elements.
Soil Sci. 145:215-221.
Lindsay, W.L., and W.A Norvell. 1978. Developmentof DTPA test for zinc, iron, manganese,and
copper.Soil Sci. Soc. Am. 1. 42:421-428.
Martini, J.A, and RG. Mutters. 1985. Effect of lime rates on nutrient availability, mobility, and
uptake during the soyean-growingseason:1. Aluminum, manganese,and phosphorus.Soil
Sci. 139:219-226.
Mascagni,HJ. Jr., and F.R. Cox. 1988. Residualeffect of manganesefertilization. Soil Sci. Soc. Am.
J. 52:434-438.
682 GAMBRELL
Martens, D.C., and W.L. Lindsay. 1990. Testing soils for copper, iron, manganese,and zinc. p.
229-264.In RL. Westerman(ed.) Soil testing and plant analysis,third edition. SSSA Book
Ser. 3. SSSA,Madison,WI.
McBride, M.B. 1978.Transitionmetal bondingin humic acid: An ESR study. Soil Sci. 126:200-209.
Neilsen,D., P.B. Hoyt, B.G. Drought,and G.H. Neilsen.1990.Manganesesoil testsfor both deficient
and toxic levels in appleorchards.Can. J. Soil Sci. 70:503-507.
Nelson,L.E. 1983. Tolerancesof 20 rice cultivars to excessAI and Mn. Agron. J. 75:134-138.
Norvell, W.A. 1984. Comparisonof chelatingagentsas extractantsfor metalsin diversesoil materi-
als. Soil Sci. Soc. Am. J. 48:1285-1292.
Ritchie, G.S.P. 1989. The chemicalbehaviourof aluminum, hydrogen,and manganesein acid soils.
In Soil acidity and plant growth. Acad. Press,Australia.
Schnitzer,M., and E.H. Hansen.1970. Organo-metallicinteractionsin soils: VIII. An evaluationof
methodsfor the determinationof stability constantsof metal-fulvic acid complexes.Soil Sci.
109:333-340.
Sharpe,RR, and w.L. Parks. 1982. A comparativeevaluationof three tests for determiningplant-
availablemanganesein soils. Agron. 1. 74:785-788.
Sherman,G.D., J.S. McHargue,andW.S. Hodgkins.1942.Determinationof active manganesein soil.
Soil Sci. 54:253-257.
Stevenson,FJ. 1986.The micronutrientcycle. In Cyclesof soil: Carbon,nitrogen, phosphorous,sul-
fur, and micronutrients.JohnWiley & Sons,New York.
Sumner, M.E., and W.P. Miller. 1996. Cation-exchangecapacity and exchangecoefficients. p.
1201-1229. In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.
U.S. EnvironmentalProtectionAgency. 1986. Test methodsfor evaluatingsolid waste.Vol. 1A. Lab-
oratory manual,physicaVchemicalmethods.SW-846.USEPA,Office Solid WasteEmergency
Response,Washington,DC.
Verloo, M., and A. Cottenie.1972. Stability and behaviourof complexesof Cu, Fe, Mn, and Pb, with
humic substancesof soils. Pedologie22:174-184.
Warden,B.T., and H.M. Reisenauer.1991. Fractionationof soil manganeseforms importantto plant
availability. Soil Sci. Soc. Am. J. 55:345-349.
Watson,M.E., and R.A. Isaac. 1990. p. 691-740.Analytical instrumentsfor soil and plant analyses.
p. 691-740.In R.L. Westerman(ed.) Soil testingand plant analysis.3rd ed. SSSABook Ser.
3. SSSA,Madison,WI.
Willard, H.H., and L.H. Greathouse.1917. The colorimetric determinationof manganeseby oxida-
tion with periodate.J. Am. Chern. Soc. 39:2366-2377.
Zasoski, RJ., and R.G. Burau. 1988. Sorption and sorptive interaction of cadmium and zinc on
hydrousmanganeseoxide. Soil Sci. Soc. Am. J. 52:81-87.
Published 1996
Chapter 25
Chromium
RICHMOND J. BARTLETT, University o/Vermont, Burlington, Vermont
BRUCE R. JAMES, University of Maryland, College Park, Maryland
Chromium is unique amongheavy metals.It exists in soil environments,natural

waters,and othergeologicand biological systemsin two valencestateswith op-
posing ionic charge,solubility, toxicity, and mobility. These differencesin the
valencestatesof Cr require specialconsiderationsduring extraction,speciation,
and analysisfor the purposesof assessingpotential hazardsor mobility in soil-
water-plantsystems.
Trivalent Cr [Cr(lll)] is cationic in the pH range of most soils, similar to
Fe(III) andAl(III), and it is olatedand precipitatedby OH- at pH values>4 to 5.5.
It forms thermodynamicallystableand kinetically inert complexeswith organic
and inorganicligandsthat may be both soluble and insoluble,dependingon type
of ligand, the molecularweight, and soil pH (Cotton & Wilkinson, 1988). A sol-
uble Cr(I1I)-picolinic acid speciesor a similar complex activates insulin, and
therefore is essentialfor human health and sugar metabolism. Most Cr(I1I) is
insoluble in near neutral soils, as paracrystallineCr(OHh or more crystalline
forms, such as Cr203; but some forms of Cr(III) may oxidize to more soluble
Cr(VI) forms in soils by Mn(III, IV) (hydr)oxides.
HexavalentCr [Cr(VI)] is similar to orthophosphate or sulfatein that it may
exist as the solublebichromateor chromateanion, it may be adsorbedby sesqui-
oxide colloidal surfacesor be precipitatedby certain cations, such as Ba2+ or
Pb2+. Although addedCr(IlI) may be oxidized to Cr(VI) in soils; in the same soils
and at the sametime, Cr(VI) may be reducedto Cr(III) by a variety of soilborne
compoundscontainingreducingequivalentsin variouscompoundsof organicC,
Fe, S, or partially reducedreactiveoxygen species.
Tenth metal in abundancein the earth'scrust, Cr ranks in the top five in
economicimportance.We encounterit mainly where we have put it-into steel
and industrial liquors, into leatherand tannery waste, as a coating on metalsto
protect and beautify, and into the environmentas a sometimestoxic and carcino-
genic pollutant, and sometimesas a nonhazardousmaterial similar to iron and
aluminum oxides. It is so widely used in modern industrial societiesthat it has
becomea commonconstituentof diversewastematerials,and is found in vary-
Book Seriesno. 5.
683
684 BARTLETT & JAMES
ing concentrationsat approximatelyone-third of Superfundsites in the USA

(Anonymous,1993).
Most of the Cr of interestto soil and environmentalchemistsis wasteCr,
originally having beenbroughtto the USA from southernMrica or from Russia
as chromiteore, having the characteristicspinel structure,FeO • Cr203. After an
alkaline roasting processconverts Cr(lll) in chromite ore to Cr(Vl), residual
Cr(lll) and Cr(VI) becomewasteforms of Cr in soil-watersystems.Generallythe
Cr in soils is in the Cr(III) form, tightly boundto both organicand inorganiclig-
ands.Boundin mineralandorganicforms, Cr(lll) is analogousto AI(III) (Bartlett
& James,1988). HexavalentCr also occursoccasionallyin high concentrations
in soils from point sources of pollution and in ground or surface waters
(USEPA,1984).Soluble Cr(lll) also may be found in waters as low molecular
weight organic complexes.In nature, Cr may be found substitutedfor small
amountsof octahedrallycoordinatedAI in clay minerals,and in rocks anduncon-
solidatedmaterials,as in chromite.
Making assumptionsabout the form and valencestate of Cr presentin a
given soil environmentis a prevalentpractice that is extremely risky from the
point of view of environmentalassessment. Therefore,analytical methodsthat
speciateCr by valencestateand form in soils are neededand have beendevel-
oped (Vitale et aI., 1994; Jameset aI., 1995). Fortunately,measurementof Cr
speciesin soils is not difficult, and actualanalysescan disproveor supportinnu-
merable hypothesesregarding valence state and solubility of Cr in soils.
Empirical analytical studieselucidating chemical behavioror Cr in soils have
beenreportedby Bartlett and Kimble, 1976a,b;Bartlett and James,1979; James
and Bartlett, 1983a,b,c,1984a,b;Ross et aI., 1981; Amacherand Baker, 1982;
and Bartlett, 1981, 1985, 1986, 1987,1988; Richard and Bourg, 1991; James,
1994; Losi et aI., 1994; Milacic and Stupar,1995; and Fendorf, 1995.
SOIL HANDLING
The Problem
Chemicalmethodsfor measuringaccuratelyany parametersin soils or sed-

iments affectedby redox require that the soil samplesbe field moist (Bartlett &
James,1980). This is especiallytrue where Cr solubility and redox transforma-
tions are involved or identification of Cr speciesand valenceis required.Before
Bartlett and James(1979) demonstratedCr(III) oxidation in moist soils, they and
other earlier workers-usingdried and stored samples-hadnot been able to
show the occurrenceof Cr(III)-to-Cr(VI) oxidation in soils, regardlessof level of
Cr(III) added,original pH, lime or phosphorustreatments,or aeration,moisture,
time, and temperatureconditions imposed (Bartlett & Kimble, 1976a,b).
Researchers assumedthat thermodynamicsgoverningsoil organicmatteroxida-
tion and Cr(VI) reduction precludedthe formation or persistenceof Cr(VI) in
soils.
Drying soils altersorganicand mineral redox and acidity characteristicsso
that behaviorof a dried samplewill changemarkedlywith time during dry stor-
age and while it is returning to its metastablemoist stateif water is addedback
CHROMIUM 685
to the dry soil (Bartlett & James,1980). Rarely will stored dried soil samples
[designated"lab dirt" for emphasis(Bartlett & James,1979)] oxidize added
Cr(lll). And rarely will a field moist soil samplefail to oxidize soluble salts of
Cr(III). After drying, a soil will readily reduceCr(VI) when it is remoistened;
whereas,in a continuouslymoist soil, Cr(VI) may persistin the presenceof sta-
ble humic and fulvic acids for periodsof monthsto years,especiallyat circum-
neutral pH's. Drying, puddling, acidifying, freezing and thawing, or stimulating
production of electron donating microbial byproductsby the addition of fresh
unhumifiedplant or animal residues-allcan causereductionof Cr(VI) in a soil.
Using the USEPAacid acetateextractant(Fed. Reg., 1980)for Cr(VI) may cause
reductionof soluble forms of Cr(VI) in a soil sample,ensuringthat Cr(VI) will
not be found.
Method
Comparedwith handling of dried samples;sieving, mixing, and storing
moist soil samplesis inconvenientin laboratory experimentalwork, but neces-
sary for correct identification of Cr redox statusand reactions.Moist soil sam-
ples are most suitablefor handling if nearfield capacitymoisture(approximate-
ly -10 kPa water potential).It is difficult to get moist samplesto passthrough a
2-mm sieve, but a 4-mm polyethylenesieve generally is suitable and has the
advantageof preservingsomeof the soil crumb structureand aerationstatusof
field soils during storage beforeanalysis.Samplesshouldbe mixed individually
before sieving. Sampleswill remain most stable if they are stored in double,
25-~m thick polyethylenebagsin a refrigeratorat 4°C. The processof freezing
destroysmany microbial and small root cells, and freezerstoragecausesdrying
in different partsof a sample.Freeze-driedsampleschangealmost as rapidly as
air-dried samples,following exposureto the air. Double-baggingwith moist
papertowels betweenthe two bags stabilizesmoisture contentsof samplesby
greatly lesseningwater vapor losses.Oxygen and carbondioxide (C02) readily
passthrough thin polyethylenefilm (Bartlett, 1965). Microbial activity appears
to becomestabilizedafter two or three mo (-8 or 12 wk) at 1°C, and most soil
samplesseemto reachan internal, metastableequilibrium.
Additional redox changesare slow comparedwith the changesthat would
be occurringin a previouslydried sample(Bartlett, 1986).Added Cr(VI) will not
be reducedin such a moist, stablesample,even if flooded for weeks. However,
underthe beststorageconditions,a soil samplegradually will lose a major part
of its Cr(III) oxidizing ability, eventhoughtitration with Fe2+ showsthat most of
the Mn(III, IV) (hydr)oxidesare still present(Bartlett, 1986). It is safe at this
time to transfersamplesto tightly sealed,heavy-walledplastic bagsor garbage
containerswith lids to minimize further moistureloss. Samplesshouldbe kept in
darknessor in diffuse room light, but temperaturesas high as 10 to 12°C can be
toleratedin thesemoist soils.
Well-mixed subsamplesmay be weighed for analysesafter determining
moistureon separatesamples.For many determinations,a volume measurement,
later correctedfor dry weight, is more convenient,althoughsomewhatlessaccu-
rate. A packedand leveledteaspoonof soil is 5 cm3 and is approximately5 g dry
weight for many granularA horizon soil samples.
686 BARTLETI & JAMES
DIGESTION METHODS FOR TOTAL CHROMIUM

IN SOILS AND PLANTS
Wet oxidation with combinationsof nitric (HN02), sulfuric (H2S04), per-

chloric acids (HCI04), and sulfuric acidlhydrogenperoxide (H20 2) all have
provenuseful for nearly 100% recoveryof Cr from soil samples(Gorsuch,1970;
Reisenauer,1982). Dry ashing below 550°C is reliable, but recoverieshave
sometimesbeendepressedin the absence of ashingaidsor at highertemperatures
(Gorsuch, 1970). Chromyl chloride (Cr02CI2), boiling at 117°C, is readily
decomposedby H20, and thereforewill not persist in aqueousmedia and be
availablefor loss of Cr during digestion.Treatmentwith hydroflouric acid (HF)
is essentialto preventloss of Cr by occlusionin silica residue(Cary & Rutzke,
1983).
McCarthy and Ellis (1991) reported a microwave digestion that gives
resultsfor Cr in shellfish comparableto wet digestion methods,and somewhat
higher than dry ashingat 450°C. Bowman (1988) reporteda rapid wet chemical
digestionwith H2S04, H20z, and HF, without HCI04, employingonly a hot plate
for completesoil digestionin a short time. Condron's(1991) modificationof this
method, using HN03 in place of H2S04 also is recommendedas describedin
"Digestion of Soil Materialsfor ChromiumAnalysis."
We also recommendthe method for Cr oxidizable by hypochlorite (see
"Chromium Oxidizableby Hypochlorite") for usewhereabsolutevaluesof total
Cr are not required. For practical purposes,this simple hypochlorite method,
modified from Amacherand Baker (1982), extractsand measuresnearly all of
the Cr in a soil sample,even though the sampleis not dissolved.Hypochlorite
readily oxidizesandextractskinetically inert forms of Cr(lll), suchas in chrome-
tannedleather,chromite-containingsoils, chromiteores,slags,and otherchromi-
um mineralwastesand Cr(lll) previouslyaddedto soils in wastessuchassewage
sludge,tannerywastes,or sewagelagoonsediments(Amacherand Baker,1982).
For many practical purposes,the Cr measuredcan be considered"total Cr." It
also hasbeenusedfor analyzingCr in feedsand feces(Suzuki & Early, 1991).
s-DIPHENYLCARBAZIDE METHOD FOR DETERMINING

SOLUBLE CHROMIUM(VO
ReactionMechanismand Conditions
In usesincethe tum of the century(Cazeneuve,1990),colorimetric analy-

sis of Cr(VI) with s-diphenylcarbazide(DPC) is sensitive,highly specific, and
simple. It is particularly applicableto aqueousextractsof soils, plants, and nat-
ural watersin which Cr(IlI) and Cr(VI) may be presentin micromolarconcen-
trations in samplescontaininghigh concentrationsof plant nutrients and envi-
ronmentalpollutants.The DPC method measuresonly Cr(VI) in solutionsthat
alsomay containCr(III), but following oxidativedigestionpretreatmentthat con-
verts soluble Cr(III) to Cr(VI), the methodcan be used to measuresoluble Cr
(Saltzman,1952; Bryson & Goodall, 1981).Analytical speciationof Cr(VI) and
CHROMIUM 687
solubleCr(IlI) organiccomplexesin soilleachates,naturalwaters,and plant saps

is neededbecausecertain organic Cr(IlI) complexesare essentialfor human
nutrition, and Cr(VI) is toxic to living cells and must not exceed0.1 mgIL in
drinking water (Goldhaber& Vogt, 1989; Katz, 1991).
The molar absorptivityof the complex is approximately43 000 Llmollcm
at 540 to 546 nm, providing an absorbanceof approximately1.0 unit with 23 ~
Cr(VI) (1.2 mgIL) in a 1-cm cuvette and a detectionlimit of 0.1 ~ (0.0052
mgIL) or less, dependingon the purity of the DPC and the age of the mixed
reagent(Marczenko,1986, p. 233-242).The limit of quantitation(3 times limit
of detection)for the methodis approximately0.3 ~ (Am. Chern. Soc., 1983).
The limits of quantitationfor total solubleCr for flame atomic absorption(AA),
graphite furnace AA, and optical emission spectroscopyare approximately2,
0.01, and 0.3 ~, respectively(Baker & Suhr, 1982; Soltanpouret aI., 1982).
Hexavalentand trivalent Cr havedifferent sensitivitiesin both flame and graph-
ite furnaceAA (Perkin-ElmerCorp., 1984).
The red-violet complex results from the coupling of a rapid oxidation-
reduction reactionwith formation of a chromophoricCr(III)-organic complex,
but addedCr(lll) salts will not react directly with DPC at low pH (Pflaum &
Howick, 1956).Addition of the DPC reagent to a Cr(VI)-containingsolution at
pH 1 to 2 in H2S04 or phosphoricacid (H3P04) results in oxidation of DPC to
diphenylcarbazonecoupledto reductionof Cr(VI) to unhydratedcr3+. A Cr3+-
diphenylcarbazonecomplex then forms instantaneously(Marchant, 1964), pre-
sumably forestalling hydration of cr3+, a very slow reaction at room tempera-
tures(Cotton & Wilkinson, 1988, p. 686).
The stoichiometryof the Cr-diphenylcarbazone complex is believedto be
two moles Cr(III) to three moles of ligand, in accordancewith the oxidation-
reductionreaction,in which eachmole of DPC losestwo electrons(and protons)
and each mole of Cr(VI) gains three electrons.Thus, Cr(III) addeddirectly to
diphenylcarbazonedoes form a complex with an absorbancemaximum at 540
nm. But divalent Cr does not react with DPC, and Cr(VI) does not react with
diphenylcarbazone,providing further evidencefor the specificity of DPC for
Cr(VI).
Thus, in acidic solutions,DPC is highly specific as a reductant-complex-
ing agentfor Cr(VI). However,underalkalineconditions,slow oxidationof DPC
in air occursspontaneously,as indicatedby increasinglyyellow discolorationof
the storedreagent.To preventair-oxidation,the DPC reagentmust be mixed and
stored in acetone(C3H60) or ethanol (CzHsOH), with or without H2S04 or
H3P04 added(Pilkington & Smith, 1967). Urone (1955) found that an acidified
95% ethanol solution could be used effectively for 2 wk or longer, although
100% acetoneas the solventpreservedDPC appreciablylonger. Lower toxicity
and volatility make ethanol a better choice. Either HCI04 or hydrochloric acid
(HCI) may interfere with the redox reaction central to the measurementof
Cr(VI).
Addition of acid to Cr(VI)-containingsamplesbeforeDPC, recommended
in the method of the American Public Health Association(1985, p. 201-204),
should be avoided. Reducingagents,such as fulvic acids, H20 2 (Pettine et aI.,
1988), Fe(II), and sulfides, could reduce Cr(VI) under the acidic conditions
before addition of OPC(Bloomfield & Pruden,1980). Addition of OPC in acid

results in the simultaneousreduction of Cr(VI) and complexation of newly
formed Cr(III) requiredfor the accuratemeasurementof Cr(VI) in the original
samplesolution. Bloomfield and Pruden(1980) were not correct in faulting the
OPC methodby equatingreductionof Cr(VI) in extractsthey treatedwith acid
alonewith expectedreductionin OPC-treatedsamples.Certainly, water and soil
samplesin which Cr(VI) will be measuredshould neverbe acidified for preser-
vation, as hasbeenrecommended(Am. Publ. Health Assoc.,1985, p. 201-204).
HexavalentCr can be reducedby reducingagentsin the soil, in water or evenin
the addedacid [e.g., nitrite (N02) in HN03 exposedto light, (Pavelet aI., 1985;
Stollenwerk& Grove, 1985)].
Interferencein s-DiphenylcarbazideChromium (VI) Test
Positive interferencescould result from Fe, V, Mo, Cu, Ni, and Hg inner-
spherecomplex formation with diphenylcarbazone,but thesecomplexesform
only at concentrationsof 0.1 to 60 roM, dependingon the element,and they are
destabilizedif the pH is less than four (Pflaum & Howick, 1956; Bryson &
Goodall, 1981). Negativeinterferencescould result from Fe(III)-complexes,but
the interferenceof up to 200 mg L-l in the presenceof 1 mg Cr(VI) L-l is elim-
inatedby H3P04 as a componentof the OPC reagent(Pilkington & Smith, 1967).
Ferric iron interferencecan be eliminatedby precipitationof Fe(III) hydroxide
following oxidation of Cr(III) during digestionof aqueoussamples(Marczenko,
1986), and Hg interferencemay be masked by addition of sodium chloride
(NaCl) before the DPC. High concentrations(>1 mM) of hypochlorite(CIO,n,
z),
nitrite (NO permanganate(MnO,n, and other oxidizing agentsinterfere with
color development. Permanganateinterference is eliminated with NaN3
(Saltzman, 1952), and that of nitrite with sulfanilamide (James & Bartlett,
1984a).
OXIDATIVE PREPARATIONOF SAMPLES

FOR s-DIPHENYLCARBAZIDE TEST
Oxidation of OrganicLigands
Soluble organic complexesof Cr(III) with well-characterizedligands and

with fulvic acids may be stable in soils for months (James& Bartlett, 1983b),
and certain Cr(III)-complexes(e.g., with picolinic acid) are nutritionally impor-
tant and bioavailable to humans (Fisher, 1990). Conversion of such soluble
Cr(lll) compoundsto Cr(VI) for inclusion with Cr(VI) in the OPC method
requiresoxidation of the organic ligands to CO2 before oxidation of Cr(III) to
Cr(VI) by Ce(IV) (Bryson & Goodall, 1981). Theseresearchersreportedinter-
ferencesfrom oxalate, tartrate, citrate, quinoline sulfate, and ethylenediamine
tetraaceticacid (EDTA) in the concentrationrange0.5 to 2.2 mM in the Ce(IV)
oxidation procedurefor Cr(lll). To oxidize solubleorganiccomplexingligands,
volatilize F, and releaseCr3+ into solution in aquatedforms, we recommend
either a low temperatureHN03 digestion(James& Bouldin, 1986) or a HNO}
CHROMIUM 689
H20 2 digestion procedure (see "Nitric AcidNacuum Oven" and "Nitric

Acid/HydrogenPeroxideMethod, respectively).
Oxidation of Chromium (III) to Chromium (VI)

The Cr(III) held in low molecular weight organic acid complexeswas
shownto be more readily oxidized by naturalsoil Mn (III, IV) (hydr)oxidesthan
were the organicacidsholding it (Bartlett & James,1988).However,freshly pre-
cipitated Cr(OH)3 addedto a soil was more quickly oxidized than was Cr(III)-
citrate (James& Bartlett, 1983b).Sections"Nitric AcidNacuumOven Method"
and "Nitric Acid/Hydrogen Peroxide Method" outline methodsfor oxidizing
organic ligandsbinding Cr(III), a first step requirementfor oxidizing the Cr(IlI)
extractedby the section"Extraction of Labile Chromium(III)" citrate extraction
method.The secondstepis the Ce(IV) oxidation of the inorganicCr(III) product
to Cr(VI), which then can be determinedby meansof the DPC test. In a study of
oxidizable, inert Cr(III) in leather and sewagesludge treatedsoils, the citrate
"available" Cr(IlI) ("Extraction of Labile Chromium(III),,) predicted(r2 = 0.78)
the quantity of Cr(VI) formed during a long-termequilibrationwith Mn and cit-
rate [section on "Oxidizability of Inert Soil Chromium(III)," (Bartlett, 1985,
1986)].
ALTERNATIVE METHODS FOR CHROMIUM(lII),

CHROMIUM (VI), AND TOTAL CHROMIUM
IN SOLUTION
While the diphenylcarbazidemethod is the most reliable and simplest

method for measuringCr(VI) and total soluble Cr (following oxidative diges-
tion) in aqueousextracts from soils, natural waters, biological samples,and
wastes,numerousother methodsare availablefor particular applications(Table
25-1). If rapid measurementsof total soluble Cr are needed,flame AAS and
inductively coupled plasma methods [ICP-atomic emission spectrophotome-
try(AES)] are convenient,but lesssensitivethan the DPC method.Graphitefur-
naceatomic absorptionspectrophotometry(GFAAS) providesa somewhatmore
sensitive,but lessconvenient,methodthan DPC for total soluble Cr.
RECOMMENDED PROCEDURESFOR DETERMINATION

OF CHROMIUM
Chromium(VI) Standards
Dissolve0.2263g of K2CrZ07, previously dried at 140°C, in 1.0 L of dis-
tilled, deionized water (Saltzman, 1952). This stock solution contains 80 mg
Cr(VI)/L, and is used to prepare more dilute working standards daily.
Commerciallyavailable,certified 1000 mg Cr(VI)/L stocksolutionsalso may be
used. Becauseordinary (not deionzied) distilled water may contain reducing
Table 25-1. Alternative methodsto diphenylcarbazidecolorimetry for quantificationof solubleCr(lIl,VI) species.
Valence(s) Detectiont
i
Method category Specific methodcombination detection limit· Matrix:!: Reference
J.1g/L
Diphenylcarbazidecolori- Reductionof Cr(VI) by diphenylcarbazidecoupledto b 5 efgi Bartlett & James,1996,this chapter
metric method complexationof newly formed, unhydratedcr3+
with diphenylcarbazone
Liquid chromatography Complexwith 8-hydroxyquinolinehigh-pressureliquid a 100 g Lopez et aI., 1991
chromatography(HPLC) separation/visiblelight detection
Comparisonof DPC, chelatingion-exchangechromatography, b 0.3-30 f Milacic et aI., 1992
and ion-pairing HPLC
Ion chromatographylDPC detection b 1 efg Dionex Corp., 1990,p. 1-7; Arar et
aI., 1991; USEPA, 1991;
Trojanovicz& Pobozy,1992
Ion chromatography/ICP-mass
spectrometry b g Arar et aI., 1992
Anion exchange/columnreduction/graphitefurnaceatomic c 0.01 e Hiraide and Mizuike, 1989
absorptionspectrophotometry
(GFAAS)
Ion chromatography/conductimetric
detection b 92 gi Mehra & Frankenberger,1989;
Salehet aI., 1989
Ion exchangeseparationand pre-concentration/GFAAS c 0.02 e Johnson,1990
Complexwith 8-hydroxyquinoline/resincollection/GFAAS c 1 e Isshiki et aI., 1989
Gel chromatographyseparationof Cr(lIl) and Cr(VI) c 1 e Osaki et aI., 1983
Ion-pairing HPLC/AAS c 40--80 e Syty et aI., 1988
Ion chromatography/ICP-AES c 1 eg Urasa& Nam, 1989; Prokischet aI.,
1994; Tomlinsonet aI., 1994
Ion chromatography/AAS c 1000 e Fong & Wu, 1991; Milacic &
Stupar,1994 ~
Ion chromatography/chemiluminescence c 0.1 e Williams et aI., 1989; Beere&
Jones,1994
Cation exchangecolumns c e Collins et aI., 1988 ~
Ion-exchange/flowanalysisof Cr(VI) b 0.5ng e Yoshimura,1988 Ro
Reversedphaseion-pair HPLC c 0.4-1.6ng e Jenet aI., 1993
i
t"'l
I'll
Table 25-1. Continued. ("l
Valence(s) Detectiont ==
Method category Specific methodcombination detection limit Matrix:\: Reference ~
::
mgIL ~
-
AAS and ICP-AES§ Review of AAS for Cr d efghi Rubio et aI., 1992
::
Coprecipitation/dissolution/GFAAS d h Cary & Rutzke, 1983
Pentamethylene dithiocarbamate/methylisobutyl
ketone d 0.4 e Babu & Naidu, 1991
(MIBK) extraction/AASextraction
Ammonium pyrrolidinecarbodithioate-MIBKextraction/ c 0.3 e Subramanian,1988
GFAAS detection
ChelatingresinllCP-AESdetection c 12 e Miyazaki & Barnes,1981
GFAAS d 0.05 e Apte et al.,1991
CoprecipitationlflamelessGFAAS d 0.05 Cranston& Murray, 1978
Coprecipitation/GFAAS a 0.3 eg Vos, 1985
Colorimetry Complexwith EDTA a e Rengasamy& Oades,1977;
DenBoefet aI., 1960
Complex with hydroxyamidines b 100 e Golwelker et aI., 1990
Complex with basic xanthenedye b 1 e Liu & Wang, 1991
Oxidation of hydroxylamine/diazotization b 40 e Rathore& Tarafder,1992
Flow injection for total Fe and Cr(VI) b 200 e Araujo et aI., 1989
Voltammetry Differential pulse voltammetryof Cr-diphenylcarbazone b 0.02 e Elleouet et aI., 1992
Differential pulse polarography b 10 e Crosmun& Mueller, 1975
Chemiluminescence Ion chromatography/chemiluminescence a 15 gi Marina-Sanchezet aI., 1992
Luminol chemiluminescence a 0.2 e Changet aI., 1980
Gaschromatography Solventextraction/electroncapturedetection c 0.1 e Lovett & Lee, 1979
Precipitation Coprecipitationwith barium sulfate(BaSo4)/dissolution/ b 0.02 e Yamazaki,1980
DPC analysis
t Letters correspondto the following: a =Cr(IlI) only; b =Cr(VI) only; c =Cr(III) and Cr(VI) measuredseparately;d =total soluble Cr measuredwithout valencesep-
aration.
:\: Letters correspondto the following matrixes: e =natural waters;f =soil extracts;g =wastematerial extracts;h =plant digests;and i =soil digests.
§ Atomic absorptionspectrometry/inductivelycoupledplasma-atomicemissionspectroscopy.
...$
agentsfor Cr(VI), and glassstoragevesselsmay sorb Cr(VI) anionsfrom solu-

tion, high-purity water(with conductivity<1 or 0.1 dim) and plastic storage ves-
selsshouldbe usedfor standardsolutions.
ChromiumStandards
Pipet 5.0 mL of stock Cr(VI) solution (80 mgIL) into a beaker,and add
about 15 mg of Na2S03 to reduce the Cr(VI), and add 0.5 mL 16 M HN03
(Saltzman,1952). Evaporateto drynesswith low heat. High heatmay reoxidize
the Cr(III). Add 0.5 mL of HN03, and evaporategently to drynessto destroy
excesssulfite. Dissolve in 1.0 mL HN03 and bring to 200 mL volume with dis-
tilled, deionizedwater. Verify the absenceof Cr(VI) with DPC reagent.This pri-
mary standardcontains2 mg Cr(lII)/L and is stablefor a few days. OtherCr(III)
standardsmay be preparedfrom dried Cr(N03)3 • 6H20 salt.
Digestionof Soil Materialsfor ChromiumAnalysis

1. Place0.5 g of dry soil ground to passa 60-mesh(=300mm)sieve in a
Teflon beakerand add3.0 mL 16 M HN03. Heaton a hot plate at 100°C
for 1 h, remove,and cool.
2. Add 2 mL HN03 and return to hot plate. While graduallyincreasingthe
heatto 200°C, add 8.8 M (30%) H20 2 in six O.5-mL aliquots (perform
in hood and weargoggles).
3. Heat gently to dryness,add 5 mL concentratedHF, and evaporateat
200°C. Repeat the addition and evaporationof HF (hood, goggles,
gloves).
4. Dissolvethe ashin 5 mL of 6 M HCl, and heatuntil boiling. Filter into
a 100-mL volumetric flask, and dilute to volume.
5. Determine Cr according to the sections on "Oxidation of Inorganic
Chromium(lII) with Cerium(IV)" and"Determinationof Chromium(VI)
in Solutions"or by AA, ICP, or othermethodsfor total solubleCr in soil
digest.
ChromiumOxidizableby Hypochlorite
1. Add 40 mL of undiluted bleach solution (5.25% or 0.7M NaOCl),
adjustedto pH 9.5, to 1.0 g dry soil in a 100-mLglasstesttube,and mix
using a vortex mixer.
2. Placetube in a boiling water bath for 20 min (hood and goggles),mix
again,and obtain a clearextractby centrifuginga portion of the suspen-
sion. Determinetotal Cr oxidized in the extract by AA using an air-
acetyleneflame and standardscontaining the same concentrationsof
NaOCI as the unknowns,or, after a lOx dilution, by ICP.
Determinationof Exchangeableand Total Chromium{V1) in Soils

Extract exchangeableCr(VI) from soil at 3:1, solutiOn/soil ratio, basedon
dry weight of soil or on a packedspoonvolume, bygentle5-min shakingwith 10
mM K2HPOJKH2P04,buffered at pH 7.2 (James& Bartlett, 1983c). Filter or
CHROMIUM 693
centrifuge and measureCr(VI) by the DPC method (see "Determination of

Cr(VI) in Solutions").
The following alkaline digestionmethodis recommendedfor measurement
of total Cr(VI) in soils or wastematerials,including soluble,exchangeable,and
nonexchangeable forms (Jameset ai., 1995):
1. Add 50 mL of 0.28M Na2C03in 0.5 M NaOH in triplicate, 2.5-g sam-
ples of soil in 250-mL Pyrex beakersweighedto the nearest10 mg.
2. Swirl the soil suspensions on arotary shakerfor 1 h and then transferthe
beakersto an unheatedhot plate. With a plate temperatureof 200°C,
heat the suspensionsat 90 to 95°C liquid temperaturefor 60 ± 2 min.
The suspensiontemperaturesremain just below the boiling point, and
the suspensionsshouldbe swirled periodically during the heatingperi-
od.
3. After allowing the suspensions to cool, add distilled water by weight to
give a total solution of 100 g in eachbeaker.Centrifugethe suspensions
(25 000 x g, 20 min, 20oq, and measuresoluble Cr(VI) by the DPC
method (Bartlett & James,1979) using 1.0 mL of DPC reagent (in
H3P04 and ethanol)addedto 0.5 mL of alkaline digestdiluted to 10 mL
with distilled H20. At least a 1:5 dilution of the alkaline digestateis
neededto allow acidification to pH <2 and color developmentof the Cr-
diphenylcarbazone complex.
Extractionof Labile Chromium(III)

Shakeintermittently 1 g, dry-weightbasis,of moist soil contaminatedwith
a Cr sourcesuch as tannery sludgewith 50 mL of 10 mM K 2H-citrate for 72 h
and determinetotal Cr extractedaccordingto sectionson "Nitric AcidNacuum
Oven Method," "Nitric Acid/Hydrogen Peroxide Method," and "Oxidation of
InorganicChromium(III) with Cerium(IV)," or by AA or ICP-AES.
Determinationof Chromium(VI) in Solutions

1. s-diphenylcarbazide reagent.Prepareby adding 120 mL of 85% H3P04,
diluted with 280 mL distilled water, to 0.38 g of DPC previously dis-
solved in 100 mL of 95% ethanol.Store in an amberglassbottle in the
dark at 4°C.
2. DetermineCr(VI) by adding 1 mL DPC reagentto 8 mL of extract or
water, mix, and let stand20 min and comparecolor with that in stan-
dards(0.5-50 uM) at 540 nm.
3. Organicmattermay presentpositive interferencewith absorbancemea-
surements,but it doesnot interferewith developmentof the color com-
plex. If cloudinessfrom precipitatedorganic or mineral matter is pre-
sent, it is easily removedfollowing color developmentusing a syringe
and 0.2-Jlm filter. However,filtration removessomeDPC complexthat
becomesadsorbedto colloids. A preferred method, if the amount of
backgroundturbidity is small, uses a "blank reagent" added to un-
knownsfor backgroundcorrection.This reagentcontainsH3P04, ethan-
ol, and water, but no DPC. The absorbanceat 540 nm of the unknowns
694 BARTLETI' & JAMES
treatedwith the blank reagentis subtractedfrom that of the cloudy sam-

ples treatedwith the DPC-containingreagent.
Oxidation of OrganicLigandsBinding Chromium(III)
Nitric Acid/VacuumOven Method

1. Add 1.0 mL colorless16 M HN03 (Le., free of discolorationby N02
formed during storage in the light) to 2.50 mL of sample (filtered
througha O.4-llm polycarbonatemembrane)in a 50-mL Teflon or glass
beakerweighedto the nearest10 mg. Evaporateto drynessin a vacuum
oven at 70 to 80°C undera vacuumof about50 kPa.
2. Add a secondaliquot of HN03 and evaporateagain to drynessunder
vacuum.After a third aliquot of HN03 hasbeenevaporated,dissolvethe
ashin a fourth I-mL aliquot ofHN03 and bring to 25.00-mLvolume by
weight with distilled deionizedwater. The final digest is 0.6 M HN03
(James& Bouldin, 1986).
Nitric Acid/HydrogenPeroxideMethod
1. Evaporateslowly to dryness5.0 mL of filtrate in a graduated,25- by
200-mm Pyrex digestiontube in an aluminum digestionblock on a hot
plate at a temperatureof 130 to 150°C.
2. Then add 2 mL 16 M HN03 and 1 mL 8.8 M (30%) H20 2 and again
evaporateto dryness.
3. Add 1 mL 16 M HN03 to dissolve ash and bring to 25.00 mL volume
with distilled water (James& Aschmann,1992).
4. Underthe conditionsof this digestion,Cr in the ashis in the Cr(lll) form
becauseperoxide reducesCr(VI) under acid conditions (Pettine et aI.,
1988).
Oxidation of InorganicChromium(1m with Cerium(IV)

1. Transfer1.0 mL of samplecontainingup to 4 mg Cr(lll, VI)/L to a test
tube, add 3.8 mL distilled water, 0.05 mL 16 M HN03, and 0.15 mL 50
mM Ce(S04)2in 1.0 M HN03.
2. Mix by vortexing and allow to stand.Under theseconditions,oxidation
of Cr(lll) to Cr(VI) is completewithin 4 min for standards.
3. In the presenceof >2 mM phosphate,a Ce(IV) phosphateprecipitates
and interfereswith Cr(III) oxidation by Ce(IV). To preventthis, replace
the HN03 with 0.1 mL 18 M H2S04, and allow at least1 h for the diges-
tion. A sulfatoceratecomplex forms, allowing oxidation to proceed
more slowly than in the presenceof HN03, but without precipitationof
Ce(IV). If phosphateconcentrationsare as high as 100 mM' add 0.2 mL
of H2S04 and allow up to 16 h for digestion.
4. After oxidation of Cr(lll), add 0.01 mL 0.77 M NaN3, and mix by vor-
texing to reduceexcessCe(IV), the yellow color and oxidant properties
of which interferewith the subsequentanalysisof Cr(VI) by DPC.
5. Immediatelyadd 1.0 mL of DPC reagentcontainingH3P04, vortex, and
measure absorbance at 540 nm. Sodium azide does not reduceCr(VI),
CHROMIUM 695
but if digestscontainhigh concentrationsof Pe(III), NaN3 will reduceit

to Pe(II), and subsequentreductionof Cr(VI) may occur if DPC is not
addedimmediately(methodof Bryson & Goodall, 1981).
METHODS FOR STUDYING SOIL REDOX

USING CHROMIUM AS A REDOX TOOL
Most of the proceduresthat follow servedual functions by characterizing

behaviorof Cr in soils, sedimentsand watersand at the sametime characterizing
behavior in soils of other redox substancesand parameters.Relatively easy to
measure,a speciestransformationin Cr in soil always is tied to a speciestrans-
formation of anotherredox substance.Practically any procedurefor measuring
Cr(VI) can be useful for estimating the potential of a soil either to oxidize
reducedsubstances or, alternatively,to reduceoxidized substances.
StandardChromiumNet Oxidation Test
1. Shake2.5 g of soil (dry weight basisor 2.5-cm3 packedvolume) for 15

min with 25 mL of 1 mM CrC13•
2. Add 0.25 mL of 1 MpH 7.2 phosphatebuffer (see"Determinationof
Exchangeableand Total Chromium(VI) in Soils"), shake 15 s longer,
and then filter or centrifuge.
3. Determine Cr(VI) in the extract using DPC (see "Determination of
Chromium(VI) in Solutions").
A portion of the Cr oxodizedduring the courseof this test is not measured
as Cr(VI) becauseit is reducedalmostas fast as it is oxidized. Dependingon the
availability of easily oxidizableorganicmatter,someor evenmost of the Cr(VI)
formed is reducedduring the 15-min period. With dried and storedlab dirt sam-
ples, the reduction frequently equalsthe oxidation and no Cr(VI) is measured,
eventhoughsomeCr may havebeenoxidized during the test by remainingman-
ganeseoxidesnot reducedby the drying.
Thus,net Cr oxidation is measured.The net oxidation testvalue is an over-
all soil redox measurementin that it is the result of both oxidation and reduction
by a soil sample.But, in fact, it characterizesoxidation only to the extentthat it
exceedsreduction,and doesnot representeither parameterif they are equal and
cancelone another.Soils are poisedredox systems.A soil that oxidizes Cr gen-
erally hasthe potentialto reducesomeof it. Leachinga sample,moist or dry, will
remove some of the low molecularweight reducing organics and thereby will
increasethe Cr(VI) quantity measured(Bartlett, 1981).
Soil ManganeseElectron Demand

1. As a direct titration of the reduciblemanganeseoxidesin a soil sample,
the ManganeseElectron Demand(MED) determinationis a direct way
of estimatingthe Cr oxidizing capability by the manganeseoxides in a
soil without regardto the Cr(VI) reductionthat could take place under
the conditionsof oxidation measurement (Bartlett, 1988).
2. Shakeintermittently for 18 h, 2 g, dry weight basis,or 2 cm3 moist soil

with 12 mL of pH 4.0 NH40Ac, 0.6 M with respectto ammonium,and
4 mL of 0.2 M potassiumiodide (KI). Add four dropsof starchsolution
(0.3 g potatostarchboiled with 50 mL water), and titrate to a colorless
endpointwith 2 mM Na2S203.Millimoles of thiosulfateper unit of soil
are equivalentto millimoles of electronsor plus chargeof Mn.
Total Chromium(VI) ReducingCapacity
This is the classicWalkley-Black(1934)"total" soil organicmattermethod.

[See "Walkley-Black Method" from Chapter34, Nelson & Sommers,1996.] It
measuresthe portion of organic matter that is oxidizable by Cr(VI) in the pres-
enceof concentratedH2S04, using the heat of dilution. The Cr(VI) not reduced
is titrated with ferrous iron.
Available ReducingCapacityfor Chromium(VI)
1. Shakeintermittently 2.5 g, dry weight basis,or 2.5 cm3 of moist soil 18

h with 25 mL of 0.1, 0.5, 2.5, or 10 mM as K2Cr207 in 10 mM H3P04,
filter or centrifuge,and determineCr(VI) (DPC method,see"Determin-
ation of Cr(VI) in Solutions")not reducedin the extract.
2. Begin with the lowest concentrationof Cr(VI). If all of the Cr(VI) is
reduced,repeatwith increasedconcentrationsuntil the Cr(VI) remaining
is measurablebut below 0.1 mM.
Soil ReducingIntensity for Chromium(VI)
1. Shakeintermittently 2.5 cm3 of moist soil 18 h with 20 mL of pH 4.0

NH40Ac, 0.6 M with respectto ammonium,containing0.1, 0.5,or 2.5
mMK2Cr207·
2. Filter or centrifuge,and determineconcentrationof Cr(VI) remaining
(see "Determination of Chromium(VI) in Solutions"). If all of the
Cr(VI) is reduced,repeatwith increasedconcentrationsuntil the Cr(VI)
remainingis measurable.One mole of Cr is equivalentto six moles of
Mn plus charge.
3. By omitting Cr(VI) from the equilibratingsolution,this sametest [or the
test for "Available ReducingCapacityfor Chromium(VI)"] canbe used
to measurethe tendencyof a soil to reduceCr(VI) that is presentin the
soil as contamination.
4. To measurethe tendencyof a particularoxidizableorganicsubstanceto
reduceCr(VI), repeatthis test [or use the test for "Available Reducing
Capacityfor Chromium(VI)"] after adding the organicsubstanceto the
soil sample.
QualitativeOxidizability of Inert Soil Chromium(1m

Oxidizability of forms of soil Cr(III) generallyconsideredto be inert is test-
ed in a wetlandsimulatedin a plastic beaker.This is feasibleat a water/soiloxi-
CHROMIUM 697
dizing interface where optimum conditions of aeration and freshnessof man-

ganese-oxidesurfacesare provided along with a soluble chelatingagentsuch as
citrate. Conditionsfor test incubationsmimic thosepossiblyoccurringin natural
wetlandsoils.
1. Placeinto a 400-mL polyethylenebeaker,a SO-mL volume of moist, fer-
tile, and friable soil, nearneutral in pH, that hasbeenspiked with up to
half its volume with an "inert" form of Cr suchas tannerywaste,leather
dust, chromeore, stainlesssteelslag, high chromesoil, or soil fertilized
with high-chromiumsewagesludge.The soil will fill the beakerto about
1.S-cmdepth.Add 100 mL of distilled water containing1 mmol eachof
manganesesulfate (MnS04) and Krcitrate. Stir occasionallyfor a day
or two and then storethe beakers outof direct light without disturbance,
replacingwater as it evaporates.
2. Alternatively, the samesuspensions can be placedin conical flasks with
50 mL of additional water. Stopperthe flask loosely with a foam plug
and place on a rotating shakerproviding vigorous horizontal swirling
motion. DetermineChromium(VI) by the DPC method(see"Determin-
ation of Chromium(VI) in Solutions") every 3 wk in the supernatant
solutionsin the beakersor in clear filtrates from the conical flasks.
Great patienceis required. Positive tests may not appearuntil a month's
time or more haselapsed,and then the appearance of Cr(VI) is ephemeral.Levels
of Cr(VI) in solution are usually low, in the rangeof 1 to 5 uM. Suchfleeting and
low Cr(VI) levels probably are not to be consideredhazardousin a natural wet-
land. Nevertheless,the merepresenceof Cr(VI) indicatesa highly oxidative inter-
face, and oxidation of Cr presentsitself as a valuabletool for characterizingoxi-
dation statusin a wetland.
Millions of tons of leatherprotein are cross-linkedby Cr(I1I) eachyear to
become nearly nonbiodegradablethrough the chrome tanning process. Soil
organicmattercan be "chrome tanned"as well when it is contaminatedwith Cr
waste.Both leatherand soil organic mattercan be detannedthrough removal of
Cr from the leatherwith oxalateor citrate. In essence,this procedureis a method
for detanningand quantifying Cr boundin inert forms by soil organicmatter,tan-
nery wasteor other high-chromiumbiosolids.
Long-termincubationat field capacitymoistureof soil samplescontaining
inert Cr(I1I) bound in tannery sludge sometimeswill result in detanningof the
leatherscrapsafter 1 to 4 yr, resulting inextractableCr(VI) (Bartlett, 1986). From
a Cr(VI) toxicity standpoint,it is fortunatethat detanningis a slow process,even
though it meansour civilization is becomingsatiatedwith well worn and out-of-
fashion old leather.
REFERENCES
Amacher,M.e., and D.E. Baker. 1982. Redox reactionsinvolving chromium, plutonium, and man-
ganesein soils. DOE/DP/04515-1.Inst. Res. Land Water Resourc.,PennsylvaniaStateUniv.
and U.S. Dep. Energy,Las Vegas,NY.
AmericanChemicalSociety.1983. Principlesof environmentalanalysis.Anal. Chern.55:2210-2218.
American Public Health Association. 1985. Chromium. StandardMethods for the examinationof
water and wastewater.16th ed. APHA, Washington,De.
Anonymous.1993. Chromium. HazardousWasteConsultant.11:2.25-2.30.

Apte, S.C., S.D.W. Comber,M.I. Gardner,and AM. Gunn. 1991. Direct determinationof chromium
in estuarineand coastalwatersby electrochemicalatomic absorptionspectrometry.1. Anal.
At. Spectros.6:169-172.
Arar, E.I., S.E. Long, T.D. Martin, and S. Gold. 1992. Determinationof hexavalentchromium in
sludgeincineratoremissionsusing ion chromatographyand inductively coupledplasmamass
spectrometry.Environ. Sci. Technol. 26:1944-1950.
Arar, E.I., S.E. Long, andJ.D. Pfaff. 1991.Determinationof dissolvedhexavalentchromiumin drink-
ing water, groundwater,and industrial effluents by ion chromatography.Method 218.6. Rev.
3.0. USEPA Environ. Monit. Sys. Lab., Cincinnati, OH.
Araujo, AN., J.L.F.C. Lima, AO.S.S. Rangel, 1. Alonso, J. Bartoli, and R. Barber. 1989. Simul-
taneousdeterminationof total iron and chromium (VI) in wastewaterusing a flow injection
systembasedon the sandwichtechnique.Analyst 114:1465-1471.
Babu, D.R., and P.R. Naidu. 1991. A solventextraction-atomicabsorption technique for the simulta-
neousdeterminationof low concentrationsof iron, nickel,chromium,and manganese in drink-
ing water. Talanta38:175-179.
Baker, D.E., and N.H. Suhr. 1982. Atomic absorption and flame emission spectrophotometry.p.
13-28. In AL. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9.
ASA and SSSA,Madison,Wi.
Bartlett, R.I. 1965. A biological method for studying aeration status of soil in situ. Soil Sci.
100:403-408.
Bartlett, R.I. 1981. Oxidation-reductionstatus of aerobic soils. p. 77-102. In R.H. Dowdy (ed.)
Chemistry in the soil environment.ASA Spec. Publ. 40. ASA, CSSA, and SSSA, Madison,
WI.
Bartlett, R.I. 1985. Chromium. p. 49-62. In D.E. Baker (ed.) Criteria and recommendations for land
application of sludgesin the Northeast.NortheastRegional Publ. PennsylvaniaState Univ.
Bull. 851.
Bartlett, R.I. 1986. Soil redox behavior. p. 197-207.In D.L. Sparks(ed.) Soil physical chemistry.
CRC Press,Inc., Boca Raton, FL.
Bartlett, R.J. 1987. Chromium oxidation in soils and water: Measurementsand mechanisms.p.
310-330. In D. Serrone(ed.) Proc. ChromiumSymp. 20-21 May 1986.Update.Indust. Health
Found.,Pittsburgh,PA
Bartlett, R.J. 1988. Manganeseredox reactionsand organic interactionsin soils. p. 59-73. In R.D.
Graham et al. (ed.) Manganese insoils and plants. Kluwer Acad. Publ., Dordrecht, the
Netherlands.
Bartlett, R., and B.R. James.1979. Behaviorof chromium in soils: III. Oxidation. J. Environ. Qual.
8:31-35.
Bartlett, R.I., and B.R. James.1980. Studyingdried, storedsoil samples-some pitfalls. Soil Sci. Soc.
Am. J. 44:721-724.
Bartlett, R.I., and B.R. James.1988. Mobility and bioavailability of chromium in soils. p. 267-304.
In J.O. Nriagu and E. Nieboer (ed.) Chromium in the natural and humanenvironments.John
Wiley & Sons,Inc., New York.
Bartlett, R.J., and 1.M. Kimble, J.M. 1976a. Behavior of chromium in soils. I. Trivalent forms. J.
Bartlett, R.I., and J.M. Kimble. 1976b. Behavior of chromium in soils. II. Hexavalent forms. J.
Beere, H.G., and P. Jones. 1994. Investigation of chromium(lll) and chromium(VI) speciation in
water by ion chromatography with chemiluminescencedetection. Anal. Chim. Acta
293:237-243.
Bloomfield, C., and G. Pruden. 1980. The behaviourof Cr(VI) in soil under aerobicand anaerobic
conditions.Environ. Pollut. Ser. A 23:103-114.
Bowman, RA 1988.A rapid method to determinetotal phosphorusin soils. Soil Sci. Soc. Am. J.
52:1301-1304.
Bryson,W.G., and C.M. Goodall. 1981. Improvedspectrophotometricdeterminationof chromium in
animal tissuedigestswith diphenylcarbazide.Anal. Chim. Acta 124:391-401.
Cary, E.E., and M. Rutzke. 1983. Electrochemicalatomic absorptionspectroscopicdeterminationof
chromium in plant tissues.J. Assoc. Off. Anal. Chern. 66:850-852.
Cazeneuve,P. 1900.Sur la diphenylcarbazide,reactiftressensiblede quelquescomposesmetalliques:
cuivre, mercure,fer au maximum,acide chromique.Bull. Soc. Chim. Paris.23:701-706.
Chang,C.A, H.H. Patterson,L.M. Mayer, and D.E. Bause.1980. Determinationof trivalent chromi-
um in seawaterby chemiluminescence. Anal. Chern.52:1264-1266.
CHROMIUM 699
Collins, K.E., P.S.Bonato,C. Archundia,M.E.L.R. deQueiroz,and C.H. Collins. 1988. Column chro-
matographic speciation of chromium for Cr(VI) and several species of Cr(III).
Chromatographia26:160-162.
Condron,M. 1991. Soils with spodiccharacteristicson the EasternShoreof Maryland. M.S. thesis.
Univ. of Maryland, College Park.
Cotton, FA, and G. Wilkinson. 1988. Advancedinorganic chemistry. 5th ed. JohnWiley & Sons,
Inc., New York. p. 686.
Cranston,R.E., and 1.W. Murray. 1978. The determinationof chromium speciesin natural waters.
Crosmun,S.T., and T.R. Mueller. 1975. The determinationof chromium(VI) in natural watersby dif-
ferential pulse polarography.Anal. Chim. Acta 75:199-205.
DenBoef, G., W.J. Dejong, G.C. Krijn, and H. Poppe. 1960. Spectrophotometricdeterminationof
chromium(ill) with EDTA. Anal. Chim. Acta 23:557-564.
Dionex Corporation.1990. Determinationof Cr(VI) in water, wastewater,and solid waste extracts.
Tech. Note 26. Dionex Corp., Sunnyvale,CA
Elleouet, e., F. Quentel,and e. Madec. 1992. Determinationof trace amountsof chromium(VI) in
water by electrochemicalmethods.Anal. Chim. Acta 257:301-308.
FederalRegister.1980.45:33084.(May 19).
Fendorf,S.E. 1995. Surfacereactionsof chromium in soils and waters.Geoderma67:55-71.
Fisher,J.A 1990. The chromiumprogram.Harperand Row, New York.
Fong,w.L., andJ.C.G.Wu. 1991.Chromiumspeciationusingion chromatography-atomic absorption
systemwith on-line preconcentration.Spectros.Lett. 24:931-941.
Goldhaber,S., and C. Vogl. 1989. Developmentof the reviseddrinking water standardfor chromium.
Sci. Tot. Environ. 86:43-51.
Golwelker, A, K.S. Patel, and R.K. Mishra. 1990. Extraction-spectrophotometric determinationof
chromium (VI) with hydroxyamidines.Bull. Chern. Soc. Jpn. 63:605-608.
Gorsuch,T.T. 1970. The destructionof organicmatter. PergamonPress,Oxford.
Hiraide, M., and A. Mizuike. 1989. Separationand determinationof chromium (VI) and chromium
(III) associatedwith negatively-chargedcolloids in river water by sorption on DEAE-
sephadexA-25. Fres.1. Anal. Chern. 335:924-926.
Isshiki, K., Y. Sohrin, H. Karatani, and E. Nakayama.1989. Preconcentrationof chromium(lII) and
chromium(VI) in seawater by complexationwith quinolin-8-01 and adsorptionon macrop-
orous resin. Anal. Chim. Acta 224:55-64.
James,B.R. 1994. Hexavalentchromium solubility and reduction in alkaline soils enriched with
chromite ore processingresidue.1. Environ. Qual. 23:227-233.
James,B.R, and S.G. Aschmann.1992. Solublephosphorusin a forest soil Ap horizon amendedwith
municipal wastewatersludgeor compost.Commun.Soil Sci. Plant Anal. 23:861~75.
James,B.R., and RJ. Bartlett. 1983a. Behavior of chromium in soils: V. Fate of organically-com-
piexed Cr addedto soil. J. Environ. Qual. 12:169-172.
James,B.R., and R.J. Bartlett. 1983b. Behavior of chromium in soils: VI. Interactionsbetweenoxi-
dation- reductionand organiccomplexation.J. Environ. Qual. 12:173-176.
James,B.R, and R.I. Bartlett. 1983c. Behaviorof chromium in soils: VII. Adsorption and reduction
of hexavalentforms. J. Environ. Qual. 12:177-181.
James,B.R., and R.I. Bartlett. 1984a.Nitrification in soil suspensions treatedwith chromium (III, VI)
saltsor tannerywastes.Soil BioI. Biochem. 16:293-295.
James, B.R., and R.I. Bartlett. 1984b. Plant-soil interactions of chromium. J. Environ. Qual.
13:67-70.
James,B.R., and D.R. Bouldin. 1986. A cathodicstripping voltammetricmethodfor nanomolarcon-
centrationsof labile and total iron and zinc in soil solutions. Commun.Soil Sci. Plant Anal.
17:1185-1201.
James,B.R., J.C. Petura, R.I. Vitale, and G.R. Mussoline. 1995. Hexavalentchromium extraction
from soils: A comparisonof five methods.Environ. Sci. Technol. 29:2377-2381.
Jen, J-E, G-L. Ou-Yang, C-S. Chen, and S-M. Yang. 1993. Simultaneousdeterminationof chromi-
um(III) and chromium(VI) with reversed-phase ion-pair high-performanceliquid chromatog-
raphy. Analyst 118:1281-1284.
Johnson,e.A. 1990. Rapid ion-exchangetechniquefor the separationand preconcentrationof chro-
mium (VI) and chromium (III) in fresh waters.Anal. Chim. Acta 238:273-278.
Katz, S. 1991. The analytical biochemistryof chromium. Environ. Health Perspect.92:13-16.
Liu, S.P., and Ee. Wang. 1991. A highly sensitivecolour reaction for chromium (VI) with iodide
basic xanthenedye-PVA system.Talanta38:801-804.
700 BARTLETT & JAMES
Lopez, A., T. Rotunno, F. Palmisano,R. Passino,G. Tiravanti, and P.G. Zambonin. 1991. Simul-
taneousdeterminationof chromium(III), aluminum (III), and iron (II) in tannerysludgeacid
extractsby reversed-phase high-performanceliquid chromatography.Environ. Sci. Technol.
25:1262-1266.
Losi, M.E., C. Amrhein, and W.T. Frankenberger,Jr. 1994. Environmentalbiochemistryof chromi-
um. Rev. Environ. Contam.Toxicol. 136:91-121.
Lovett, RJ., and G.F. Lee. 1979. Analysis of chromium in natural waters by gas chromatography.
Marczenko,Z. 1986. Separationand spectrophotometricdeterminationof elements.Ellis Horwood
Ltd., Chichester,England.
Marchant,H. 1964. Uber die reaktion von chrom mit diphenylcarbazidund diphenylcarbazon.Anal.
Chim. Acta 30:11-17.
Marina-Sanchez,M.A., M.E. Diaz-Garcia,and A. Sanz-Medel.1992. Simultaneousdeterminationof
cobaltand chromiumby ion chromatographywith chemiluminescence detectionand its appli-
cation to glassanalysis.Mikrochim. Acta 106:227-234.
Mehra, H.C., and W.T. Frankenberger.1989. Single-columnion-chromatographicdeterminationof
chromium(VI) in aqueoussoil and sludgeextracts.talanta36:889-892.
Milacic, R, and 1. Stupar. 1994. Simultaneousdetermination of chromium(III) complexesand
chromium(VI) by fast proteinanion-exchangeliquid chtomatography-atomic absorptionspec-
trometry.Analyst 119:627-{)32.
Milacic, R, and J. Stupar. 1995. Fractionationand oxidation of chromium in tannery waste- and
sewagesludge-amended soil. Environ. Sci. Technol.29:506-514.
Milacic, R, J. Stupar,N. Kozuh, and J. KoroSin. 1992. Critical evaluationof three analytical tech-
niquesfor the determinationof chromium(VI) in soil extracts.Analyst 117:125-130.
Miyazaki, A., and R.M. Barnes.1981. Differential determinationof chromium (VI)-chromium (III)
with poly(dithiocarbamate)chelating resin and inductively coupledplasma-atomicemission
spectrometry.Anal. Chern.53:364-366.
Nelson, D.W., and L.E. Sommers. 1996. Total carbon, organic carbon, and organic matter. p.
Osaki,S., T. Osaki, M. Setoyoma,and Y. Takashima.1983. Gel chromatographicbehaviourof trace
amounts of chromium (VI) and hydrolysed Cr(III) in aqueoussolution. J. Chromatogr.
257:180--184.
Pavel,1., J. Kliment, S. Stoerk,and O. Suter. 1985. Preservationof tracesof chromium(VI) in water
and wastewater samples.Fres. Z. Anal. Chern. 321:587-591.
Perkin-Elmer Corporation. 1984. Analytical protocol for atomic absorption. Perkin-Elmer Corp.,
Norwalk, CT.
Pettine, M., T. LaNoce, A. Liberatori, and L. Loreti. 1988. Hydrogen peroxide interferencein the
determination of chromium (VI) by the diphenylcarbazidemethod. Anal. Chim. Acta
209:315-319. .
Pflaum,RT., andL.C. Howick. 1956.The chromium-diphenylcarbazide reactions.J. Am. Chern.Soc.
78:4862-4866.
Pilkington, E.S., and P.R Smith. 1967. Spectrophotometricdeterminationof chromium in ilmenite.
Prokisch,1., B. Kovacs,Z. Gyori, and J. Loch. 1994. Interfacingion chromatographywith inductive-
ly coupledplasmaatomic emissionspectrometryfor the determinationof chromium(1I1)and
chromium(VI). J. Chromatogr.A683:253-260.
Rathore,D.P.S.,and P.K. Tarafder.1992. Spectrophotometric determinationof chromium in geologi-
cal samples.Anal. Chim. Acta 257:129-133.
Reisenauer,H.M. 1982. Chromium. p. 337-346.In A.L. Pageet al. (ed.) Methodsof soil analysis.
Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Rengasamy,P., and J.M. Oades. 1977. Spectrophotometricdetermination of monomeric plus
oligomeric and polymeric hydroxy speciesof chromium (III) in aqueoussolutions. Aust. J.
Chern. 30:1383-1385.
Richard, F.C., and A.C.M. Bourg. 1991. Aqueousgeochemistryof chromium:A review. Wat. Res.
25:807-816.
Ross, D.S., R.E. Sjogren, and RJ. Bartlett. 1981. Behavior of chromium insoils: IV. Toxicity to
microorganisms.J. Environ. Qual. 10:145-148.
Rubio, R, A. Sahuquillo,and G. Rauret.1992.Determinationof chromiumin environmentaland bio-
logical samplesby atomic absorptionspectroscopy:A review. Int. J. Environ. Anal. Chern.
47:99-128.
CHROMIUM 701
Saleh,EY., J.H. Huang, and R.Y. Lewis. 1989. Ion Chromatographyof soluble chromium (III) and
chromium (VI). J. Chromatogr.Sci. 27:480-484.
Saltzman,B. 1952. Microdeterminationof chromium with diphenyIcarbazideby permanganateoxi-
dation. Anal. Chern.24:1016-1020.
Soltanpour,P.N., J.B. JonesJr., and S.M. Workman. 1982. Optical emissionspectrometry.p. 29-66.
In A.L. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA, Madison,WI.
Stollenwerk,KG., and D.B. Grove. 1985. Reductionof hexavalentchromium in water samplesacid-
ified for preservation.J. Environ. Qual. 14:396-399.
Subramanian,KS. 1988. Determinationof chromium (III) and chromium (VI) by ammoniumpyrro-
Iidinecarbodithioate-methylisobutyl ketone furnace atomic absorption spectrometry.Anal.
Chern. 60:11-15.
Suzuki, E.Y., and R.I. Early. 1991. Analysis of chromic oxide in small samplesof feeds and feces
using chlorine bleach.Can. J. Anim. Sci. 71:931-934.
Syty, A., R.G. Christensen,andT.e. Rains. 1988. Determinationof addedchromium(III) and chromi-
um (VI) in natural water by ion-pairing high-performanceliquid chromatographywith detec-
tion by atomic absorptionspectrometry.J. Anal. At. Spectrom.3:193-197.
Tomlinson, M.I., J. Wang, and J.A. Caruso.1994. Speciationof toxicologically important transition
metalsusing ion chromatographywith inductively coupledplasmamassspectrometricdetec-
tion. J. Anal. At. Spec.9:957-965.
Trojanowicz, M., and E. Pobozy. 1992. Speciationof chromium by ion-pair chromatographywith
postcolumnspectrophotometricdetection.Anal. Lett. 25:1373-1387.
Urasa,I.T., and S.H. Nam. 1989. Direct determinationof chromium(III) and chromium (VI) with ion
chromatographyusing direct current plasma emission as element selective detector. J.
Chromatogr.Sci. 27:3~37.
Urone, P.E 1955. Stability of colorimetric reagentfor chromium,s-diphenyIcarbazide, in varioussol-
vents. Anal. Chern.27:1354-1355.
u.S. EnvironmentalProtectionAgency. 1984. Health assessment documentfor chromium. USEPA
Rep. 600/8-83-014EU.S. Gov. Print. Office, Washington,De.
U.S. EnvironmentalProtectionAgency. 1991. Determinationof dissolvedhexavalentchromium in
drinking water, groundwaterand industrial wastewatereffluents by ion chromatography.
Method 218. U.S. Gov. Print. Office, Washington,DC.
Vitale, R.I., G.R. Mussoline, J.e. Petura,and B.R. James.1994. Hexavalentchromium extraction
from soils: Evaluationof an alkaline digestionmethod.J. Environ. Qual. 23:1249-1256.
Vos, G. 1985. Determinationof dissolvedhexavalentchromium in river water, seawater, and waste
water. Fres. Z. Anal. Chern. 320:556-561.
Walkley, A., and LA. Black. 1934. An examinationof the Degtjareff method for determiningsoil
organic matter and a proposedmodification of the chromic acid titration method. Soil Sci.
37:29-38.
Williams, T., P. Jones, and L. Ebdon. 1989. Simultaneousdeterminationof chromium (III) and
chromium (VI) at ultratracelevels using ion chromatographywith chemiluminescence detec-
tion. J. Chromatogr.482:361-366.
Yamazaki, H. 1980. Preconcentrationand spectrophotometricdeterminationof Cr(VI) in natural
watersby coprecipitationwith barium sulfate.Anal. Chern. 113:131-137.
Yoshimura, K 1988. Application of ion-exchanger phase absorptiometry to flow analysis.
Determinationof trace amountsof chromium (VI) in water. Analyst 113:471-474.
Published 1996
Chapter26
Copper and Zinc
S. T. REED, Florida A&M University, Tallahassee, Florida
D. C. MARTENS, Virginia Polytechnic Institute and State University,

Blacksburg, Virginia
Occurrencesof Cu and Zn deficiencieshaveled to the needfor laboratoryproce-

duresto predict the plant availablelevelsof thesetraceelementsin soils. In 1914,
Maze providedthe evidencethat Zn was needed byplantsand, in 1928, Sommer
and Lipman demonstratedthe plant requirementfor Cu (Stout, 1956). Sincethen,
numerousreportshaveshown that Cu and Zn deficienciesoccur in plants under
field conditions(Martens& Westermann,1991). The small amountsof Cu and
Zn required to correct thesedeficienciesover a numberof yearswould not be
expectedto lead to accumulationof phytotoxic concentrationsin soils (Martens
& Westermann,1991). Phytotoxic soil concentrationsare most probablewhere
wasteswith high Cu and Zn levels are used as soil amendments,e.g., where
municipal sludge,wastefrom the smeltingindustry, and manurefrom the poultry
and swine industriesare applied to soils.
Formsand Concentrationsof Copperand Zinc in Soils
Copperand Zn exist in the following forms in soils: (i) as free and com-
plexed ions in soil solution, (ii) as nonspecifically and specifically adsorbed
cations,(iii) as ions occludedmainly in soil carbonatesand hydrousoxides, (iv)
in biological residuesand living organisms,and (v) in the lattic structureof pri-
mary and secondaryminerals(McLaren & Crawford, 1973; Iyengaret aI., 1981;
Neilsen et aI., 1986; Liang et aI., 1991). The distribution of Cu and Zn in these
forms varieswidely in soils as a result of differencesin mineralogy,parentmate-
rials, and organic matter content(Iyengar et aI., 1981; Hazra & MandaI, 1987;
Baker, 1990; Liang et aI., 1991).
Nonspecificadsorptionof Cu and Zn occursin soils from ionic bond for-
mation, e.g., by electrostaticinteractionof the cation with the chargethat arises
from isomorphoussubstitutionin phyllosilicates(Sposito, 1981). A nonspecifi-
cally adsorbedcation usually is referredto as an exchangeablecation. Specific
SegoeRd., Madison,WI 53711,USA. Methods of Soil Analysis. Part 3. Chemical Methods--SSSA
Book Seriesno. 5.
703
704 REED & MAKI'ENS
adsorptionof Cu and Zn takesplacein soils as thesecationsreactwith electron

pair donors(e.g., 0 and N in functional groups)to form bondswith relatively
high covalency.In soils, Cu and Zn are specificallyadsorbedby carbonates,soil
organicmatter,phyllosilicates,andhydrousoxidesof AI, Fe, andMn (Udo et al.,
1970; Schnitzer,1978; Kabata-Pendia& Pendias,1991).
Occlusionoccursin soils asa layer of AI, Fe, or Mn hydroxideprecipitates
over Cu and Zn that are complexedby organicmatteror adsorbedby carbonate,
hydrousoxide, or phyllosilicatesurfaces.The occludedcation is not in contact
with soil solution and, hence,cannotdissociateinto soil solution. Occlusionof
Cu and Zn also proceedsin soils by coprecipitationreactions.
Uncontaminatedsoils generallycontainfrom 6 to 60 mg total Cu andfrom
17 to 160 mg total Zn kg-1 (Iyengar et al., 1981; Kabata-Pendias& Pendias,
1985).A low portion of the total soil Cu and Zn existsin soil solution.The ionic
andcomplexedCu andZn in soil solutionrangedfrom 0.004to 0.039mg Cu L-l
for 20 calcareoussoils (Hodgsonet al., 1966)andfrom 0.032to 0.172mg Zn L- 1
for severalacid soils (Hodgsonet al., 1965). Although small in amountas com-
paredwith solid forms, Cu and Zn in soil solution playa vital role in supplying
the plant requirementfor thesetraceelements. Thatis, most of the plant require-
ment of Cu and Zn is suppliedby root absorptionfrom soil solution.
Available Copperand Zinc forms in Soils

A labile nutrientrefersto the amountof dissolvednutrientper unit volume
of soil plus the amountof surface-boundnutrient per unit volume of soil that is
in rapid equilibrium with the dissolvednutrient (Corey, 1990). Hence,a labile
ion is an ion in soil solutionor on the solid phasethat may exchangewith an ion
of the samekind on the solid phaseor in soil solution, respectively(Barber,
1984).Labile Cu and Zn consistmainly of free andcomplexedCu andZn in soil
solution, which provide the intensity of soil to supply thesenutrientsto plants,
and the nonspecificallyadsorbedCu and Zn, which provide the capacityof soil
to replenishthesenutrientsinto soil solution. Copperand Zn deficienciesdevel-
op wheresoils havean inherentlylow level of Cu andZn in soil solutionor a low
capacityto replenishthe Cu and Zn removedfrom soil solution.
Nonlabile Cu and Zn exist in forms that are tightly bondedto soil surfaces
which are in contactwith soil solution suchas a portion of the Co and Zn com-
plexedby organicmatterand in forms that are not in contactwith soil solution,
e.g.,the occludedCu and Zn, and the Cu and Zn in biological materialsand liv-
ing organismsand in the lattice structureof primary and secondaryminerals.
Labile Cu and Zn are in a metastableequilibria with nonlabile forms in soils.
Therefore,portionsof the labile forms revert to nonlabileforms with time and,
conversely,portionsof the nonlabileforms revertto the labile forms uponweath-
ering. Overall, Cu and Zn reversionfrom nonlabileto labile forms is very slow
and rarely would supply crop needsduring a growing season.
Soil PropertiesVersusDeficienciesof Copperand Zinc

Copper deficienciesare most prevalent in organic soils. Complexation
reactionsbetweenCo and functional groupsof organicmattercausethe Cu defi-
COPPER& ZINC 705
cienciesin peatsand mucks (Martens& Westermann,1991). Zinc deficiencies

also occur on organicsoils, but to a lesserdegreethan Cu deficiencies,because
Cu is complexedmore tightly than Zn by functional groups in organic matter
(Alloway, 1990). Copperand Zn deficienciesare more likely to occur in sandy
soils than in either silty or clayey soils. Low amountsof Cu and Zn in parent
material, quartz, accountfor the low amountof availableCu and Zn in sandy-
textured soils(Krauskopf, 1972).
Copperand Zn deficienciesfrequently result from managementpractices
usedduring crop production. Overliming acid soils may causeCu and Zn defi-
cienciesbecausecrop uptakeof the trace metalsdecreasesas soil pH increases
(Martens& Westermann,1991). Low Cu and Zn availabilitiesat higher levelsof
soil pH reflect decreases in dissolutionof soil mineralswith releaseof Cu and Zn
as well as increasesin Cu and Zn complexationby organicmatter,adsorptionon
surfacesof inorganic soil components,and occlusion by soil hydroxides and
oxides,High levels of availablesoil P, often from fertilization, induce Zn defi-
ciencies(Mengel & Kirkby, 1987). Phosphorus-induced Zn deficienciesoften
occur on near-neutralto alkaline soils with marginal amountsof available Zn
(Martens& Westermann,1991). Zinc deficienciesalso are commonon exposed
subsoil where surfacesoil was removedby erosion, land leveling, or terracing
(Frye etaI., 1978; Gerwing et aI., 1982).The Zn deficiencieson exposedsubsoil
refle~t removalof availableZn in the surfacesoil, i.e., removalof Zn c~mplexed
by soil organicmatteror adsorbedby mineral surfacesin the topsoil.
Developmentof Copperand Zinc Soil Tests
Developmentof soil tests to predict the availabilities of Cu and Zn to

plantsgenerallyrequiresthe following threesequentialsteps:(i) extractantselec-
tion, (ii) greenhouseevaluation,and (iii) field calibration.The extractantshould
be selectedto solubilize amountsof Cu and Zn that are proportionalto amounts
that will be absorbedby plantsduring a single growing season,and also should
be effective over a wide rangeof soil types.Greenhouseevaluationis undertak-
en to determineif a relationshipexistsbetweenthe amountof extractableCu and
Zn and the quantitiesabsorbedby plantsand, hence,to ascertainif it is desirable
to completefield calibration. It is reasonedthat, if extractableCu and Zn are
unrelated.toCu and Zn uptakesunder controlledgreenhouseconditions,then a
suitablerelationshipwill not be obtainablein harsherfield environments.Field
calibrationis necessaryto determinecritical levels of extractableCu and Zn that
separatesoils into deficient and sufficient categories.
Critical Cu and Zn levels could be obtainedlessexpensivelyundergreen-
housethan underfield conditions.However, a critical level for a Cu or Zn soil
test usually differs undergreenhouseand field conditions.The following factors
causedissimilar Cu and Zn uptakesand, hence,lead to differencesin the critical
levels undergreenhouseand field conditions(Logan & Chaney,1983; Mortvedt,
1977):
1. Use of higher levels of NHt in the greenhousecauseslower pH levels
in pots than in field soils.
706 REED & MARrENS
2. Use of higher levels of fertilizers in the greenhousecausehigher solu-

ble salt levels in pots than in the field.
3. Greater root-to-soilcontactoccursfrom plant root confmement inpots
as comparedwith field conditions.
4. Abnormal light, humidity, and moisture regimes occur in the green-
houseas comparedwith field conditions.
5. Plantsseldomaregrown to maturity in the greenhouse.Extrapolationof
critical nutrient levels from the vegetativegrowth stage in the green-
houseto matureplantsin the field overlooksthe needfor additionalCu
and Zn replenishmentof the soil solution in the field.
Use of the threesequentialstepsin the orderof extractantselection,green-
houseevaluation,and field calibration is highly suitablefor developmentof Cu
and Zn soil tests.This sequentialprocedurewould not be suitable for develop-
mentof soil testsfor all essentialnutrients.For example,Tierney (1981) was not
able to obtain a growth increaseof soybean[G/ysine max (L.) Merr.] plantsfrom
Mn applicationin the greenhouseon eight soilson which severeMn deficiency
occurredin the field. Increasesin Mn availability from the unavoidablereducing
conditionsdue to watering precludedthe developmentof Mn deficiency in the
greenhouseon the eight soils. Thus, only the extractionselectionand calibration
stepswould be usedfor soil testdevelopmentwheregreenhouseproceduresdras-
tically affect nutrient availability.
Overview of Copperand Zinc Soil Tests
Initially single elementavailability tests,such as the 0.1 M HCI extractant

for Zn, were developedfor estimatingthe availability of soil Cu or Zn (Wear &
Sommer,1948). Later, simultaneousextraction of micronutrientswas initiated
with the developmentof the DTPA-TEA (diethylenetriaminepentaacetic acid-tri-
ethanolamine)extractantfor Cu, Fe, Mn, and Zn (Lindsay & Norvell, 1969).
Currently, soil test developmentis directed toward simultaneousextraction of
micronutrients and macronutrientsin a single method, as is the case for the
Mehlich-III and DTPA-AB (diethylenetriaminepentaacetic acid-NH4HC03) pro-
cedures(Mehlich, 1984; Soltanpour,1991). Simultaneousextractionand deter-
mination of micronutrientsand macronutrientsis desirablefrom the standpoint
of rapid conveyanceof soil test datato crop producersat a reasonablecost.
Extractants for current Cu and Zn availability tests include chelating
agents,inorganicacids,and a combinationof a chelatingagent,acids, and salts.
Amounts of extractableCu and Zn solubilized from soil by these extractants
dependon concentrationof extractionsolutioncomponents,extractiontime, soil-
extraction solution ratio, extraction temperature,type of extraction vessel and
shaker, and shaker speed(Sorensenet aI., 1971; Mehlich & Bowling, 1975;
Soltanpour etaI., 1976; Lindsay & Norvell, 1978). Variations in the extraction
conditionslead to wide differencesin amountsof Cu or Zn solubilizedby a spe-
COPPER& ZINC 707
cific soil test. Thus, calibrationdatafor a soil test apply solely to soil test values
obtainedby the extractionconditionsusedduring the calibration.
Soil testshave beencalibratedto determineif soils contain adequateCu
and Zn for normal plant growth (Martens & Lindsay, 1990). It would be desir-
able to usethe samesoil testto determinedeficientand toxic levelsof Cu and Zn
from the standpointof rapid conveyanceof data to the crop producerat a rea-
sonablecost. Thus far, soil test calibration data are not available to determine
phytotoxic levels of soil Cu and Zn (Sims & Johnson,1991).
Soil Sampleand ExtractantPreparation
Copper and Zn contaminationof soil samplesand extraction solution

should be avoided to ensureaccurateextractableCu and Zn data. Galvanized
containers,cast iron mortars, rubber stoppers,brassscreens,and other metal
utensilsmay causeCu and Zn contaminationand, hence,shouldbe avoideddur-
ing preparationof soil samplesfor extractableCu and Zn analyses(Eik &
Gelderman,1988).Extractionsolutionsshouldbe preparedin acid-washedglass-
ware or plasticwareto prevent Cu and Zn contamination.Reagentsand water
usedduring extractionsolution preparationmay be a sourceof Cu and Zn cont-
amination.BakerandAmacher(1982)indicatedthat highly puredeionizedwater
shouldbe usedfor extractableCu and Zn analysesto preventcontamination.
Drying temperatureaffectsthe amountsof Cu and Zn extractedfrom soils
(Leggett & Argyle, 1983). Copper and Zn are releasedduring drying due to
reduction of Mn in hydrous oxides and to alteration of functional groups in
organic matter (Hammes& Berger, 1960). Grinding to a smaller particle size
increasessurfaceareaand, therefore,increasesamountsof extractableCu and Zn
(Seversonet aI., 1979;Soltanpouret aI., 1979).The differencesin grinding force,
samplevolume, and time causechangesin soil particle size. Variationsin drying
and grinding conditionslead to large differencesin amountsof Cu and Zn solu-
bilized by a specificsoil test. Thus,calibrationdatafor a soil test apply solely to
soil test valuesobtainedby the drying and grinding proceduresusedduring cal-
ibration of the soil test.
acid-TriethanolamineMethod
The DTPA-TEA soil test was developedby Lindsay and Norvell (1969,
1978) to identify near-neutraland calcareoussoils with insufficient levels of
available Cu, Fe, Mn, and Zn. This method previously was referred to as the
DTPA method,but is called the DTPA-TEA methodhereinto preventconfusion
with the DTPA-AB method, which was developedby (Soltanpour& Schwab,
1977). The chelatingagent,DTPA, was selectedfor the soil test becauseit has
the most favorablecombinationof stability constantsfor simultaneouscomplex-
ation of Cu, Fe, Mn, and Zn (Lindsay & Norvell, 1978).
Lindsay and Norvell (1978) describedthe theoreticalbasisfor the DTPA-
TEA method. The DTPA moleculesform water-solubleCu and Zn complexes
and, thereby,decreaseCu2+ and Zn2+ activities in soil solution. In response,Cu
and Zn desorb from soil surfacesto replenish soil solution Cu2+ and Zn2+.
708 REED & MARTENS
Amountsof chelated Cu and Zn that accumulatein solution during the extraction

are functions of the Cu2+ and Zn2+ activities in the soil solution (intensityfactor)
and the ability of the soil to replenishCu and Zn (capacityfactor). Plant uptake
of Cu and Zn increaseswith increasesin the amountsof Cu and Zn in soil solu-
tion and with the capacityof soil solid phasesto replenishCu and Zn depleted
from soil solution, respectively(Corey, 1990).
SinceZn deficienciesare prevalenton calcareoussoils, the extractantwas
designedto avoid excessivedissolutionof CaC03 with releaseof occludedZn.
This precautionis necessarybecausethe occludedZn (or Cu) in CaC03 is nor-
mally not available for absorption by plant roots. Excessive dissolution of
CaC03 is preventedby inclusion of solubleCaz+ as CaClz in the extractionsolu-
tion and by buffering the solution at pH 7.3 with TEA [HOCH2CH2)3N].
Equipment
The amountof DTPA-TEA extractableCu and Zn in solution is common-
ly measuredby atomic absorptionor inductively coupledplasmaemissionspec-
troscopy.
Reagents
The DTPA-TEA extraction solution consistsof 0.005 M DTPA, 0.01 M
CaCI2, and 0.1 M TEA adjustedto pH 7.3 with HCl. To prepare10 L of this solu-
tion, dissolve 149.2 g of reagentgrade TEA, 19.67 g of DTPA, and 14.7 g of
CaClz·2H20 in approximately200 mL of deionizedwater. Allow sufficient time
for DTPA to dissolve, and dilute to approximately9 L with deionizedwater.
Adjust the pH to 7.3 :!:: 0.05 with 1.0M HCI while stirring, and dilute to 10 L with
deionizedwater. This solution is stablefor severalmonths.
Copper and Zn stock solutions with concentrationsof 1000 mg L-I are
availablefrom commercialsources.To preparea stocksolutionwith 1000mg Cu
or Zn L-l, weigh 1000 mg of Cu or Zn metal, respectively,into a 250-mL coni-
cal flask, add 10 mL of deionizedwater and 5 mL of concentratedHN03, warm
gently to completedissolutionof Cu or Zn, and boil to expel oxides of N (Am.
Public Health Assoc., 1989). Then evaporateto almost dryness,transfer to a
1000-mLvolumetric, and dilute to volume with DTPA-TEA extractionsolution.
Working standardsshould be preparedby diluting the stock solution to bracket
the amountsof DTPA-TEA extractableCu and Zn in the samples.The DTPA-
TEA extractionsolution should be used for dilutions during preparationof the
working standards.
Procedure
Grind air-driedsoil to passthrougha 1.0 mm stainlesssteelsievein prepa-
ration for the DTPA-TEA extraction.Weigh 10 g of the sievedsoil into a 125-
mL conical flask, add 20 mL of the DTPA-TEA extractionsolution, cover each
flask with stretchableParafilm,andsecurethe flask upright on a horizontalshak-
er with a stroke length of 8.0 cm and a speedof 120 cycles min-I. After a 2-h
shaking period, filter the suspensionsthrough Watman no. 42 filter paper.
COPPER& ZINC 709
Determinethe level ot DTPA-TEA extractableCu andZn in the filtrate by atom-

ic absorptionor inductively coupledplasmaemissionspectrometry.
Calculation
If the filtrate for a samplecontains1.0 mg Cu or Zn L-l, the soil contains
2.0 mg kg-1 ofDTPA-TEA extractableCu or Zn, respectively,as shownbelow
(1.0 mg metal L- 1)(0.020 L) 1

- - - - - - - - - =2.0 mg metal kg- .
0.010 kg
Hence,the numericalvalue for DTPA-TEA extractableCu or Zn expressedas

milligrams per kilogram of soH canbe obtainedby multiplying the milligrams of
the metalper liter in filtrate by two.
Comments
Amountsof DTPA-TEA extractableCu in 75 agriculturalsurfacesoils col-
lectedin Coloradorangedfrom 0.26 to 2.6 mg kg-1 (Lindsay & Norvell, 1978).
A tentativelevel of <0.2 mg DTPA-TEA extractableCu per kilogram was pro-
posedasinadequatefor normalcrop growth on near-neutralandcalcareoussoils.
Amountsof DTPA-TEA extractableZn in 77 agriculturalsurfacesoils collected
in Colorado rangedfrom 0.17 to 11.5 mg kg-1 (Lindsay & Norvell, 1978). A
level of<0.8 mg DTPA-TEA extractableZn perkilogram in near-neutralandcal-
careoussoils indicatedinadequateZn for com (Zea mays L.) production.There
is evidencethat the DTPA-TEA procedurecould be usedto evaluatethe avail-
able Cu and Zn statusof acidic soils by considerationof soil pH along with the
level of DTPA-TEA extractablemetal (Lindsay & Norvell, 1978).
acid-NH4HC03 Method
The DTPA-AB soil test was developedto identify near-neutraland cal-

careoussoils with insufficient levelsof availablemicronutrientsCu, Fe, Mn, and
Zn and macronutrientsN03-N, P, and K (Soltanpour& Schwab,1977; Soltan-
pour et aI., 1982; Soltanpour,1991). Like the DTPA-TEA method(Lindsay &
Norvell, 1969),the extractionsolutionfor the DTPA-AB methodcontains0.005
M DTPA and is alkaline. Hence,the theoreticalbasisfor useof DTPA in an alka-
line solution for extractionof labile soil Cu2+ and Zn2+ from soils by the DTPA-
AB methodis the sameas thoseunder"Diethylenetriaminepentaacetic acid-tri-
ethanolamineMethod" for the DTPA-TEA method(Soltanpour,1991).
SinceZn deficienciesare prevalenton calcareoussoils, the extractantwas
designedto avoid excessivedissolutionof CaC03 with releaseof occludedZn.
This precautionis necessarybecausethe occludedZn (or Cu) in CaC03 normal-
ly is not availablefor absorptionby plant roots. Excessivedissolutionof CaC03
is preventedby inclusion of solubleHCOj" as NH4HC03 in the extractionsolu-
tion and by adjustingthe solution to pH 7.6 with NH40H.
Activated charcoaloriginally was addedto soil prior to extractionby the
DTPA-AB method (Soltanpour& Schwab,1977). The activatedcharcoalwas
710 REED & MARTENS
usedto removeorganic compoundsthat interferedwith the N03" determination

by the chromotropic acid procedure. Researchby Soltanpourand Workman
(1979) indicatedthat use of activatedcharcoalduring the DTPA-AB extraction
should be eliminatedbecausethe charcoaladsorbsmetal chelatesand, thereby,
decreaseslevels of extractablemetals. Following this research,the use of acti-
vatedcharcoalwas discontinuedfor the DTPA-AB method(Soltanpour,1991).
Equipment
The amountof DTPA-AB extractableCu and Zn in solution is commonly
measuredby atomic absorptionor inductively coupled plasma emissionspec-
trometry.
Reagents
The DTPA-AB extractionsolution consistsof O.OOS M DTPA and 1.0 M
NH4HC03 adjustedto pH 7.6 with NH40H. To prepare1.0 L of this solutiondis-
solve 1.87g of DTPA in about800 mL of deionizedwater. Add 2 mL of 1:1 con-
centrated NH40Hldeionized water (v/v) solution to facilitate dissolution of
DTPA and to prevent effervescenceduring addition of HC03". Shake until the
DTPA is dissolved,and then add 79.06g of NH4HC03 and stir gently until com-
plete dissolution of the NH4HC03• After the DTPA and NH4HC03 are com-
pletely dissolved,adjustthe pH of the solution to 7.6 with either HCI or NH40H
and dilute to 1.0 L with deionizedwater. The solution is unstablewith respectto
pH and, hence,the solution eithershouldbe prepareddaily or storedunderabout
3 cm of mineral oil. The solution pH remainsquite stablefor 2 wk when stored
underthe oil.
Copper and Zn stock solutions with concentrationsof 1000 mg L -1 are
or Zn L -1, weigh 1000 mg of Cu or Zn metal, respectively,into a 2S0-mL coni-
cal flask, add 10 mL of deionizedwater and S mL of concentratedHN03, warm
lO00-mL volumetric, and dilute to volume with DTPA-AB extraction solution.
Working standardsshould be preparedby diluting the stock solution to bracket
the amountsof DTPA-AB extractableCu and Zn in the samples.The DTPA-AB
extractionsolution should be usedfor dilutions during preparationof the work-
ing standards.
Procedure
Grind air-driedsoil to passthrougha 2.0-mmstainlesssteelsievein prepa-
ration for the DTPA-AB extraction.Weigh 10 g of the sievedsoil into a 2S0-mL
Erlenmeyerflask, add 20 mL of the DTPA-AB extraction solution, securethe
mixture on a reciprocalshaker,and shakein an openflask at 180 cycles min-I.
After a IS-min shakingperiod, filter the suspensionsthrough Whatmanno. 42
filter paper.Determinethe level of DTPA-AB extractableCu and Zn in the fil-
trate by atomic absorptionor by inductively coupledplasma emission spectrom-
etry.
COPPER& ZINC 711
Calculation
If the fIltrate from a samplecontains1.0 mg Cu or Zn L- t, the soil contains
2.0 mg kg-1 of DTPA-AB extractableCu or Zn, respectively,as shown below
(1.0 mg metal L-l)(0.020 L) 1

-------'-'----'- =2.0 mg metal kg- .
0.010 kg
Hence,the numericalvaluefor DTPA-AB extractableCu or Zn expressedas mil-

ligramsper kilogram of soil canbe obtainedby multiplying the milligrams of the
metal per liter in filtrate by two.
Comments
A "crossflow" type of nebulizerclogs when Cu and Zn in high salt solu-
tions are determined by inductively coupled plasma emission spectrometry
(Soltanpouret aI., 1982).Cloggingis preventedby shakinga 2-mL aliquot of the
sampleextractwith 0.25 mL of concentratedHN03 for 10 min in an openves-
sel prior to the Cu and Zn analyses.This proceduredestroysthe COj--HC03"
matrix in the sampleextract.
Amountsof DTPA-AB and DTPA-TEA extractableCu in 400 near-neutral
and calcareousagriculturalsoils in Coloradowere compared bySoltanpourand
Workman(1979). The level of DTPA-AB extractableCu was about0.6 mg kg-1
for soils with 0.2 mg kg-1 of DTPA-TEA extractableCu. Soltanpour(1991) pro-
posedguidelinesfor interpretationof DTPA-AB extractableCu levels for crop
productionas follows: $0.2 mg Cu kg-I, low; 0.3 to 0.5 mg Cu kg-I, medium;
and >0.5 mg Cu kg-I, high.
Amountsof DTPA-AB and DTPA-TEA extractableZn in 400 near-neutral
and calcareousagriculturalsoils in Coloradowere comparedby Soltanpourand
Workman(1979). The level of DTPA-AB extractableZn was about 1.4 mgkg-1
for soils with 0.8 mgkg-1 of DTPA-TEA extractableZn. Soltanpour(1991) pro-
posedguidelinesfor interpretationof DTPA-AB extractableZn levels for crop
productionas follows: $0.9 mg Zn kg-I, low; 1.0 to 1.5 mg Zn kg-I, medium;
and>1.5 mg Zn kg-I, high.
Mehlich-I (DoubleAcid) Method
The Mehlich-I methodwasoriginally developedto estimatelevelsof avail-

able P in soils (Nelson et aI., 1953). By 1974, this extractantalso was used in
many soil testing laboratoriesin the southernregion to measureavailable Ca,
Mg, and K (Sabbe& Breland(ed.), 1974).The Mehlich-I extractantwasfirst cal-
ibratedto determinethe level of an availablemicronutrientin soil by Cox (1968).
He showedthat soils could be separatedinto Mn deficient and sufficient cate-
goriesby a function of Mehlich-I extractableMn and soil pH. Researchby Wear
and Evans(1968) and by Perkins(1970) with Atlantic CoastalPlain soils led to
the use of the Mehlich-I methodas an availableZn test. Wear and Evans(1968)
showedthat Mehlich-I extractableZn correlatedmore closely with Zn uptakeby
com plants than did either 0.1 M HCI or 0.05 M EDTA (ethylenediaminete-
712 REED & MARTENS
traaceticacid) extractableZn, and Perkins(1970) reporteda significant correla-

tion betweenZn in leaf bladesof com plantsand Mehlich-I extractableZn.
Since the extractionsolution for the Mehlich-I methodconsistsof 0.05 M
HCI and 0.0125M H2S04, this procedurehasbeenreferredto as the doubleacid,
dilute doubleacid, and dilute HCI-H 2S04 methodin the literature.StructuralZn,
which is not in contactwith soil solution Zn, may be extractedby shakingsoil
with acid solution, such as the Mehlich-I extractant(Martens& Lindsay, 1990).
The shortextractionperiod of 5 min (Cox, 1968) decreasesextractionof nonla-
bile Zn by the acidic Mehlich-I extractionsolution.
Soil sampleswere shakenin the presenceof charcoalfor the original mul-
tielementMehlich-I extraction(Cox, 1968). The charcoalwas addedto remove
organic componentswhich interfered with the P determinationby the vanado-
molybdophosphoricacid method.Use of charcoalhasbeendiscontinuedbecause
the P in the extraction solution is now being determinedby colorimetric and
instrumentalmethodsnot subject to the interference(Counc. Soil Test. Plant
Anal., 1980).
Equipment
The amountof Mehlich-I extractableZn in solutionis commonlymeasured
by atomic absorptionor inductively coupledplasmaemissionspectrometry.
Reagents
The Mehlich-I extractionsolution consistsof 0.05 M HCI and 0.0125 M
H2S04, To prepare1 L of Mehlich-I extractionsolution, add 4.17 mL of con-
centratedHCI (12M) and 0.70 mL of concentratedH2S04 (17.8M) to about800
mL of of deionizedwater, and dilute to 1 L with deionizedwater.
Zinc stocksolutionswith concentrationsof 1000mg L -1 are availablefrom
commercialsources.To preparea stock solution with 1000 mg Zn L -1, weigh
1000mg of Zn metal into aZ50-mLconical flask, add 10 mL of deionizedwater
and5 mL of concentratedHN03, warm gently to completedissolutionof Zn, and
boil to expel oxides of N (Am. Public Health Assoc., 1989). Then evaporateto
almost dryness,transfer to a 1000-mL volumetric, and dilute to volume with
Mehlich-I extractionsolution. Working standardsshouldbe preparedby diluting
the stocksolution to bracket theamountof Mehlich-I extractableZn in the sam-
ples. The Mehlich-I extraction solution should be used for dilutions during
preparationof the working standards.
Procedure
This Mehlich-I procedurewas adaptedfrom Cox and Wear (1977). Grind
air-dried soil to passthrough a 2.0-mmstainlesssteelsievein preparationfor the
Mehlich-I extraction. Weigh 5 g of the sieved soil into a 50-mL Erlenmeyer
flask, add 20 mL of the Mehlich-I extractionsolution, and shakethe mixture at
180 cycles min-Ion a reciprocalshakerwith a stroke length of 4.5 ern. After a
5-min shakingperiod,filter the suspensionsthroughWhatmanno. 42 filter paper.
COPPER& ZINC 713
Determinethe level of Mehlich-I extractable Znin the filtrate by atomic absorp-

tion or inductively coupledplasmaemissionspectrometry.
Calculation
If the filtrate for a samplecontains0.5 mg Zn L -I, the soil contains2.0 mg
kg-I of Mehlich-I extractableZn, as shown below
(0.5 mg Zn L-1)(0.020L)
= 2.0 mg Zn kg-I.
0.005 kg
Hence,the numericalvalue for Mehlich-I extractable Znexpressedas milligrams

per kilogram of soil can be obtainedby multiplying the milligrams of the Zn per
liter in filtrate by four.
Comments
Amounts of Mehlich-I extractableZn in soils normally range from negli-
gible to 7.6 mg kg-I (Perkins, 1970; Cox & Wear, 1977). A regional study was
conductedin southeasternUSA to determinethe critical deficiency level for the
Mehlich-I Zn test for com production. In this research,0.8 mg Zn kg-I of
Mehlich-I extractableZn separatedsoils into·Zn-sufficientand Zn-deficientcat-
egories(Cox & Wear, 1977). This critical value is applicableto soils with CEC
levels <7.5 cmole kg-I.
Mehlich-III Method
The Mehlich-III method was developedto evaluate levels of available

macronutrientand micronutrient in soils of southeasternUSA (Mehlich, 1984).
Ammonium fluoride in the Mehlich-III extractionsolution provideda betteresti-
mateof P availability in near-neutraland alkaline soils than did the acid solution
for the Mehlich-I method.The chelatingagent,DTPA, causedinterferencein the
colorimetric determinationof P and, therefore,could not be a constituentof the
Mehlich-III extractionsolution. Hence,the chelatingagent,EDTA was included
in the solution for extractionof availableCu, Mn, and Zn.
Equipment
The amountof Mehlich-III extractableCu and Zn in solution is common-
ly measuredby atomic absorptionor inductively coupledplasmaemissionspec-
trometry.
Reagents
The Mehlich-III extractionsolution consistsof 0.2 M CH3COOH, and 0.25
M NH4N03, 0.015 M NH 4F, 0.013 M HN03, and 0.001 M EDTA. To prepare1
L of Mehlich-III extractionsolution, add 11.49 mL of concentratedCH3COOH
(17.4M), 20.0 g of NH4N03 , 0.56 g NH 4F, 0.84 mL of concentratedHN03 (15.5
714 REED & MARTENS
M), and 0.29 g EDTA to about800 mL of of deionizedwater, mix, and after dis-
solution dilute to 1 L with deionizedwater.
Copperand Zn standardswith concentrationsof 1000mg L-1 are available
from commercialsources.To preparea stock solution with 1000 mg Cu or Zn
L -1, weigh 1000mg of Cu or Zn metal, respectively,into a 250-mL conicalflask,
add 10 mL of deionized waterand 5 mL of concentratedHN03, warm gently to
completedissolution of Cu or Zn, and boil to expel oxides of N (Am. Public
Health Assoc., 1989). Then evaporateto almost dryness,transferto a 1000-mL
volumetric, and dilute to volume with Mehlich-III extractionsolution. Working
standardsshould be prepared by diluting the stock solution to bracket the
amountsof Mehlich-III extractableCu and Zn in the samples.The Mehlich-III
ing standards.
Procedure
Grind air-dried soil to passthrougha 2.0-mmstainlesssteelsievein prepa-
ration for the Mehlich-III extraction.Place2.5 cm3 of the sievedsoil in a 100-
mL extraction bottle, add 25 mL of the Mehlich-III extraction solution, and
shakethe mixture at 200 cyclesmin-1 on a reciprocal shakerwith a 4-cm stroke
length. After a 5-min shakingperiod, filter the suspensionthroughWhatmanno.
42 filter paper.Determinethe level of Mehlich-III extractableCu and Zn in the
filtrate by atomic absorptionor inductively coupledplasmaemissionspectrome-
try.
Calculation
If the filtrate for a samplecontains0.1 mg Cu or Zn L -1, the soil contains
1.0 mg dm-3 of Mehlich-III extractableCu or Zn, respectively,as shownbelow
0.1 mg metal L-1)(0.025 L) ) (1000 cm3)

( -'---=--2-.5-cm----'3'--'-----'- -3
dm3 = 1.0 mg metal dm .
Hence, the numerical valuefor Mehlich-III extractableCu or Zn expressedas

milligrams per cubic decimeterof soil can be obtainedby multiplying the mil-
ligrams of the metal perliter in filtrate by 10.
If the filtrate for a samplecontains0.1 mg Cu or Zn per liter and if the vol-
ume weight of soil is 1.25 g cm-3, the soil contains 0.8 mg of Mehlich-III
extractableCu or Zn kg-I, respectively,as shownbelow
( 0.1 mg metal L-1)(0.025 L») (1000 g) -1

- - = 0.8 mg metal kg .
3.125 g kg
Hence, the numerical value for Mehlich-III extractableCu or Zn expressedas

milligrams per kilogram of soil can be obtainedby multiplying the milligrams of
COPPER& ZINC 715
Mehlich-III extractablemetal per cubic decimeterby 0.8. The 0.8 factor is valid
for soil with a volume weight of 1.25 g cm-3•
Comments
The Mehlich-III methodis basedon a volume of soil ratherthan weight of
soil. Use of a volume of soil gives a constantsoil volume to soil extractingsolu-
tion ratio (Mehlich, 1972). Critical Mehlich-III extractablemetal levels usedfor
North Carolinasoils are 0.5 mg Cu and 1.0 mg Zn dm-3 (Mehlich, 1984).
Dilute HydrochloricAcid Method
The 0.1 M HCI extractableZn method has been used much longer than
other testsfor separatingsoils into adequateand inadequateavailable Zn cate-
gories. Use of the 0.1 M HCI methodto determinethe needfor Zn fertilization
of corn on slightly acid, sandy-texturedAlabamasoils was reportedby Wear and
Sommer (1948). This extractant currently is being used in the north central
region of the USA to evaluatethe available Zn statusof neutral and acid soils
(Whitney, 1988). The method does not provide a satisfactoryestimateof Zn
availability in calcareoussoils becausethe 0.1 M HCI dissolvesCaC03 with
releaseof occludedZn, which under normal conditions,is inaccessiblefor plant
uptake (Trierweiler & Lindsay, 1969; Lauer, 1971). Furthermore, variationsin
CaC03 contentsin sampleslead to dissimilar degreesof neutralizationof the 0.1
M HCl extractant solutions and, hence,to differencesin solubilizationof soil Zn
(Nelsonet aI., 1959).
The 0.1 M HCI method has undergonenumerousmodifications since its
first use by Wear and Sommer(1948). Thesemodificationsincludedchangesin
soil mass-to-extraction solution ratio, shakingvesseland time, and type of shak-
ing (Wear & Sommer,1948; Nelson et aI., 1959; Whitney, 1988). The currently
usedversionof the 0.1 M HCl method,which is usedonly for extractionof Zn,
will be discussedherein(Brown et aI., 1971; Whitney, 1988). A higher molarity
of HCI hasbeenusedas the extractionsolutionfor determinationof the available
Cu statusof organicsoils (Lucas, 1948; Whitney, 1988).
Equipment
The amountofO.1M HCI extractableZn in solution is commonlymeasured
by atomic absorptionor inductively coupledplasmaemissionspectrometry.
Reagents
To prepare1 L of 0.1 M HCI extractionsolution, add 8.33 mL of concen-
trated HCI (12 M) to about 800 mL of of deionizedwater dilute to 1 L with
deionizedwater, and mix.
Zinc stocksolutionswith concentrationsof 1000mg L -1 are availablefrom
commercialsources.To preparea stock solution with 1000 mg Zn L -1, weigh
1000mg of Zn metal into a 250-mL conical flask, add 10 mL of deionizedwater
716 REED & MARTENS
and5 mL of concentratedHN03, warm gently to completedissolutionof Zn, and

boil to expel oxidesof N (Am. Public Health Assoc., 1989). Then evaporateto
almostdryness,transferto a 1000-mLvolumetric, and dilute to volume with 0.1
M HCl. Working standardsshould be preparedby diluting the stock solution to
bracketthe amountof 0.1 M HCI extractableZn in the samples.The 0.1 M HCI
ing standards.
Procedure
Grind air-dried soil to passthrougha 2.0-mmstainlesssteelsievein prepa-
ration for the 0.1 M HCI extraction.Place4.25 cm3 (or 5 g) of the sieved soilin
a 50-mL Erlenmeyerflask, add 20 mL of the 0.1 M HCI extractionsolution, and
shakethe mixture on a reciprocal shakerat 180 cycles min-I. After a 30-min
shaking period, filter the suspensionsthrough Whatman no. 42 filter paper.
Determine the amount of 0.1 M HCI extractableZn in the filtrate by atomic
absorptionor inductively coupledplasma emissionspectrometry.
Calculation
If the filtrate for a samplecontains0.2 mg Zn L-I, the soil contains0.94
mg of 0.1 M HCI extractableZn per cubic decimeter,as shown below
0.2 mg Zn L-I)(0.020 L)} (1000 cm3 ) _

( -'----=--4.-2-5-cm~3---'- -3
dm3 - 0.94 mg Zn dm .
Hence, the numerical value for 0.1 M HCI extractableZn expressedas mil-
ligrams per cubic decimeterof soil can be obtained by multiplying the mil-
ligrams of Zn per liter in filtrate by 4.7.
The 4.25-cm3 volume is assumedto contain 5 g of samplefor soils of the
north central region of the USA, i.e., a volume weight of 1.18 g cm-3 (Peck,
1988). If the filtrate for a sample contains 0.2 mg Zn L-I and if the volume
weight of soil is 1.18 g cm-3, the soil contains 0.8 mg L-I of 0.1 M HCI
extractableZn, as shown below
( 0.2 mg Zn L-I)(0.020 L)} ( 1000 g ) = 0.8 mg Zn kg-I.

5.0 g kg
Hence, the numerical value for 0.1 M HCI extractableZn expressedas mil-
ligramsper kilogram of soil canbe obtainedby mUltiplying the milligrams of 0.1
M HCI extractableZn per cubic decimeterby 0.85. The 0.85 factor is valid for
soil with a volume weight of 1.18 g cm-3.
Comments
Researchhas shown that, for the above test, a 0.1 M HCI extractableZn
level of <2.0 mg kg-l indicatesthat Zn applicationis neededin com and sorghum
COPPER& ZINC 717
[Sorghum bicolor (L.) Moench.] production (Whitney, 1980). The calibration

datadiffers for modificationsof the 0.1 M HCI extractableZn procedureand for
soils with different chemicaland physicalproperties.For example,wheresouth-
easternsoils were shakenas above,exceptfor a 1:10 soil-to-extractionsolution
ratio and for 15 ratherthan 30 min, the critical level for the soil test for com pro-
duction was 0.5 mg Zn kg-1 (Cox & Wear, 1977). The southeasternsoils under
study had CEC levels <7.5 cmolc kg-I.
TOTAL COPPERAND ZINC
Total eu and Zn determinationsrequire solubilization of thesetrace ele-

ments from the solid phaseof soils. Solubilization of soil Cu and Zn requires
either completedestructionof the inorganic and organicsoil fractions or partial
destructionand then Cu and Zn extraction.Initial proceduresusedfor Cu and Zn
solubilizationby destructionof inorganic and organicsoil componentswere the
HF-HN03-HCI04-H2S04 digestion and Na2C03 fusion methods (Jackson,
1958). Later, Bernas(1968) developeda methodwherebyCu and Zn were dis-
solved during digestion of a soil samplewith an aqua regia-HF mixture under
pressureat a temperatureof 120°C. The sampleis placedin a Teflon container
surroundedby a metal support (Parr bomb) during the digestion. Sridhar and
Jackson(1974) used widemouth polypropylenecontainers(250-mL capacity)
rather than Parr bombs,for the aqua regia-HF digestion of soil samples.Either
polycarbonateor polysulfone may be more suitable, than polypropylenecon-
tainers,for use during the aquaregia-HFdigestion(Farrahet aI., 1980).
A H20z-HNOrHCI reflux method(USEPA Method 3050) was developed
to solubilize soil metals(USEPA, 1986). Simplicity and adaptabilityas a routine
procedurehaveled to the frequentuseof this methodfor extractionof Cu and Zn
from contaminatedsoils. There is incompletedissolution of soil Cu and Zn by
the USEPA reflux method(Bakhtaret aI., 1991) and, hence,the Cu and Zn con-
centrationsdeterminedby this procedureshould not be referred to as total soil
Cu or Zn.
The HF-HNOr HCI04-H2S04 digestionmethodof Cu and Zn solubiliza-
tion for total Cu and Zn analyseswill be discussedherein. Long-termsatisfacto-
ry resultsandsimplicity havecontributedto the commonusageof the HF-HNOr
HCl04-H2S04 digestionmethod.Precautionsmust be followed during the diges-
tion becauseHCI04 reactsviolently and explosively during oxidization of easi-
ly oxidizablecompounds(Assoc.Official Anal. Chern.,1970; Am. Public Health
Assoc.,1989; Sulcek& Povondra,1989). Furthermore,contactof HCI04 with a
dehydrating agent may result in formation of explosive anhydrous HCl04
(Assoc. Official Anal. Chern., 1970). The user should have thoroughfamiliarity
with safety precautionsfor eachacid prior to use of the digestion.
Hydrofluoric, Nitric, Perchloric, andSulfuric Acid Method
This methodsolubilizes Cu and Zn through destructionof inorganic and

organicsoil fractionsby action of the HF, HN03, HCI04, and H2S04 in the pres-
718 REED & MARTENS
enceof heat.The HF decomposessilicatesby reactionof F with Si to form SiF4,

which is volatile when heated in the presenceof strong acids. The HCI04
becomesa strongoxidizing agentin the presenceof heatand oxidizessoil organ-
ic matter. Prior to addition of HCI04, easily oxidizable organic in the sampleis
oxidizedby HN03 at an elevatedtemperature.This HN03 oxidationstepis need-
ed to preventan explosivereactionbetweenthe easily oxidizableorganic matter
and HCI04. Soil hydroxidesand oxides are decomposedby reactionwith H30+
from the acids in the digest. Much of the excess HF, HN03, and HCI04
volatilizes from the sampleas the temperatureof the digest exceedsthe boiling
points of thesethree acids but remainslower than the boiling point of H2S04,
SpecialApparatusor Equipment
1. Platinumcrucible, 30-mL capacityor more.
2. Perchloricacid hood.
The amountof total Cu and Zn in solutionis commonlymeasuredby atom-
ic absorptionor inductively coupledplasmaemissionspectrometry.Instrumental
adaptationsare requiredwhen HF is in the matrix used fortotal Cu and Zn deter-
minationsby inductively coupledplasmaemissionspectrometry.
Reagents
2. Nitric acid, 70%.
3. Perchloricacid, 70-72%.
4. Sulfuric acid, 95 to 97%.
Copper and Zn stock solutions with concentrationsof 1000 mg L-l are
or Zn L-l, weigh 1000 mg of Cu or Zn metal, respectively,into a 250-mL coni-
cal flask, add 10 mL of deionizedwater and 5 mL of concentratedHN03, warm
1000-mLvolumetric, and dilute to volume with deionizedwater.
Procedure
This total Cu and Zn method was adaptedfrom Lim and Jackson(1982)
and Reisenauer(1982). Grind air-dried soil to passthrougha 0.150-mmstainless
steel sieve in preparationfor the total Cu or Zn determination.Transfera 1.0-g
sampleof the finely groundsoil into a 30-mL platinumcrucible,wet the soil with
a few drops of deionizedwater, add 3 mL of HN03, and heat gently until effer-
vescenceceases.Add 2 mL of HCI04, 5 mL of HF, and 1 mL of H2S04, Heat the
soil-acid mixture at low heat (8~90°C) on a hot plate in a perchloricacid hood
until completeevolution of the dark brown fumes from reactionof HN03 with
organicmatter.Then washdown particlesfrom the crucible sideswith 1.0 mL of
HCI04, and heatfor an additional20 min. Placethe crucible in a sandbath,cover
about 90% with a platinum lid, and heat at a higher temperatureuntil the com-
plete evolution of white HCI04 fumes and the initial appearanceof H2S04
COPPER& ZINC 719
fumes. Allow the crucible to cool and add 5 mL of deionizedwater. If the sam-
ple does not dissolve completely, add 5 mL of HF and heat again until H2S04
fumes appear.When the residuecompletely dissolves,transfer the sampleto a
25-mL volumetric flask, dilute to volume with deionized water, and filter
through WhatmanNo. 42 filter paper.Working standardsare preparedby appli-
cation of the appropriateamountsof the stock solution to blanks. The same
digestion,dilution, and filtration proceduresshould be usedfor the samplesand
blanks. Amounts of total Cu and Zn in solution for samplesand standardsare
measuredby atomic absorptionor inductively coupled plasma emission spec-
troscopy.
Calculation
If a digestedsamplecontains0.2 Cu or Zn L- 1 and if 1.0 g of soil is used
in the digestion,the soil contains5.0 mg of total Cu or Zn kg-I, respectively,as
shown below
(0.2 mg metal L- 1)(0.025 L) 1

'------"'-----~-- = 5.0 mg metal kg-
0.001 kg
Hence,the numericalvalue for total Cu or Zn expressedas milligrams per kilo-

gram of soil can be obtainedby multiplying the milligrams of the metal per liter
in filtrate by 25.
Comments
A 250-mL teflon beakerwith a lid can substitutefor the platinum crucible
(Lim & Jackson,1982). The teflon beakeris heat resistantto 260°C and is inert
to all materials except molten metals. The procedureis the same with use of
either Teflon beakersor platinum crucibles.
Amounts of total Cu in uncontaminatedsoils usually are within the range
of 6 to 60 mg kg-1 with the highest amountsin ferralitic soils and the lowest
amountsin sandy-texturedand organicsoils (Kabata-Pendias & Pendias,1985).
Amounts of total Zn in uncontaminatedsoils usually are within the rangeof 17
to 160 mg kg-1 with the highestamountsin clayey-texturedsoils and the lowest
amountsin sandy-texturedand organic soils (Iyengar, 1981; Kabata-Pendias&
Pendias,1985).
REFERENCES
Alloway, B.l. 1990. Soil processesand behaviourof metals. p. 7-28. In B.1. Alloway (ed.) Heavy
metalsin soils. John Wiley & Sons,New York.
wastewater.17th ed. Am. Public Health Assoc.,Washington,DC.
Associationof Official Analytical Chemists. 1970. Official methodsof analysis. 11th ed. Assoc.
Official Anal. Chern.,Washington,DC.
Baker, D.E. 1990. Copper. p. 151-176.In B.1. Alloway (ed.) Heavy metalsin soils. John Wiley &
Sons,New York.
720 REED & MARTENS
Baker, D.E., and M.e. Amacher.1982. Nickel, copper,zinc, and cadmium.p. 323-336.In A.L. Page
et al. (ed.) Methods of soil analysis. Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,
Madison,WI.
Bakhtar,D., G.R Bradford, A.L. Page,and JA Frampton.1991. Comparisonof five soil digestion
methodsin determiningelementlevels in contaminatedsoils. p. 238. In Agronomy abstracts.
ASA, Madison,WI.
Barber,SA 1984. Soil nutrient bioavailability. John Wiley & Sons,New York.
Brown, A.L., J. Quick, and J.L. Eddings. 1971. A comparisonof analytical methodsfor soils zinc.
Corey, RB. 1990. Physical-chemicalaspectsof nutrient availability. p. 11-24.In RL. Westermanet
al. (ed.) Soil testingand plant analysis.3rd ed. SSSA,Madison,WI.
Council on Soil Testing and Plant Analysis. 1980. Handbookon referencemethodsfor soil testing.
1980. Univ. Georgia,Athens,GA.
Cox, F.R 1968.Developmentof yield responsepredictionand manganesetest interpretationfor soy-
beans.Agron. J. 60:521-524.
Cox, F.R, and J.1. Wear (ed.). 1977. Diagnosisand correctionof zinc problemsin com and rice pro-
duction. North CarolinaStateUniv. South. Coop. Ser. Bull. 222.
Eik, K., and R.H. Gelderman. 1988. Soil sample preparation. p. 2-4. In w.e. Dahnke (ed.)
Recommended chemicalsoil test proceduresfor the north central region. North DakotaAgric.
Exp. Stn. Bull. 499, rev.
Farrah,RF., S.A. Matthes, andAJ. Mackie. 1980.A simple low-costmethod fordissolutionof metal
and mineral samplesin plastic pressurevessels.Bur. Mines RI-8489. Bur. Mines, Pittsburgh,
PA.
Frye, w.w., H.F. Miller, L.w. Murdock, and D.E. Peaslee.1978. Zinc fertilization of com in
Kentucky. Kentucky Coop. Ext. Servo Agron. 11:1-4.
Gerwing, J., P. Fixen, and R. Gelderman.1982. Zinc rate and sourcestudies.South DakotaExp. Stn.
Prog. Rep. 13.
Hammes,J.K., and K.C. Berger. 1960.Chemicalextractionand crop removalof manganese from air-
dried and moist soils. Soil Sci. Soc. Am. Proc. 24:361-364.
Hazra,B.M., and L.N. MandaI. 1987. Distribution of zinc fractions and their transformationin sub-
mergedrice soils. Plant Soil 104:175-181.
Hodgson,J.F., H.R. Geering,and W.A. Norvell. 1965. Micronutrient cation complexesin soil solu-
tion: Partition betweencomplexedand uncomplexedforms by solvent extraction. Soil Sci.
Soc. Am. Proc. 29:665-669.
Hodgson,J.F., W.L. Lindsay, and J.F. Trierweiler. 1966. Micronutrient cation complexing in soil
solution: II. Complexingof zinc and copperin displacedsolution from calcareoussoils. Soil
Sci. Soc. Am. Proc. 30:723-726.
Iyengar, S.S., D.e. Martens,and w.P. Miller. 1981. Distribution and plant availability of soil zinc
fractions. Soil Sci. Soc. Am. J. 45:735-739.
Kabata-Pendias, A., and H. Pendias.1985.Traceelementsin soils and plants.CRC Press,Inc., Boca
Raton, FL.
Kataba-Pendias, A., and H. Pendias.1991.Traceelementsin soil and plants.2nd ed. CRC Press,Inc.,
Boca Raton, Fl.
Krauskopf, K.B. 1972. Geochemistryof micronutrients. p. 7-40. In lJ. Mortvedt et ai. (ed.)
Micronutrientsin agriculture.SSSA,Madison,WI.
Lauer, D.A. 1971.Evaluationof plant availableZn by the DTPA soil test, 0.1 M HCI extraction,and
labile Zn measurements.Ph.D. diss. Colorado State Univ., Fort Collins, (Diss. Abstr. 31-
6157B).
Legget,G.E., and D.P. Argyle. 1983. The DTPA-extractableiron, manganese,copper,and zinc from
neutral and calcareoussoils dried under different conditions. Soil Sci. Soc. Am. J.
47:518-522.
Liang, J., J.W.B. Stewart,and R.E. Karamanos.1991. Distribution and plant availability of soil cop-
per fractions in Saskatchewan. Can. J. Soil Sci. 71:89-99.
Lim, e.H., and M.L. Jackson.1982. Dissolution for total elementalanalysis.p. 1-12. In A.L. Page
Madison,WI.
Lindsay, w.L., and W.A. Norvell. 1978. Developmentof a DTPA micronutrientsoil test. p. 84. In
Agronomy abstracts.ASA, Madison,WI.
COPPER& ZINC 721
Lindsay,w.L., and W.A Norvell. 1978. Developmentof a DTPA test for zinc, iron, manganese, and
copper.Soil Sci. Soc. Am. J. 42:421-428.
Logan,T.1., and R.L. Chaney.1983. Utilization of municipal wastewaterand sludgeon land-met-
als. p. 235-323.In AL. Page(ed.) Utilization of municipal wastewaterand sludgeon land.
Univ. California, Riverside,CA
Lucas,R.E. 1948. Chemicaland physicalbehaviorof copperin organicsoils. Soil Sci. 66:119-129.
Martens, D.C., and w.L. Lindsay. 1990. Testing soils for copper, iron, manganese,and zinc. p.
229-264. In R.L. Westermanet al. (ed.) Soil testing and plant analysis. 3rd ed. SSSA,
Madison,WI.
Martens,D.C., and D.T. Westermann.1991.Fertilizer applicationsfor correctingmicronutrientdefi-
ciencies.p. 549-592.In J.J. Mortvedt et al. (ed.) Micronutrientsin agriculture.2nd ed. SSSA,
Madison,WI.
Mclaren, R.G., and D.V. Crawford. 1973. Studieson soil copper: I. The fractionationof copperin
soils. J. Soil Sci. 24:172-181.
Mehlich, A 1972. Uniformity of expressingsoil test results:A casefor calculatingresultson a vol-
ume basis.Commun.Soil Sci. PlantAnal. 3:471-424.
Mehlich, A 1984. Mehlich 3 soil test extractant:A modification of Mehlich 2 extractant.Commun.
Mehlich, A, and S.S.Bowling. 1975. Advancesin soil test methodsfor copperby atomicabsorption
spectrophotometry.Commun.Soil Sci. Plant Anal. 6:113-128.
Mengel, K., and E.A. Kirkby. 1987. Principles of plant nutrition. 4th ed. Int. Pot. Inst., Bern,
Switzerland.
Mortvedt, J.J. 1977. Micronutrient soil test correlationsand interpretations.p. 99-117.In T.R. Peck
et al. (ed.) Soil Testing: Correlatingand interpretingthe analytical results.ASA Spec.Publ.
29. ASA, CSSA, and SSSA,Madison,WI.
Neilsen, D., P.B. Hoyt, and A.E MacKenzie. 1986. Distribution of soil Zn fractions in British
Columbiainterior orchardsoils. Can. J. Soil Sci. 66:445-454.
Nelson,W.L., A Mehlich, and E. Winters. 1953.The development,evaluation,and use of soil tests
for phosphorusavailability. p. 153-188.In W.H. Pierreand AG. Norman(ed.) Soil and fer-
tilizer phosphorusin crop nutrition. Agron. Monogr. Acad. Press,New York.
Nelson, J.L., L.C. Boawn, and EG. Viets. 1959. A method for assessingzinc statusof soils using
acid-extractablezinc and "titratable alkalinity" values.Soil Sci. 88:275-283.
Peck,T.R. 1988.Standardsoil scoop.p. 4-5. In w.e. Dahnke(ed.) Recommended chemicalsoil test
proceduresfor the north central region. North DakotaAgric. Exp. Stn. Bull. 499, rev.
Perkins,H.E 1970.A rapid methodof evaluatingthe zinc statusof coastalplain soils. Commun.Soil
Sci. Plant Anal. 1:35-42.
Reisenauer,H.M. 1982.Chromium.337-346.In A.L. Pageet al. (ed.) Methodsof soil analysis.Part
2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Sabbe,W.E., and H.L. Breland.(ed.). 1974.Proceduresusedby soil testinglaboratoriesin the south-
ern region of the United States.South.Coop. Ser. Bull. 190.
Schnitzer,M. 1978. Humic substances:Chemistryand reactions.p. I-M. In M. Schnitzerand S.U.
Khan (ed.) Soil organicmatter.ElsevierSci. Publ. Co., New York.
Severson,R.e., J.M. McNeal, and J.1. Dickson. 1979. Effects of soil preparation on DTPA-
extractableelementsin soils of the northerngreatplains. Soil Sci. 128:70-79.
Sims,J.T., and G.V. Johnson.1991. Micronutrient soil tests.p. 427-476.In J.J. Mortvedt et al. (ed.)
Micronutrientsin agriculture.2nd ed. SSSA,Madison,WI.
Soltanpour,P.N. 1991. Determinationof nutrient availability and elemental toxicityby AB-DTPA
soil test and ICPS. Adv. Soil Sci. 16:165-190.
Soltanpour,P.N., J.B. Jones,Jr., and S.M. Workman. 1982.Optical emissionspectrometry.p. 29-65.
In AL. Pageet al. (ed.) Methodsof soil analysis.Part2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA,Madison,WI.
Soltanpour,P.N., A Khan, and w.L. Lindsay. 1976. Factorsaffecting DTPA-extractableZn, Fe, Mn,
and Cu from soils. Commun.Soil Sci. Plant Anal. 7:797-821.
Soltanpour,P.N., A. Khan, and AP. Schwab.1979. Effect of grinding variableson the NH4HCOr
DPTA soil test valuesfor Fe, Zn, Mn, Cu, P, and K. Commun.Soil Sci. Plant Anal. 10:903-
909.
Soltanpour,P.N., and A.P. Schwab.1977. A new soil test for simultaneousextractionof macro-and
micro-nutrientin alkaline soils. Commun.Soil Sci. Plant Anal. 8:195-207.
Soltanpour,P.N., and S.M. Workman. 1979. Modification of the N~HC03-DTPA soil test to omit
carbonblack. Commun.Soil Sci. Plant Anal. 10:1411-1420.
722 REED & MARTENS
Sorenson,R.C., D.D. Oelsligle,and D. Knudsen.1971.Extractionof Zu, Fe, and Mn from soils with
0.1 N hydrochloricacid as affectedby soil properties,solution:soilratio, and length of extrac-
tion period. Soil Sci. 111:352-359.
Sposito,O. 1981. The operationaldefinition of the zero point of chargein soils. Soil Sci. Soc. Am.
J. 45:292-297.
Sridhar, K., and M.L. Jackson.1974. Layer charge decrease by tetrahedralcation removal and sili-
con incorporationduring natural weatheringof phlogopite to saponite.Soil Sci. Soc. Am.
Proc. 38:847-S51.
Stout, P.R. 1956. Micronutrientsin crop vigor. J. Agric. Food Chern.4:1000-1006.
SuIcek,Z., and P. Povondra.1989. Methodsof decompositionin inorganicanalysis.CRC Press,Inc.,
Boca Raton,FL.
Tierney, e.E. 1981. The effect of the numberand the timing of foliar Mn applicationupon soybean
yield. M.S. thesis. Virginia Polytech. Inst. StateUniv., Blacksburg,VA
Trierweiler, J.F., and w.L. Lindsay. 1969. EDTA-ammonium carbonatesoil test for zinc. Soil Sci.
Soc. Am. Proc. 33:49-54.
Udo, E.1., H.L. Bohn, and T.C. Tucker. 1970 Zinc adsorptionby calcareoussoils. Soil Sci. Soc. Am.
Proc.34:405--407.
U.S. EnvironmentalProtectionAgency. 1986. Acid digestionof sediment,sludgeand soils. In Test
methodsfor evaluatingsoil wasteSW-846. USEPA, Cincinnati, OH.
Wear, J.I., and AL. Sommer. 1948. Acid-extractablezinc of soils in relation to the occurrenceof
zinc-deficiency symptoms in com: A method of analysis. Soil Sci. Soc. Am. Proc.
12:143-144.
Whitney, D.A 1980. Micronutrient soil tests-zinc,iron, manganeseand copper.p. 18-21.In w.e.
Dahnke(ed.) Recommended chemicalsoil test proceduresfor the north centralregion. North
DakotaAgric. Exp. Stn. Bull. 499.
Whitney, D.A. 1988. Micronutrientsoil testsfor zinc, iron, manganeseand copper.p. 20-22.In W.e.
Dahnke(ed.) Recommendedchemicalsoil test proceduresfor the north central region. North
DakotaAgnc. Exp. Stn. Bull. 499, rev.
Published 1996
Chapter 27
Molybdenum and Cobalt
J. L. SIMS, University of Kentucky, Lexington, Kentucky
The molybdenum(Mo) and cobalt (Co) portions of Chapter27 in (Kuboto &

Cary, 1982) the agronomy monographMethods of Soil Analysis (Page et aI.,
1982) have beenrevisedand updated.The readeris referredto both the above-
mentionedchapterand to Chapters74 (Reisenauer,1965) and 76 (Beesonet aI.,
1965) in Methods of Soil Analysis (Black et aI., 1965) for additional background
information and procedures.The reviews of the Mo literature of Jarrell et a1.
(1980) and of Guptaand Lipsett (1981) are excellent.
Analysesof plants and soils for Mo and Co are of interestfrom both ani-
mal and plant nutritionstandpoints.High Mo concentrations(10-20 mg Mo kg-t
tissue)in plants are responsiblefor Mo toxicity (Mo-inducedCu deficiency) in
ruminantanimals.The diseaseknown asmolybdenosisoccursmost often on high
pH soils typical in the westernUSA. Molybdenumis an essentialnutrient of ani-
mals but a deficiency has neverbeenobservedin cattle (Gupta& Lipsett, 1981).
Molybdenumalso is an essentialelementof plantsbut, in contrastto animals,Mo
deficienciesrather than Mo toxicities are more likely to occur in plants.
In addition to areascontainingalkaline soils, Mo toxicity in ruminantani-
mals is of concernin soils contaminatedwith Mo due to mining activities(Boon,
1984; Soltanpour, 1991) and lands receiving sewagesludgesfrom municipal
waste-watertreatmentplants (CAST, 1976; Jarrell et aI., 1980; Soon & Bates,
1985;Pierzynski& Jacobs,1986a).Dependingon the source,nature,and amount
of sludge applied, considerableMo can be added to soil. Jarrell et a1. (1980)
reportedthat the mean Mo concentrationin sewagesludgefrom 16 USA cities
was 15.1 mg kg-t and the rangewas 1.2 to 40 mg kg-to Similarly, the meanand
range of Mo concentrationof 15 small city sludgesin New York State were
reportedto be 11.0 and 1.7 to 29.2 mg kg-I, respectively(Mumma et aI., 1988).
Additional considerationsare the facts that (i) application of some sludgescan
increasesoil pH markedly (Pierzynski & Jacobs,1986b), thus increasingthe
availability of Mo to plants, and (ii) the plant availability of sludge-borneMo
may be higher than that of other heavy metalssuchas Cu and Co (Soon& Bates,
1985). Both situationscould causeincreasedMolCu ratios in foragesand result
in greateroccurrenceof molybdenosis.
The Mo contentof soils is dependenton the Mo contentof parentmaterial
(Masseyet aI., 1967). Total concentrationsin soil averagefrom 1 to 2 mg Mo
Book Seriesno. 5.
723
724 SIMS
kg-l but may rangeup to 30 mg kg-l. Kubota (1976) reporteda medianvalue of

6 mg Mo kg-l for severalsoils of the westernUSA but soils from the eastern
USA hada medianof only 0.5 mg kg-l. Molybdenumin the soil solutionat water
pH >5.0 existsprincipally asthe anion MoO~- and is greatly affectedby soil pH.
For eachunit increasein soil pH above pH 5.0, the soluble Mo concentration
increases100-fold (Lindsay, 1972). At high pH, Mo is thought to be associated
with Ca and as suchis readily availableto plants. Hydrousoxidesof Fe and Al
provide positively chargedabsorptionsitesfor Mo in acidic soils (Reisenaueret
aI., 1962).Soils high in iron oxidesmay renderaddedMo in fertilizer unavailable
underacidic conditions(Sims et aI., 1975).The amountof Mo adsorbedalso has
beenshownto be closely relatedto soil organicmattercontent(Karimian & Cox,
1978).
The total accumulationof Mo by plantsis lower than for any other miner-
al nutrient and in plants Mo servesprimarily as cofactor for nitrogenaseand
nitrate reductaseenzymes(Marschner,1986). Hence,Mo is requiredfor both N2
fixation in biological systemsand for N03 reduction in nonlegumes.The Mo
requirementby plantsvaries widely amongcrop speciesand sufficiency levels
for plant tissuesaregenerallytwo to threetimeshigherfor legumes(>0.5 mg Mo
kg-l) than for nonlegumes(0.1--0.4 mg Mo kg-l) (Haweset al., 1976; Sims &
Atkinson, 1976; Gupta & Lipsett, 1981). Plant absorptionof Mo is greatly en-
hancedby solubleP andto a lesserextentby Cl- and NO)", but is greatly lowered
by SO~- (Stout et aI., 1951; Sims et aI., 1979; Gupta & Lipsett, 1981; Eivazi et
aI., 1983).After measuringa greaterrateof Mo uptakeat plant Mo concentrations
above1 mg Mo kg-l thanbelow this concentration,Eivazi andco-workers(1984)
proposedthat two mechanismsof Mo absorptionexist, dependenton Mo level in
the root zone. At low levels of Mo, Mo is absorbedprimarily by metabolically
active means;at high Mo levels in the rooting media,Mo is absorbedby passive
means.Work of Jablonski et a1. (1985) support this contention. Kannan and
Ramani (1978) had previously noted that metabolic inhibitors suppressedMo
uptakeby roots,suggestingthe presenceof a metabolicmechanism.Positivecrop
responseto addedMo on deficientsoils hasbeenwidely reportedin the literature
(Gupta& Lipsett, 1981).
Ruminantanimals have a requirementfor Co and problemswith lack of
appetite,poor growth, and poor reproductionin cattle and sheepattributableto
Co deficiency have beenrecognizedin parts of the easternUSA since colonial
times (Kubota & Allaway, 1972). Theseproblemsoccur when foragesdeficient
in Co are fed but can be greatly alleviatedtoday due to the easeof providing Co
to animalsin salt licks or animal supplements.The legumeroot nodulebacteria
also have a requirementfor Co (Ahmed & Evans, 1960; Lowe & Evans,1962)
and legumegrowth responsesto Co havebeenrecordedin Australia (Gladstones
et aI., 1977).Accordingto Dilworth et a1. (1979),Co deficiencyleadsto impaired
protein synthesis, depressedcell division, and reduced leghemoglobin in
Rhizobium, and ultimately to reducedN2 fixation in legumes.
Soil contentsof Co vary widely but range from 0.5 to 3 mg Co kg-l in
sandysoils to 20 to 30 mg kg-l and higherin clay soils (Beesonet aI., 1965).Soil
Co appearsto be associatedclosely with soil oxides, particularly reducibleMn
oxides (McLaren et aI., 1986; Jarvis, 1984). This associationwith Mn or Fe
MOLYBDENUM & COBALT 725
oxides (Kubota, 1965) would be consistentwith increasedplant availableCo in

water logged soils (Adams & Honeysett,1964). Forageplants from areasof the
USA having low soil Co had Co concentrationsrangingfrom 0.04 to 0.07 mg Co
kg-1 (Kubota & Cary, 1982). Jarvis(1984) reportedlevels of 0.11 mg Co kg-1 in
dry forage as being adequatefor ruminant animals. Becauseof the many indus-
trial usesof Co and the application of waste water and sewagesludge to land,
considerableconcernexists that soil levels of Co may becomeelevated(Purves,
1977; Pinkerton& Brown, 1985; Soon & Bates,1985).
GENERAL PRINCIPLES
The filtrates obtainedfrom carbonatefusion of soils (Lim & Jackson,1982,

p. 9-11) may be usedfor the analytical determinationsof total soil Mo and Co.
Generally,0.5 to 1 g of well-mixed soil ground to passa 250 meshsieve (250-
mmz openings)is ignited to 900°C in a platinum crucible, first without NazC03
and then with NazC03. The silica is removedand the residueis taken up in water
and conc HCI. Aliquots of the HCl solutionscan then be transferreddirectly to
separatoryfunnels for complexing and solvent extraction.Becauseof the low
concentrationsof Mo and Co in soil, both are concentratedby solventextraction
to facilitate analyticaldeterminationand to reduceinterfering elements.
A note of caution: the solvent used to separatelyextract and concentrate
both Mo and Co is CCl4 (carbontetrachloride).Although usedwidely in labora-
tories in the past, CCl4 can causepoisoningin humansby inhalation, ingestion,
or skin absorptionand could be a carcinogen (Budavariet aI., 1989). Hencethe
chemicalshould be usedwith care e.g., use under a hood with adequateventila-
tion while wearingglovesand a nasalmask.
The basic equipmentfor complexing and solvent extraction are Squibb-
type separatoryfunnels with Teflon stopcocks,a laboratory shaker,and shaker
racks to hold the separatoryfunnels. Descriptionsof shakerracks designedto
increasethe efficiency of routine analyseshave beengiven previously by Doran
and Martens (1970) and Kubota and Cary (1982). The rotary rack describedby
Doran and Martens(1970) appearsparticularly attractive in that it will hold 20,
60-mL funnels that can fit undera hood and be shakensimultaneously.
All glasswaremust be thoroughly cleaned,rinsed with hydrochloric acid
(HCl), and rinsedagain with distilled water. Double deionizedor deionizedand
distilled water shouldbe usedthroughoutall procedures.
Working standardsolutions canbe preparedfrom commercially available
Mo and Co solutionsor preparedusing saltsof Mo and Co.
TOTAL MOLYBDENUM
Principles
The thiocyanate-stannous chloride (SnClz • 2H20) methodhas been used

extensivelyin the pastto determinetotal Mo in soils. The methodpresentedhere
is that of Johnsonand Arkley (1954) with modifications. In the procedure,Mo
726 SIMS
reactswith thiocynateand excessFe, in the presenceof SnCl2 • 2H20, to form a

coloredcomplex.The complexis extractedfrom the aqueousphasewith isoamyl
alcohol dissolvedin CC4. The amountof Mo present isdeterminedby compari-
son of the absorbanceof the samplewith appropriatestandards.
Method
SpecialApparatus
1. Separatoryfunnels,60-mL Squibb-typewith shortenedtips and marked
at 45 mL.
2. Rack for separatoryfunnels.
Reagents
1. Hydrochloric acid, reagentgrade.
2. Ferric chloride hexahydrate(FeCI3 • 6H20) (requiredfor standardsolu-
tions, plant samples,and reagentblanks, but not for soils): Add 0.5 g
FeCl3 • 6H20 to 560 mL concentratedHCl and dilute to 1 L with dis-
tilled water.
3. Isoamyl alcohol-CC~: Mix equalvolumesof American ChemicalSo-
ciety reagentgradeCC4 and reagentgradeisoamyl alcohol. The specif-
ic gravity of the resulting mixture is 1.19. Another note of caution: the
vapor from this solution is toxic (follow precautionsand suggestions
under"GeneralPrinciples."
4. Ammonium thiocyanate(NH4SCN): Dissolve40 g N~SCN in distilled
water and make to 100 mL.
5. Stannouschloride: Suspend40 g SnCl2 • 2H20 in 20 mL concentrated
HCI. Add distilled waterto dissolve,while heating(keepbelow boiling),
andmaketo 100mL. Filter the solutionif turbid, andpreparefresh daily.
6. Standard Mo solution: Dissolve 0.150 g of molybdenum trioxide
(M003) in 10 mL of 0.1 M sodium hydroxide (NaOH), make slightly
acid with HCl, then maketo 1 L with distilled water. Use 100 mL of this
solution and dilute to 1 L for a working standardhaving 10 mg Mo L-l.
Alternatively, a working standardhaving 10 mg Mo L-l can be prepared
from a certified reference.To preparecalibrationstandards,dilute 0, 0.5,
1.0,2.0,4.0,and 6.0 mL of the 10 mg Mo L-l working standardto 250
mL with 1.5 M HCI. To establisha calibrationcurve,25-mL ali-quotsof
the calibrationstandardscontaininga totalof 0, 0.5, 1.0, 2.0, 4.0, and 6.0
J..lg Mo are carriedthrough the extractionprocess.
Procedure
Quantitativelytransferthe solution of an unknown soil samplefollowing
carbonatefusion (or standardaliquot) to a separatory funnelusing small amounts
of distilled water. Add in sequencelO-mL ferric chloride solution (to blanks,
standards,and plantssamplesonly), water to dilute to 45 mL, and 3-mL organic

solventmixture to saturatethe sample.After eachaddition,shakefor 2 min, then
allow to stand 10 min. To settle small droplets of extractantsuspendedat the
organic-aqueous interface,swirl the contentsof the funnel, placesharply into the
rack, and allow 10 min for phasesto separate.Discardthe organicsolventphase
locatedin the lower part of the funnel. Invert the funnel and dry the tip and stem
of the funnel with a pipe cleaner.Add 1 mL NH4SCN solution, 1 mL SnClz •
2HzO solution, 2 mL organic solvent(add 3-5 mL if Mo contentis greaterthan
2 mg kg-I) and shake2 min after eachaddition. Allow 15 min for phasesto sep-
arate.Discardtwo dropsof the organicphaselocatedin the lower part of the fun-
nel, then transfer the remainder into a spectrophotometercell. Measure the
absorbanceof the solution at 470 nm using water as a reference.Read results
from the standardcurve preparedby running the standardsthroughthe same pro-
cedureand plotting absorbance(y axis) vs. Mo concentration(x axis).
Comments
Molybdenumin soil andplant extractshasbeendeterminedsuccessfullyby

other colorimetric methods (Purvis & Peterson,1956; Kubota & Cary, 1982;
Gupta& MacKay, 1965). However,colorimetric methods,including the one out-
lined above,are tediousand time consuming.Recently,advantagehasbeentaken
of advancesin atomic absorption,plasmaemission,and automatedinstrumenta-
tion to measureMo that offers great improvementsin easeand numberof analy-
ses. Unfortunately,few of the reportedmethodshave beentestedadequatelyon
soil extractsand many suffer one or more weaknesses, particularly when the Mo
concentrationof the material to be measuredis less than 1 mg Mo kg-I. Hence,
the methodand instrumentof choice may dependgreatly on the Mo concentra-
tion of the extracts to be analyzedand the intended use of the data obtained.
Plants deficient in Mo are likely to have concentrationsless than 0.5 mg kg- 1
whereasplants growing on contaminatedsoils, or thoseproducingforagestoxic
to animals,have much higher concentrations.
For extractslow in Mo, graphite furnace atomic absorptionspectroscopy
(GFAAS) as proposedby Henning and Jackson(1973) and modified by Wilson
(1979) and Curtis and Grusovin(1985) may be useful. The limitations of matrix
interferenceand nonatomicabsorptionpresentin the earlier method appearto
have been alleviated in the Wilson (1979) and Curtis & Grusovin (1985) meth-
ods. The catalyticeffect of Mo on the Kl (potassiumiodide)-HzOz (hydrogenper-
oxide) reaction(Quin & Woods, 1979; Eivazi et aI., 1982; Sims & Crutch-field,
1992) hasbeenusedwith successfor Mo determinationin plant materials(and to
a limited extent for soils; seeLiu, 1995) in the author'slaboratory.The sensitiv-
ity of the automatedprocedureis <0.1 mg kg- l in plant materialand 20 determi-
nationsper hour of digestedsamplescan be made.However,maximumsensitiv-
ity of the methodis obtainedin 0.125 M HCl solutionswhich is much lower than
the 1.0 to 1.5 M HCl of the final solution commonly found for soil digests(and
necessaryfor thiocynate-Mocomplex development) for total Mo. This methodis
betteradaptedto anion resin extractsand other extractantsusedfor availableMo
than for total Mo (see"Availability Indexesfor Molybdenum").
728 SIMS
For extractshigh in Mo, atomic absorptionor Ar plasmaemissionspec-

troscopy,eitherdirect currentor inductively coupled,has beenused(Khan et aI.,
1979; Soltanpouret aI., 1982; Soltanpour,1991; Pierzynskiet aI., 1986).The list-
ed detectionlimits for atomic absorption(about 0.03 mg L-1) and interferences
from other elementsdecreaseits usefulnessfor determiningMo. Similarly, en-
hancementof Mo readingsat the low concentrationscommonly found in plants
and soils has limited the usefulnessof plasmaemission.Pierzynskiet a1. (1986)
suggestedadding Ca and Mg to the Mo standardsat concentrationsexpectedin
the samplesolutionsto alleviate the enhancementproblem while Woodis et aI.,
(1980) used K addition during analysis for Mo in fertilizers. Becauseof the
potential usefulnessof plasmaemissioninstrumentsto analyzefor Mo, further
researchin this areais warranted.
AVAILABILITY INDEXES FOR MOLYBDENUM
Principles
A good test for availablesoil nutrientsshould extractamountsof nutrients

that correlateclosely to plant uptakeof the samenutrients.Additionally, the test
shouldbe rapid, reproducible,and accurate.Meeting thesecriteria for soil Mo is
more difficult than for most nutrients since Mo is presentin soil and plants in
extremely small quantities and is affected greatly by soil propertiesand seed
reserves (Gupta & Lipsett, 1981). Among differing typesof extractantsproposed
for available Mo and usedwith varying successin the past include ammonium
oxalate, pH 3.3 (Grigg, 1953), hot water (Lowe & Massey, 1965), anion-
exchangeresin (Dowex 1-X4, J.T. Baker Chem.Co., Phillipsburg,NJ) (Bhella &
Dawson,1972), and ammoniumbicarbonate-diethylenetriamine-pentaacetic acid
(AB-DTPA) (Soltanpouret aI., 1982; Boon & Soltanpour,1983).
Generally,the amountof Mo extractedby ammoniumoxalateis many fold
greaterthan amountsby the other methodsand less well correlatedto plant up-
take of Mo. Reports in the literature showing significant correlationsbetween
ammoniumoxalate(pH 3.3) soil Mo and Mo uptake by plants at some(but not
all) locationsinclude Haley and Melsted(1957), Andersonand Mortvedt (1982),
Lombin (1985), and Wang et a1. (1994). In contrast,acid ammoniumoxalatesoil
Mo was not related to plant Mo in a numberof reports(Lowe & Massey,1965;
Pathaket aI., 1969; Little & Kerridge, 1978; Karimian & Cox, 1979; Cox, 1987;
Burmesteret aI., 1988). In many of the latter studies,plant Mo was more closely
relatedto somesoil property other than Mo, such as soil pH or somefraction of
soil Fe.
Recently,Liu (1995) found significantcorrelations(r =0.81**) for soil Mo
extractedwith a solution of ammoniumoxalate(pH 6.0) in a group of Kentucky
soils and Mo uptake by tobacco(Nicotiana tabacum L.) growing in the green-
house.Similar correlationsfor the strongly bufferedextractantof pH 3.3 were not
statisticallysignificant. The improvedcorrelationsfor extractsof higher pH like-
ly occurredbecause(i) the pH 6.0 extractantsolution was poorly buffered and
becamebuffered by soil during extraction,and (ii) the pH of the soil-extractant
mixture during extraction was closer to the pH of soil solutions in which the
plantswere grown. Also, the pH 6.0 extractsand filtrates are cleanerand much
lower in extractableFe that may interferewith analytical determinationsof Mo.
Theseresultsfor onestudyshow promisefor acid soils andwarrantfurther inves-
tigation.
Anion exchangeresinshavebeenusedwith successto extractMo in a num-
ber of studies (Bhella & Dawson, 1972; Dawson & Bhella, 1972; Jarrell &
Dawson,1978; Ritchie, 1988). Becauseof theselatter studies,we will outline a
modified anionexchangeresinmethodfor useon acid soils. The methodis essen-
tially that of Jarrell and Dawson (1978) with modifications from Jacksonand
Meglen (1975). We alsowill outline the methodof Soltanpouret al. (1982) (also
seeSoltanpour,1991) for use on alkaline and contaminatedsoils. This method
hasnot beenwidely testedbut hasproducedMo valuesin soil that were correlat-
ed well with plant uptakeof Mo (Boon, 1984; Pierzynski& Jacobs,1986a;Wang
et ai., 1994). Furthermore,the extractantextractsmultiple nutrientswhich canbe
measuredsimultaneouslywith plasma emission (ICP), and hence offers the
potentialfor measuringMo over wide geographicalareasduring routine analysis
of soils.
Method
SpecialApparatus
1. Reciprocatingshaker,top loading.
2. Sieve,60 mesh(0.25 mm), brass.
3. Hot plate.
4. Peristalticpump.
Reagents
1. AG1-X4 anion exchangeresin!, CI saturated.
2. Sodiumchloride (NaCl), reagentgrade.
3. Sodiumhydroxide,reagentgrade.
4. Sodiumnitrate (NaN03), reagentgrade.
5. Hydrogenperoxide,30% reagentgrade.
6. Sulfuric acid (H 2S04), reagentgrade.
Procedure
AG1-X4 anion exchangeresin (chloride saturated)is used to extract Mo
from soil suspensions.The resinshouldbe preconditionedto avoid decreasingthe
pH of soil suspensions.Alternately rinse the resinwith 2 M NaOH and2 M NaCl
solutions,followed by distilled water.After seiving,the resinfraction> 0.25-mm
(60-mesh)size is storedin moist form until used.Grind 10 g air-dried soil to pass
a 0.25-mm(60-mesh)sieveand placein a 2.0- (Ld.) by 20-cm glasstube. Add 5
g AG l-X4 resin and 20 mL double-distilled water to each tube. Stopperand
1 Bio-Rad Laboratories,Melville, NY 11747.

730 SIMS
shakethe soil-resinmixture for 18 h at 25°C and separatethe resin from the soil
by washingwith distilled water on a 0.25-mm (60-mesh)brassscreen.Transfer
the resin to a glassfunnel with a 1.5-cm (o.d.) stem insertedin a lO-mL dispos-
able pipet tip. Prior to addition of resin, glasswool shouldbe placednearthe tip
of eachpipet. Connectthe tip of the pipet to a peristalticpump with small tubing
and add30 mL 2 M NaN03 to the funnel. Allow the resin and NaN03 to equili-
brate45 min prior to displacingthe NaN03 solution and Mo from the resin with
the pump over an additional 45-min period. Evaporatethe extractantNaN03
solution from a 50-mL Erlenmeyerflask to drynesson a hot plate at low heat
(80°C) to avoid loss of sampleby splattering.Mter cooling, add 2 mL of 30%
H20 2, 8 mL of 2 M HCl, and two glassbeadsto eachflask. Heat, uncovered,on
a hot plate at 80°C until dry. If necessary,repeatthe H20 2 portion of the organic
matter destructionstep to renderthe residuecolorless.Dissolve the residuein 2
M HCl and dilute to 25 mL. Analyze for Mo by the thiocynateprocedure(see
"Procedure" under "Total Molybdenum"). For optimum developmentof the
molybdenum-thiocynatecomplex, the fmal acid concentrationof the solution
shouldbe about 1 M.
Comments
Modifications made in the procedureof Jarrell & Dawson(1978) include

useof a reciprocatingshaker,useof a funnel and larger diameterleachingtubes,
a peristalticpump, and NaN03 to replaceMo from the resin. Severaltubescon-
tainedin styrofoamor othershakingrack canbe shakenon their sidesat onetime.
The funnel and tubeswith a slightly largerdiameterpermit easiertransferof resin
from shakingtube to leachingtube and the peristaltic pump ensuresmeasured,
more even dispensingof the NaN03 extraction solution of multiple samples.
BecauseLombin (1985) reportedhigh Cl- concententrationtendedto depress and
sometimesenhancethe Mo signal in the carbonrod flamelesstechnique,NaN03
was usedin placeof NaCl. The resin extractsaboveshouldlend themselveswell
to analysisof Mo by methodsotherthan colorimetricwhen the Mo concentration
falls within the detectionlimits of the instrument(see"Comments"sectionunder
"Total Molybdenum"above).
Method
SpecialApparatus
2. Inductively coupledplasmainstrument.
Reagents
acid.
3. Ammonium hydroxide(NH40H).
4. Nitric acid (HN03).
5. Ammonium bicarbonate-diethylenetriaminepenataacetic
acid solution.
A 1 M NH4HCOT O.OO5 M DTPA solution may be madeby adding1.97
g of DTPA to 800 mL of distilled-deionizedwater (DDW). To aid dis-

solution and to preventeffervescence,add approximately2 mL of 1:1
NH40H. After most of the DTPA is dissolved,add 79.06g of NH4HC03
and stir gently until dissolved.Adjust the pH to 7.6 with either HCI or
NH 40H and dilute the solution to 1.0 L with DDW. The pH of the
extractingsolution may changeunlessstoredunderabout3 cm of min-
eral oil. However,use of a fresh solution is preferable.
6. Standards:The standardsmust be made in the AB-DTPA extracting
solution. Becauseof the linearity of ICP standardcurves,only two stan-
dardsusually are usedto standardizean element.The low standardfor
Mo containsonly AB-DTPA. To preparethe high standard,take 1 mL of
a certified referencestandardcontaining1 mg Mo and make to 500 mL
with AB-DTPA solution.
Procedure
To 10 g of 2-mm soil in a 125-mL conicalflask, add 20 mL of AB-DTPA
solution. Shakeon a reciprocalshakerfor 15 min at 180 cyclesmin-1 with flasks
kept open. Use Whatmanno. 42 filter paperor its equivalentto filter the extracts.
Place0.25 mL concentratedHN03 in a 1O-mL beakerand carefully add 2.5 mL
AB-DTPA extract or standardsolution to the beaker.Mix on a rotary shakerfor
15 min to eliminatecarbonatespecies.DetermineMo on an ICP spectrometer.
Comments
Nebulizers,devicesusedfor the aspirationof the sampleinto the plasma,
often clog when high-salt solutions are aspirated.The HN03 step above is de-
signedto eliminate the problem. The HN03 treatmentmay be eliminated if the
ICP spectrometeris equippedwith a high-salt nebulizer(Soltanpour,1991).
The AB-DTPA-ICP test may be usedfor multiple elementanalysis,in addi-
tion to Mo. Becauseof the close relationshipbetweensoil pH and availableMo,
inclusion of soil pH and Mo soil test valuesin multiple regressionequationsmost
often improves thepredictionof Mo uptakeby plantsand interpretationof results
(Pierzynski& Jacobs,1986a).Similar findings for pH havebeennotedprevious-
ly in conjunction with other soil Mo measurements(Lowe & Massey, 1965;
Bhella & Dawson,1972; Sims & Atkinson, 1974; Mortvedt & Anderson,1982).
TOTAL COBALT
Principles
The basic procedureis that of Kubota and Cary (1982). Cobalt in solution
is complexedwith dithizone in CCI4, the CCl4 phaseis drained off, CCL4 is
volatilized, and the chelateis destroyedby oxidation. After taking to dryness,the
residueis dissolvedin dilute acid, and the Co is determinedby atomic absorption
spectrophotometry.Pleaseuse caution: CCL4 can causepoisoning, follow sug-
gestionsunder"GeneralPrinciples."
732 SIMS
Method
SpecialApparatus
1. Racksfor separatoryfunnels.
3. Atomic absorptionspectrophotometer.
4. Separatoryfunnel, 4 L.
Reagents
1. Hydrochloric acid, reagentgrade:Dilute as needed.
2. Ammonium hydroxide,reagentgrade:Dilute as needed.
3. Carbontetrachloride,reagentgrade.
4. Chloroform (CHCI3), reagentgrade.
5. Phenolphthalein:Dissolve1 g of phenolphthaleinin 100 mL of reagent-
grade,95% ethanol.
6. Dithizone (diphenylthiocarbazone): Dissolve0.2 g of dithizone in 75 to
100 mL of CHCI3• Filter into a 4-L separatoryfunnel containing2.5 to
3.0 L of 0.02 M NH40H, and extract the dithizone into the aqueous
phaseby shakingthe funnel. Discardthe CHCl3 phase.Shakethe aque-
ous solution with 50-mL portionsof reagent-gradeCCI4, discardingthe
CCl4 phasein eachcase.Repeatthe washingwith CCl4 until the sepa-
rated CCl4 phaseis a pure green color. Add 1 L of CCI4, and acidify
slightly the aqueousphasewith 6 M HCI. (The desiredpH is indicated
by a light orangecolor that will changeto blue-blackwith the addition
of two to three drops of 6 M HCI; avoid excessacidity since this will
causethe dithizone to precipitatefrom the aqueousphase.)Extract the
dithizone into the CCl4 phaseby shakingthe funnel. Discard the aque-
ous phase.Store the dithizone in a cool, dark place,preferablya refrig-
erator.
7. Ammonium citrate solution, 40%: Dissolve BOO g of citric acid in 600
mL of distilled water, and while stirring, slowly add 900 mL of concen-
trated NH40H. The reaction is exothermic,and care must be taken to
preventsplattering.Adjust the pH to B.5, if necessary.Dilute the solu-
tion with water to a volume of 2 L, and extract it with 25-mL portions
of dithizone in CCl4 until the aqueousphasestaysorangeand the CCl4
phaseremainspredominantlygreen.Then extractthe solution with CCl4
until all the orangecolor is removed.
B. Cobalt standards,stock standard,0.1 g of Co per liter: Heat cobaltous
sulfateheptahydrate(CoS04 • 7H20) in an oven at 250 to 300°Cto con-
stantweight (~h). Dissolve0.2630g of the CoS04 in 50 mL of redis-
tilled waterand 1 mL of concentratedreagent-grade H2S04. Transferthe
sblution to a 1-L volumetricflask, anddilute it to volumewith redistilled
water. Prepareworking standardswith 0.5, 1.0, 1.5,2.0,and 4mg Co per
liter by diluting stock standardwith deionizedwater.
Procedure
Dithizone Extraction. Transferto a 125-mL separatoryfunnel an aliquot
of the soil extractcontaining0.5 to 1.5 /J.g of Co. Add 10 mL of 40% ammonium
citrate for each25-mL aliquot or lessof the soil solution. Add one drop of phe-
nolphthalein,and adjustthe pH to 8.5 with 7 M NH40H. If a precipitateforms at
this stage,addadditionalammoniumcitrate.Add 10 mL of dithizonein CC4, and
shake the mixture for 5 min. Draw off the CCl4 phaseinto a 50-mL beaker.
Repeatthe extractionas many times as necessary,using 5-mL quantitiesof dithi-
zone solution and shaking the mixture for 5 min each time. (The extraction is
completewhen the aqueous phase remainsorangeand the CC4 phaseremains
predominantlygreen.)Then add 10 mL of CC4, shakethe mixture for 5 min, and
draw off the CCl4 phaseinto the samebeaker.The final 10 mL of CCl4 shouldbe
pure green. If not, the extraction with dithizone was incomplete and must be
repeated.Add 0.5 mL of perchloricacid (HCI04) to the combinedCC4 extracts,
and digestthe contentuntil it is colorless.Cool. Transferthe solution to a 15-mL
graduatedcylinder (flat-bottom with ground-glassstopper)and make to 5 mL
with deionizedwater.
To avoid using corrosiveHCI04 in the atomic absorptionspectrophotome-
ter, continueheatingandevaporatethe combinedCC4 solutionsto dryness,cool,
then dissolvethe residuein 5 mL 1 M HCI.
Atomic AbsorptionAnalysis
Consult operationsmanual for instrumentalsettings.With the 4 mg L- 1
working standard,adjustthe burnerheight and flow rate for optimum instrumen-
tal setting.Detectionlimit for practicalpurposesis about0.5 mg mL-1 of Co, but
somelower concentrationscan be determinedwhen the concentrationmode and
scaleexpansionare adjusted.
Blanksand Standards
Each determinationrequires an appropriateset of standardsand reagent
blank, including a proportionatevolume of filtrate from carbonatefusion. Zero
the atomic absorptionspectrophotometer using the zero standardCo with added
acid (equivalentto amountin the unknowns),and obtain readingsfor the other
Co standards.
Comments
The determinationof Co in most U.S. soils should be possibleby atomic

absorptionfollowing the procedureoutlined. Cobalt may not be detectablein
soils having as little as 0.2 mg kg-I Co and the useof GFAAS, if available,may
be necessary.Otherwise,increasethe initial samplesize to raisethe Co present in
the final solution.
The o-nitrosocresolmethodrecommendedearlier (Beesonet aI., 1965) is
appreciablymore sensitive(0.01 ± O.Olllg of Co), but its use has beenlimited,
becausethe method is exacting and tedious and requires a larger number of
reagents.
Solutionsremainingafter Co is determinedcan be usedto determinecon-
centrationsof Cu, Zn, and Pb, all of which are presentin appreciablylarger
amountsand are quantitativelyextractedby the dithizoneprocedure.
734 SIMS
AVAILABILITY INDEX FOR COBALT
Becauseattemptsto measureplant available Co in soils have had limited

success,an availability index will not be outlined here.Alban and Kubota (1960)
modified the original aceticacid Co procedureby Mitchell et al. (1957) by com-
bining dithizone with acetic acid. Alban and Kubota found one relationship
betweenacetic acid-dithizoneextractablesoil Co and plant content for well-
drainedsoils and anotherfor poorly drainedsoils of the southeasternU.S. Black
gum (Nyssa sylvatica Marsh var. biflora), a Co accumulatorplant, was usedas
the test plant. McLaren et al. (1986) reportedthe Co extractedwith acetic acid,
ethylenediaminetetraacetic acid (EDTA), pyrophosphate,or hydroxylamineto be
derivedfrom easily reducibleMn oxidesandconcludednoneof the reagentswere
likely to give good estimatesof the quantity or intensity factors of soil Co supply
to plants, as defined by isotopic exchangedeterminations.The amountsof Co
extractedby aceticacid and EDTA were greatly affectedby the temperatureand
length of extractionperiod. Jarvis (1984) concludedthat significant proportions
of the Co in soil was associatedwith Mn oxidesand would show the samesensi-
tivity to changesin acidity and redox potential as Mn.
ACKNOWLEDGMENTS
The investigationreportedin this chapter(no. 91-3-231) is in connection

with a project of the Kentucky Agricultural ExperimentStation and is published
with approvalof the Director.
REFERENCES
Adams, S.N., and J.L. Honeysett.1964. Someeffects of soil waterloggingon the cobalt and copper
statusof pastureplantsgrown in pots. Aust. J. Agric. Res. 15:357-367.
Ahmed, S., and HJ. Evans. 1960. Cobalt: A micronutrientelementfor the growth of soybeanplants
undersymbiotic conditions.Soil Sci. 90:205-210.
Alban, AL., and J. Kubota. 1960.A study of extractableCo in soils of the southeasternUnited States.
Anderson,O.E., and J.1. Mortevedt. 1982. Soybeans:Diagnosisand correction of manganeseand
molybdenumproblems.Univ. GeorgiaSouthernCoop. Ser. Bull. 281.
Beeson,K.C., J. Kubota, and V.A Lazar. 1965. Cobalt. p. 1064-1076.In C.A Black et al. (ed.)
Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Bhella, H.S., and M.D. Dawson.1972.The useof anion exchangeresin fordeterminingavailablesoil
molybdenum.Soil Sci. Soc. Am. Proc. 36:177-178.
Black, C.A, D.D. Evans,J.L. White, L.E. Ensminger,and F.E. Clark. 1965. Methodsof soil analysis.
Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Boon, D. Y. 1984. The ammoniumbicarbonate-DTPAsoil test (AB-DTPA) for determinationof plant
availablePb, Cd, Ni, and Mo in mine tailings and contaminatedsoils. Proc. 6th High Altitude
RevegetationWorkshop,ColoradoStateUniv. Inform. Ser. 53.
Boon, D.Y., and P.N. Soltanpour.1983.The ammoniumbicarbonate-DTPAsoil test for determination
of plant availablelead,cadmium,and molybdenumin mine tailings and contaminatedsoils. p.
29. In Agronomy abstracts.ASA, Madison,WI.
Budavari,S., M.J. O'Neil, A Smith, and P.E. Heckelman.1989.The Merck Index: An encyclopedia
of chemicals,drugs,and biological materials.11th ed. Merck and Co., Rahway,NJ.
Burmester,C.H., J.E Adams,and J.w. Odom. 1988. Responseof soybeansto lime and molybdenum
on ultisols in northernAlabama.Soil Sci. Soc. Am. J. 52:1391-1394.
Council for Agricultural Scienceand Technology.1976. Application of sewagesludge to cropland:

Appraisalof potential hazardsof the heavy metalsto plants and animals.Rep. no. 64. CAST,
Ames, IA.
Cox, ER. 1987. Micronutrient soil tests:Correlationand calibration. p. 97-117.In 1.R. Brown (ed.)
Soil testing: Sampling,correlation,calibration,and interpretation.SSSA, Madison,WI.
Curtis, P.R., and1. Grusovin. 1985. Determinationof molybdenumin plant tissueby graphitefurnace
atomic absorption spectrophotometry (GFAAS). Commun.Soil Sci. Plant Anal.
16:1279-1291.
Dawson,M.D., and H.S. Bhella. 1972. Subterraneanclover yield and nutrient contentas influenced
by molybdenumstatus.Agron. J. 64:308-311.
Dilworth, MJ., A.D. Robson,and D.L. Chatel. 1979. Cobalt and nitrogenfixation in Lupinus angus-
lifolius L. II. Nodule formationsand functions. New Phytol. 83:63-79.
Doran,1.W., and D.C. Martens.1970. Rotary separatoryfunnel rack and shakerfor determinationof
molybdenumin plants and soils. J. Assoc. Off. Anal. Chern. 53:228-234.
Eivazi, E, J.L. Sims, M. Casey,G.D. Johnson,and J.E. Leggett. 1983. Growth and molybdenumcon-
centrationsof burley tobaccoas influencedby potassium,molybdenum,and chloride in trans-
plant fertilizer solutions.Can. J. Plant Sci. 63:531-538.
Eivazi, E, J.L. Sims, and J.D. Crutchfield. 1982. Determinationof molybdenumin plant materials
using a rapid, automatedmethod.Commun.Soil Sci. Plant Anal. 13:135-150.
Eivazi, E, lL. Sims, and J.E. Leggett. 1984. Phosphorusand molybdenuminteractioneffectsduring
accumulationof molybdenumby burley tobacco.J. Plant Nutr. 7:1075-1092.
Gladstones,J.S.,J.E Loneragan,and N.A. Goodchild. 1977. Field responsesto cobalt and molybde-
num by different legume specieswith interferenceson the role of cobalt in legume growth.
Aust. J. Agric. Res. 28:619-628.
Grigg, J.L. 1953. Determinationof availablemolybdenumof soils. N.Z.J. Sci. Technol. 34:405-414.
Gupta,U.C., andJ. Lipsett. 1981. Molybdenumin soils, plants,and animals.Adv. Agron. 34:73-115.
Gupta, U.c., and D.C. MacKay. 1965. Determinationof Mo in plant materialsusing 4-methyl 1, 2
dimercaptobenzene (dithiol). Soil Sci. 99:414-415.
Haley, L.E., and S.w. Melstad. 1957. Preliminary studiesof molybdenumin Illinois soils. Soil Sci.
Soc. Am. Proc. 21:316-319.
Hawes,R.L., J.L. Sims, and K.L. Wells. 1976. Molybdenumconcentrationof certain crop speciesas
influencedby previousapplicationsof molybdenumfertilizer. Agron. J. 68:217-218.
Henning, S., and T.L. Jackson.1973. Determinationof molybdenumin plant tissue using flameless
atomic absorption.At. Absorpt. Newsl. 12:100-101.
Jablonski,P.P.,J.E. Leggett,and J.L. Sims. 1985.The absorptionof molybdenumby excisedtobacco
(Nicotiana tabacum L. roots). J. Plant Nutr. 8:277-287.
Jackson,D.R., and R.R. Meglen. 1975. A procedurefor extraction of molybdenumfrom soil with
anion exchangeresin. Soil Sci. Soc. Am. Proc. 39:373-374.
Jarrell, W.M., and M.D. Dawson. 1978. Sorptionand availability of molybdenumin soils of Western
Oregon.Soil Sci. Soc. Am. J. 42:412-415.
Jarrell, W.M., A.L. Page,and A.A. Elseewi. 1980. Molybdenum in the environment.ResidueRev.
74:1-43.
Jarvis, S.C. 1984. The associationof cobalt with easily reducible manganesein someacidic perma-
nent grasslandsoils. J. Soil Sci. 35:431-438.
Johnson,C.M., and T.H. Arkley. 1954. Determinationof molybdenumin plant tissue. Anal. Chern.
26:572-574.
Kannan,S., and S. Ramani. 1978. Studieson molybdenumabsorptionand transportin beanand rice.
Plant Physiol. 62:179-181.
Karimian, N., and ER. Cox. 1978. Adsorption and extractability of molybdenumin relation to some
chemicalpropertiesof soil. Soil Sci. Soc. Am. J. 43:757-761.
Karimian, N., and ER. Cox. 1979. Molybdenumavailability as predictedfrom selectedsoil chemical
properties.Agron. J. 71:63-65.
Khan, S.U., R.O. Cloutier, and M. Hidiroglou. 1979. J. Assoc. off. Anal. Chern. 62:1062-1064.
Kubota, J. 1965. Distribution of total and extractableforms of cobalt in morphologically different
soils of easternUnited States.Soil Sci. 99:166-174.
Kubota, J. 1976. Molybdenum statusof United Statessoils and plants. p. 555-581.In W. Chappell
and K. Peterson(ed.) The geochemistry,cycling and industrial usesof molybdenum,Vol. 2.
Marcel Dekker, Inc., New York.
Kubota, J., and W.H. Allaway. 1972. Geographicdistribution of traceelementproblems.p. 525-554.
In 1.1. Mortvedt et al. (ed.) Micronutrientsin agriculture.SSSA,Madison,WI.
736 SIMS
Kubota,1., and E.E. Cary. 1982. Cobalt, molybdenum,and selenium.p. 485-500.In AL. Pageet al.
(ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA, Madison,
WI.
Lim, C.H., and M.L. Jackson.1982. Dissolutionfor total elementalanalysis.p. 1-12. In AL. Pageet
al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,
WI.
Lindsay,w.L. 1972. Inorganicphaseequilibria of micronutrientsin soil. p. 41-57.In J.J. Mortvedt et
al. (ed.) Micronutrientsin agriculture.SSSA,Madison,WI.
Little, I.P., and P.C. Kerridge. 1978. A laboratory assessmentof the molybdenum status of nine
Queenslandsoils. Soil Sci. 125:102-106.
Liu, D. 1995. An index for plant available molybdenum in Kentucky soils. M.S. thesis. Univ.
Kentucky, Lexington, KY.
Lombin, G. 1985. Micronutrient soil testsfor the semi-aridSavannahof Nigeria: Boron and molyb-
denum.Soil Sci. Plant Nutr. 31:1-1l.
Lowe, R.H., and HJ. Evans. 1962. Cobalt requirementfor the growth of rhizobia. J. Bacteriol.
83:210-21l.
Lowe, R.H., and H.F. Massey.1965. Hot water extraction for available soil molybdenum.Soil Sci.
100:238-243.
Marschner,H. 1986. Mineral nutrition of higher plants.Acad. Press,New York.
Massey,H.F., R.H. Lowe, and H.H. Bailey. 1967. Relation of extractablemolybdenumto soil series
and parentrock in Kentucky. Soil Sci. Soc. Am. Proc. 31:200-202.
McLaren, R.G., D.M. Lawson, and R.S. Swift. 1986. The forms of cobalt in some Scottishsoils as
determinedby extractionand isotopic exchange.J. Soil Sci. 37:223-234.
Mitchell, R.L., J.W.S. Reith, and I.M. Johnston.1957. Trace elementuptake in relation to soil con-
tent. J. Sci. Food Agric. 8:51-59.
Mortvedt, lJ., and O.E. Anderson.1982. Foragelegumes:Diagnosisand correctionof molybdenum
and manganeseproblems.Univ. GeorgiaSouthernCoop. Ser. Bull. 278.
Mumma, R.O., K.A. Rashid, D.C. Raupach,B.S. Shane,1.M. Scarlet-Kranz,C.A Bache, W.H.
Gutenmann,and DJ. Lisk. 1988. Mutagens,toxicants, and other constituentsin small city
sludgesin New York State.Arch. Environ. Contamin.Toxicol. 17:657-663.
Page,A.L., R.H. Miller, and D.R. Keeney. 1982. Methods of soil analysis. Part 2. 2nd ed. Agron.
Pathak, AN., H. Shankar. and R.Y. Misra. 1969. Correlation of available molybdenum values
obtained by different methods to molybdenum uptake by alfalfa. J. Ind. Soc. Soil Sci.
17:151-153.
Pierzynski,G.M., S.R. Crouch,and L.W. Jacobs.1986. Use of direct-currentplasmaspectrometryfor
the determinationof molybdenumin plant tissuedigestsand soil extracts.Commun.Soil Sci.
Plant Anal. 17:419-428.
Pierzynski,G.M., and L.w. Jacobs.1986a. Extractability and plant availability of molybdenumfrom
inorganicand sewagesludgesources.J. Environ. Qual. 15:323-326.
Pierzynski,G.M., and L.W. Jacobs.1986b.Molybdenumaccumulationby com and soybeansfrom a
molybdenum-richsewagesludge.J. Environ. Qual. 15:394-398.
Pinkerton,B.W., and K.w. Brown. 1985. Plant accumulationand soil sorptionof cobalt from cobalt-
amendedsoils. Agron. 1. 77:634-638.
Purves,D. 1977.Trace-elementcontaminationof the environment.Fundamentalaspectsof pollution
control and environmentscience:I. Elsevier Sci.Publ. Co., New York.
Purvis, E.R., and N.K. Peterson.1956. Methodsof soil and plant analysesfor molybdenum.Soil Sci.
81:223-228.
Quin, B.F., and P.H. Woods. 1979.Automated catalytic method for the routine determinationof
molybdenumin plant materials.Analyst 104:552-559.
Reisenauer,H.M. 1965.Molybdenum.p. 1050-1058.In C.A Black et al. (ed.) Methodsof soil analy-
sis. Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Reisenauer,H.M., AA. Tabikh, and P.R. Stout. 1962. Molybdenum reactionswith soils and the
hydrousoxidesof iron, aluminum,and titanium. Soil Sci. Soc. Am. Proc. 26:23-27.
Ritchie, G.S.P. 1988. A preliminary evaluation of resin extractable molybdenum as a soil test.
Sims,J.L., andW.O. Atkinson. 1974. Soil andplant factorsinfluencing accumulationof dry matterin
burley tobaccogrowing in soil madeacid by fertilizer. Agron. J. 66:775-778.
Sims, J.L., and W.O. Atkinson. 1976. Lime, molybdenum,and nitrogen sourceeffects on yield and
selectedchemicalcomponentsof burley tobacco.Tob. Sci. 20:181-184.
Sims, J.L., w.o. Atkinson, and Chirtchart Smitobol. 1975. Mo and N effects on growth, yield, and
Mo compositionof burley tobacco.Agron. J. 67:824-828.
Sims, J.L., and J.D. Crutchfield. 1992. Determinationof molybdenumin plants. p. 61...Q6. In e.0.
Plank (ed.) Plant analysisreferenceproceduresfor the southernregion of the United States.
Univ. GeorgiaSouthernCoop. Ser. Bull. 368.
Sims, J.L., J.E. Leggett, and U.R. Pal. 1979. Molybdenumand sulfur interactioneffects on growth,
yield, and selectedchemicalconstituentsof burley tobacco.Agron. J. 71:75-78.
Soltanpour,P.N. 1991.Determinationof nutrientavailability and elementaltoxicity by AB-DTPA soil
test and ICPS. p. 165-190.In B.A. Stewart(ed.) Advancesin soil science.Vol. 16. Springer-
Verlag, Inc., New York.
Soltanpour,P.N., J.B. Jones,Jr., and S.M. Workman. 1982. Optical emissionspectrometry.p. 245.
In A.L. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and
SSSA,Madison,WI.
Soon, Y.K., and T.E. Bates. 1985. Molybdenum, cobalt, and boron uptake from sewage-sludge
amendedsoils. Can. J. Soil Sci. 65:507-517.
Stout, P.R., W.R. Meagher,G.A. Pearson,and C.M. Johnson.1951. Molybdenum nutrition of crop
plants.I. The influenceof sulfate and phosphateon the absorptionof Mo from soils and solu-
tion cultures.Plant Soil 3:51-87.
Wang, L., K.J. Reddy. and L.e. Munn. 1994. Comparisonof ammoniumbicarbonate-DTPA,ammo-
nium carbonate,and ammonium oxalate to assessthe availability of molybdenum in mine
spoils and soils. Commun.Soil Sci. Plant Anal. 25:523-536.
Wilson, D.O. 1979.Determinationof molybdenumin wet-asheddigestsof plant materialusingflame-
lessatomic absorption.Soil Sci. Plant Anal. 10:1319-1330.
Woodis, T.C., Jr., J.H. Holmes,Jr., J.D. Ardis, and F.J. Johnson.1980. Argon plasmaemissionspec-
trometric determinationof molybdenum in mixed fertilizers. J. Assoc. Off. Anal. Chern.
63:1245-1247.
Published 1996
Chapter 28
Nickel, Cadmium, and Lead
MICHAEL C. AMACHER, USDA-FS, Logan, Utah
Backgroundlevels of Ni, Cd, and Pb in soils are generally quite low (Adriano,
1986)exceptin areaswherethe parentmaterialhashigh levels of theseelements.
Increasedlevels of theseelementsin soils also can occur as a result of air pollu-
tion and land disposalof industrial and municipal wastes(Adriano, 1986).
Nickel, Cd, and Pb havebeengroupedtogetherin this chapterbecause they
are not essentialfor plant growth and development,are regarded primarilyaspol-
lutant metals,and may reachpotentiallytoxic levelsin soils. New digestionmeth-
ods havebeendevelopedsincethe previousedition and the sectionson extraction
methodshave beenexpandedbecauseof the continuing interestin their use.
SAMPLING AND SAMPLE PREPARATION

For details onthe variousapproachesto soil sampling,refer to Petersenand
Calvin (1986). Seealso Petersenand Calvin (1996, Chapter1) of this edition for
problemsassociatedwith soil samplingand storage.Generally,soil samplesare
air dried and groundto passa 2-mm sievebefore analysis.Air drying may affect
somesoil propertiesand alter the resultsof someof the selectiveextractionmeth-
ods(Bartlett & James,1980; Rapin et aI., 1986).If this is the case,it may be more
appropriateto analyze moist samples,which may be stored by refrigeration at
less than 4°C. Resultsfrom extractionand analysisof moist samplesare typical-
ly reportedon a dry-weight basis.The water contentof the samplecan be deter-
mined gravimetrically (Gardner,1986). For samplesthat are to be analyzedfor
total Ni, Cd, and Pb, oven drying the samplesat 105°C is appropriate.It should
be specifiedwhetherthe samplewas moist, air dried, or oven dried when report-
ing the resultsof analysis.
Generally, the less than 2-mm size fraction of a soil sample is analyzed.
Digestion methodsare more effective and precisionis improved if smallerparti-
cle sizes are used. Samplescan be ground to passsmaller size fraction sieves
(e.g., 0.5, 0.25, 0.125 mm, etc.) if necessaryto improve digestionefficiency and
precision.In general,as samplesizedecreases,particle size also must decreaseto
achieve reproducible results. For sample sizes of 200 mg or less, the sample
should be ground to passa 0.125-mmsieve.
SegoeRd., Madison,WI 53711,USA. Methods of Soil Analysis: Chemical Methods, Part 3-SSSA
Book Seriesno. 5.
739
740 AMACHER
Sometimesthe analystwill want to determinethe amountsof metalsasso-

ciated with specific size fractions (e.g., coarsesand, medium sand, fine sand,
very fine sand,coarsesilt, fine silt, coarseclay, fme clay; Horowitz, 1991). In
suchcases,particle-sizeseparationswill needto be done prior to any digestions
or extractions(Kunze & Dixon, 1986; Gee & Bauder,1986).
Grinding and sievingcan potentially introducemetalcontaminantsinto the
sample, which will give erroneouslyhigh results when analyzed.The use of
grinders with ceramicor other nonmetalgrinding parts and plastic sieveswill
eliminatethis sourceof contamination.
TOTAL NICKEL, CADMIUM, AND LEAD
Principles
Although methodsexist for direct determinationof the Ni, Cd, and Pb con-
tent of soils such as x-ray fluorescencespectrometry(XRF) and solid sample
introduction in atomic absorption(AAS) and inductively coupledplasmaatom-
ic emissionspectrometry(ICPAES) (Jacksonet al., 1991),most analyticaldeter-
minations require sample dissolution prior to analysis. Sulcek and Povondra
(1989) havepresentedmany of the methodsfor inorganicsampledecomposition
including fusion and acid digestions.Seealso Hossner(1996, Chapter3) of this
publicationfor dissolutionmethodsfor total elementalanalysis.
Lithium metaboratefusion is commonly used, but this method is more
applicableto major elementsin soils. Becauseof the small sampleamountsused
in this method (typically0.1--0.2g), analyticalsensitivity for trace elementscan
be a problemat low levels in the presenceof large concentrationsof major ele-
mentssuch as Si, AI, and Fe.
Various combinations of acids including hydroflouric acid (HF),
hydrochloricacid (HCI), nitric acid (HN03), perchloricacid (HCI04) and sulfu-
ric acid (H2S04) and digestiontechniquesincluding openbeakerson hot plates,
digestion tubes in block digesters,and digestion bombs in conventionaland
microwaveovens have beenused to dissolve sampleswith varying degreesof
success(e.g., Baker & Amacher,1982; Burau, 1982; Bettinelli et aI., 1987; Hee
& Boyle, 1988).
Microwave digestiontechniquesare amongthe newestmethodsfor digest-
ing samplesof soils, sediments,and other geologic materials(Lamothe et aI.,
1986; Kingston & Jassie,1988; Bettinelli et aI., 1989; Millward & Kluckner,
1989; Hewitt & Reynolds,1990).Specialdigestionbombsand microwaveovens
are now commerciallyavailable(e.g., CEM Corp., Matthews,NC). Advantages
of microwavedigestionin sealedcontainersinclude less time for samplediges-
tion (min vs. h or d in some cases),avoidanceof contaminationproblems
becausethe digestionis done in a closedcontainer,more completedigestionfor
somesamplesbecauseof higherpressures,and avoidanceof sampleloss and ele-
ment volatilization in somecases.With pressurerelief valvesand properdiges-
tion bomb capping techniques,safety is maximized and sample loss is mini-
mized. The National Institute of Standardsand Technology(NIST) in conjunc-
NICKEL, CADMIUM & LEAD 741
tion with the U.S. EnvironmentalProtectionAgency (USEPA) has developed

and evaluateda microwavedigestionmethod(USEPA SW-846, Method 3051)
that shows great promise for routine analysis of soils (Rasberry,1991). This
methodmay becomethe methodof choicein time.
More traditional digestionmethodsusing beakersand hot platesor diges-
tion tubesand blocks continueto be widely used.Three digestionmethodsare
presentedhere including Method 1-USEPASW-846, Method 3050 [digestion
with HN03, hydrogenperoxide (H20 2), and HCl in a conical beakeron a hot
plate; USEPA, 1986); Method 2--digestion with HN03, HCI04 and HCl in
digestiontubesin a block digester(Burau, 1982); and Method3--digestionwith
H20 2, HF, HN03, HCI04, and HCl in Teflon beakerson a hot plate (Baker &
Amacher, 1982). Thesemethodshave the advantagesof relatively inexpensive
and simple equipmentneedsand have been extensively evaluated.Disadvan-
tagesinclude somewhatlengthy digestiontimes in somecases,increasedpossi-
bility of samplecontaminationand loss, and potential explosion hazardswhen
HCl04 is used.
United StatesEnvironmentalProtectionAgency SW-846, Method 3050
will not dissolve silicate mineralsso that the total amountsof metals in a soil
sample will not be extractedwith this method. The digestion method does
destroy organic matter by oxidation with H20 2 and probably dissolvesmost
metal oxides and other mineral phasesexceptsilicatesso that near total extrac-
tion can be achievedin many cases.The methodmay be applicableto extraction
of near total amountsof metals from anthropogenicsourcesin soils, and thus
may be suitablefor routine assessment of metal contaminationof soils.
Method 2, which usesa block digester,is essentiallythe digestionpart of
the methodgiven by Burau (1982). A HCI04 fume hood with wash-downcapa-
bility mustbe usedto avoid possibleexplosionhazardsfrom HCl04• Heatingthe
sampleabove210°C must be avoidedto minimize the HCl04 explosionhazard.
Small funnels insertedin the open tops of the digestiontubesproducea reflux-
ing action during digestion.This methodand the use of a block digestergive a
more precisecontrol over heating and evaporationof acids. Most explosions
occurwith sampleswherethe easily oxidizableorganicmatteris not completely
oxidized or that have a high fat content. Prolongedrefluxing in HN03 will
destroythe easily oxidizableorganicmatter.Samplesalso may be dry ashedin a
muffle oven to destroythe organicmatterbeforedigestion(Page& Ganje,1970).
Optimal dry ashingtimes and temperaturesmay have to be determinedfor dif-
ferent typesof samples,but typically temperaturesrangefrom 450 to 550°C and
times rangefrom 2 h to overnight. Method 2 gives a more completedigestion
than Method 1, but resistantsilicatesalso are not destroyedby this method.Near
total recoveryof metalsis possiblein most cases.
Method 3 is similar to the digestionmethodgiven by Baker and Amacher
(1982), but H2S04 hasbeeneliminatedfrom the digestionprocedurebecauseof
possiblelow recoveryof Pb from precipitationof PbS04 and potential interfer-
encesin the subsequentanalysisfor Pb (Burau, 1982).Method3 destroyssilicate
mineralsbecauseHF is includedin the digestionsequenceandprovidesthe most
completesampledissolution and metal recovery. Care must be taken to avoid
samplecontaminationor loss,which is more likely with this methodbecausethe
742 AMACHER
digestion is done in open beakers.A sand bath can provide for more uniform
heatingof the Teflon beakers.
To minimize contaminationproblems in each of the methods outlined
below, all glass and plastic laboratory ware must be acid washed. Overnight
soakingin a dilute HN03 bath followed by rinsing with deionizedwater is ade-
quate.The useof rubberor metalobjectsin any of the proceduresmustbe avoid-
ed. Reagentgradeacids are satisfactoryin most cases,but in caseswhere trace
levels of metals are being determinedwith ultrasensitiveanalytical methods,
acids that arespecifiedfor trace metal analysisshouldbe used.Ultrapure grade
acids are rarely necessaryexceptfor the most demandingapplications.Reagent
grade(1Ype I) watershouldbe used inall procedures(Am. Public HealthAssoc.,
Am. Water Works Assoc.,and Water Pollut. Control Fed., 1992).
Although each of the methodsoutlined below specifiesa sampleweight
and final solution volume, both the sample weight and final volumes can be
increasedor decreasedas needed.The final resultsin milligrams per kilogram of
soil are equal to the solution concentrationsin milligrams per liter multiplied by
the solution to soil ratio. Inductively coupled plasma atomic emission spec-
trophotometry,inductively coupledplasmamassspectrometry(ICPMS), or AAS
are the preferredmethodsfor determiningNi, Cd, and Pb in the final solutions
(seesectionon "Analytical Determinationof Nickel, Cadmium,and Lead").
The reliability of theseor any digestion proceduresis best evaluatedby
analysis of standardreferencematerials. Geostandards Newsletter is a major
sourceof informationon geologicalreferencematerialsandcompilationsof data.
Spiking sampleswith known amountsof elementsis useful for evaluatingpoten-
tial interferencesduring analytical determinations,but is less useful for evaluat-
ing elementrecoveriesfrom digestionprocedures.Added elementsoften areonly
weakly adsorbedto the soil matrix and are not in the form of the elementsin the
original sample.
United StatesEnvironmentalProtectionAgency SW-846,Method3050:

Digestionwith Nitric Acid and HydrochloricAcid
in a Conical Beakeron a Hot Plate
Equipment
1. 125-mL conical beakers.
2. Watch glassesor small glassfunnels for refluxing.
3. Hot plate.
4. Quantitativeacid-washedfilter paper(Whatmanno. 41 or equivalent),
filter funnels, and racks.
5. 50-mL volumetric flasks.
Reagents
1. HN03, concentratedand 1:1 (voVvol conc. HNOYH20).
2. H20 2, 30%.
3. HCI, concentratedand 1:100 (voVvol concentratedHCVH20).
Procedure
1. Add 1.000g of preparedsoil sampleto a 125-mL conical beaker.
2. Add 10 mL of 1:1 HN03, mix the slurry, andcoverwith a watch glass.
3. Heat on a hot plate to 95°C and reflux for 10 to 15 min without boil-
ing.
4. Cool, add 5 mL of concentratedHN03, coverwith a watch glass,and
reflux for 30 min. Repeatthe addition of concentratedHN03 and
refluxing.
5. Evaporatethe solution to 5 mL without boiling. Do not allow the solu-
tion to go to dryness.
6. Cool, add 2 mL of deionizedwater and 3 mL of 30% H20 2, and cover
with a watch glass.
7. Heat until reactionwith H20 2 subsidesand then cool.
8. Continueto add 30% H 20 2 in I-mL aliquots and heat until reaction
with H20 2 is minimal or sampleappearsunchanged.Do not add more
than 10 mL of 30% H20 2 •
9. Add 5 mL of concentratedHCI and 10 mL of deionized water, and
cover with a watch glass.Reflux for 15 min.
10. Cool andfilter throughquantitative,acid-washedfilter paperinto a 50-
mL volumetric flask. Rinse the conical beakerand filter paperwith
small volumes of 1:100 HCI, dilute to volume, and mix. Retain the
solution for analysisof Ni, Cd, and Pb by one of the analytical meth-
ods of sectionon "Analytical Determinationof Nickel, Cadmium,and
Lead".
Comments
Small glassfunnels placedin the necksof the conical beakershave been
found to be a satisfactorysubstitutefor watch glassesfor refluxing the acidsdur-
ing the digestionsteps.
Digestionwith Nitric Acid and PerchloricAcid in a Block Digester
Equipment
1. Block digester [Tecator DS-40 (Tecator AB, Hoganas,Sweden) or
equivalent]or hot plate and heatingblocks. Aluminum heatingblocks
may be machinedfrom aluminum stock into ll-cm diam. cylinders.
Four cavities,3.7 em in diameterare drilled into the top of eachcylin-
der for the glassdigestiontubes.Two block heightsare required.The
short block is 7.5 cm high, and the test tube cavities are drilled to a
5.5-emdepth.The tall block is 15 cm high, and the cavitiesare drilled
to a 13-cm depth.A thermometerwell is drilled in the centerof the
blocks to the samedepth as the test tube cavities.
2. Digestion tubesor test tubes(3.5-cm o.d., 20 cm in length).
3. Small funnels (35-mm Ld.) for refluxing.
744 AMACHER
4. Quantitativeacid-washedfilter paper(Whitman no. 40 or equivalent),

filter funnels,and racks.
5. Fifty-milliliter volumetric flasks.
Reagents
1. HN03, concentrated.
2. HCI04, 70%.
3. HCI, 1:10 and 1:100 (vol/vol concentratedHCl/H20).
Procedure
1. Add 1.000g of preparedsoil sampleto a digestiontube.
2. Add 10 mL of concentratedHN03, placea funnel in the mouth of the
tube,andheatat 80 to 90°C overnight.Use the shortblock for this step
if the hot plate and AI heatingblock systemis used.
3. Heat at 125 to 130°C to dryness.Use the tall block for this step if hot
plate and AI heatingblock systyemis used.
4. Cool and add1 mL of concentratedHN03 and 4 mL of HCl04.
5. Heat at 205°C until fumes of HCI04 appear.Continueheatingto near
dryness.Do not exceed210°C becausethe potential for explosionof
HCI04 will increase.
6. Cool, add 10 mL of 1:10 HCI, heatto 70°C for 1 h.
7. Filter samplethrough quantitativeacid-washedfilter paperinto a 50-
mL volumetric flask. Rinse the digestion tube and filter paper with
small volumes of 1:100 HCI, dilute to volume, and mix. Retain the
solution for analysisof Ni, Cd, and Pb by one of the analyticalmeth-
odsof sectionon "Analytical Determinationof Nickel, Cadmium,and
Lead."
Comments
A HCI04 fume hood with wash-down capability is essential for this
method.Sampleswith high levels of organicmattermust be dry ashedin a muf-
of from 450 to 550°Cand
fle ovenprior to the digestion procedure. Temperatures
times of from 2 h to overnightmay be useddependingon the degreeof treatment
neededto destroythe organicmatter.
Digestionwith HydrogenPeroxide,Hydroflouric Acid, Nitric Acid,

and PerchloricAcid in Teflon Beakerson a Hot Plate
Equipment
1. 150-mL Teflon beakers.
2. Hot plate.
3. Quantitative acid-washed filter paper(Whatmanno. 40 or equivalent),
filter funnels, and racks.
Reagents
2. Hydroflouric acid, 48%.
3. Nitric acid, concentrated(vol/vol concentratedHNO:fH20).
5. Hydrochloric acid, 1:10 and 1:100(vol/vol concentratedHCl!H 20).
Procedure
1. Add 1.000g of preparedsoil sampleto a Teflon beaker.
2. Add 10 mL of 30% H20 2 and heatat 80 to 90°C to dryness.Cool and
repeatHzOz treatmentuntil organicmatteris destroyed.
3. Add 10 mL of HF and heat to dryness.Cool and repeatthe HF treat-
ment oncemore.
4. Add 15 mL of concentratedHN03 and 5 mL of HCI04. Heat until
fumes of HCI04 appearand continueheatingto neardryness.Do not
exceed 210°C becausethe potential for explosion of HCI04 will
increase.
5. Cool, add 10 mL of 1:10 HCI, and heat to 70°C for 1 h.
6. Filter through quantitativeacid-washedfilter paperinto a 50-mL vol-
umetric flask. Rinsethe beakerand filter paperwith small volumesof
1:100HCI, dilute to volume, and mix. Retain the solution for analysis
of Ni, Cd, and Pb by one of the analytical methodsof section on
"Analytical Determinationof Nickel, Cadmium,and Lead."
Comments
Sampleswith high levels of organic matter require numeroustreatments
with HZ0 2 to destroy the organic material. Such samplescan be dry ashedin a
muffle oven at 450 to 550°C for 2 h to overnight dependingon the degreeof
treatmentneededto destroythe organic matter prior to the digestionprocedure.
Sampleswith large amountsof carbonateminerals also may be pretreatedby
adding up to 25 mL of 1:10 HN03 to the sampleand evaporatingto dryness
before proceedingwith the digestion procedure.If a HCI04 fume hood with
wash-downis not available, then HCI04 should be omitted from the digestion
procedure.Near completerecoveryof metals is still possibleif HCI04 is omit-
ted.
SELECTIVE EXTRACTION OF NICKEL, CADMIUM,

AND LEAD FROM SOILS
Principles
The objectiveof selectiveextractionmethodsis to determinethe amounts

of metalsassociatedwith variousphases(e.g., carbonates,organicmatter,metal
oxides,silicate minerals)in soils and sediments.Both instrumentaland selective
extraction methodshave been used to determinechemical partitioning of trace
746 AMACHER
elementsin sediments(Horowitz, 1991), and thesemethodsare applicableto

soils as well.
Instrumentalmethodsthat have beenusedto study chemicalassociations
of elementswith specificmineralphasesin geologicalmaterialsincludethe elec-
tron microprobe, scanningelectron microscopy with energy dispersive x-ray
analysis (SEMJEDAX), x-ray photoelectronspectroscopy(XPS), ultraviolet
photoelectronspectroscopy,Auger electronspectroscopy(AES), secondaryion
mass spectroscopy(SIMS), and ion scatteringspectroscopy(ISS) (Horowitz,
1991). Most of thesetechniquesare relatively new and have the advantageof
allowing, in many cases,for nondestructivequantitativeanalysisof mineralsin
earth surface materials without prior separation or preconcentration.
Disadvantages,aside from the large capital outlay for instrumentation,include
problemsassociatedwith detectionlimits and the surfacetextureof the material.
Despitethe problemsinherentwith the instrumentalmethods,they are useful for
studyinga limited numberof mineralcomponentswith well-definedchemistries
(Horowitz, 1991). For recent reviews on the use of instrumentalmethodsfor
studying geological materials refer to Mogk (1990) and Jacksonet al. (1989,
1993,1995).
In comparisonwith instrumentalmethods,selective extractionmethods
offer a relatively simpleand inexpensivemeansof chemicallyfractionatingsoils.
Selectiveextractionmethodsare labor intensiveand time consuming,however,
negatingsomeof the expenseadvantage.
The principal limitations of selectiveextraction methodsare the lack of
selectivityof mostextractantsfor the particularphaseof interest,the potentialfor
elementreadsorptionby other soil componentsduring the extractionsteps,and
other artifactsintroducedduring extractionprocedures.For thesereasons,selec-
tive extraction methods,as with most analytical methodsfor soils, are opera-
tionally defined.
Considerablecontroversyhas developedover the accuracyof selective
extractionmethodsand thesemattersare probably still largely unresolved(e.g.,
Tipping et aI., 1985; Kheboian & Bauer, 1987; Tessierand Campbell,1988;
Bauer & Kheboian, 1988; Gruebelet aI., 1988; Belzeile et aI., 1989). Many of
the evaluationstudieshavebeenconductedwith modelsystemsof syntheticmin-
eral phasesdopedwith trace elements.Theseartificial systemsmay not be an
accuraterepresentationof naturalsoils and sediments.Furthermore,addedtrace
elementsare often only weakly sorbedto mineral surfacesand are likely not in
the sameform as the native traceelementsin the soil. Samplepreparationmeth-
ods such as drying can introduceartifacts into the extractionmethods(Rapin et
aI., 1986). Despiteall the limitations of selectiveextractionmethodsthey offer
the best meansof evaluatingchemical associationswith amorphousor poorly
crystallinesoil phasesthat are not readily studiedby other means.
Pickering (1981), Chao (1984), and Salomonsand Forstner (1984) have
reviewed the use of selectivefractionationsmethods.Most of the applications
havebeenwith sedimentsamples.Someexamplesof the useof selective extrac-
tion of trace elementsin soils include the work of Harrison et a1. (1981),
Schalschaet al. (1982), Spositoet al. (1982), Miller and McFee(1983), Shuman
(1985) and Miller et al. (1986).
The selective extraction methodspresentedhere are based on methods

developedby Tessieret al. (1979), Chao(1972), Chaoand Zhou (1983), Shuman
(1982, 1983, 1985), and Miller et al. (1986) and have beenextensivelyusedfor
soils. The extractionscan be done sequentiallyor in somecasesindividually, if
desired,dependingon the objectivesof the study.
The methodpresentedherefor exchangeable metalsusesMg(N03)z. Some
sequentialextractionmethodsusechloridesaltsfor the exchangeable phase(e.g.,
Tessieret aI., 1979). Chloride will complex significant amountsof Cd and thus
extract more Cd from the soil than will nitrate or perchloratesalts (Fujii et aI.,
1983). Some sequentialextraction methods use buffered salts such as pH 7
NH40Ac for the exchangeablephase (e.g., Salomons & Forstner, 1984).
However, amountsof exchangeablemetals,particularly Pb, extractedfrom acid
soils and sedimentsare likely to be quite low using salts buffered to pH levels
substantiallyabovethoseof the soils and sediments.Neutral saltswith noncom-
plexing anionsare preferredfor extractingexchangeablemetalsat the pH of the
soil or sediment.
The method presentedhere for carbonate-boundmetals uses pH 5, 1 M
NaOAc (Tessieret aI., 1979). An alternatemethod usespH 4.5, 1 M NH40Ac
(Chao, 1984). Some dissolution of amorphousmetal oxides may occur with
buffered acetatesolutions and some readsorptionof metalsextractedfrom car-
bonatesonto metal oxidesis likely.
Extractionof metalsassociatedwith organicmatter is particularly trouble-
some.Hydrogenperoxideand sodium pyrophosphatewill extractmetalsassoci-
atedwith organicmatterbut alsowill extractmetal oxides(Shuman,1983; Miller
et aI., 1986). Changingthe orderof extractantscan partially overcomethis prob-
lem (Miller et aI., 1986). Manganeseoxidesmustbe extractedwith pH 2 NHzOH
• HCI (Chao, 1972) prior to extractionof organically bound metalsby H20 2 or
Na4P207·
Organicmatterand sulfide mineral oxidation by H20 2 using the Tessieret
al. (1979) methodor similar methodsis a problemwith samplescontaininglarge
amountsof thesephases.Incompleteoxidationand excessivereactionwith H20 2
leadingto expulsionof samplefrom the centrifugetube can occur. Furthermore,
HzOz solutionsdo not producea cleanseparationbetweensolid and liquid phas-
es on centrifuging. Fine clay particlesremain suspendedin solution by the bub-
bling action of the H20 2 . One approachto theseproblemsis to use one of the
digestionmethodsof the section"Total Nickel, Cadmium,and Lead" in the final
stepof the extractionsequenceto extractmetalsassociatedwith the organicmat-
ter, sulfide, and residualphases.Of course,this doesnot permit partitioning the
metalsbetweenthe organicmatterand residualphases.
Sodium hypochloritewill selectivelyoxidize organicmatterwith the least
damageto carbonate,metal oxide, and silicate clay phasesin the soil (Anderson,
1963; Lavkulich & Wiens, 1970). Unfortunately,at the high pH of the extraction
(pH 8.5 used by Shuman, 1983), readsorptionof metal ions, particularly Pb,
releasedduring oxidation of organicmatterby the inorganicphasesremainingis
likely. This problem is only partly overcomeby extractingthe residueafter the
NaOCI oxidation step with NaOAc buffered to pH 5. This is the samereagent
usedin the carbonateextractionstepand will not interferewith subsequentsteps.
748 AMACHER
An alternateapproachthat can be usedto estimateamountsof metalsread-

sorbed during NaOCI extraction is to extract duplicate samplesomitting the
NaOCI oxidation step with one of the samplesand including it with the other.
The differencesin metal ion concentrationsbetweenthe two samplesthat are
extractedby the subsequentmetal oxide extractantsgive the amountsof read-
sorbed metals previously associatedwith the organic matter. This approach
assumesthat the metal oxide extractantswill not removemetalsassociatedwith
organic matter. Unfortunately,none of theseextractantsis completelyselective
so this assumptionmay not be valid.
Three methodsfor extracting metals associatedwith organic matter are
presentedhere: NaOCI (Shuman,1983), Na4P207(Miller et aI., 1986),and H20 2
(Tessieret aI., 1979). None of the methodsis fully satisfactory.The choice will
dependsomewhaton study objectivesand propertiesof the soils or sedimentsto
be extracted.The NaOCI methodshouldbe usedprior to any of the metal oxide
extractions.The Na4P207methodshould be usedafter the Mn oxide extraction
step, and perhapsafter all the metal oxide extraction steps.The H20 2 method
shouldbe usedafter all the metal oxide extractionsteps.
Amorphousand crystalline iron oxides can be distinguishedby selective
extractionmethods.Acid ammoniumoxalate in the dark (Shuman,1982; Chao
& Zhou, 1983) or 0.25 M NH20H· HCI + 0.25 M HCI at 50°C (Chao & Zhou,
1983) will extract amorphousiron oxides and associatedmetals while acid
ammoniumoxalate with ascorbicacid (Shuman,1982) will extract crystalline
iron oxides and associatedmetals.Crystalline iron oxides also can be extracted
with acid ammoniumoxalate under ultraviolet light or with dithionite, but the
ultraviolet (UV) light method is inconvenient and dithionite may precipitate
metal sulfidesleadingto incompleterecoveries(Shuman,1982).
ResidualNi, Cd, and Pbare bestdeterminedby the digestionmethodgiven
in the section "Digestion with Hydrogen Peroxide,Hydroflouric Acid, Nitric
Acid, and PerchloricAcid in Teflon Beakerson a Hot Plate" , althoughthe diges-
tion methodsin the section "United StatesEnvironmentalProtection Agency
SW-846,Method 3050: Digestionwith Nitric Acid and HydrogenPeroxidein a
Conical Beakeron a Hot Plate" and "Digestion with Nitric Acid and Perchloric
Acid in a Block Digester"also are useful if completemetal recoveryis not essen-
tial. Alternatively, determineresidualNi, Cd, and Pb by summingthe metal con-
centrationsfrom the sequentialextractionstepsand subtractingthesefrom total
Ni, Cd, and Pb (see"Total Nickel, Cadmium,and Lead").
Equipment
1. 50-mL graduatedplastic centrifugetubesand racks.
2. Vortex mixer.
3. Reciprocatingshaker.
4. Shakingwater bath.
5. 1-L glassbeakerfor useas a boiling water bath.
6. Hot plate.
7. Evaporatingdishes.
8. Centrifuge.
9. Membranefilters and syringe filter units or quantitativefilter paper,

funnels, and filter racks.
10. Steamplate or drying oven.
11. Muffle oven.
Reagents
Prepareeither Reagent3, 4, or 5 and 6 dependingon which organicmatter
extractionmethodis selected.
1. 0.1 M Mg(N03h: Dissolve25.64g of Mg(N03h • 6H20 in deionized
water and dilute to 1 L.
2. pH 5, 1 M NaOAc: Dissolve 82.03 g of sodium acetatein deionized
water, adjust to pH 5.0 with acetic acid, and dilute to 1 L.
3. pH 8.5, 5.25% NaOCI: Adjust 5.25% NaOCI to pH 8.5 by dropwise
addition of 1:1 HC!. Preparefresh for eachuse.
4. 0.1 M Na4P207:Dissolve 26.50 g of Na4PZ07in deionizedwater and
dilute to 1 L.
5. pH 2, 30% HzOz: Adjust 30% H20 2 to pH 2.0 with 1:10 HN03.
6. 3.2M NH40Ac in 1:5 HN03: Dissolve247 g ofNH 40Ac in deionized
water, add 200 mL of concentratedHN03, and dilute to 1 L.
7. pH 2, 0.1 M NH 20H • HCl in deionizedwater. Adjust to pH 2.0 with
1:10 HCl and dilute to 1 L.
8. 0.25 M NHzOH • HCl + 0.25 M HCI: Dissolve 17.37 g of NH20H •
HCI in deionizedwater, add 20.8 mL of concentratedHCI and dilute
to 1 L.
9. pH 3, 0.2 M ammoniumoxalate+ 0.2 M oxalic acid + 0.1 M ascorbic
acid: Dissolve 28.42 g of (NH4hCz0 4 • HzO plus 25.21 g of HZC20 4
• 2HzO plus 17.61 g of ascorbicacid in deionizedwater. Adjust to pH
3.0 with 1:10 NH 4 0H, and dilute to 1 L. Preparefresh for eachuse.
10. 95% ethanol.
11. Acetic acid.
12. Nitric acid, concentrated,and 1:10 (vol/vol concentrated HNOYHzO).
13. Hydrochloric acid, concentrated,1:1, and 1:10 (vol/vol concentrated
HClIHzO).
14. NH 40H, 1:10 (vol/vol concentratedNH 40H/HzO).
Procedure
Omit Step 2 if the sampledoesnot contain carbonates.Use either Step 3,
5, or 8 dependingon which organicmatterextractionmethodis chosen.
1. Exchangeablephase:Add 0.500g of preparedsoil sampleto a 50-mL
graduatedplastic centrifugetube. Add 10.0 mL of 0.1 M Mg(N03)z'
Shakefor 2 h at 180 cycles/min. Centrifuge for 10 min at 1500 x g.
Decantsupernatant,filter if necessaryto removefloating organicmat-
ter, and retain for analysis. Wash residue three times with 5 mL of
ethanol,centifuging each time for 5 min at 1500 x g to separatethe
residue,discard ethanolwashes,and evaporateethanol in residueto
dryness.
750 AMACHER
2. Carbonatephase(omit if carbonates are not present):Add 10.0 mL of

pH 5, 1 M NaOAc to the residuefrom the previousstep. Vortex mix
and shakefor 5 h at 180 cycles/min.Centrifugefor 10 min at 1500 x
g. Decantsupernatant,filter if necessary,and retain for analysis.Wash
residuethree times with 5 mL of ethanol,and evaporateto dryness.
3. Organicmatter phaseusing NaOCI (omit if Step 5 or 8 is used):Add
10 mL of pH 8.5, 5.25% NaOCI to the residuefrom the previousstep.
Vortex mix, and place in a boiling water bath for 15 min. Centrifuge
for 10 min at 1500 x g and decantthe supernatantinto an evaporating
dish. Repeat the NaOCI treatments until the organic matter is
destroyedas signified by the appearanceof a pink color (perman-
ganateion) in the supernatant(Anderson& O'Connor,1972).Add the
supernatantsto the evaporating dish after each treatment.Wash
residuethreetimes with 5 mL of pH 5, 1 M NaOAc adding thesuper-
natant each time to the evaporatingdish. Wash residue three times
with 5 mL of ethanol,and evaporateto dryness.Evaporatethe NaOCI
solution to dryness,dissolvethe residuein 1:10 HN03, quantitatively
transferthe solution with rinsing to a 25-mL volumetric flask, dilute
to volume, and retain for analysis.
4. ManganeseOxide phase:Add 25.0 mL of pH 2, 0.1 M NH20H • HCI
to the residuefrom the previousstep.Vortex mix andshakefor 30 min
at 180 cycles/min.Centrifugefor 10 min at 1500 x g. Decantsuper-
natantand retain for analysis.Wash residuethree times with 5 mL of
ethanol,and evaporateto dryness.
5. Organicmatterphaseusing Na4P207(omit if Step3 or 8 is used):Add
25.0 mL of 0.1 M Na4P207to the residue from the previous step.
Vortex mix and shakeovernight at 180 cycles/min.Centrifugefor 10
min at 1500 x g. Decant supernatantand retain for analysis. Wash
residuethree times with 5 mL of ethanol,and evaporateto dryness.
6. AmorphousFe Oxide phase:Add 25.0 mL of 0.25 M NH20H • HCI +
0.25 M HCI to the residue from the previous step. Vortex mix and
shakefor 30 min at 50°C and 180 cycles/min. Centrifugefor 10 min
at 1500 x g. Decantsupernatantand retain for analysis.Wash residue
three times with 5 mL of ethanol,and evaporateto dryness.
7. Crystallineiron oxide phase:Add 25.0 mL of pH 3,0.2M ammonium
oxalate+ 0.2 M oxalic acid + 0.1 M ascorbicacid to the residuefrom
the previousstep.Vortex mix and place in a boiling water bath for 30
min. Centrifugefor 10 min at 1500x g and decantthe supernatantinto
an evaporating dish. Wash residue three times with 5 mL of the
oxalateextractingsolution, and add the supernatantsto the evaporat-
ing dish. Wash residuethree times with 5 mL of ethanol,and evapo-
rate to dryness.Evaporatethe oxalatesolution to dryness,ignite the
residuein a muffle oven at 450°Cfor 4 h, dissolvethe residuein 1:10
HN03, quantitativelytransferthe solutionwith rinsing to a 25-mL vol-
umetric flask, dilute to volume, and retain for analysis.
8. Organic matter phaseusing HzO z (omit if Step 3 or 5 is used):Add

25.0 mL of pH 2, 30% HzO z to the residuefrom the previous step.
Placein a boiling waterbathfor 30 min. Centrifugeuntil a clearsuper-
natantis obtainedand filter the supernatantinto a 100-mL volumetric
flask. Repeatthe HzO z treatmentsup to two more times if necessary
to destroy the organic matter, and add the supernatantsto the volu-
metric flask eachtime. Wash residuethree times with 5 mL of 3.2 M
NH40Ac in 1:5 HN03 adding the supernatanteachtime to the volu-
metric flask, dilute to volume, and retain for analysis.Wash residue
three times with 5 mL of ethanol,and evaporateto dryness.
9. Residualphase:Add 5 mL of ethanolto the residuefrom the previous
step and vortex mix to resuspendthe residue.Quantitatively transfer
the residuefrom the centrifugetube to an appropriatedigestionvessel
by rinsing with small amountsof ethanol, and evaporateto dryness.
Carry the residuethrough one of the digestion methodsgiven in the
section on"Total Nickel, Cadmium,and Lead."
Comments
1. Exchangeablephase:Mg(N03h is substituted for MgCl2 that was
used in the original Tessieret al. (1979) method. Chloride will com-
plex Cd and increasethe amountof Cd extracted.If only exchangeable
metalsare to be extracted,then a noncomplexinganion must be used.
2. Carbonatephase:This step is omitted if carbonatesare not presentin
the soil sample.
3. Organicmatter phase:Washingthe residuewith pH 5, 1 M NaOAc is
included in the NaOCI methodto partially extract metals readsorbed
during organic matteroxidation, which is likely at the high pH used.
The samesolution from the carbonateextractionis usedbecauseit is
not as likely to interfere with subsequentextractions.Alternatively,
duplicatesamplesmay be carried through the extraction sequencein
which one sampleis carriedthrough the extractionsequenceomitting
the organicmatterdigestionstepand the othersampleis digestedwith
NaOCI to destroyorganic matter. Differencesin the amountsof met-
als removedby subsequentmetal oxide extractionsbetweenthe two
samplesgive the amountsof readsorbedmetalspreviously associated
with organicmatter.
4. Metal oxide fractions: All the metal oxides may be dissolvedby the
oxalateextractantin a single extractionstep if selectiveextractionof
the metal oxidesis not desired.Destructionof the oxalateis necessary
for flame AAS analysisof the oxalatesolution in Step 6. Otherwise,
clogging of the nebulizer occurs. The oxalate solution may be ana-
lyzed directly by ICPAES, although more frequent cleaning of the
torch is necessary.In this casecombine the oxalate extract with the
oxalatesolution washesin Step6 in a 50-mL volumetric vlask, dilute
to volume, and retain the solution for analysis.
5. Residualphase:One of the digestionmethodsof the sectionon "Total
Nickel, Cadmium,and Lead" may be useddependingon the proper-
752 AMACHER
ties of the sampleand the desireddegreeof digestion.Metals in the

residual phasealso may be determinedas the differencebetweenthe
sum of all the extractedphasesand total metals.
6. Inductively coupledplasmaatomic emissionspectrometry(sectionon
"Analytical Determinationof Nickel, Cadmium, and Lead.") is the
methodof choice for determiningNi, Cd, Pb, and other metalsin the
extracts,but AAS (section on "Analytical Determinationof Nickel,
Cadmium, and Lead") also may be used. The concentrationof each
metal in milligrams per kilogram of soil is obtainedby multiplying the
solution concentrationof metal in milligrams per liter by the solution
to soil ratio. Different sampleweightsandsolutionvolumesthan those
specifiedmay be selectedas needed.
7. In severalof the extractionsequences reportedin the literature,a water
wash is used between extraction steps to prevent carry-over of
entrainedsolution. However, water and soil suspensionsoften do not
separatesatisfactorily by centrifugation.The ethanol washesused in
the methodpresentedheregive a good separationof soil and solution
during centrifugationand entrainedethanolis easily evaporatedby air
drying prior to subsequentextractionsteps.
PLANT AVAILABLE FRACTIONS OF NICKEL, CADMIUM,

AND LEAD
Principles
Many approacheshave developedfor measuringthe fraction of the total

quantity of an elementin soils that is availablefor uptakeby plant roots. For a
descriptionof someof the concepts,procedures,and interpretationsof soil test-
ing for plant availableelements,consult someof the many references available
on this subject(e.g., Counc. Soil Test. Plant Anal., 1980; Brown, 1987; Jones,
1989; Westerman,1990).
In general,three approachesto measuringplant availability of elements
can be described.In the first approach,concentrationsof elementsin the soil
solution are measuredand are used to calculateionic activities (Adams, 1971;
Amacher,1984; Baham,1984; Sposito,1984). Ionic activities (intensity factor)
are assumedto be a measureof elementbioavailability to plant roots (Baker &
Low, 1970; Khasawneh,1971; Sparks,1984).
Numerousmethodshave beenused to extract the soil solution from soils
(Table 28-1) including saturationextracts, miscible and immiscible displace-
ment, centrifugation,and pressureor vacuum extraction of soil samplesin the
laboratory or soil water samplers(lysimeters)in the field. All of the methods
haveadvantagesand disadvantages, but the major problemto be overcomeis to
obtain a representativesoil solution extractwithout altering its compositiondur-
ing the extractionprocedure.
In addition, chelateequilibria methodshave beenusedas a meansof cal-
culating ionic activities and hence element bioavailability in soils (Norvell &
Lindsay, 1982; Fujii et aI., 1983; Baker & Amacher,1981; Amacher,1984).
Table 28-1. Methodsfor extractingsoil solutions.

Method Reference
Saturationextract Rhoades,1982
Immiscible displacement-centrifugation Mubarak& Olsen, 1976
Whelan& Barrow, 1980
Kinniburgh & Miles, 1983
Miscible displacement-vacuum
extraction Wolt & Graveel,1986
High-speedcentrifugation Elkhatib et aI., 1987
Pressure filtration Ross& Bartlett, 1990
In the secondapproach,the amountsof plant availableelementsin the soil

are determined(quantity factor, labile pool) by isotopicdilution (Fujii & Corey,
1986) or with somesuitableextractantin which the extractableamountscorre-
late with the quantitiesof elementsremovedfrom the soil by plants.This is the
approachmost commonly used in routine soil testing. A list of some of the
extractantsusedto measureplant availableNi, Cd, andPb is given in Table28-2.
Many of the successfulusesof theseextractantswere obtainedundercontrolled
greenhouseconditions and with elevatedlevels of metals added to the soils.
Table 28-2. Someextractantsusedto estimateplant availableNi, Cd, and Pb in soils.

Extractants References
Nickel
Neutral salts Sauerbeck& Hein, 1991
N140Ac Misra & Pande,1974
HOAc Misra & Pande,1974; Haq et aI., 1980
HCI Misra & Pande,1974
Double acid Korcak & Fanning,1978
Acid ammoniumoxalate Misra & Pande,1974
DTPA Korcak & Fanning,1978; Valdareset aI., 1983
AB-DTPA Barbarick& Workman, 1987
Cadmium
NH40Ac Johnet aI., 1972; Haghiri, 1973
HOAc Haq et aI., 1980
NH4N03 Symeonides& McRae,1977
HN03 Bidwell & Dowdy, 1987
HCI Joneset aI., 1973b,1975
Double acid Korcak & Fanning,1978
Sodiumpyrophosphate Latterell et aI., 1978
DTPA Binghamet aI., 1975; Korcak & Fanning,1978,Streetet aI., 1977;
Browne et aI., 1984,Singh & Narwal, 1984; Bidwell & Dowdy,
1987
AB-DTPA Barbarick& Workman,1987
Lead
MgCI2 Miller et aI., 1975
BaCI2 Joneset aI., 1973a
NH40Ac John,1972; Misra & Pandey,1976
HN03 John,1972
HCI Miller et aI., 1975
Bray 1 and 2 Miller et aI., 1975
Acid ammoniumoxalate Misra & Pandey,1976
EDTA Joneset aI., 1973a;Miller et aI., 1975
754 AMACHER
Extractantstend to be lesssuccessfulat predictingplant-availablequantitiesof

metalsunderfield conditions.
Nickel and Cd can be readily takenup and accumulatedby plantsdepend-
ing on soil conditions and especiallyfrom soils containing elevatedlevels of
theseelements(Adriano, 1986). Lead on the otherhandtendsnot to be takenup
and accumulatedby plants, and where uptakeoccursmost of the Pb is located
within the root systemof the plant (Adriano, 1986).It is probablyfor this reason
that attemptsto measureplant availablePb with soil extractantsaregenerallynot
very successful,particularlyunderfield conditionsandat higherpH levelswhere
Pb is not very soluble(Santillan-Medrano& Jurinak,1975).
The third approachis relatedto the secondin that the plant availablefrac-
tion of an elementin soils is determined,but in this approachan ion exchange
resin is usedto mimic plant roots by extractinga portion of plant availableele-
mentsfrom the soil (Yang et aI., 1991).As with the secondapproach,correlation
of resin extractableamountsof elementswith quantitiestakenup by plantspro-
videsthe bestmeansof assessing the resinextractionmethod.The processof ele-
ment removalfrom the soil by ion exchangeresinsis probably more similar to
the processof ion removalfrom soil by plant roots than are chemicalextraction
methods(Yang et aI., 1991). Resin extractionmethodsalso do not causesharp
pH changesand excessivemineral dissolutionas do chemicalextractionmeth-
ods. Unfortunately,resin extraction methodshave not as yet beenextensively
usedto measureplant availablefractionsof Ni, Cd, and Pb.
Becauseof the many soil testingapproachesavailabletoday and the vary-
ing degreesof successof theseapproacheswith different soils and plants, it is
not possibleto recommenda universalapproachthat will meet all the require-
mentsof a given study. Resultswill vary with plant speciesand soils. Instead,
five commonly usedextractionmethodsare presented.Thesemethodsare suit-
able for a wide rangeof soil types,but for properevaluationin any given appli-
cationthey mustbe correlatedagainst plantresponse.Changesin extractantcom-
position, solution to soil ratios, shakingtimes and speeds,and other extractant
conditions will produce different results making comparisonsdifficult if not
impossible.Precautionsregardingthe avoidanceof samplecontaminationgiven
in the sectionon "Total Nickel, Cadmiumand Lead" also are applicable with
these methods. Inductively coupled plasma atomic emissionspectrometryor
AAS are the analyticalmethodsof choicefor determiningNi, Cd, and Pb in the
extracts.
Acid Method
The DTPA methodwasoriginally developedto extractMn, Fe, Cu, andZn

from slightly acidic to alkaline soils (Lindsay & Norvell, 1978), but in some
casesit can be usedfor other metalssuch as Ni, Cd, and Pb and for acid soils
provided soil pH and perhapsother soil propertiesare usedin interpretingthe
results(Haq & Miller, 1972; Korcak & Fanning,1978; Singh & Narwal, 1984).
The DTPA is usedto extractsoil metalsby forming chelateswith free metalsin
solution,which lowerstheir ionic activitiesso that additionalmetalquantitiesare
releasedfrom the soil until equilibrium is attained.In practice,extractionsare

done for a fixed time interval becauseequilibrium may not be attainablewithin
a reasonablyshort time period. The degreeof chelationof the various metals is
a function of the magnitudesof the metal-DTPA stability constantsand also
varies with pH (Sommers& Lindsay, 1979). Triethanolamine(TEA) is used to
buffer the extractingsolution to pH 7.3 in an attemptto optimize the extraction
of variousmetals.Calciumchloride is includedin the extractingsolution to min-
imize dissolutionof calcium carbonateminerals.
Norvell (1984) and O'Connor(1988) discussedmany of the successfuland
unsuccessfulapplicationsof the DTPA soil test. Unsuccessfulusesof the test
may be related to unjustifiable method alteration, use in strongly acidic soils,
excessivemetal loadingsin the soil that exceedthe capacityof DTPA to chelate
the metals, and greenhousevs. field experiments.Norvell (1984) showedthat
DTPA, ethylenediaminetetracetic acid (EDTA), and hydroxyethylenediaminetri-
acetic acid (HEDTA) buffered at pH 5.3 were effective extractantsfor several
metals including Ni and Cd from metal-enrichedsoils. Effective extraction of
metals from metal-enrichedsoils may require increased concentrationsof
chelates,higher extractantto soil ratios, or multiple extractions(Norvell, 1984).
Equipment
1. One hundredtwenty-five-milliliter Erlenmeyerflasks.
2. Reciprocatingor orbital shaker.
3. Quantitativeacid-washedfilter paper(Whatmanno. 40 or equivalent),
filter funnels, receivingflasks, and racks.
Reagents
The pH 7.3, 0.005M DTPA extractionsolution: Dissolve 1.96 g of DTPA,
14.92 g of TEA, and 1.47 g CaCl2 • 2H20 (calcium chloride dihydrate) in 950
mL of deionizedwater in a l-L beaker.Adjust to pH 7.3 ± 0.05 using 6 M HCl
while stirring. Quantitativelytransferthe solution with rinsing to a l-L volumet-
ric flask, dilute to volume, and mix. The solution is stable for severalmonths.
The final concentrationof eachreagentis 0.005 M DTPA, 0.1 M TEA, and 0.01
MCaCI2·
Procedure
Add 20 mL of DTPA-extractingsolution to 10 g of preparedsoil in a 125-
mL Erlenmeyerflask. Shakefor 2 h on a reciprocatingor orbital shakerat 180
cycles/min.Filter the suspensionthroughquantitativefilter paperand analyzethe
filtrate for Ni, Cd, and/orPb usingoneof the analyticalmethodsgiven in the sec-
tion on "Analytical Determinationof Nickel, Cadmium,and Lead". The results
in milligrams per kilogram of soil are calculatedby multiplying the filtrate con-
centrationsin milligrams per liter by two (extractantvolume in milliliters divid-
ed by soil sampleweight in grams).For expressionof resultson a volume basis
or in other units refer to the Council on Soil Testing and Plant Analysis
Handbook(1980).
756 AMACHER
Ammonium Bicarbonate-Diethylenetriaminepentaacetic
Acid Method
The ammonium bicarbonate-diethanolaminetetraaceticpentaacetic acid

(AB-DTPA) solution was developedas a multielement extractantfor alkaline
soils and gives similar resultsto the DTPA method fortrace elements(Barbarick
& Workman, 1987; Soltanpour,1991). It has the potential to extract plant avail-
able amountsof Na, K, Mo, Mn, Fe, Ni, Cu, Zn, Cd, B, Pb, N03, P, As, Se, and
perhapsotherelementsfrom alkaline soils, and is most advantageous when used
with ICPAES.
Equipment
SeeDTPA methodin "Diethylenetriaminepentaacetic
Acid Method".
Reagents
The NH4HC03-DTPA extractingsolution: Dissolve1.97g of DTPA in 800
mL of deionizedwater plus 2 mL of 1:1 NH40H in a 1-L beaker.Add 79.06g of
NH4HC03 and stir to dissolve. Adjust to pH 7.6 using dilute NH40H or HCI
solution while stirring. Quantitativelytransferthe solution with rinsing to a 1-L
volumetric flask, dilute to volume, and mix. Use the freshly preparedsolution
promptly or store undermineral oil becausethe solution pH changeswith time.
Procedure
Add 20 mL of AB-DTPA solution to 10 g of preparedsoil in a 125-mL
Erlenmeyerflask. Shakefor 15 min on a reciprocatingor orbital shakerat 180
cycles/min.Filter the suspension throughquantitativefilter paperandanalyzethe
filtrate for the elementsof interestby ICPAES or othersuitableanalytical meth-
ods. For ICPAES analysisneutralizethe excess bicarbonate by adding 0.25 mL
of concentrateHN03 for each2 mL of filtrate. Calculationof the amountof each
extractableelementis the sameas for the DTPA method.
One-TenthMolar Hydrochloric Acid Method
Although this methodwasoriginally developedfor extractingZn from acid

soils (Nelsonet aI., 1959), it is applicablefor extractingother heavy metalsfrom
acid soils. Becauseof the many variationsin proceduresusedwith the 0.1 M HCI
extractant, results between different labs using different soil/solution ratios,
shakingtimes, etc., may not be comparable.Two variationsare presentedhere:
the original sequentialextractionprocedureand a single extractionwith a 1:10
soil/solution ratio that is more advantageous for routine use. A single extraction
with a 1:4 soil/solution ratio also is commonly used (Counc. Soil Test. Plant
Aoal., 1980).
Equipment
SeeDTPA methodin "DiethylenetriaminepentaaceticAcid Method." Also
need50-mL plasticcentrifugetubesand racks,centrifuge,and 100-mL volumet-
ric flasks if sequentialextractionprocedureis used.
Reagents
One-tenthmolar HCI extracting solution: Dilute 8.3 mL of select-grade
concentrateHCI to 1 L with deionizedwater in a volumetricflask. Commercially
available0.1 M HCl solutionsare acceptableif low in traceelementcontent.
Procedure
1. Sequentialextraction procedure:Add 20 mL of 0.1 M HCI to 2 g of
preparedsoil in a 50-mL plastic centrifugetube. Shakefor 5 min on a
reciprocatingshakerat 180 cycles/min.Centrifugeuntil a clear super-
natantis obtainedand decantthe supernatantinto a 100-mL volumet-
ric flask. Repeatthe extractiontwo or more times until the solution pH
is less than 2.0, adding the supernatantto the volumetric flask each
time. Dilute the flask contentsto volume and mix. Analyze the solu-
tion for Ni, Cd, and/or Pb using one of the analytical methods in
"Analytical Determination of Nickel, Cadmium, and Lead." The
resultsin milligrams per kilogram of soil are equalto the solutioncon-
centration of metal in milligrams per liter multiplied by 50 (solu-
tion/soil ratio).
2. Single extractionprocedure:Add 20 mL of 0.1 M HCl to 2 g of pre-
paredsoil in a 125-mL Erlenmeyerflask. Shakefor 30 min on a reci-
procatingor orbital shakerat 180 cycles/min.Filter through quantita-
tive filter paper and analyze the filtrate for the elementsof interest
using one of the analytical methodsof "Analytical Determinationof
Nickel, Cadmium,and Lead." The resultsin milligrams per kilogram
of soil are equal to the solution concentrationof metal in milligrams
per liter times 10.
DoubleAcid (Mehlich I) Method
This methodwas originally developedfor extractingexchangeablecations

and P from acid soils, but also is applicablefor extracting heavy metals from
soils (Korcak & Fanning,1978).
Equipment
Acid Method."
Reagents
1. 6 M HCI: Dilute 500 mL of select-gradeconcentrated HCIto 1 L with
deionizedwater in a volumetric flask.
758 AMACHER
2. 0.5 M H2S04: Add 28 mL of select-gradeconcentratedH2S04 to about

500 mL of deionizedwater, cool, and dilute to 1 L in a volumetric
flask.
3. 0.05 M HCL-0.0125M H2S04extractingsolution: Dilute 8 mL of 6 M
HCI and 25 mL of 0.5 M H2S04 to 1 L with deionizedwater in a vol-
umetric flask.
Procedure
Add 20 mL of double acid solution to 5 g of preparedsoil in a 125-mL
cycles/min.Filter throughquantitativefilter paperand analyzethe filtrate for the
elements of interest using appropriate analytical methods (see "Analytical
Determinationof Nickel, Cadmium,and Lead"). The results in milligrams per
kilogram are obtainedby multiplying the solution concentrationsin milligrams
per liter by four (solution/soilratio).
Mehlich III Method
Although developedfor extractionof K, Mg, Ca, Mn, Fe, Cu, Zn, B, and
P from acid soils (Mehlich, 1984), this methodis applicableto other heavy met-
als.
Equipment
Acid Method."
Reagents
1. NH4F-EDTA solution: Dissolve 138.9g of NH4F and 73.5 g of EDTA
in deionizedwater and dilute to 1 L.
2. 1 M HN03 : Dilute 62.5 mL of select-gradeconcentratedHN03 to 1 L
3. Mehlich III extractingsolution: Dissolve 80 g of NH4N03 in 3 L of
deionizedwater.Add 16 mL of NH4F-EDTA solution,46 mL of acetic
acid, and 13 mL of 1 M HN03. Dilute to 4 Lwith deionizedwater.The
final pH shouldbe 2.0 ± 0.1.
Procedure
Add 20 mL of Mehlich III solution to 2 g of preparedsoil in a 125-mL
cycles/min.Filter throughquantitativefilter paperand analyzethe filtrate for the
elementsof interestusing the analytical methodsof "Analytical Determination
of Nickel, Cadmium,and Lead." The resultsin milligrams per kilogram of soil
are obtainedby multiplying thesolutionconcentrationsin milligrams per liter by
10 (solution/soil ratio).
ANALYTICAL DETERMINATION OF NICKEL, CADMIUM,

AND LEAD
Numerousanalytical techniquesare availablefor determiningthe concen-

trations of Ni, Cd, and Pb in soil digestion and extraction solutions including
flame atomic absorption spectrophotometry(FAAS), graphite furnace atomic
absorption spectrophotometry(GFAAS), ICPAES, ICPMS, colorimetry, ion
chromatography,and electrochemicalmethods.For recentreviews on the appli-
cationof eachof thesetechniquesfor the determinationof Ni, Cd, and Pb in geo-
logical materialsand water refer to Jacksonet al. (1991) and MacCarthy et al.
(1991).
The standardcolorimetric method for the analysis of Ni in water is the
dimethylglyoxime methodwhereasdithizone methodsare standardfor Cd and
Pb (Am. Public Health Assoc.,Am. Water Works Assoc.,Water Pollut, Control
Fed., 1992). Thesemethodsare applicableto soil solutionsand someextractsas
well. Their usewith more complexextractsmay be difficult becauseof the pres-
enceof potentialinterferences.Spiking extractswith known amountsof the ele-
mentsof interestwill provide an evaluationof any interferences.
BastaandTabatabai(1990) presentedan ion chromatographicmethodwith
postcolumn reactions for determining total metals in soil digestion solutions
including Ni, Cd, and Pb. However, recoveryproblemswere noted with Cd and
Pb becauseof incompleteextractionof the dithizone complexesfrom the aque-
ous digestionsolutionswith chloroform.
Electrochemicalmethods (e.g., anodic stripping voltammetry) are most
suitedfor analysisof watersamples(e.g., Figura & McDuffie, 1980),but also are
applicableto soil solutionsand someextractsaswell (Opydo, 1989).Chemically
modified electrodesthat preconcentratethe analyte offer improved selectivity
along with the sensitivity of electrochemicaltechniques.Baldwin et al. (1986)
useddimethylgloxime-coatedgraphiteto concentrateNi from a sedimentdiges-
tion solution for quantitationby differential pulsevoltammetry.
Of all the techniquesavailablefor quantifying Ni, Cd, And Pb, the most
common are AAS and ICPAES and theseare emphasizedhere. For additional
information on the use of thesetechniquesin soil analysisrefer to Wright and
Stuczynski(1996) and Soltanpouret al. (1996) in this publication.
Atomic Absorption Spectrophotometry
Atomic absorptionspectrophotometrytechniquesinclude both flame and

flameless (graphite furnace) methods.Flame AAS techniquescan suffer from
matrix effects resultingfrom the presenceof large concentrationsof salts in soil
digestionor extractionsolutions.Direct determinationof Ni, Cd, and Pb in soil
digestionor extractionsolutionsis often only possibleif backgroundcorrection
and standardadditions techniquesare qsed. Background correction methods
include deuteriumarc, Zeeman,and Smith-Hieftje dependingon the particular
instrumentmanufacturerand model. Matching the matrix of samplesand stan-
dardsis good practice.If there is any doubt aboutthe effect of samplematrix on
760 AMACHER
Table 28-3. Analytical conditionsfor flame AAS analysisof Ni, Cd, and Pb.
Optimum
Element Wavelength Flamegases Detectionlimit Sensitivity concentrationrange
nm mgIL
Ni 232.0 Air acetylene 0.02 0.15 0.3-10
Cd 228.8 Air acetylene 0.002 0.025 0.05-2
Pb 283.3or Air acetylene 0.05 0.5 1-20
217.0
the analytical results,then standardadditionsmethodsshouldbe usedto evalu-

ate potential interferences.Kane (1988) conducteda detailedstudy on the opti-
mizationof flame parametersand characterizationof interferencesfor the simul-
taneousdeterminationof severalelementsincluding Ni, Cd, and Pb in geologi-
cal materials.Analytical conditionsfor flame AAS analysisof Ni, Cd, and Pbare
presentedin Table 28--3 (Am. Public Health Assoc.,Am. Water Works Assoc.,
Water Pollut. Control Fed., 1992). Actual lower limits of quantitation will
dependon the samplematrix. The methodsmanualavailablefor the particular
instrumentin usealso shouldbe consulted.
Graphite furnace AAS offers substantially lower detection limits than
flame AAS, but is not without its problems.Matrix effectsin soil digestionand
extractionsolutionscanbe severe.Backgroundcorrectionis mandatorywith this
technique. Improved graphite furnace design offers isothermal atomization,
which has eliminatedmany of the problemsassociatedwith older instruments.
Matrix modification andstandard additions may still be necessaryin many cases
howeverto achievesatisfactoryresults.Varma(1990) haswritten a handbookon
furnace AAS that contains numerous referenceson applications of GFAAS
analysisto geological materials.Analytical conditions for GFAAS analysisof
Ni, Cd, and Pb are presentedin Table 28-4 (Am. Public Health Assoc., Am.
Water Works Assoc.,Water Pollut. Control Fed., 1992). The actual lower limits
of quantitationwill dependon the samplematrix. The methodsmanualavailable
for the particularinstrumentin usealso shouldbe consulted.
Analysis of soil and sedimentslurriesis possiblewith GFAAS. Epsteinet
al. (1989)usedGFAAS with Zeemanbackgroundcorrectionandautomatedslur-
ry sampleintroduction into the furnace to determinePb and other elementsin
river sediment.Hoenig et al. (1989) determinedseveral elementsby GFASS
including Ni, Cd, and Pb in soils and sedimentsby slurried sampleintroduction.
Hinds and Jackson(1987,1990)usedslurry injection onto a L'vov platform with
Pd and Mg as matrix modifiers to determinePb while Rygh and Jackson(1987)
usedslurry injection into a Delvescup to determineCd.
Table 28-4. Analytical conditionsfor GFAAS analysisof Ni, Cd, and Pb.
Estimated Optimum
Element Wavelength detectionlimit concentrationrange Matrix modifier
lim -------~g!L--------
Ni 232.0 1 5-100 None
Cd 228.8 0.1 0.5-10 (Nl4)zHP04
Pb 283.3 or 217.0 1 5-100 (Nl4)sMo,~ • 4H20 or H~4
NICKEL, CADMIUM, & LEAD 761
Vapor or hydride generationAAS also can be usedfor the analysisof Pb

in soils. Li et al. (1990) usedoxalic acid, ammoniumcerium(IV) nitrate, potas-
sium tetraborate,and NaOH to generatelead hydride to determinePb in soils.
Inductively CoupledPlasmaAtomic EmissionSpectrophotometry
This method has rapidly gained favor as the method of choice for multi-
elementanalysisof soil digestionand extractionsolutions.Thompsonand Walsh
(1989) edited a handbookon ICP spectrometrythat containsinformation on the
applicationof ICPAES to analysisof geologicalmaterialsincluding soils. Matrix
effectsare largely absentwith this technique,the dynamicrangeis wide, and the
techniqueis better for the determinationof refractory elements(e.g., AI, Mo)
than AAS, but spectralinterferencescan be severe.Winge et al. (1985) give tab-
ular and graphical information on spectralline intensitiesand interferencesfor
most elementsand particularly major elementsin geologicalmaterials.Interele-
ment and backgroundcorrectionsare now routinely possibleusing today'sfully
automatedinstrumentsin which instrumentoperationis controlledby a comput-
er programthat performsall corrections.
Direct analysisof aqueoussuspensionsof soils and other geological mate-
rials is possible,but sampleuniformity and particle size are important consider-
ations (Jacksonet aI., 1991).
Separationand ConcentrationMethod for Atomic Absorption

Spectrometryor Inductively CoupledPlasmaAtomic Emission
Spectrometry
If the soil extract containsexcessivelevels of interfering ions or too low

levels of Ni, Cd, or Pb for accuratequantitationby flame AAS or ICPAES, then
separationand concentrationof theseelementswill extend the lower limits of
quantitationas well as eliminate some interferences.Methodsof separationand
concentrationinclude ion exchange,precipitation,and chelation-solventextrac-
tion. The latter approachhas beenwidely used in the analysisof soils and other
geological materials(Page& Ganje, 1970; Balraadjsing,1974; Kinrade & van
Loon, 1974; 0ien & Gjerdingen, 1977; Viets, 1978; Viets et aI., 1984;
Donaldson, 1989). Additional information on preconcentrationmethods to
enhanceanalyticalsensitivity and eliminate matrix interferencescan be found in
a new volume by Alfassi and Wai (1991).
An ammonium pyrrolidine dithiocarbamate(APDC) and chloroform
extraction method is presentedhere becauseit is suitable for severalelements
including Ni, Cd, and Pb, and it can be adaptedto routine analysisusing AAS or
ICPAES (USEPA, 1976; Bradford & Bakhtar, 1991). Alternatively, for flame
AAS analysis,chelation by APDC and extraction into methyl isobutyl ketone
(MIBK) can be used (Am. Public Health Assoc., Am. Water Works Assoc.,
Water Pollut. Control Fed., 1992).
Equipment
1. Glassor teflon separatoryfunnels and funnel rack.
762 AMACHER
2. Separatoryfunnel shaker.
3. 100-mL glassbeakers.
4. Hot plate.
5. pH meter,combinationpH electrode,and pH 4 and 7 buffers.
Reagents
1. 3% APDC solution: Dissolve 3 g of APDC in 100 mL of deionized
water. Purify this solution by extractingat least three times with 10-
mL aliquots of chloroform. Preparefresh solution as needed.
2. Hydrochloric acid, 1:1 and 1:10 (vol/vol HCl/H20).
3. Ammonium hydroxide, 1:1 and 1:10 (vol/vol NH40H/H20).
4. Chloroform, high purity.
5. Nitric acid, concentratedand 1:1 (vol/vol HN03/H20).
Procedure
1. Transfersamplesolution to a separatoryfunnel and dilute to 100 mL
with deionized water. The sample volume used will dependon the
concentrationfactor desiredas well as the volume availablefor analy-
sis.
2. Adjust the pH to 2.5 by adding dilute NH40H or HCI.
3. Add 5 mL of APDC solution and mix. After 5 min add 10 mL of chlo-
roform and shakevigorously for 2 to 3 min. Allow the phasesto sep-
arateand drain the chloroform layer into a 100-mL glassbeaker.
4. Repeatthe extractionwith 5 mL of ADPC solution and 10 mL of chlo-
roform until the chloroform layer is colorless.Drain the chloroform
layer into the beakerafter eachextraction.
5. Evaporatethe chloroform in the beakerto drynesson a hot plate at low
temperature.Do not char the residue.
6. While holding the beakerat a 45° angle, slowly add 2 mL of concen-
trated HN03. Rotate and tilt the beakerto contact all of the residue
with the acid.
7. Evaporatethe HN03 to dryness over low heat. Add 2 mL of 1:1
HN03, and heatto dissolvethe residue.
8. Cool, quantitativelytransferthe solution to a lO-mL volumetric flask
with rinsing, dilute to volume with deionizedwater, and mix. Retain
the solution for analysisby AAS or ICPAES.
9. A blank, standards,and spiked samplesshould be carried through the
extraction procedureto monitor recoveriesand potential contamina-
tion.
Comments
In an earlier version of this method pyrrolidine dithiocarbamic acid
(PDCA) in chloroform was used insteadof APDC (USEPA, 1976). However,
Bradford and Bakhtar(1991) found that APDC dissolvedmore readily in HN03
than PDCA, and did not cause instability problems within the plasma for
ICPAES analysis.In our analyticalenvironmentwe found that APDC gavemore
complete recovery of Ni, Cd, and Pb than PDCA. We routinely obtain 90 to

100% recovery of these three elementsusing ADPC vs. 80 to 90% recovery
using PDCA.
Aqueoussolutions may also be buffered at a fixed pH and still achieve
maximum extraction efficiency for most trace elements.Bradford and Bakhtar
(1991) bufferedtheir extractionsat pH 5.0 with 1 M ammoniumacetate.For Ni,
Cd, and Pb, the pH rangesfor optimum extractionare 2 to 4 for Ni, 1 to 6 for Cd,
and 0.1 to 6 for Pb (Am. Public Health Assoc.,Am. Water Works Assoc.,Water
Pollut. Control Fed., 1992).
The numberof extractionstepsneededfor maximum trace elementrecov-
ery will dependon their concentrationsin the aqueoussolutions.Extractionscan
be continueduntil the chloroform layer is colorlessindicating that trace element
extractionis complete.
Large concentrationsof Fe candecreaserecoveriesof othertraceelements.
Iron can be removedfrom the aqueoussolutionsprior to extractionof the other
traceelementsby extractingwith acetylacetoneand chloroform(Burau, 1982)or
HCI and MIBK (Brooks et aI., 1985; Zhou et aI., 1985).
ACKNOWLEDGMENTS
This paperwas written and preparedby U.S. governmentemployeeson

official time, and thereforeis in the public domainand not subjectto copyright.
The use of trade namesis for the convenienceof the readeronly and does not
constituteendorsementby the USDA ForestService.
REFERENCES
Adams, F. 1971. Ionic concentrationsand activities in soil solutions. Soil Sci. Soc. Am. Proc.
35:421-426.
Adriano, D.C. 1986. Trace elementsin the terrestrialenvironment.Springer-Verlag,New York.
Alfassi, Z.B., and C.M. Wai. 1991. Preconcentrationtechniquesfor traceelements.CRC Press,Boca
Raton,FL.
Amacher,M.e. 1984. Determinationof ionic activities in soil solutionsand suspensions:Principal
limitations. Soil Sci. Soc. Am. J. 48:519-524.
American Public Health Association, American Water Works Association, and Water Pollution
Control Federation.1992. Standardmethodsfor the examinationof water and wastewater.
18th ed. Am. Public Health Assoc.,Am. WaterWorks Assoc.,and Water Pollut. Control Fed.,
Washington,De.
Anderson,J.U. 1963. An improved pretreatmentfor mineralogicalanalysesof samplescontaining
organicmatter. Clays Clay Miner. 10:380--388.
Anderson,J.U., and G.A. O'Connor.1972. Productionof permanganateion by sodium hypochlorite
treatmentto removesoil organic matter. Soil Sci. Soc. Am. Proc. 36:973-975.
Baham,J. 1984. Predictionof ion activities in soil solutions: Computerequilibrium modeling. Soil
Sci. Soc. Am. 1. 48:525-531.
Baker, D.E., and M.C. Amacher.1981.The developmentand interpretationof a diagnosticsoil test-
ing program.PennsylvaniaAgric. Exp. Stn. Bull. 826.
Baker, D.E., and M.e. Amacher.1982. Nickel, copper,zinc, and cadmium.p. 323-336.In A.L. Page
Madison,WI.
Baker, D.E., and P.F. Low. 1970. Effect of the sol-gel transformationin clay-watersystemson bio-
logical activity: II. Sodium uptakeby com seedlings.Soil Sci. Soc. Am. Proc. 34:49 56.
764 AMACHER
Baldwin, R.P.,J.K. Christensen,and L. Kryger. 1986. Voltammetricdeterminationof tracesof nick-

el(II) at a chemicallymodified electrodebasedondimethylglyoxime-containingcarbonpaste.
Anal. Chern. 58:1790-1798.
Balraadjsing,B.D. 1974. The determinationof total lead in soil. Commun. Soil Sci. Plant Anal.
5:25-37.
Barbarick,K.A, and S.M. Workman. 1987.Ammonium bicarbonate-DTPA and DTPA extractionsof
sludge-amended soils. J. Environ. Qual. 16:125-130.
Bartlett, R.J., and B.R James.1980. Studyingdried, storedsoil samples-some pitfalls. Soil Sci. Soc.
Am. J. 44:721-724.
Basta,N.T., and M.A Tabatabai.1990. Ion chromatographicdeterminationof total metal sin soils.
Soil Sci. Soc. Am. J. 54:1289-1297.
Bauer,C.F., and C. Kheboian.1988. Responseto commentson testingof the accuracyof an extrac-
tion procedurefor determining the partitioning of trace metals in sediments.Anal. Chern.
60:1477.
Belzile, N., P. Lecomte,and A Tessier.1989. Testing readsorptoinof trace elementsduring partial
chemicalextractionsof bottom sediments.Environ. Sci. Technol.23:1015-1020.
Bettinelli, M, U. Baroni, and N. Pastorelli.1989. Microwave oven sampledissolutionfor the analy-
sis of environmentaland biological materials.Anal. Chim. Acta 225:159-174.
Bidwell, AM., and RH. Dowdy. 1987. Cadmiumand zinc availability to com following termination
of sewagesludgeapplications.J. Environ. Qual. 16:43842.
Bingham,F.T., AL. Page,R.J. Mahler, and T.J. Ganje. 1975. Growth and cadmiumaccumulationof
plants grown on a soil treated with cadmium-enrichedsewagesludge. J. Environ. Qual.
4:207-211.
Bradford, G.R, and D. Bakhtar. 1991. Determinationof trace metals in saline irrigation drainage
waterswith inductively coupledplasmaoptical emissionspectrometryafter preconcentration
by chelation-solventextraction.Environ. Sci. Technol. 25:1704-1708.
Brooks, R.R, A Siriwardena,and J. Lee. 1985.A plasmaemissionmethodfor determiningelemen-
tal constituentsof geologicalmaterialswith high iron content.J. Chern. Geol. 53:31-35.
Brown, J.R (ed.). 1987. Soil testing: Sampling, correlation,calibration, and interpretation.SSSA
Spec.Pub. 21. SSA, Madison,WI.
Browne, C.L., Y.-M. Wong, and D.R Buhler. 1984.A predictivemodel for the accumulationof cad-
mium by container-grownplants.J. Environ. Qual. 13:184-188.
Burau, R.O. 1982. Lead. p. 347-365.In AL. Pageet al. (ed.) Methodsof soil analysis.Part 2. 2nd
ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Chao,T.T. 1972. Selectivedissolutionof manganeseoxides from soils and sedimentswith acidified
hydroxylamine hydrochloride.Soil Sci. Soc. Am. Proc. 36:764-768.
Chao,T.T. 1984. Use of partial dissolutiontechniques ingeochemicalexploration.J. Geochem.Expl.
20:101-135.
Chao, T.T., and L. Zhou. 1983. Extraction techniquesfor selectivedissolution of amorphousiron
oxides from soils and sediments.Soil Sci. Soc. Am. J. 47:225-232.
Council on Soil Testing and Plant Analysis. 1980. Handbookon referencemethodsfor soil testing.
Counc. Soil Test. Plant Anal., Athens,GA
Donaldson,E.M. 1989. Determinationof coibalt, nickel, lead,bismuth,and indium in ores,soils, and
relatedmaterialsby atomic absorptionspectrometryafter separationby xanthaneextraction.
Talanta36:543-548.
Elkhatib, E.A., lL. Hem, and T.E. Staley. 1987. A rapid centrifugationmethod for obtaining soil
Epstein,M.S., G.R Carnrick, W. Slavin, and N.J. Miller-Ihli. 1989. Automatedslurry sampleintro-
duction for analysisof a river sedimentby graphite furnace atomic absorptionspectropho-
tometry. Anal. Chern. 61:1414-1419.
Figura, P., and B. McDuffie. 1980. Determinationof labilities of solubletrace metal speciesin aque-
ous environmentalsamplesby anodic stripping voItammetry and Chelex column and batch
methods.Anal. Chern. 52:1433-1439.
Fujii, R, and RB. Corey. 1986. Estimationof isotopically exchangeablecadmiumand zinc in soils.
Soil Sci. Soc. Am. J. 50:306-308.
Fujii, R., L.L. Hendrickson,and RB. Corey. 1983. Ionic activities of trace metalsin sludge-amend-
ed soils. p. 179-180.In E.A. Jenneand RE. Wildung (ed.) Biological availability of trace
metals. Chemical estimation,ecological, and health implications. Proc. Hanford Life Sci.
Symp., 21st, Richland, WA 4-8 Oct. 1981. Sci. Total Environ., Elsevier Scientific Publ.,
Amsterdam.
Gardner,W.H. 1986. Water content.p. 493-544.In A Klute (ed.) Methodsof soil analysis.Part 1.
2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Gee, G.w., and 1.w. Bauder.1986. Particle-sizeanalysis.p. 383-411.In A Klute (ed.) Methodsof
Gruebel,KA, J.A. Davis, and J.O. Leckie. 1988.The feasibility of usingsequentialextractiontech-
niquesfor arsenic andseleniumin soils and sediments.Soil Sci. Soc. Am. 1. 52:390 397.
Haghiri, E 1973. Cadmiumuptakeby plants.1. Environ. Qual. 2:93-96.
Haq, AU., and M.H. Miller. 1972.Predictionof availablesoil Zn, Cu, and Mn usingchemicalextrac-
tants. Agron. 1. 64:779-782.
Haq, AU., T.E. Bates,and Y.K Soon.1980.Comparisonof extractantsfor plant-availablezinc, cad-
mium, nickel, and copperin contaminatedsoils. Soil. Sci. Soc. Am. J. 44:772-777.
Harrison,R.M., D.P.H. Laxen, and S.I. Wilson. 1981. Chemicalassociationsof lead, cadmium,cop-
per, and zinc in streetdustsand roadsidesoils. Environ. Sci. Technol. 15:1378-1383.
Hee, S.S.Q.,andJ.R. Boyle. 1988.Simultaneousmultielementalanalysisof someenvironmentaland
biological samples by inductively coupled plasma atomic emission spectrometry.Anal.
Chern. 60:1033-1042.
Hewitt, AD., and C.M. Reynolds.1990. Microwave digestion of soils and sedimentsfor assessing
contaminationby hazardouswaste metals. Spec. Rep. 90-19. U.S. Army Corps Eng. Cold
RegionsRes. Eng. Lab., Hanover,NH.
Hinds, M.W., and KW. Jackson. 1987. Lead atomization from soil by slurry introduction elec-
trothermalatomizationatomic absorptionspectrometry:Part 1. Effects of matrix components
on the absorbanceversustime profile. 1. Anal. At. Spectrom.2:441-445.
Hinds, M.W., and KW. Jackson.1990. Comparisonof palladium nitrate and chloride as a chemical
modifier for the determinationof lead in solutionsand soil slurriesby electrothermalatomic
absorptionspectrometry.J. Anal. At. Spectrom.5:199-202.
Hoenig,M., P. Regnier,and R.I. Wollast. 1989.Automatedtracemetal analysisof slurried solid sam-
ples by graphite furnace atomic absorptionspectrometrywith application to sedimentsand
suspendedmattercollectedin natural waters.J. Anal. At. Spectrom.4:631-634.
Horowitz, AJ. 1991. A primer on sediment-traceelementchemistry.2nd ed. Lewis Publ., Chelsea,
MI.
Hossner,L.R. 1996. Dissolution for total elementalanalysis. p. 49-64. In D.L. Sparkset al. (ed.)
Madison,WI.
Jackson,L.L., P.A Boedecker,T.L. Fries, and PJ. Lamothe. 1993. Geologicaland inorganic materi-
als. Anal. Chern.65:12R-28R.
Jackson,L.L., P.A Boedecker,T.L. Fries, and P.I. Lamothe. 1995. Geologicaland inorganicmateri-
als. Anal. Chern. 67:71R-85R.
Jackson,L.L., T.L. Fries, J.N. Grossman,B.-S.W. King, and P.J. Lamothe. 1991. Geological and
inorganicmaterials.Anal. Chern. 63:33R-48R.
John, M.K 1972. Lead availability related to soil propertiesand extractablelead. J. Environ. Qual.
1:295-298.
John, M.K, CJ. VanLaerhoven,and H.H. Chuah. 1972. Factorsaffecting plant uptake and phyto-
toxicity of cadmiumaddedto soils. Environ. Sci. Technol. 6:1005-1009.
Jones,J.B., Jr. 1989. Soil testing and plant analysislaboratory manual. Benton Lab., Inc., Athens,
GA.
Jones,L.H.P., S.C. Jarvis, and D.W. Cowling. 1973a. Lead uptakefrom soils by perennialryegrass
and its relation to the supply of an essentialelement(sulphur). Plant Soil 38:605-619.
Jones, R.L., T.D. Hinesley, and E.L. Ziegler. 1973b. Cadmium content of soybeansgrowing in
sewagesludgeamendedsoil. J. Environ. Qual. 2:351-353.
Jones,R.L., T.D. Hinesley,E.L. Ziegler, andJ.J.Tyler. 1975. Cadmiumand zinc contentsof corn leaf
and grain producedby sludgeamendedsoil. 1. Environ. Qual. 4:509-514.
Kane,J.S. 1988. Optimizationof flame parametersfor simultaneousmultielementatomic absorption
spectrometricdeterminationof trace elemtnsin rocks. J. Anal. At. Spectrom.3:1039-1045.
Khasawneh,EE. 1971. Solution ion activity and plant growth .Soil Sci. Soc. Am. Proc. 35:426-436.
Kheboian,C., and e.E Bauer.1987.Accuracyof selectiveextractionproceduresfor metal speciation
in model aquaticsediments.Anal. Chern. 59:1417-1423.
Kingston, H., and L.B. Jassie(ed.). 1988.Introductionto microwavesamplepreparation:Theory and
practice.Am. Chern. Soc.,Washington.
soils and rocks. Environ. Sci. Technool. 17:362-368.
Kinrade,J.D., andJ.e.van Loon. 1974.Solventextractionfor usewith flame atomicabsorptionspec-
trometry. Anal. Chern.46:1894-1898.
766 AMACHER
Korcak, R.F., and D.S. Fanning.1978. Extractability of cadmium,copper,nickel, and zinc by double
acid versusDTPA and plant contentat excessivesoil levels. J. Environ. Qual. 7:506-512.
Kunze, G.W., and J.B. Dixon. 1986. Pretreatment formineralogicalanalysis.p. 91-100.In A. Klute
(ed.) Methodsof soil analysis.Part 1. 2nd ed. Agron. Monogr. 9. ASA and SSSA, Madison,
WI.
Lamothe,PJ.,T.L. Fries, and J.1. Consul. 1986. Evaluationof a microwaveoven systemfor the dis-
solution of geologicsamples.Anal. Chern.58:18811886.
Latterell, 1.1., R.H. Dowdy, and W.E. Larson. 1978. Correlation of extractablemetals and metal
uptake of snap beans grown on soil amended with sewage sludge. J. Environ. Qual.
7:435-440.
Lavkulich, L.M., and J.H. Wiens. 1970. Comparisonof organicmatterdestructionby hydrogenper-
oxide and sodiumhypochloriteand its effectson selectedmineral constituents.Soil Sci. Soc.
Am. Proc. 34:755-758.
Li, 1., Y. Liu, and T. Lin. 1990. Determinationof lead by hydride generationatomic absorptionspec-
trometry: Part 1. A new medium for generatinghydride. Anal. Chim. Acta 231:151-155.
Lindsay, W.L., and W.A Norvell. 1978. Developmentof a DTPA soil test for zinc, iron, manganese,
and copper.Soil Sci. Soc. Am. 1. 42:421-428.
MacCarthy,P., R.W. Klusman,and S.w. Cowling. 1991.Wateranalysis.Anal. Chern.63:301R-342R.
Mehlich, A 1984. Mehlich No.3 soil test extractant:A modification of Mehlich No.2. extractant.
Commun.Soil Sci. Plant Anal. 15:1409--1416.
Miller, J.E., 1.1. Hassett,and D.E. Koeppe. 1975. The effect of soil propertiesand extractablelead
levels on lead uptakeby soybeans.Commun.Soil Sci. Plant Anal. 6:339--347.
Miller, w.P., and w.w. McFee. 1983. Distribution of cadmium,zinc, copper, and lead in soils of
industrial northwesternIndiana.J. Environ. Qual. 12:29--33.
Miller, w.P., D.e. Martens,and L. W. Zelazny. 1986. Effect of sequencein extractionof tracemetals
from soils. Soil Sci. Soc. Am. J. 50:598-{)01.
Millward, e.G., and P.O. K1uckner. 1989. Microwave digestiontechniquefor the extractionof min-
erals from environmentalmarine sedimentsfor analysis by inductivelky coupled plasma
atomic emissionspectrometryand atomic absorptionspectrometry.J. Anal. At. Spectrom.
4:709--713.
Misra, S.G.,and P. Pande.1974.Evaluationof a suitableextractantfor availablenickel in soils. Plant
Soil 41:697-700.
Misra, S.G., and G. Pandey.1976. Evaluationof suitableextractantfor availablelead in soils. Plant
Soil 45:693--Q96.
Mogk, D.W. 1990. Application of Auger electronspectroscopyto studiesof chemical weathering.
ReviewsGeophys.28:337-356.
Mubarak,A, and R.A Olsen. 1976. Immiscible displacementof the soil solutionby centrifugation.
Soil Sci. Soc. Am. J. 40:329--331.
Nelson,J.L., L.C. Boawn, and F.G. Viets, Jr. 1959. A methodfor assessingzinc statusof soils using
acid-extractablezinc and "titratable alkalinity" values.Soil Sci. 88:275-283.
Norvell, W.A 1984. Comparisonof chelatingagentsas extractantsfor metalsin diversesoil materi-
als. Soil Sci. Soc. Am. J. 48:1285-1292.
Norvell, W.A, and w.L. Lindsay. 1982. Estimationof the concentrationof Fe3+ and the (Fe3+)(OH'-
)3 ion product from equilibria of EDTA in soil. Soil Sci. Soc. Am. 1. 46:710-715.
O'Connor,G.A 1988. Use and misuseof the DTPA soil test. 1. Environ. Qual. 17:715-718.
0ien, A, and K. Gjerdingen.1977. Determinationof cadmiumand lead in soils by meansof solvent
extractionand atomic absorption.Acta Agric. Scand.27:67-70.
Opydo,1. 1989.The determinationof Zn, Cd, Pb, and Cu in soil extractsby anodicstripping voltam-
metry. Water Air Soil Pollut. 45:43-48.
Page,AL., and TJ. Ganje. 1970. Accumulationsof lead in soils for regionsof high and low motor
vehicle traffic density.Environ. Sci. Technol.4:140-142.
Petersen,R.G., and L.D. Calvin. 1986. Sampling.p. 33-51.In A Klute (ed.) Methodsof soil analy-
sis. Part 1. 2nd ed. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Petersen,R.G., and L.D. Calvin. 1996. Sampling.p. 1-17. In D.L. Sparkset al. (ed.) Methodsof soil
Pickering, W.F. 1981. Selectivechemical extraction of soil componentsand bound metal species.
CRC Cril. Rev. Anal. Chern. 2:233-266.
Rapin, F., A. Tessier,P.G.e.Campbell,and R. Carigan.1986. Potentialartifactsin the determination
of metalpartitioningin sedimentsby a sequentialextractionprocedure. Environ.Sci. Technol.
20:836-840.
Rasberry,S. 1991. Referencematerials.Am. Environ. Lab. 3:36.
Rhoades,1.D. 1982. Solublesalts.p. 167-179.In A.L. Pageet al. (ed.) Methodsof soil analysis.Part
2. 3rd ed. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Ross, D.C., and R.I. Bartlett. 1990. Effects of extractionmethodsand samplestorageon properties
of solutionsobtainedfrom forestedspodosols.J. Environ. Qual. 19:108-113.
Rygh, G., and K.w. Jackson.1987. Direct determinationof cadmium in soil slurries by microsam-
piing cup atomic absorptionspectrometry.J. Anal. At. Spectrom.2:397-400.
Salomons,w., and U. Forstner.1984. Metals in the hydrocycle.Springer-Verlag,Berlin.
Santillan-Medrano,1., and1.1. Jurinak. 1975.The chemistryof lead andcadmiumin soil: Solid phase
formation. Soil Sci. Soc. Am. Proc. 39:851-856.
Sauerbeck,D.R., and A. Hein. 1991. The nickel uptake from different soils and its prediction by
chemicalextractions.Water Air and Soil Pollut. 57-58: 861-871.
Schalscha,E.B., M. Morales, I. Vergara, and A.C. Chang. 1982. Chemical fractionation of heavy
metalsin wastewater-affected soils. 1. Water Pollut. Contr. Fed. 54:175-180.
Shuman,L.M. 1982. Separatingsoil iron- and manganese-oxide fractions for microelementanalysis.
Soil Sci. Soc. Am. J. 46:1099-1102.
Shuman,L.M. 1983. Sodiumhypochloritemethodsfor extractingmicroelementsassociatedwith soil
organicmatter. Soil Sci. Soc. Am. J. 47:656-660.
Shuman,L.M. 1985. Fractionationmethodfor soil microelements.Soil Sci. 140:11-22.
Singh, B.R., and R.P. Narwal. 1984.Plantavailability of heavymetalsin sludge-treated soil: II. Metal
extractability comparedwith plant metal uptake.J. Environ. Qual. 13:344-349.
soil test and ICPS. p. 165-190. In BA Stewart (ed.) Advances in soil science. Vol. 16.
Soltanpour,P.N., G.W. Johnson,S.M. Workman, lB. Jones,Jr., and R.O. Miller. 1996. Inductively
coupledplasmaemissionspectrometryand inductively coupledplasma-mass spectrometry.p.
91-139. In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemicalmethods.SSSA
Sommers,L.E., and W.L. Lindsay. 1979. Effect of pH and redox on predictedheavy metal-chelate
equilibria in soils. Soil Sci. Soc. Am. 1. 43:39-46.
Sparks, D.L. 1984. Ion activities: An historical and theoretical overview. Soil Sci. Soc. Am. J.
48:514-518.
Sposito, G. 1984. The future of an illusion: Ion activities in soil solutions. Soil Sci. Soc. Am. J.
48:531-536.
Sposito,G., L.I. Lund, and A.c. Chang.1982.Trace metal chemistryin arid-zonefield soils amend-
ed with sewagesluge:I. Fractionationof Ni, Cu, Zn, Cd, andPb in solid phases.Soil Sci. Soc.
Am. J. 46:26G--264.
Street,J.J., W.L. Lindsay, and B.R. Sabey. 1977. Solubility and plant uptake of cadmium in soils
amendedwith cadmiumand sewagesludge.J. Environ. Qual. 6:72-77.
Sulcek,Z., and P. Povondra.1989.Methodsof decompositionin inorganicanalysis.CRC Press,Boca
Raton, FL.
Symeonides,C., and S.G. McRae. 1977. The assessmentof plant-availablecadmium in soils. J.
Environ. Qual. 6:12G--123.
Tessier,A., and P.G.c. Campbell. 1988. Commentson the testing of the accuracyof an extraction
procedure for determining the partitioning of trace metals in sediments.Anal. Chern.
60:1475-1476.
Tessier,A., P.G.C. Campbell,and M. Bisson. 1979. Sequentialextractionprocedurefor the specia-
tion of particulatetrace metals.Anal. Chern. 51:844-851.
Thompson,M., and J.N. Walsh (ed.).1989.Handbookof inductively coupledplasmaspectrometry.
Routledge,Chapman,& Hall, New York.
Tipping, E., N.B. Hetherington.H. Hilton. D.W. Thompson,E. Bowles, and J. Hamilton-Taylor.
1985. Artifacts in the use of selectivechemicalextractionto determinedistributionsof met-
als between oxidesof manganeseand iron. Anal. Chern. 57:1944-1946.
United StatesEnvironmentalProtectionAgency. 1976. Methodsfor chemicalanalysisof water and
wastes.EPA-625-/6-74-003a.USEPA, Cincinnati, OH.
United StatesEnvironmentalProtectionAgency. 1986. Test methodsfor evaluatingsolid wastes.
USEPA SW 846. U.S. Gov. Print. Office, Washington,DC.
Valdares,J.M.A.S., M. Gal., U. Mingelgrin, and A.L. Page.1983. Someheavy metalsin soils treat-
ed with sewagesludge, their effects on yield, and their uptake by plants. J. Environ. Qual.
12:49-57.
Varma,A. 1990.CRC handbookof furnaceatomic absorptionspectroscopy.CRC Press,Boca Raton,
FL.
768 AMACHER
Viets, J.G. 1978. Determinationof silver, bismuth, cadmium, copper, lead, and zinc in geological
materialsby atomic absorptionspectrometrywith tricapylylmethylammoniumchloride.Anal.
Chern.50:1097-1101.
Viets, J.G., R.M. O'leary, and J.R. Clark. 1984. Determinationof arsenic,antimony, bismuth, cad-
mium, copper,lead, molybdenum,silver, and zinc in geologicalmaterialsby atomic absorp-
tion spectrometry.Analyst 109:1589-1592.
Westerman,R.L. (ed.). 1990. Soil testingand plant analysis.3rd ed. SSSA,Madison,WI.
Whelan,B.R, and N.J. Barrow. 1980. A study of a methodfir displacingsoil solution by centrifug-
ing with an immiscible liquid. J. Environ. Qual. 9:315-319.
Winge, RK., V.A. Fassel,V.J. Peterson,and M.A. Floyd. 1985. Inductively coupledplasma-atomic
emissionspectroscopy:An atlasof spectralinformation. Elsevier,Amsterdam.
Wolt, J., and J.G. Graveel. 1986. A rapid routine method methodfor obtyainingsoil solution using
vacuumdisplacement.Soil Sci. Soc. Am. J. 50:602-605.
Wright, RJ., andT. Stuczynski.1996.Atomic absorptionandflame emissionspectrometry.p. 65-90.
In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSA Book
Ser. 5. SSSAand ASA, Madison,WI.
Yang, J.E.,E.O. Skogley,SJ.Georgitis,B.E. Schaff,andA.H. Ferguson.1991.Phytoavailabilitysoil
test: Developmentand verification of theory. Soil Sci. Soc. Am. J. 55:1538-1365.
Zhou, L., T.T. Chou,and RE Sanzolone.1985. Removalof iron interfgerencesby solventextraction
for geochemicalanalysisby atomic absorptionspectrometry.talanta32:475-478.
Published 1996
Chapter29
Mercury
J. G. CROCK, u.s. Geological Survey, Denver Federal Center,

Denver, Colorado
Mercury hasbeena focus of scientific studyfor at leastthe past2500yr, because

of its uniquechemicalandphysicalpropertiesamongthe metallic elements.His-
torical recordsindicate that Hg was usedin the Mediterraneanareaas early as
500 B.C. for extractingpreciousmetalsfrom ores by amalgamation.Alchem-
ists, for more than 1000yr, thoughtthat Hg was the key to the transformationof
basemetalsinto Au. Mercury is namedafter the messengerof the Romangods
due to its greatmobility, as seenin both the gaseousand liquid elementalstate.
The symbol,Hg, is derivedfrom the Latin word "hydragyrum"or "liquid silver"
(Greenwood& Earnshaw,1984, p. 1395-1422).Contemporaryinterest in Hg
remainshigh, not becauseof its unique room-temperatureliquid form, but be-
causeof its extremetoxic natureto most life forms and its widespreadoccur-
rence. Keen interestis demonstratedby the recent internationalconferenceon
Hg as a worldwide pollutant(Lindqvist, 1991).
A summaryof the generalchemistryof Hg is presentedin Greenwoodand
Earnshaw(1984, p. 1395-1422)and its geochemistryis summarizedin Roseet
al. (1979). Mercury is a relatively rare elementwith an averagecrustal abun-
danceof only 0.08 mg kg-I (Roseet aI., 1979). Mercury is presentin most geo-
logical materialsas a trace constituentbut doesform its own sulfide mineral,
cinnabar(HgS). Mercury is normally associatedwith the chalcophile(S-loving)
elements(e.g., As, Bi, Sb, Se, Ag, Au, Zn, and Pb) (Roseet aI., 1979).Average
abundances for commongeologicalmaterialsare (Roseet aI., 1979): (i) igneous
rocks-O.OO4mg kg-I, ultramafic; 0.01 mg kg-I, mafic; 0.04 mg kg-I, granitic;
(ii) sedimentaryrocks-O.04mg kg-I, limestone;0.03 mg kg-I, sandstone;0.02
to 0.40 mg kg-I, shale;(iii) soils-O.056mg kg-I; (iv) plant ash-O.01mg kg-I;
(v) fresh water-O.07~g L- I , with 2 ~g L- I as the safe drinking water health
limit establishedby the u.s. EnvironmentalProtectionAgency (USEPA); and,
(vi) air-3 ng/m3.
Mercury has found a wide diversity of usesbasedon its electricalproper-
ties, toxicity, high vapor pressureat room temperature,and unique liquid-metal
form. These uses include laboratory equipmentfor temperatureand pressure
measurements, electrodes,ultraviolet lamps, diffusion pumps, dental prepara-
Copyright <ill 1996 Soil ScienceSociety of America and American Society of Agronomy, 677 S.
Book Seriesno. 5.
769
770 CROCK
tions, batteries,explosives,catalysts,and preciousmetal recoveryby amalgama-

tion. Mercury also is usedextensivelyin antiseptics,slimicides,fungicides,and
antifouling paints(Varma, 1984).
Mercury'srelatively high vaporpressureat room temperatures (1.22 x 10-3
mm at 20°C) resultsin appreciableamountsof atmosphericHg from both natur-
al and anthropogenicsources(Adriano, 1986, p. 298-328).The anthropogenic
input of Hg into the environmenthas been estimatedby Nriagu and Pacyna
(1988) to be as high as 6 x 106 kg/yr. Whereasthis amountof Hg seemslarge,
the estimatedtotals over all historic time for Hg producedand releasedinto the
environmentby both anthropogenicand natural sourcesare even more impres-
sive. Thesehavebeenestimatedat 638 x 1Q6 kg Hg produced,and 741 x 106 kg
releasedinto the atmosphere,118 x 106 kg releasedin water, and 806 x 106 kg
releasedinto the soil (Andren & Nriagu, 1979). Combustionof fossil fuels and
volcanic activity account for most of the Hg releasedinto the atmosphere.
Weatheringof soils and rocks accountfor most Hg releasedinto water andsoils.
Various industrial inputs tend to be localized, but can be very important to the
total Hg releasedover a small area.The large input of Hg into the environment
has resultedin its widespreadoccurrencein the entire food chain, and it now is
considereda healthhazardfor many areas,especiallywhereindustrial input is a
point sourceof significantHg contamination.
The analyticalchemistis faced with severalchallengeswhen determining
Hg in soils and sediments.Theseinclude: (i) widespreadcontaminationof the
atmosphereleading to possiblesignificant blank levels and samplecontamina-
tion (Welz, 1985), especiallyfrom reagentsand laboratoryglasswares.Mercury
is ubiquitousin most laboratoryenvironments,due to the use of thermometers,
barometers,electrodes,and other equipment. Significant contaminationmay
occurif any unprotectedsamplesor labwaresare exposedto a laboratoryatmos-
pherefor any length of time (Williams & Funston,1987); (ii) volatility of Hg or
its compoundsduring the grinding or digestingof the sample;(iii) wide rangeof
valuespossible,ranging from the low nanogramper gram in backgroundareas
to hundredsof milligrams per kilogram in contaminatedareas;and, (iv) sample
inhomogeneity.All of these problems must be addressedfor the successful
determinationof Hg in soils and sediments.
Many analyticalinstrumentsand methodshavebeenappliedto the analy-
sis of soils and sedimentsfor the determinationof Hg. However,for most stud-
ies, the method should have a detectioncapability in the nanogramper gram
range(Williams & Funston,1987). Currentreviewsfor the determinationof Hg
in soils and sedimentsand similar matricesare presentedin the analyticalchem-
istry literature,for example,Geological and Inorganic Materials (Jacksonet al.,
1991) and Environmental Analysis (Clement et aI., 1991). Neutron activation
analysis(Van Delft & Vos, 1988), inductively coupledplasma-atomicemission
spectrometry(ICP-AES), inductively coupledplasma-massspectrometry(ICP-
MS), and inductively coupled plasma-atomicfluorescencespectrometry(ICP-
AFS) (Montaser& Golightly, 1992) plus many other instrumentalmethodshave
beenapplied to the determinationof Hg. Of thesemethods,ICP-MS offers the
ability to measurethe isotopic abundances of Hg and, as a result, isotopic label-
ing for studiesis possible.However,its detectionlimits (severalmg L-1) may be
MERCURY 771
too high for useful studiesat this time. If the matrix problemsthat are common
to ICP-AFS can be eliminated,this methodmay becomea preferredmethodfor
the analysisof soils and sediments.Mercury has been determinedby graphite
furnace-atomicabsorption spectrometrywith Zeeman backgroundcorrection
(Grobenskiet aI., 1985),with solventextraction(Sanzolone& Chao, 1983), and
with a gold-coatedgraphite tube (Lee et aI., 1989). Mudroch and Kokotich
(1987) describeda methodusing a gold-film analyzerto determineHg. The gold
film's electricalresistancechangesproportionatelyto the amountof Hg amalga-
mated.
The most successfuland widely used method of Hg determinationstill
remains cold vapor-atomic absorptionspectrometry(CV-AAS) basedon the
original work of Hatch and Ott (1968). This is the acceptedmethodof analysis
by the USEPA (Lobring & Potter, 1992). The broad acceptanceof CV-AAS for
the determinationof Hg was demonstratedat the recentinternationalconference
on Hg as a global contaminantwhere CV-AAS was the method of choice for
most of the investigationspresented(Lindqvist, 1991). Significant advancesin
CV-AAS have centeredaroundautomationof the determinationas demonstrat-
ed by the continuous-flowsystemsand more recentlyby the flow-injection meth-
ods (Burbuera,1989; Welz et aI., 1992) and various new approachesin sample
digestions.Microwave digestionsof difficult samplesis becomingthe preferred
method.On-line microwave digestion for Hg determinationhas recently been
reported(Welz et aI., 1992). Review of sampledigestionmethodsfor Hg are pre-
sentedby Chaoand Sanzolone(1992) and Van Delft and Vos (1988).
TOTAL MERCURY IN SOILS AND SEDIMENTS

Introduction
The low-temperatureand pressuregeochemicalcycle of Hg has receiveda

great deal of attention in recent years [e.g., Andersson, 1979; D'ltri, 1990;
Kataba-Pendias& Pendias, 1984; Lindqvist, 1991; Moore & Ramanoorthy,
1984; and, Williams & Funston,1987]. The common theme in the literature is
the complexity and diversity observedin the controlsof Hg in the surficial envi-
ronment.Due to Hg's complex natureand the scopeof this chapter,no attempt
will be madeto presenta comprehensivereview of Hggeochemistry.
Severalfactors controlling the occurrence,distribution, and abundanceof
Hg in soils and sedimentsare: (i) the high volatility of Hg at room temperature,
both as the free metal and as organiccompounds;(ii) the chalcophilic natureof
Hg to form very strong chemicalbondswith S, which exists as either the inor-
ganic sulfide ion or associatedwith organic matter; (iii) the formation of rela-
tively stableorganomercurycompounds,such as methylmercuryand ethylmer-
cury; (iv) the ability of oxides,hydroxides,and clays to adsorbinorganicallyand
organicallycomplexedHg from the aqueousphase(Schuster,1991); and, (v) the
redox and pH controls of Hg, especiallyas influencedby bacteriaand fungi in
both anaerobicand aerobicenvironments.As a result of volatility, Hg tendsto be
a ubiquitouscontaminantfrom both anthropogenicand naturalsources.The great
772 CROCK
affinity of Hg for S explainsthe strongbinding of Hg to soil organicmaterialand

the stability of HgS.
Somegeneralizationsabout Hg in soils, basedon the controlling factors,
are: (i) Hg tendsto concentratein the surfacehorizons where organic matter is
generally the highest. Humic substancesin water, soils, and sedimentsvery
strongly adsorbHg. As a result, the top layer of soil or sedimentact as a barrier
in the mobility of Hg in thesesystems(Johannsonet aI., 1991). (ii) Below the
surface,Hg can be mobile due to the formation of stable organomercurycom-
pounds.It is mainly the fractionationof the soil's organicmatter(dissolutionvs.
adsorption) thatdeterminesthe behaviorand distribution of Hg in the lower soil
profile. (iii) In general,for acidic soils (soil pH < 7), Hg follows the organic C
profile; and for alkaline soils (soil pH > 7) Hg follows the iron oxides/hydrox-
ides and clay profile, with adsorptionbeing the most importantcontrol. The de-
sorption of Hg from soils and sedimentstends to be a kinetically slow process
posing a problem long after the source of Hg is eliminated (Moore & Rama-
noorthy, 1984). (iv) Mercury will precipitateas a hydroxide in alkaline soil and
as a sulfide in reducing,acidic soils. Metallic mercurymay form if the soil is suf-
ficiently reducing(Adriano, 1986, p. 298-328).Mercury also can precipitateas
a phosphateor carbonateif the conditionsare proper.
One of the most important controls of Hg in soils and sedimentsis the
methylation of Hg into its toxic and mobile/volatile phases.This methylation
processis usually controlled by bacteria,fungi, or plants, in either aerobic or
anaerobicenvironments.Mercury is an enzymeand protein inhibitor. Its toxici-
ty tendsto be nonspecificand its accumulationnonreversible.
The normal, commonlyobservedlevel of Hg in soils and sedimentsis less
than0.1 mg kg-l., but concentrationsof hundredsof milligrams per kilogram have
beenmeasuredin contaminatedsurfacesoils. The Hg contentof soils is a func-
tion of the parentmaterial as well as of additionsfrom the atmosphereand other
sources,such as agricultural practices(applicationof sewagesludgeand chemi-
cal fertilizers, treatedseed,etc.). Theseadditionsresult in the Hg contentof soils
being highly variable.The phasesof Hg in soils are quite varied also, including
inorganic, organic, and adsorbedspecies.However, most of the Hg phasesin
most soils and sedimentsremain unidentified(Fang, 1978).
BecauseHg can exist in volatile phases,inorganic precipitates,organic
matter,or free metal in soils and sediments,the analystmust tailor the procedure
for the determinationof Hg to the expectedforms presentin the samples.If total
Hg is the desiredanalyte,great care must be taken not to volatilize metallic or
organic phases,and the sample preparationand digestion must be vigorous
enoughto insure completedissolution of the phasescontainingHg. Precaution
must be taken to minimize sample,reagent,and labwarecontaminationwith Hg.
Principlesof Total Mercury Analysis
Ever since its introduction by Hatch and Ott (1968), CV-AAS has
remainedthe preferredmethodof Hg analysisin soils and sediments(Risser&
Baker, 1990; Viets & O'Leary, 1992). It also has beenthe acceptedmethod by
MERCURY 773
the USEPA for determining Hg in solids (USEPA, 1986, Method 7471). The
methodhas undergonefew changessince its introduction,with the exceptionof
automationby continuousflow systems(e.g., Sturman,1985; Kennedy& Crock,
1987; Chan& Bina, 1989). Varma (1984) presentsan extensivebibliographyon
the analysisof Hg in mostmatricesand highlightsCV-AAS. The basicchemistry
of the methodhas remainedrelatively the samethroughoutthesechanges.
The CV-AAS method can be broken down into four basic steps:sample
decomposition,reduction of mercuric ion, phaseseparationof Hg vapor, and
AAS measurement.The samplemust be digestedto convertall the forms of Hg
presentinto Hg(I1). The solution then must be quantitatively reducedto form
HgO, which is strippedfrom the liquid phase.The Hgo vapor is then transported
into a suitablecell for measurementby AAS. At eachof thesesteps,great care
must be exercisedto eliminateloss throughvolatilization or adsorption,and gain
through contamination.
Summariesof applicabledigestionproceduresare given by Chaoand San-
zolone (1992), Van Delft and Vos (1988), and Welz (1985). All digestionproce-
durestend to be very oxidative with strongmineral acids, either singularly or in
mixturesof HN03, H2S04, HCl, and perchloricacid. Also usedwith theseacids
are strong oxidants,including hydrogenperoxide,potassiumor sodiumdichro-
mate, potassiumpermanganate,or vanadium pentoxide (V Z0 5) (Korunova &
Dedina, 1980). Most digestions use elevated temperaturesand/or pressureto
insure the completedissolutionof the Hg in the sample.Digestionsusing direct
heatingon a hot plate, autoclaveheating,or microwave heatingwith closed or
openvesselshaveall beenusedwith success.Theserigorous,strongly oxidizing
conditionsare requiredto preventpossibleloss of Hg.
Methodology using microwave heating in a closed vesselhas gained ap-
proval by the USEPA(USEPA, 1990, Method 3051). Its advantageis that no
lossesare reportedfor the volatile elements(Van Delft & Vos, 1988). Commer-
cially availablesystemsmake this an attractivemethodof digestionwhen diffi-
cult or a limited numberof samplesare to be analyzed.In general,microwave
digestionsare fasterfor a small numberof samples;but, if large numbersof sam-
ples are to be analyzedroutinely, closedvesselsheatedin a heating block on a
hot plate is the preferred,most efficient method(Van Delft & Vos, 1988).
A closedvesselis usually preferredat elevatedtemperaturesand pressures
to minimize potentiallossesof mercury. If elevatedtemperaturesor pressuresare
not used,incompletedigestionand oxidation are observed.In the digestionstep,
it is critical for the Hg to be quantitatively convertedto Hg(II). Forms of Hg
other than Hg(II), especiallythe organomercuryforms, will either be volatilized
or will not be convertedto Hgo in the next reducingstep. This volatilization or
nonconversionare important becauseHg is commonly associatedwith organic
material, especiallyin the surfacesoil horizonsand sedimentsas statedearlier.
A total decompositionof the silicate matrix, as with hydrofluoric acid
(HF), is usually not required, especially if the sample is sufficiently ground.
Mercury doesnot form any natural silicate mineralsdue to its large ionicradius
(Greenwood& Earnshaw,1984, p. 1395-1422).An exceptionmay be the case
where the Hg-containing phase is occluded and dispersedwithin a silicate
matrix.
774 CROCK
Stannouschloride (SnCI2) or sodiumborohydride(NaB14) are most com-

monly usedto reduceHg(II) to Hgo. Stannouschloride is the most commonly
usedreductant,but NaB14 is becomingmore popular. Sodiumborohydrideis a
strongerreductantcapableof reducingorganomercuryforms to Hgo and gener-
ateslarge amountsof H2 gas,thus eliminating the needof an auxiliary gasflow.
A disadvantageof NaBH4 is a less stable baselineresulting from the H2 gas
evolved. It is in this reduction step where most of the chemical interferences
occur for the CV-AAS method.Elementalinterferenceshave been noted from
the preciousmetals (including Au, Pd, Pt, and Ag), Sb, As, Bi, Cu, I, and Se
(Bartha & Ikrenyi, 1982; Pratt & Elrick, 1987). Interferencesfrom these ele-
mentsdo not post a significant problemfor most soils and sedimentsbecauseof
their low concentration.If theseelementsare presentat interfering levels, they
may precipitateandlower the Hg contentby coprecipitationor adsorption.These
chemicalinterferencestend to be more severewhen NaBH4 is usedas the reduc-
tant becauseof its greaterreducingpower.
The reducedHgo is transportedto a phaseseparatorand then transported
to a cell for measurement (Tuncel & Ataman, 1980).To avoid any potentialloss
by adsorption,the surfacecontactedby the analytical gas flow containingHgo
shouldbe as small aspossibleand shouldbe madeof glassor quartz.The useof
plastic tubing should be avoided. Frequentmonitoring by blank and standard
solution measurementsare required. The distribution coefficient of elemental
Hgo betweenthe gaseous phase and the liquid analyticalstreamhas beenfound
to be relatively independentof most variables tested (Koirtyohann & Khalil,
1976).
To improve the sensitivity of the CV-AAS method, the analytical gas
streamcan be passedover a gold or silver gauzefor preconcentrationby amal-
gamation(Huffman et aI., 1972; Welz et aI., 1984). After the samplehas been
aeratedto liberatethe Hgo vapor,the precious-metalgauzeis rapidly heated,usu-
ally by induction, to rapidly releasethe Hgo. This preconcentrationstephasmin-
imal kinetic interferences,as long as the collection time is sufficient; but it does
not eliminatethe chemicalinterferencesmentionedabove.An increaseof tenfold
in sensitivity is commonly seenwith this amalgamationstep,with the increase
usually limited by contaminationor blanks.
The CV-AAS methodis basedon the absorbanceof radiationat the 253.7-
om resonancewavelengthof Hg asthe elementalHgo vaporpassesthrougha cell
in the optical pathof an atomicabsorptionspectrometer.Atomic absorptionspec-
trometry hasexcellentsensitivity for Hgo; the instrumentationis easyto use,it is
costeffective,and it readily adaptsto routine, automatedanalysisof a large num-
ber of samples.There are virtually no spectralinterferenceswith the CV-AAS
method. Possibleinterferencesfrom water vapor condensingin the cell or the
tubing are eliminatedby either drying the gasstreamor keepingthe cell and the
transmissiontubing warm at about40°C. FreeCl 2 gasmay absorbat this wave-
length, but this interferenceis eliminatedby the addition of hydroxylaminesul-
fate.
The methodspresentedhere rely on a closed-vesseldigestion using con-
centratedHN03 and sodium dichromate.The automated,continuous-flowsys-
tem is the recommended procedure[Crock et aI., 1987; Kennedy& Crock, 1987],
MERCURY 775
even if a small numberof samplesare to be analyzed.Greatercontrol of conta-

mination, easeof operation,lesschemicalinterference,and easystandardization
of the method are the main advantages.The manual method presentedis given
relatively unchangedfrom the previousedition of this book (Stewart& Bettany,
1982). A similar manualmethodis approvedby the EPA (USEPA, 1986).
Total Mercury Method

The following is a combinationof two previously reported methodsfor
total Hg in soils. The manualmethodis from Stewartand Bettany (1982) and is
relatively unchangedfrom their presentation.The automatedmethod is from
Crock et al. (1987) and Kennedyand Crock (1987) and is the recommendedpro-
cedure.Automatedmethodsare more rapid and reproducible,and less prone to
operatorvariationsand, therefore,give more reliable data.
SpecialApparatus
1. Digestion vessel.thick-walled, 30-ml Teflon round vials (30-mm o.d.,
66-mm height) with threaded caps (#G-08936-50, Cole-Palmer
InstrumentCompany,Niles, IL, or an equivalent).!
2. Aluminum heatingblock. 2.5 cm thick by 25 cm wide by 50 cm long,
with 34-mm holesdrilled through in a 5-row by lO-column matrix.
3. Hot plate. standardlaboratorywith at least a 30- by 60-cm heatingsur-
face, with capability of lO5°C surfacetemperature.
4. Heat lamps. GeneralElectric Chil ChaserDeluxe Infrared Heat Lamp.
Position aroundthe flow-through cell and the phaseseparator.!
5. Argon gas. Welding grade, cylinder with standardpressureregulating
valve.
6. Opensystem,cold-vaporapparatus(manualmethod),takenin its entire-
ty from Stewartand Bettany (1982).
...The apparatus(shown schematicallyin Fig. 29-1) is designedso that Hg
can be partitionedbetweena samplesolution and a fixed volume of air for a
set period time before venting through an absorptioncell for measurement.
The reductionflask is essentiallya 2S0-ml Erlenmeyerfitted with a 20-ml
reservoir to allow introduction of the reducing agent to the sealedsystem.
Alternatively the reductantmay be quickly added through the neck of the
flask and the flask immediately connected to the apparatus,therebyprevent-
ing loss of Hg vapor. A tight-fitting neoprenestopperor ground-glassstop-
per, throughwhich are passedPyrex gasdelivery tubes,sealsthe flask that is
connectedto the opencirculationsystemthroughnylon three-waystopcocks.
There is an empty Pyrex U-tube in line with the flask as a safetytrap to pre-
vent solution enteringthe absorptioncell.
The cylindrical absorptioncell is madeof UV transparentmaterial with flat
optical faces(Hatch and Ott, 1968)and may be readily obtainedfrom instru-
ment supply companies.Silicon rubbertubing shouldbe usedfor all connec-
tions, and onceassembledthe apparatusshouldbe checkfor gas leaksand if
necessarysealedwith silicone sealant.Either Ar or N2 may be usedas a car-
rier gas,which is introducedto the systemthrough a flow meter and needle
I Any use of tradenamesor productsis for descriptivepurposesonly and doesnot imply endorse-
ment by the U.S. GeologicalSurvey.
776 CROCK
By-Pass
I \
To Oit Hel Cylindrical AbsOfption Cell i \
in Beaker (l00mm path)
==~ rr===~
"--, -~ \
- E3---~- ---- Photo I
Flowmeter
(1-12 Vmin.)
Hollow
Cathode Mulbplier
U-l\Jbe
Trap Reduction Flask
(25Oml Erienmeyer)
Fig. 29-1. Open systemcold-vapor apparatusto be used in the manual method [see section on
"Mercury Determination(Manual),,]. Arrows indicateflow patternsduring analysis.
valve assemblyadjustedto give a flow rate of 3 L min-I. (Argon is recom-

mended.)
The absorptioncell is usually securelymountedandoptically alignedon the
burnerstemof an atomic absorptionspectrophotometer. A 40-W lamp bulb
is arrangedto shineon the absorptioncell to preventcondensationof mois-
ture inside the cell. Sincethe sensitivity of the methodis extremelygood, a
simple single beam instrument with chart recorder is sufficient. Modem
instrumentswith microprocessorcircuitry do not needa recorderif they can
be set to measurepeak height. The absorptionof Hg vapor is measuredat
253.7nm....
Apparatussimilar to that describedabove may be obtainedin a com-
plete kit from various instrument suppliers. Also a proven manual
method for determining Hg is presentedas USEPA Method 7471,
Mercury in Solid or Semisolid Waste (Manual Cold-Vapor Technique)
(USEPA, 1986).
7. Open-systemcold-vapor apparatus(automatedsystem), taken in its
entirety from Crock et al. (1987) and Kennedy and Crock (1987): auto
sampler,Bran and Luebbe,Inc. (Buffalo, IL) (formerly Technicon,Inc.,
Tarrytown, NY)1 auto samplerAAlI modified by replacing sampling
probe with a glass tube (3 by 100 mm); pump, Gilson Medical Elec-
tronic (Middleton, WI) Model Minipulse 21, eight-channel,variable-
speedperistalticpumpequippedwith standardTygon pump tubing; spe-
cial glassware,see Figs. 29-2 and 29-3 for the phaseseparatorand
flow-through cell used in this method. Thesehave been describedby
Crock et al. (1987) and Skougstadet al. (1979). Mixing coils1 are avail-
able from Bran and Luebbe, Inc. (Buffalo, IL) (formerly Technicon,
Inc., Tarrytown, NY); flow manifold, see Fig. 29-4 for the continuous
flow-through manifold for Hg determination.The flow-through cell is
positionedin the light path of a standardAAS spectrophotometer with
recorder.The standard2S3.7-nmwavelengthwith no backgroundcor-
rection and full-scale responseare usedwith a standardHg hollow cath-
MERCURY 777
r;:::===::J T~
~e
To Flow·Through Cell
Liquid to Waste
E
E
~
Mercury Phase Separator
Fig. 29-2. Phaseseparatorusedin the determinationof Hg. The whole separatoris madefrom Pyrex
glass.
ode lamp. The silver trap shown is a quartz tube (20 by 150 mm),
neckedand dimpled at both endsafter about5 g of 36-gauge(0.2 mm)
silver wire has been positioned inside. This trap will minimize the
amountof Hg vapor ventedinto the laboratory.
Reagents
Care must be taken to preventcontaminationof all reagentsand labware
used in the determinationof Hg. In general,most reagentscontain finite, mea-
surableamountsof Hg and labwarewill adsorbHg from the atmosphere.Check
To Silver·WireTrap
Anaytical Stream from
Phase Separator Smml.O.
~~
IN ~~===========!~ Ll
~ ~1'----------------l~mm --------------~·I T -
Mercury Flow Through Cell
Fig. 29-3. The flow-through cell usedfor the determinationof Hg. The two 16-mm end-windowsof
this cell are quartz,and the remainderof the cell is Pyrex glass.
IAll flow ratesasmVmin. I
Fig. 29-4. Manifold usedfor the automatedgenerationof Hg vapor.
all reagentsfor a Hg blank. It may be necessaryto selectdifferent vendorsor

manufacturinglots of reagentsto insure the lowest Hg blank. Heat all unused
labwareat 105°Covernightto drive off adsorbedHg. Soakingglasswarein 16 M
HN03 overnightalso will removeadsorbedHg.
Reagentsfor the automateddeterminationmethod [Crock et aI., 1987;
Kennedy& Crock, 1987] and for the manualmethod(Stewart& Bettany,1982):
1. 25% (weight/volume,WN) sodium dichromatesolution: dissolve500
g reagentgradeNa2Cr207• 2H20 in sufficient demineralizedwater to
bring the volume to 2.0 L.
2. Nitric acid wash: dilute 40 mL of 16 M Baker Instra-Analyzedgrade
HN031 to 4.0 L with demineralizedwater.
3. Stannouschloride solution (for the automatedmethod):dissolve100.0
g SnCI2 • 2H20 (Baker,suitablefor Hg determinationgrade)in 100 mL
concentrated(12 M) Baker Instra-Analyzedgrade HCI.I Let the solu-
tion stand for 20 to 30 min until the SnCl2 • 2H20 totally dissolves.
Dilute to 1.0 L with demineralizedwater. This solution is stable for
about 1 wk with refrigeration.
4. Complexing-reducingsolution (for automatedmethod):dissolve30.0 g
hydroxylaminehydrochlorideand 30.0 g NaCI in about 500 mL dem-
ineralized water. Add very slowly 100.0 mL concentrated(18 M)
H2S04, BakerInstra-Analyzedgrade.I When the solution is cool, dilute
to 1.0 L with demineralizedwater.
5. Reducingsolution (for manualmethod): prepareby adding 100 mL of
concentratedH2S04, 5 g NaCI, 10 g of hydroxylaminesulfate,and 20 g
of stannoussulfate (SnS04)to 600 mL of demineralizedwater. Cool,
bring to 1 L with demineralizedwater, and filter.
MERCURY 779
6. StandardHg solution: use commerciallyavailable1000 mgIL Hg solu-

tions and serially dilute to the working calibrationstandardsof 1, 5, 10,
and 20 ~g L-l; or place 1.354g of HgClzin a 1-L volumetric flask con-
taining 2oo mL concentratedHN03 and 5 g sodium dichromate.Mix
and dilute to volume with distilled water. Cool under tap water while
mixing. The working calibration standardsshould be 3.2 M HN03 and
0.5% (WN) NaZCrZ07(20 mL 16 M HN03 and0.5 g NaZCrZ07per 1oo
mL final volume.
Procedure
Preparation of Soil Digest for Total Mercury. The following digestion
procedureis from Kennedyand Crock (1987) and is applicableto most soils and
sediments.
1. Soil samplesshould be dry and groundto passa 100-mesh(0. 149-mm)
sieve.Weigh 0.100g of sampleinto a clean,thick-walled,30-mL Teflon
vessel. The threadsof the vessel should be wrappedpreviously with
Teflon tape to insure a tight sealwhen capped.Larger sampleweights,
up to 0.500g, may be usedif Hg is expectedto be low «0.1 mg kg-1)
and the organic materialis less than 1%.
2. Add 2.0 mL 16 M HN03 and 0.50 mL of the sodium dichromatesolu-
tion to the sample.Swirl gently to wet the entire sample.
3. Cap the vessel and place in the aluminum heating block for 3 h at
105°e. Removefrom the heatingblock and allow to cool completely.
(Overnightcooling is acceptable.)
4. Uncap and rinse lid with demineralizedwater into the vessel.transfer
entire contentsinto an empty, tared, 16- by 100-mm disposableflint
glasstest tube (or othersize,dependingon the autosamplerused).Bring
contentsto massof 12.00g with demineralizedwaterusing a disposable
big-bulb pipetteand a top-loadingbalancecapableof O.Ol-g sensitivity.
Mix with a vortex mixer. Do not filter the sample-letsettle or cen-
trifuge.
5. If the sampleis thoughtto be mineralizedor if the samplehasmore than
10 mg kg-1 Ag or Se, transfer the vesselcontentsto a 100-mL volu-
metric flask after digestion.Add 20 mL 16 M HN03 and 2.0 mL of the
sodium dichromatesolution and dilute to volume with demineralized
water. Use an aliquot of this solutionfor analysis.If the samplecontains
elevatedSe concentrations(>500 mg kg-1 ), use a three-pointmethod
or standardadditions (unspikedsample and two samplesspiked with
different Hg contents)to establishthe Hg content.
6. Prepareduplicate standardreferencematerials and duplicate samples
for each set of samples(usually less than 40). Replicatesof samples
totaling at least 5% of the samples should be routinely analyzed.
Aqueousstandardsof 5, 10, and 20 ~g L-l Hg are usedfor the calibra-
tion curve with eachday'sanalyses.
7. If the samplecontainslarge amountsof organic material, add approxi-
mately 50 mg of vanadiumpentoxide(V Z0 5) in Step 2 aboveto aid in
780 CROCK
the rapid oxidation of the sample.Add the V 205 to a separatereagent

blank to measurepossiblecontamination(Korunova & Dedina, 1980).
Mercury Determination(Manual). This section is taken directly with

only minor changesfrom Stewartand Bettany(1982).
.. .Transferthe diluted digestsolutioncontainingnot morethan 150 ng of Hg in the
reductionflask, dilute to about30 mL, and connectthe flask to the analyticalsys-
tem (Fig. 29-1) with the three-waynylon stopcocksarrangedso that Ar is bypass-
ing the flask andthe flask is, therefore,a closedsystem.In samplescontaininghigh
chloride,briefly purgethe systemwith Ar beforeaddingthe reducingsolution.Add
a lO-mL aliquot of the reducingsolution through the side reservoir, and stir the
solution for 2 minutesat a constantrate. The total volume of the solution in the
flask mustbe constantto maintainthe samesurfacearea/volume ratio.Discontinue
stirring, and divert the Ar flow through the reductionflask by openingthe stop-
cocks and thus venting the Hg vapor through the absorptioncell. The absorption
peaksobtainedfrom Hg vaporat 253.7nm are plottedby the recorderor displayed
directly on instrumentswith microprocessorcircuitry, enablingpeakheight to be
displayed.Finally, bubblethe exhaustedHg vaporthroughdilute Hel in a 500-ml
beakerdirectly underan exhaustvent. This analyticalsystemshouldprovide a lin-
earcurvein the rangeof 1 to 400 ng of Hg. This representsa rangeof 0.01 to 4 mg
kg-1 of soil (assuminga 0.1-g soil sampleis used),which shouldbe adequatefor
analysisof soil and sedimentsamples.The detection limit can be lowered by
increasingthe soil samplesize....
Mercury Determination(Automated).This section is condensedfrom
Crock et al. (1987) and Kennedyand Crock (1987).
After the sampledigesthasbeentransferredto a glasstest tube, placein a
carouselof an auto sampler and introduce to the continuous-flow system as
shown on Fig. 29-4. The sampleis mixed with air, a complexing-reducingsolu-
tion of hydroxylamine hydrochloride-NaCI,and finally a SnCl2 solution. The
samplestreampassesinto a phaseseparatorwhere the liquid goesto wasteand
the Hgo vapor passesinto an absorptioncell positionedin the light path of an
automaticabsorptionspectrometer.The absorptionsignal is recordedon a strip
chart recorderwhere the peak heights are measuredand comparedto standard
aqueousHg solution peakheightsfor calculation.
Comments
Both manual and automatedmethodshave been shown to give accurate
and precisemeasurements of total Hg in soils and relatedsamplematrices.The
automatedsystemis preferredbecauseof its simplicity, easeof operation,small-
er risk or contamination,and speedof analysis.
Short-termprecisionof the automatedmethodwas determinedby analyz-
ing sixteen10 Jlg HgIL solutionsand was found to be 1.2% RSD (relative stan-
dard deviation). For u.s. Geological Service (USGS) referenceshale SGR-l,
containing0.17 mg kg-1 Hg, the precisionwas less than 5% RSD. The sensitiv-
ity of the method is 0.21 mg Hg L -1, which produces a signal of 0.0044
absorbance(1 % absorption).The methodhasan in-samplelimit of determination
(basedon 2 standarddeviations)of 0.02 mg kg-1 Hg using the describeddiges-
tion procedure(basedon an O.I00-g sample)and operatingconditions.
MERCURY 781
If a sampleof elevatedHg content(>10 mg kg-I Hg) is analyzedundilut-

ed, the systemmust be dismantled,all Tygon tubing replaced,and all the glass-
ware soakedin 4 M HN03 overnightto preventcarry-overcontamination. The
systemthen is reassembledand the manifold flushed with reagentsuntil a zero,
stablebaselineis achieved.This stabilizationmay take as long as 8 h, depending
on the Hg contaminationof the system.It is often advisableto replacethe glass
mixing coils to reducethe clean-outtime. Alternatively, if the systemhasminor
contamination,pump a 30% (weight/volume)KOH (potassiumhydroxide)solu-
tion through the system for approximately 2 min after the system has been
flushedwith demineralizedwater.If the baselinedoesnot returnto zero after this
treatment,soakthe glasswarein hot (100°C) 30% KOH solution for 2 h.
The phaseseparatorand the absorptioncell should be warmed to 35 to
40°C using infrared heatlamps.This helpsminimize samplecarryoverallowing
the baselineto be reachedbetweensamples.The heatingalso preventsexcess
condensationof water in the absorptioncell.
Table 29-1 showsthe elementstestedthat do not interferewith the deter-
mination of Hg usingthe given manifold systemat the in-solution concentration
shown.Elementsat the less than 1 mg kg-l level in the sampledigest that sig-
nificantly reducethe observedHg signal includeAg, Au, Pt, Te, and Se. With the
given digestionand normal occurrencesof theseelementsin geological materi-
als, only Se may posea significantproblemfor most nonmineralizedor contam-
inated soils or sediments.This also has been confirmed by Pratt and Elrick
(1987) for natural water samples.For most geologicalmaterials,when an ele-
ment is presentat interferingconcentrationlevels,dilute the sampledigestor use
the methodof standardadditions.
Glasstubesratherthan plastic must be usedto preservethe Hg in solution
(Feldman,1974). The sampledigestsolutionsstoredin glasstubesare stablefor
Table 29-1. Elementsinvestigatedshowingno interferenceon the determinationof Hgt by CV -AAS.

Maximum concentration
Element testedin solution
mgL-l
Zn 1000
Pb 1000
Cu 500
Cd 50
Bi 50
Co 100
Mn 1000
U 100
Mo 100
V 100
Ni 100
Fe 500
As 30
Sb 10
t A solution of 10 I1g Hg L- 1, 3.2 M HN03 and 2 g L-l Na2Cr207·2H20.
782 CROCK
Table 29-2. Recommended(t) and proposedvaluesfor Hg in selectedreferencematerials,modified

from Govindaraju(1989).
Referencematerial Sampledescription Hg content Source*,Reference§
Ilg kg-1
GSS-2 Soil 1S+/-2t IGGE (Certificate)
GSR-6 Limestone 16+/-1 IGGE (Certificate)
10-1 Granodiorite 16t GS]
GSS-8 Soil 16.6+/-1.7t IGGE (Certificate)
SO-3 Soil 1?+/-7 CCRMP (Certificate; A)
GSD-1 Streamsediment 18+/-2t IGGE (Certificate)
AGV-1 Andesite 20+/-9 USGS(B)
FER-2 Iron formation 20 CCRMP
SO-1 Soil 21+/-3t CCRMP(A)
JB-1 Basalt 28t GS]
GSS-1 Soil 32+/-3t IGGE (Certificate)
SO-4 Soil 32+/-10 CCRMP (Certificate; A)
GSD-2 Streamsediment 40+/-St IGGE (Certificate)
GSD-8 Streamsediment 42+/-3t IGGE (Certificate)
GSD-4 Streamsediment 44+/-St IGGE (Certificate)
GSD-6 Streamsediment 4S+/-St lOGE (Certificate)
G-2 Granite 49+/-13t USGS(B)
GSD-7 Streamsediment S3+/-9t IGGE (Certificate)
GSD-12 Sediment S6+/-4t IGGE (Certificate)
NBS1646 Estuarinesediment 6Y/-12 NBS (Certificate)
GXR6 Soil 68+/-14 USGS(C)
GSS-6 Soil 72+/-St lOGE (Certificate)
GSD-ll Sediment 72+/-6t IGGE (Certificate)
SO-2 Soil 82+/-12t CCRMP (Certificate; A)
GSD-9 Sediment 83+/-6t IGGE (Certificate)
G-1 Granite 90+/-30 USGS(B)
GSD-S Streamsediment 100+/-10 IGGE (Certificate)
GXR-4 Coppermillhead 110+/-30 USGS(C)
BCSS-1 Marine sediment 129t NRCC
at least 24 h. The sampledigest solutionsstoredin the teflon vesselsare stable

for more than 48 h. A memoryeffect is possiblewith Teflon vesselsfor samples
containingmore than 20 mg kg-1 Hg. A constant12.00g weight was chosento
provide sufficient solution for the analysis.A different volume may be required
if a different auto sampleror manifold systemis used.Constantweight ischosen
becauseit is operationallyeasierto bring all samplesto a constantweight in dis-
posableglasswarethan to constant volumein a small volumetric flask.
Mercury Standard Reference Materials

Accuracyof an analyticalmethodis both a difficult and requiredmeasure-
ment in the establishmentof an analytical method.To measureaccuracy,stan-
dard referencematerials(SRMs) whoseelemental contenthas beenestablished
MERCURY 783

Referencematerial Sampledescription Hg content SourcetReference§
~g kg-1
MRG-l Gabbro 140+/-40t CCRMP (Certificate)

GXR-5 Soil 158+/-17t USGS(B)
MESS-I Marine sediment l7lt NRCC
SDO-l Marine sediment 190 USGS
W-I Diabase 210+/-70 USGS(C)
GSD-lO Sediment 280+/-18t IGGE (Certificate)
GXR-3 Deposit(hot springs) 330+/-40t USGS (C)
NBSl645 River sediment 1.1 mg kg-1 NBS (Certificate)
NBS1646 Estuarinesediment 1.44 mg kg-I NBS (Certificate)
GXR-2 Soil 2.9+/-0.7mg kg-I USGS (C)
GXR-l Jasperiod 3.9+/-0.6mg kg-I USGS(C)
PACS-1 Marine sediment 4.57 mg kg-I NRCC
SRM8406 River sediment 0.06 mg kg-I NIST (Certificate)
SRM8407 River sediment 50+/-2 mg kg-I NIST (Certificate)
SRM8408 River sediment 107+/-2 mg kg- 1 NIST (Certificate)
t Recommendedand proposed values for Hg in selected reference materials, modified from

Govindaraju(1989) (continued).... Mercury valuesare from Govindaraju(1989), exceptfor data
which includesstandarddeviation.The referencefor confidenceintervalsare in parenthesisunder
* the samplesource.
=
Source of the standardreferencematerial: GSJ Geological Survey of Japan, 1-1-3 Higashi,
Yatabe,Tuskuba,Ibaraki, 305 Japan;ASK =Analytisk SporelementKomite, Rogalandsforskning,
Postboks2503, Ullandhaug,4001 Staanger,Norway; IGGE =Inst. Geophysicaland Geochemical
Prospecting, Min. of Geology, Peoples Republic of China; CCRMP = Canadian Certified
ReferenceMaterials Project, CanadaCenter for Mineral and Energy Technology, Mines and
Resources,555 Booth Street,Ottawa,CanadaKIA OGI; NBS and NIST =Natl. Inst. of Standard
Technology,Office of StandardReferenceMaterials, Gaithersburg,MD 20899; NRCC = Natl.
Res. Council of Canada,Chemistry Div., Natl. Res. Council, Montreal Road, Ottawa, Canada;
USGS =U.S. Geol. Surv., Branchof Geochemistry(Attention of StandardsProgram),Box 25046,
Mail Stop 973, Denver,Colorado80225.
§ A =Gladneyand Roelandts(1989), B = Gladneyet al. (1983), C =Gladneyand Roelandts(1990).
by severalindependentmethodsare essential.Standardreferencematerialsare
intendedas a control to insure that different laboratoriesare analyzingthe same
homogeneous,well-characterizedmaterial. Backgroundto elevatedSRMs need
to be analyzedto establishthe completeworking rangeof a method,both in sam-
ple matrix composition and level of the analyte. Accuracy measurementsare
requiredto establishboth quality assuranceand quality control for the data pro-
duced.
A wide variety of SRMs for the determinationof Hg in soils is available.
Roelandts(1989) presentsa compilationof environmentalSRMs, datacompila-
tion, and suppliers.Table 29-2 presentssomeavailableHg SRMs and their rec-
ommendedor proposedHg content [modified from Govindaraju,1989]. These
SRMs shouldbe usedon a routine basisin the analysisof soils for Hg.
784 CROCK
TOTAL MERCURY IN SOIL EXTRACTS, LEACHATES,

AND WATER SAMPLES
Introduction
Total Hg in soils or sedimentsis not a good prediction of the bioavailable

Hg, or how much of the Hg in the systemwill eventuallymake its way into the
food chain, especiallyas the very toxic methylmercury.However,total Hg does
give an indication of the reservoirof Hg availablefor conversion.The mobility
of Hg in a soil profile dependson its speciation,and is usually influencedby its
very strongchalcophilicnature,its affinity to form organometallic bonds, and its
associationwith oxide minerals or precipitates.Methods for the quantitative
measurementof the speciationof Hg in the environmenthave yet to be estab-
lished and acceptedby the scientific community. The occurrenceof Hg can be
generalizedinto the following four categories:(i) metallic, Hgo; (ii) mercurysul-
fide, HgS; (iii) Hg associatedwith organicmatter; and, (iv) adsorbedHg.
Many partial and sequentialextraction schemesand leachingprocedures
have beendevelopedfor a variety of elementswith respectto a specific chemi-
calor physical property, or elementalor phaseassociation.In general,a mea-
surementof elementuptakeby vegetationis desired-ameasureof its bioavail-
ability. The most direct measurementof Hg bioavailability is to measurethe Hg
contentof the vegetationitself (Williams & Funston,1987).
Various common extractantshave been used to assessHg contamination
and bioavailability. These extractantsinclude: (i) ammonium acetate for ex-
changeableHg; (ii) AB-DTPA (ammoniumbicarbonate-diethylenetriaminepen-
taaceticacid) for availableHg in neutralto alkalinesoils (Soltanpour,1991);(iii)
saturatedsodium sulfide for sulfide mercury (HgS) (Revis et aI., 1989); (iv)
dilute HCl for Hg adsorbedonto clays; and,(v) hydrogenperoxideto destroythe
organicmaterial for organically bound Hg. A comprehensivesequentialextrac-
tion procedureto determine water-soluble,exchangeable,manganeseoxide-
associated,and free iron oxide-associatedHg is presentedby Harsh and Doner
(1981). They also determine available, weakly organic-bound,and strongly
bound Hg with nonsequential,selective extractions. Another comprehensive
sequentialprocedureis presentedin Eganhouseet ai. (1978). Theseextraction
proceduresare summarizedin Table 29-3. An excellentsummaryof the use of
partial and sequentialextractionsis presentedin Chao(1984).
Another method for the determinationof the phaserelationshipof Hg in
soils is the thermal evolution by the stepwiseheatingof the sample(Azzaria &
Aftabi, 1991).Unlessthis methodis combinedwith an amalgamationstep,inter-
ferencesfrom combustionproductswill occur.
Soil extractsand water samplesgenerally contain low concentrationsof
Hg. The dominantforms of Hg vary between organicand inorganiccomponents.
Watersand extractsmust be collectedand storedproperly, usually in an acidic,
oxidizing solution in flint or Pyrexglasscontainers(Feldman,1974).Watersam-
ples must be filtered through 0.45-1l membranefilters to obtain "dissolvedmer-
cury" (including organic Hg). Filtration removessuspendedmaterials, which
may contain an appreciableamountof the water'stotal Hg. If total Hg in water
MERCURY 785
Table 29-3. Common extractsand sequentialextractonschemesfor the determinationof Hg parti-

tioning among chemicalphasesin soils and sediments.
Extractant Mercury species
A. Sequentialchemicalextraction(Harsh & Doner, 1981)

1. Distilled, deionizedwater 1. Water soluble
2.0.1 M sodiumacetateat pH =7 2. Exchangeable
3. 0.1 M hydroxylaminehydrochlorideand 3. Manganeseoxide associated
0.01MHN03
4.0.15 M oxalic acid and 0.25 M ammonium 4. Iron oxide associated
oxalate
B. Sequentialchemicalextraction(Eganhouseet aI., 1978)
1. Distilled, deionizedwater 1. Water soluble
2. 1 M magnesiumchloride 2. Ion exchangeable
3. 0.005M and 0.2 M NaOH 3. Humic and fulvic acid associatedor complexed
4. 0.005 acetic acid 4. Organicbasesassociatedor complexed
5. 3% hydrogenperoxide(pH =2) at 85°C 5. Readily oxi~izable organicmatterand acid-
soluble sulfides
6. 30% hydrogenperoxide(pH =2) at 85°C 6. Remainingdegradableorganic matterand some
sulfides
7. Aqua regia at 200°C 7. Remainingsulfides
C. Nonsequentialchemicalextractions(Harsh & Doner, 1981)
1. 0.5 and 1.0 M HCI 1. Strongly adsorbedHg
2. DTPA 2. Plant available
3. 0.5 M sodium bicarbonate 3. Organicassociated
D. Other nonsequentialchemicalextractions
1. AB-DTPA (Soltanpour,1991) 1. Available
2. Saturatedsodiumsulfide (Revis et aI., 1989) 2. Cinnabarand sulfide associated
is desired,do not filter. Both water and extractsmust be digestedto convert all
the forms of Hg presentto ionic Hg to insureproperconversionto Hgo vapor for
the determinationby CV-AAS. Greatcaremust be exercisedto preventcontam-
ination or loss throughvolatilization.
Principles
Wide ranges in Hg concentrationand forms of Hg can be expectedin

extracts,leachates,or water samples.Thesesamplesmust be digestedrigorously
to insure that ionic Hg is the only form presentprior to the CV-AAS determina-
tion. It is recommendedto usethe batchmodeof digestingsamplesfor Hg, espe-
cially if one hasa large numberof samplesto do. This also allows usingonly one
manifold for the automatedcontinuous-flowmethods.
One should always matrix match the standardsolutions and run them
throughthe extractionand digestionsteps.If a new extractionmatrix is used,one
should carefully confirm recoveriesof the Hg phases,especiallyif methylmer-
cury is suspectedto be present.The methodof standard additions also helpscon-
firm recoveriesof the volatile species.All the solutions used in the method of
standardadditionsalso should be run through the extractionand digestionsteps
of a method.
786 CROCK
Insteadof constructingthe manifoldsas shownin Figs. 29-1 and 29-4, one

can obtain commercially available CV-AAS systemsthat perform equally as
well.
Manual Method for Determinationof Dissolved,Suspended,

and Total Mercury
SpecialApparatusand Reagents
The equipmentand reagentslisted previously in the sectionson "Special
Apparatus"and "Reagents"also are usedfor the manualdetermination.In addi-
tion, when manuallydigestingthe sampleson the hot plate using the batchmode
method,the following reagentsare required:
1. Concentrated(18 M) H2S04, BakerSuitablefor Mercury Determination
grade! or equivalent.
2. PotassiumPersulfateSolution: Dissolve 4.00 g K2S20 S (reagentgrade,
tested to be free from Hg contamination)in sufficient demineralized
water to bring the final volume to 1.00 L.
SamplePreparation
Stewartand Bettany(1982) define three forms of Hg associatedwith soil
extracts,leachates,and water samples."Dissolved mercury" is the samplesolu-
tion's Hg contentthat is in true solution.Thesesamplesshouldbe filtered through
a 0.45-1l filter at the time of collection and acidified with 1 mL of concentrated
HN03, and oxidized/stabilizedwith 0.5 mL of the sodium dichromatesolution
per 50 mL of the sample.All sampleand standardsolutionsshouldbe storedin
flint or pyrex glasscontainers."Suspendedmercury" is the Hg associatedwith
the solid phasesin a given volume of solution or extract.Theseare the phases
retainedon the 0.45-1l filter after filtering a given volumeof extractor water.The
material remaining on the filter must be removed, digested,and the Hg then
determined."Total mercury" for thesesamplesis the Hg contentof the unfiltered
liquid as preparedand collected or the sum of "dissolved mercury" and "sus-
pendedmercury."
DigestionProcedure
For eachof the three forms of Hg, the samplemust be digestedto insure
that all the Hg is quantitatively in the ionic form. Organomercurywill not be
reducedin the reducingstep in either the manualor automatedmethodof deter-
mination. The batch-modedigestion procedurerecommendedfor dissolvedHg
and total Hg is as follows (modified from Nygaard& Lowry, 1982):
1. Into a 125-mL wide-mouth,high-density,polyethylenebottle (or Teflon
may be used),add 40 g of the water or extract,5 mL 18 M H2S04, 0.75
mL of the potassiumpersulfatesolution, and 0.75 mL of the sodium
dichromatesolution.
2. Digest with the lid loosely cappedat 95 to 100°Con asteambathor hot-
plate for 3 h. Removeand let cool to room temperature.
MERCURY 787
3. Digest reagent blanks, standard water solutions (available from

National Institute of Standardsand Technology (NIST), USGS, and
other sources),and aqueousHg standardsmatched in matrix to the
extractionsolution.
4. Digest the filter membranein the samefashion of solid samplesas out-
lined in the section"Mercury Determination(Manual)."
AutomatedProcedurefor Determinationof Total Mercury
SpecialApparatusand Reagents
The specialapparatusand reagentsfor the automatedmethodare the same
as thoselisted in the sections"SpecialApparatus"and "Procedures."
SamplePreparationandAnalytical Procedure
Samplepreparationand analytical procedureare thosegiven in the section
"Digestion Procedure"for the digestionby the batch-modemethod.The analyt-
ical procedureis the sameas given in the section"Reagents"for the automated
total Hg method.
Comments
The automated,segmentedcontinuousflow systemsare the recommended
methodfor the determinationof Hg. Although the batch-modemethodof diges-
tion is the recommendedmethod forthe digestionof extracts,leaches,and water
samples,on-line digestion using the continuousflow system remains popular.
The recommendedmethodwould be that of Stewartand Bettany(1982). As dis-
cussedearlier, microwavedigestionsof extracts,leaches,and waters is becom-
ing a popular method. On-line microwave digestionswith the continuousflow
systemsare being currently investigated(Welz et ai., 1992).
Unfortunately,no referencematerialsfor extractableor leachableconcen-
trations of Hg in soils or sedimentsare commercially available. However, a
seriesof waterstandardsat variousHg concentrationsis availablethrough NIST
and the USGS(Water ResourcesDivision, Branch of Quality Assurance,Box
25046, Mail Stop 401, Denver FederalCenter,Denver,CO). The specific SRM
referencenumberschangedue to their stability and availability.
ORGANIC MERCURY COMPOUNDS
GeneralComments
Determiningthe "total" Hg contentof a soil or sedimentdoes not neces-

sarily define mercury bioavailability or its toxicity. Bioavailability of Hg is usu-
ally the key to understandingits toxicity. Simple groupingof Hg forms into inor-
ganic "dissolved" and "solid," "ion-exchangeable"and "total organic" Hg is a
goodstart in understandingHg in soils and sediments.It is not the purposeof this
sectionto offer a definitive procedureto determineall the forms of organic Hg
788 CROCK
becausethere is not a consensusas to methodology,nor are there any standard

referencematerialsavailablefor thesedeterminations.
Excellent summariesof organomercuryspeciesin soils are presentedby
Andersson(1979) and Wilken and Hintelmann(1990). It is the methylationthat
is the key step in the biogeochemistryof Hg. WhereasHg undergoesa number
of transformationsin soils and sediments,the primary reactionsinvolve methy-
lation and dimethylationand reduction tothe free metal (D'ltri, 1990). Mercury
is readily methylatedby certain microorganismsto either the methyl or dimeth-
ylmercury underaerobicor anaerobicconditions.In general,if Hg is in the envi-
ronment,eventuallyit will be convertedto organicforms, all of which are toxic
to life.
Becausethere exists a dynamic eqUilibrium betweenthe inorganic and
organic forms of Hg in soils and sediments,even the best analysis can only
describethe steadystateresulting from the rapid conversionfrom one form to
another(Wilken & Hintelmann,1990). Oda and Ingle (1981) describea method
to distinguish inorganic and organic Hg by selectively reducing the Hg with
SnCl2 and sodium borohydride(NaBl4), respectively.The solution is reacted
first with a SnCLz solution,which will reduceonly the inorganicHg forms. The
remaining solution is then reactedwith NaBl4 to reduce the remaining Hg.
Horvat (1991) gives various methodsof determinationof methylmercuryand
ethylmercuryin biological materialsand referencesthe existing SRMs for these
partial determinations.However, no SRMs for methylmercuryin water or soils
exist, and few laboratoriesare capableof commerciallyperformingtheseanaly-
ses(Bloom et aI., 1991). Organic mercury is determinedafter extraction into a
variety of organicsolvents,and then re-extractinginto anotherorganicphasefor
someinstrumentalseparation,usually chromatographic,and final detection.The
main problemin the detectionof organomercuryforms is in the chromatograph-
ic behaviorof the Hg compounds.Various chromatographicsystemshave been
applied but HPLC (high-pressureliquid chromatograph)offers the best results,
and a procedureis presentedby Wilken and Hintelmann(1990). Bushee(1988)
also hasinterfaceda flow injection chromatographysystemto an ICP-MS for the
sensitivedetectionof the organicforms of Hg.
In summary,the determinationof organomercuryforms in soils and sedi-
ments is a tediousand laborioustask, hamperedby the reactivity of the forms
being measured and difficulty of separatingtheseforms. Thesedeterminations
are anythingbut a routine task and usually are nonconclusivewhen completed.
REFERENCES
Adriano, D.C. 1986. Trace elementsin the terrestrialenvironment.Springer-Verlag,New York. p.

298-328.
Andersson,A. 1979. Mercury in soils. p. 79-112.In 1.0. Nriagu (ed.) The biogeochemistryof mer-
cury in the environment.ElsevierlNorth-HolIandBiomed. Press,New York.
Andren,A.Q., and1.0. Nriagu. 1979.The global cycle of mercury.p. 1-21. In 1.0. Nriagu (ed.) The
biogeochemistryof mercuryin the environment.ElsevierlNorth-HolIandBiomed.Press,New
York.
Azzaria, L.M., and A. Aftabi. 1991. Stepwisethermal analysistechniquefor estimatingmercury
phasesin soils and sediments.WaterAir Soil Pollut. 56:203-217.
MERCURY 789
Bartha, A., and K. Ikrenyi. 1982. Interfering effects on the determinationof low concentrationsof
mercury in geologicalmaterialsby cold-vapouratomic absorptionspectrometry.Anal. Chim.
Acta 139:329-332.
Bloom, N.S., C.J. Watras,and J.P. Hurley. 1991. Impact of acidification of the methyl mercury cycle
of RemoteSeepageLake. Water Air Soil Pollut. 56:447--491.
Burguera,J.L. 1989. Flow injection atomic spectroscopy.p. 139-142. Marcell-Decker,Inc., New
York.
Bushee,D.S. 1988. Speciationof mercury using liquid chromatographywith detectionby inductive-
ly coupledplasma-massspectrometry.Analyst 113:1167-1170.
Chan,C.C.Y., and S. Bina. 1989.A sensitiveautomatedmethodfor determinationof mercury in geo-
logical materialsby cold vapor atomic absorption.Geostand.Newsl. 13:181-186.
Chao, T.T. 1984. Use of partial dissolution techniquesin geochemical exploration. 1. Geochem.
Explor.20:101-135.
Chao,T.T., and R.E Sanzolone.1992. Decompositiontechniques.J. Geochem.Explor. 44:65-106.
Clement, R.E., M.L. Langhorst, and G.A. Eiceman. 1991. Environmentalanalysis. Anal. Chern.
63:270R-292R.
Crock, 1.G., P.H. Briggs, L.L. Jackson,and EE. Lichte.1987.Mercury by continuous-flow,cold-
vapor atomic adsorption.p. 13-21. In Analytical methodsfor the evaluationof streamsedi-
mentsand rocks from WildernessStudy Areas. U.S. Geol. Surv. Open-FileRep. 87-84.
D'ltri, EM. 1990.The biomethylationand cycling of selectedmetalsand metalloidsin aquaticsedi-
ments.p. 163-214.In R. Baudoet al. (ed.) Sediments:Chemistryand toxicity of in-placepol-
lutants.
Eganhouse,R.P.,D.R. Young, andJ.N. Johnson.1978. Geochemistryof mercury in PalosVerdessed-
iments.Environ. Sci. Technol. 12:1151-1157.
Fang, S.c. 1978. Sorption and transformationof mercury vapor by dry soil. Environ. Sci. Technol.
3:285-288.
Feldman,C. 1974. Preservationof dilute mercury solutions.Anal. Chern.46:99-102.
Gladney,E.S., C.E. Burns, and I. Roelandts.1983. 1982 Compilationof elementalconcentrationsin
elevenUnited StatesGeologicalSurvey rock standards.Geostand.Newslett. 7:3-226.
Gladney,E.S.,andI. Roelandts.1989. 1988Compilationof elementalconcentrationdatafor CCRMP
soils SO-1 to SO-4. Geostand.Newslett. 13:217-268.
Gladney, E.S., and I. Roelandts. 1990. 1988 Compilation of elemental concentrationdata for
CCRMP referencerock sampleSY-2, SY-3,and MRG-1. Geostand.Newslett. 14:373--458.
Govindaraju,K. 1989. 1989 Compilationof working valuesand sampledescriptionfor 272 geostan-
dards.Geostand.Newslett. 13 (Spec.Issue):1-67.
Greenwood,N.N., and A. Earnshaw.1984. Chemistryof the elements.PergamonPress,New York.
Grobenski,Z., W. Erler, and U. Voellkopf. 1985. Determinationof mercury with Zeemangraphite
furnaceAAS. Atom. Spec.6:91-93.
Harsh,J.B., and H.E. Doner. 1981. Characterizationof mercury in a riverwashsoil. J. Environ. Qual.
10:333-337.
Hatch, W.R., and W.L. Ott. 1968. Determinationof sub-microgramquantitiesof mercury by atomic
absorptionspectrophotometry.Anal. Chern. 40:2085-2087.
Horvat, M. 1991. Determinationof methylmercuryin biological referencematerials.Water Air Soil
Pollut. 56:95-102.
Huffman, C., Jr., R.R. Rahill, Y.E. Shaw,and D.R. Norton. 1972. Determinationof mercury in geo-
logic materialsby flamelessatomic absorptionspectrometry.U.S. Geol. Surv. Prof. Pap.800-
C:C203-C207.
Jackson,L.L., T.L. Fries,J.N. Grossman,B.-S.W. King, and P.J.Lamothe.1991.Geologicaland inor-
ganic materials.Anal. Chern. 63:33R--48R.
Johannson,K., M. Aastrup, A. Andersson,L. Bringmark, and Ake Iverfeldt. .1991. Mercury in
Swedishforest soils and water-assessment of critical load. WaterAir Soil Pollut. 56:267-282.
Kabata-Pendias, A., and H.K. Pendias.1984. Trace elementsin soils and plants. p. 116-125.CRC
Press,Inc., Boca Raton, FL.
Kennedy,K.R., and J.G. Crock. 1987. Determinationof mercury in geologicalmaterials bycontinu-
ous-flow, cold-vapor,atomic absorptionspectrophotometry.Anal. Lett. 20:899-908.
Koirtyohann, S.R., and M. Khalil. 1976. Variables in the determinationof mercury by cold vapor
atomic absorption.Anal. Chern.48:136-139.
Korunova, V., and J. Dedina. 1980. Determinationof trace concentrationof mercury in biological
materials after digestion under pressurein nitric acid catalyzesby vanadium pentoxide.
Analyst 105:48-51.
790 CROCK
Lee, S.H., K.-H. lung, and D.S. Lee. 1989.Determinationof mercury in environmentalsamplesby
cold vapourgenerationand atomic absorptionspectrometrywith a gold-coatedgraphitefur-
nace.Talanta36:999-1003.
Lindqvist, O. (ed.). 1991. Mercury as an environmentalpollutant. Water Air Soil Pollut. 56:1-847.
Lobring, L.B., and B.B. Potter. 1992. Method 245.5: Determinationof mercury in sedimentsby cold
vapor atomic absorptionspectrometry.p. 289-303. In C.K. Smoley (ed.) Methods for the
determinationof metals in environmentalsamples.USEPA Environ. Monitoring Systems
Lab., e.R.e.Press,Boca Raton,FL.
Montaser, A and D.W. Golightly. 1992. Inductively coupled plasmasin analytical atomic spec-
troscopy.2nd ed. VCH Pub!" Inc., New York.
Moore, 1.W., and S. Ramanoorthy.1984. Heavy metals in natural waters:Applied monitoring and
impact assessment. Springer-Verlag,New York.
Mudroch, A, and E. Kokotich. 1987. Determinationof mercury in lake sedimentsusing a gold film
mercury analyzer.Analyst 112:709-710.
Nygaard,D.D., and I.H. Lowry. 1982. Sampledigestionproceduresfor simultaneousdetermination
of arsenic,antimony,and seleniumby inductively coupledargon plasmaemissionspectrom-
etry with hydride generation.Ana!. Chern. 54:803-807.
Nriagu, 1.0., and 1.M. Pacyna.1988. Quantitative assessmentof worldwide contaminationif air,
water, and soils by trace metals.Nature (London) 333:134-139.
Oda, e.E., and I.D. Ingle, lr. 1981, Speciationof mercury by cold vapor atomic absorptionspec-
trometry with selectivereduction.Ana!. Chern.53:2305-2309.
Pratt, L.K., and K.A Elrick. 1987. Interferenceeffectsof seleniumon the determinationof mercury
in naturalwatersusing cold vaporatomic absorptionspectroscopy.At. Spectrosc. 8:170--171.
Revis, N.W., T.R. Osborne,D. Sedgley,and A King. 1989. Quantitativemethodfor determiningthe
concentrationof mercury (II) sulfide in soil and sediments.Analyst 114:823-825.
Risser,lA, and D.E. Baker. 1990.Testing soils for toxic metals. p. 275-298. In R.L. Westerman
(ed.) Soil testingand plant analysis.SSSASpec.Pub!. 3. 3rd ed. SSSA, Madison,WI.
Roelandts,I. 1989. Environmental reference materials.Spectrochim.Acta 44B:925-934.
Rose,AW., H.E. Hawkes,and1.S. Webb. 1979. Geochemistryin mineral exploration.2nd ed. Acad.
Press,New York.
Sanzolone,R.E, and T.T. Chao. 1983. Atomic-absorptiondeterminationof mercury in geological
materialsby flame and carbon-rodatomizationafter solventextractionand using co-extract-
ed silver as a matrix modifier. Analyst 108:58-63.
Schuster,E. 1991. The behaviorof mercury in the soil with specialemphasison complexationand
adsorptionprocesses-Areview of the literature.Water Air Soil Pollut. 56:667-680.
Skougstad,M.W., M.I Fishman,L.e. Friedman,D.E. Erdman,and S.S. Duncan. 1979. Methodsfor
determinationof inorganic substancesin water and fluvial sediments.Book 5. u.s. Gov.
Print. Office, Washington,De.
soil test and ICPS. p. 165-190. In B.A Stewart (ed.) Advancesin soil science.Vo!. 16.
Stewart,I.W.B., and I.R. Bettany. 1982. Mercury. p. 367-384.In AL. Page(ed.) Methods of soil
analysis.Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Sturman,B.T. 1985.Developmentof a continuous-flowhydride andmercuryvaporgenerationacces-
sory for atomic absorptionspectrophotometry. App!. Spec.39:48-56.
Tuncei, G., and O.Y. Ataman. 1980. Design and evaluatonof a new absorptioncell for cold vapor
mercury determinationby atomic absorptionspectrometry.At. Spectrosc.1:126--128.
U.S. EnvironmentalProtectionAgency. 1986. Method 7471: Mercury in solid or semi-solid waste
(manual cold-vaportechnique)p. 7471-1 to 7471-10.In Test methodsfor evaluatingsoli.ll
wastes,physicaVchemicalmethods.EPA no. SW-846.3rd ed.
U.S. Environmental Protection Agency. 1990. Microwave assistedacid digestion of sediments,
sludges,soils, andoils. Method 3051.
Van Delft, W., and G. Vos. 1988. Comparisonof digestionproceduresfor the determinationof mer-
cury in soils by cold-vaporatomic absorptionspectrometry.Ana!. Chim. Acta 209:147-156.
Varma, A. 1984. CRC handbookof atomic absorptionanalysis.Vol. 2. p. 155-187.CRC Press,Inc.,
Boca Raton,FL.
Viets, I.G., and R.M. O'Leary. 1992. The role of atomic absorptionspectrometryin geochemical
exploration.l Geochem.Explor. 44:107-138.
Welz, B. 1985.Atomic absorptionspectrometry.2nd ed. VCH Pub!., Deerfield Beach,FL.
Welz, B., M. Melcher, H. W. Sinemus, andD. Maier. 1984. Picotracedeterminationof mercuryusing
the amalgamationtechnique.At. Spectrosc.5:37-42.
MERCURY 791
Welz, B., D.L. Tsalev,and M. Sperling. 1992. On-line microwavesamplepretreatmentfor the deter-
mination of mercury in water and urine by flow-injection cold-vapour atomic absorption
spectrometry.Anal. Chim. Acta 261:91-103.
Wilken, R.D., and H. Hintelmann.1990. Analysis of mercury-species in sediments.p. 339-359.In
lA.C. Broekaert et al. (eds.) Metal speciationin the environment.Springer-Verlag,New
York.
Williams, S.E., and R.S. Funston.1987. Mercury. p. 313-323.In R.D. Williams and G.E. Schuman
(ed.) Reclaiming mine soils and overburdenin the Western United States:Analytical para-
metersand procedures.Soil Conserv.Soc. Am., Ankeny, lAo
Published 1996
Chapter 30
Selenium and Arsenic
P. M. HUANG, University of Saskatchewan, Saskatoon, Saskatchewan,

Canada
R. FUJll, U. S. Department of the Interior, Geological Survey, Sacramento,

California
Selenium(Se) is an essentialmicronutrientfor humansand animals,but if ingest-
ed in relatively large quantities,it also can be toxic. Arsenic (As) is a contami-
nant of plants, humans, and animals, and sometimesmay be concentratedin
organisms,e.g.,in plantsgrowing in As-rich soil. Arsenic also is toxic to humans
and animals.The occurrenceof Se and As in soils is quite variable,rangingfrom
tracelevels(lessthan a mg kg-I) in someuncontaminatedsoils to a few thousand
milligrams per kilogram in contaminatedsoils.
Selenium,a group VIB element,and As, a group VB element, arepresent
in soils in inorganicand organicforms. Juxtapositionof the two elementsin the
periodic table accountsfor their similar chemical properties and somewhatsimi-
lar behaviorin soils and other natural systems.Concentrationsof Se and As in
soils generally are related to their concentrationsin soil parent material. For
example,soils derivedfrom sedimentarydepositscontaininghigh concentrations
of sulfidic mineralsoften containelevatedconcentrationsof both Se and As. The
geochemicalbehaviorof both elementsis relatedto their multiple oxidationstates
in soils and natural waters.The more mobile and more predominantspeciesof
eachelementin soil and natural watersgenerally are the more oxidized forms:
selenate[Se(VI)], selenite[Se(IV)], arsenate[As(V)], and arsenite[As(III)].
Methodsto determinetotal Se and As in soil, and total Se, Se(VI), Se(IV),
total As, As(V), and As(III) in soil solutions and water extractsof soil are dis-
cussedbelow. The methodsrecommendedin this chapterare basedon common-
ly availableequipmentand instrumentationandon evaluationsof methodsreport-
ed in the literature. Use of comparableequipmentor modifications of recom-
mendedmethodsshould be thoroughly testedusing adequatequality assurance
and quality control (QNQC) measures.
Book Seriesno. 5.
793
794 HUANG & FUJII
All chemicalsusedduring the proceduresdescribedbelow shouldnot con-

tain detectablelevels of Se and As. All reagentsshouldbe reagent-gradeor bet-
ter and all acidsused shouldbe of a gradesuitablefor tracemetal analysisor bet-
ter. The demineralizedwaterusedfor preparationof reagent solutions or dilutions
should be distilled and deionizedor better and contain undetectableSe and As.
All analyticalproceduresshouldroutinely include reagentblanks, duplicatesam-
ples, analytespikes,and standardor "in-house"referencematerialsto assurethe
quality of the analysis. Standardsoils (agricultural, highly contaminated,and
moderatelycontaminated)with certified total Se and As concentrationsare avail-
able from the u.S. Departmentof Commerce,NationalInstitute of Standardsand
Technology(NIST), StandardReferenceMaterials Program.Standardsolutions
with certified total Se and As concentrationsalso are available from NIST and
the U.S. EnvironmentalProtectionAgency (USEPA). TheseONOC procedures
shouldmeetthe criteria outlined in Chapter2 (Klesta & Bartz, 1996).
Hazardouswastecriteria for SeandAs in waterandsoil arecurrentlyunder
considerationby the USEPA but none have beenadopted.For analystsworking
with SeandAs in waterandsoil, the governmenthazardous-waste criteria should
be consulted,and water and soil samplesshould be disposedof in accordance
with respectivegovernmentregulations.Similarly, any waste generatedduring
analyticalproceduresalso shouldbe evaluatedwith regardto respectivegovern-
ment hazardous-waste criteria, and handledand disposedof in accordancewith
governmentregulations.
Precautionsshouldbe takenwhen handlingSe and As compounds,and all
reagentsand acids.Arsenic is extremelytoxic, and Se also is toxic. Acids gener-
ally are hazardous,and in particular,hydrofluoric acid can adverselyaffect skin
and bone.All digestionproceduresand other proceduresthat generatepotential-
ly hazardousvapors should be done in or vented to a chemical-exhausthood.
Material safety data sheetsshould be consultedfor specific precautions,prob-
lems, treatments,etc., for all chemicalsused.
SELENIUM
Introduction
Concernfor Se in soils and the environmentis relatedto both deficiency

and toxicity of Se in animals.Deficiency problemsgenerally are relatedto low
Se concentrationsin feed and forage plants, whereastoxicity problemsusually
result from bioaccumulationand biomagnificationof Se in the food chain. Dif-
ferencesin geologic sourcesof parent materialsfrom which soils are derived
accountfor much of the observeddifferencesin Se availability. Other biogeo-
chemicalfactorsalso mustbe assessed to evaluatebioavailability and mobility of
Se in the environment.
Seleniumis an essentialnutrient for mammalsand birds and its low con-
centrationsin feed and forage plants «0.05-0.1mg kg-1 Se) (Sharma& Singh,
1983) have been implicated as the causeof white muscle diseasein livestock
(Muth et aI., 1958). Accumulationof Se in Se-indicatorplantsor "Beath indica-
SELENIUM AND ARSENIC 795
tors" (e.g., Astragalus and Stanleya spp.)(Beath et ai., 1934) has been histori-
cally associatedwith toxicity to animalsthat ingestedtheseplants. Historically,
Se toxicity also has beenreportedin humansthroughoutthe world (Rosenfeld&
Beath, 1964). More recently, deformity and mortality of waterfowl hatchlings
have been attributed to Se accumulationand toxicity at Kesterson National
Wildlife Refuge in the westernSan JoaquinValley of California resulting from
high concentrationsof Se in evaporation-pondwatersoriginating from agricul-
tural irrigation drainwater (Presser& Barnes, 1984; Ohlendorf et ai., 1986;
Presser& Ohlendorf, 1987). The USEPA (1987, 1988) classifiesSe asa priority
pollutant with an aquatic-life criteria of 5 flg L -I and a maximum contaminant
level for drinking water of 50 flg L- 1.
Concentrationsof Se in soils are quite variable, ranging from <0.1 to as
high as 8000 mg kg-1 (Lakin, 1973; Tidball et ai., 1986; Berrow & Ure, 1989).
Ure and Berrow (1982) report a Se concentrationrange of 0.03 to 2.0 mg kg-I
and a meanof 0.40 mg kg-I for 1623 soils from throughoutthe world. In gener-
al, igneousrocks are low in Se (Lakin, 1973) and thereforetend to form soils
deficient in Se (Fleming, 1980). In contrast,sedimentarydepositsdominatedby
shalescontain the highest concentrationsof Se and tend to form Se-rich soils
(Lakin, 1973; Fleming, 1980). High concentrationsof Se in sedimentarydeposits
generally are associatedwith sulfide deposits, such as pyrite and chalcocite
(Sharma & Singh, 1983), or with organic-rich shales(Lakin, 1973; Fleming,
1980). Although problems associated with deficiencyand toxicity of Se general-
ly are relatedto the relative concentrationsof Se in soils, the bioavailability and
mobility of Se also are dependenton the forms and distribution of Se in soils and
the biogeochemicalprocessescontrolling Se availability in the environment.
The geochemistryof Se in the environment(e.g., McNeal & Balistrieri,
1989) is directly relatedto its occurrencein four oxidation states.Under equilib-
rium conditions,the predominanceof inorganicSe speciesis a function of redox
potential (Eh or pe) and pH. The most oxidized and most soluble Se speciesis
selenate[Se(VI), SeO~-]. Under most environmentalconditions, selenic acid
(HZSe04,pKz = 1.97) is unprotonated(Elrashadiet ai., 1987). Under mildly oxi-
dizing conditions, selenite [Se(IV), Se01-] predominatesand seleneousacid
(HZSe03,pKl = 2.65, pKz = 8.48) (D.R. Parker,1991, personalcommunication,
after Martell & Smith, 1982) is protonatedunderacidic to neutralpH conditions.
Selenitegenerallyis lesssolublethan Se(VI) and its solubility is often controlled
by sorption and coprecipitationreactions(Leckie et aI., 1980; McNeal & Bal-
istrieri, 1989). Elemental Se (Seo) is very insoluble and predominatesunder
reducingconditions. Under very reducing conditions, selenide(Se2-) predomi-
nates.Hydrogenselenide(HzSe)is a poisonousgasand a weak acid (pKJ =3.81)
when dissolvedin water (Elrashadiet ai., 1987). Insolubility of metal selenides
(e.g., Fe, Cu) and volatility of HzSegenerallylimit the presenceof Sez- in aque-
ous environments.Selenidealso substitutesfor Sz- in organic molecules such as
amino acids (e.g., selenocysteine,selenomethionine)and occurs in methylated
forms (e.g., dimethylselenide,dimethyldiselenide)in the environment(Doran &
Alexander, 1977; Cooke & Bruland, 1987; Frankenberger& Karlson, 1989;
Masscheleynet ai., 1989; Thompson-Eagleet aI., 1989; Masscheleynet ai.
1991a),as discussedbelow.
796 HUANG & FUJII
An assessment of Se behaviormust considerthe significant reactionsand

processesaffecting Se for the particulargeochemicalenvironmentunderinvesti-
gation. Measuringtotal Se in soils generallyis a first approachusedto identify
problemsrelated to toxicity or deficiency. Analysesof soluble Se speciesand
otherlabile forms of Se provide further insightsinto Se availability and the geo-
chemical processescontrolling Se solubility. Further analysesof solid-phase
associationsof Se help to assesspotentiallong-termmobility and bioavailability
of Se in soils.
Total Seleniumin Soil
Methodsto analyzetotal Se in soil fall into two generalcategories,nonde-

structive methodsand methodsthat require sampledecompositionfollowed by
quantificationof Se in the resultingsolution. Two commonnondestructivemeth-
ods are neutronactivation analysis(NAA) and x-ray fluorescencespectrometry
(XRF). Neutronactivation analysisis an excellentmethod(Pearlman& Assaro,
1971; Helmke, 1982) but can be impracticalbecauseof the needto irradiate the
samples.X-ray fluorescencespectrometry requires relatively little sampleprepa-
ration and is readily appliedto sampleswith relatively high concentrationsof Se
(detectionlimit of about 1 mg kg-I). Becauseaccessto nuclearreactorsgeneral-
ly limits the use of NAA, and total Se concentrationsin soils commonlyare less
than the detectionlimit for XRF, other methodsgenerallyare usedto determine
total Se in soils.
The most commonmethodsusedto determinetotal Se in soil require ini-
tial decompositionof the soil followed by analysisof total Se in the resulting
solution. Methods to decomposesoil samplesinvolve a multitude of mineral
acids and various heating techniquesto digest the soil. Microwave dissolution
techniques(Kingston & Jassie,1988) may be appliedto digestionof soils for Se
analysis.Thesetechniquesmay improve the efficiency of the digestion proce-
dure describedin the section"Digestion of Soil."
Analysis of Se in solutionsresultingfrom sampledigestionand in aqueous
sampleshasbeenaccomplishedby many methods.The hydridegenerationatom-
ic absorption spectrometric(HGAAS) and fluorimetric methods, the recom-
mendedmethods,are describedin detail below. These methodswere chosen
becauseof their widespreaduse, relative sensitivities,and the generalavailabil-
ity of atomic absorptionspectrometers (AAS) and fluorimetersin most laborato-
ries. The HGAAS methodusesa continuous-flowhydride generator[e.g., Vari-
an VGA-76' (Varian, Sugarland,TX); Fig. 30--1] coupledto an AAS. The fluo-
rimetric methodusesa segmented,microcontinuousflow system[e.g., Alpkem
Se analysismanifold (Alpkem, Wilsonville, OR); Fig. 30--2] coupledto a fluo-
rimeter.
Other methodsusedto analyzeSe and Se speciesinclude hydride genera-
tion coupled to an inductively coupled plasma atomic-emissionspectrometer
(ICP-AES) (Bakhtar et aI., 1989), colorimetry (Neal et aI., 1987a;Am. Public
I The useof brand,firm, or trade namesin this chapteris for identification purposesonly and does
not imply endorsementby the U.S. GeologicalSurvey.
-=1- ~ -1018.
Penstaltic
pump
[--sam~le_ 0 mLmin- 1
1- "" -!~.. 10 ,,:}-

-~ I -
fUJj
I 10 drain
[ -----1
NaBH~ _ f ---1 0 -
mL min-1
-------+---
l =~;o~ =-J----- 40
_mL
__min- 1
_____._ _.........J
Fig_ 3{}-1. Schematicof a continuoushydride generator_(Modified from Standard Methods for the
Examination of Water and Wastewater_ 18th ed_ Copyright 1992by the Am. Public Health Assoc_,
Am_ WaterWorks Assoc_,and the WaterEnviron. Fed_ Reprintedwith permission_)
Health Assoc.,1992),and chromatographicmethodsfollowed by variousmeth-

ods of detection (Siu & Berman, 1984; Karlson & Frankenberger,1986a,b;
Goyal et aI., 1991).
Direct sampleaspirationand ICP-AES analysisalso is usedto analyzeSe
but pneumaticnebulizationprovidesa detectionlimit of 30 to 40 Ilg L-1 (Rob-
berecht& Van Grieken,1982),too high to adequatelyassessthe USEPA'saquat-
ic-life criteria of 5 Ilg L-l (USEPA, 1987, 1988). Ultrasonic nebulizationpro-
vides a detectionlimit of 1 Ilg L-1 (Robberecht& Van Grieken, 1982) and has
the potentialfor evaluatingenvironmentalsamples.
flowrale
(".Lmin-1)
peristaltic pump Reagents
fI~Orimeter
.-r-------.------+----------r--------,
./
*[*~I' .-~~~-----+----------t_--------_.
In heat bath ..---d 118 air
ffi-.DXaD--,---r-..Ji--,,-\ SO"C n + I '-.../+-+----SS-'-'-"-8+"'sa"-m-Olile----
"'-phase
separator
I 4mL 1L-__---1I--_ _ _.:...74'-f-'0""A"'Ne-._ _ __
287 cyclohexane
287 cyclohexane
waste _----I-------=38::::;S'-+d::::e::::;bu=b"'ble~-----'
organic waste _-----I-----t'f::::low:::c::::e:.:..!1ip::::u::...li-t",hr::::;u-,(O::.::.O::..:4::..S":..:IO:.<...)_ - '
sampler wash _----I------'9'-'.47'-+O;;.c..1'-CM-'-'H.:.c;C"-1- - -
organic waste _----I--------t'f.:..:ro'-Cm-"p;.;..;ha~se'-'s:c:Jep!:..::a::.::ra:.:.:to-'-'r(c:.;O...:..;OS:..o6~·
10)
L------~_-----t- --------- t - - - -__- - - - . J
+ coil, 0 = 5 turns
* air solenoid valve
'* * O.8mm teflon
* ** O.3mrn teflon
Fig. 3{}-2_ Schematicof Alpkem Se analysismanifold. (Usedwith permissionof PerstorpAnalytical

Environmental,Wilsonville, Oregon.)
798 HUANG & FUJII
Electrothermalatomic absorptionspectrometry(ETAAS) is quite sensitive

to Se with a detectionlimit of 50 pg, but the methodis complicatedby potential
chemical interferencesand volatile loss of Se (Robberecht& Van Grieken,
1982). An interlaboratorystudy conductedby R.G. Burau (University of Cali-
fornia, Davis, CA) comparedanalysisof Se in 14 referencewater sampleswith
known Se concentrationsby 24 laboratories.Resultsindicatedthat ETAAS was
clearly inferior to the othertwo methodsused, especially on the high salinity, low
Se samples(R.G. Burau, 1985, personalcommunication).The other two meth-
ods usedin the study, HGAAS and fluorimetry, providedexcellentresults.These
are the two recommendedmethodsand are describedbelow.
Flow injection analysis(FIA) techniquesapplied to atomic spectrometry
have beenextensivelyreviewed by Tyson (1991), and he cites severalanalyses
of Se and As in water, soil extracts, and other environmentalsamplesusing
HGAAS. With FIA, the sampleand reagentsare injected into a carrier stream
where selectivechemicalreactionsoccur (e.g., generationof hydrides) and the
analyte of interest is detected,usually using conventionaldetectorssuch as an
AAS. Major advantagesof the FIA techniqueinclude higher samplethroughput,
smaller sample volumes neededfor analysis, reduced reagent consumption,
greatertolerancefor high ionic strengthsolutions,and betterprecision(Zhaolun
et aI., 1986; Tyson, 1991; Chan& Sadana,1992).
Flow injection analysishas beencombinedwith HGAAS to analyzeboth
Se and As in soils, sediments,vegetation,waters,and industrial wastes(Chan &
Sadana,1992). They report detectionlimits of 0.3 ~g L -1 for both Se and As and
a throughputof 50 samplesper hour usingthis method.Zhaolunet ai. (1986)pre-
sent resultsfor combining FIA techniqueswith atomic spectrometryto analyze
Se in soils, coal fly ash,and plant tissue.They report a theoreticaldetectionlimit
of 0.06 ~g L-l and excellentprecision(1.6% relative standarddeviation) for a 4
Ilg L-l standardsolution. Resultsof analysesof National Bureau of Standards
referencematerials[wheat (triticum aestivum L.) flour, orchardleaves,and coal
fly ash] showedgood agreementbetweenmeasuredand certified values.These
recent applicationsof FIA combinedwith HGAAS to analyze Se in environ-
mental samplesare quite promising and indicate need for further consideration
for analysisof Se in soils and sediments.
Principles
Digestion of Soils. The recommendedprocedurefor decomposingsoil to
determinetotal Se involves digestionof soil in mineral acidsat elevatedtemper-
atures.This methodis a revision of the methoddescribedby Welschet al. (1990)
(E.P. Welsch, 1992, personalcommunication),which is a modification of the
methoddevelopedby Sanzoloneand Chao(1987). This methodinvolves digest-
ing soil sampleswith a mixture of nitric acid (HN03), hydrochloric acid (HCI),
perchloricacid (HCI04), sulfuric acid (H2S04), and hydrofluoric acid (HF), fol-
lowed by additionofHCI to reduceSe(Vl) to Se(IV). For samplescontainingrel-
atively high concentrationsof organicmatter(greaterthan about40 g kg-1 organ-
ic C), samplesshouldstandovernightat room temperatureafter the initial addi-
tion of HN03 and HCL (E.P. Welsch, 1992, personalcommunication).
Selenium Determination. The two recommendedmethodsfor determin-

ing total Se in the solution resultingfrom decompositionof the soil are sensitive
to only the Se(IV) species.The first methodusessodium borohydride(NaBH4)
underacidic conditionsto reduceSe(IV) andgenerateH2Se(Welschet aI., 1990)
The resultingH2Se is flushedfrom solutionwith an inert gasinto a heatedquartz

absorptioncell mountedin the light path of an AAS, where Se absorbanceis
measured.This methodwill be referredto as HGAAS.
The second method is a modification of the fluorimetric procedureof
Brown and Watkinson(1977) (R.G. Burau & A. Jacobson,1992, personalcom-
munication) and is basedon the reaction betweenSe(IV) and 2,3 diaminon-
apthalene(DAN) to form a fluorescentpiazselenolcomplex. The piazselenol
complex is extractedinto cyclohexaneand its fluorescencemeasuredusing a
fluorimeter. This methodwill be referredto as fluorimetry.
Methods
Digestion of Soil
Apparatus
1. Standardlaboratoryhotplatewith a 30- by 60-cm heatingsurface.
2. Aluminum heatingblock, 2.5 cm thick by 25 cm wide by 50 cm long,
with 34-mm holesdrilled through a 5 x 10 matrix.
3. Thick-walled,30-mL Teflon bottles(e.g.,catalognumber0201 T from
Savillex Corp., Minnetonka,MN or equivalent).
4. Perchloric-acidhood.
Reagents
1. Nitric acid, concentrated,16 M.
2. Hydrochloric acid, 1.2 M, preparedby dilution of concentratedHCI,
12M.
3. Hydrochloric acid, 6 M, preparedby dilution of concentratedHCI, 12
M.
4. Perchloricacid, concentrated, 70%,doubledistilled (availablefrom G.
Fredrick Smith, Columbus,OH), or equivalent.
5. Sulfuric acid, concentrated,18 M.
6. Hydrofluoric acid, concentrated,48%.
Procedure. Weigh a 0.250-gsample«0.18 mm) into a 30-mL Teflon bot-
tle and add9 mL of concentratedHN03 and 0.25 mL of 1.2 M HCl. If the sam-
ple containsrelatively elevatedorganic matter (greaterthan about 40 g kg-I),
allow to standovernightat room temperature.Add 2 mL of concentratedHCI04,
2 mL of concentratedH2S04, 10 mL of concentratedHF, and heat overnight at
125°C. Removethe samplefrom the heatand let it cool. Add 25 mL of 6 M HCI
and let the samplestandfor 1 h. Quantitativelytransferthe sampleto a SO-mL
volumetricflask, and bring to volumewith deionizeddistilled water for HGAAS
800 HUANG & FUJII
analysis.Samplesto be analyzedfluorimetrically needto be treatedas described

in the "Procedures"section for "Selenium DeterminationIFluorimetry"prior to
analysis.
Selenium Determination
Hydride Generation Atomic Absorption Spectrometry
APPARATUS
1. Atomic absorptionspectrometer(e.g., Perkin-ElmerModel 5100).

2. Seleniumelectrodelessdischargelamp (EDL).
3. Electrodelessdischargelamp power supply.
4. Hydride generator (e.g.,Varian VGA-76; Fig. 30-1).
5. Quartzabsorptioncell and mount.
6. Autosampier,optional.
7. Data-acquisitionsystem (e.g., strip-chart recorder, microprocessor-
basedintegrator).
REAGENTS
1. Hydrochloric acid, 7 M: Prepareby dilution of concentratedHCl, 12

M.
2. Sodiumborohydridesolution,0.6% (w/v): Preparedfresh daily by dis-
solving 2.50 g sodium hydroxide(NaOH) in deionizeddistilled water,
adding 3.00 g NaBH4' and diluting to a total volume of 500 mL with
deionizeddistilled water.
3. Selenium standard solutions:Prepare from commercially available
AAS standard(1000 mg L -1 Se) by making serial dilutions to concen-
trationsof 2, 5, 12, and 18 Ilg L -1 Se in 1.2 M HCI. Standardsolutions
shouldbe madefor eachsetof analysesand shouldbe treatedthe same
as the solutionsresultingfrom the soil digestionprocedure.
PROCEDURE.The HGAAS methodusesa continuous-flowhydride genera-

tor coupled to an AAS to measuretotal Se in the solution resulting from soil
digestion. Specific proceduresvary dependingon the particular AAS, hydride
generator,andotherequipmentused(e.g.,autosampler),andthe readeris referred
to appropriateoperationalmanualsfor detailedinstructionsfor the particularsys-
tem. The readeralso is referredto Chapter4 (Wright & Stuczynski,1996),which
provides details for analysis by AAS and/or HGAAS. Examplesof particular
HGAAS instrumentalconfigurationsand parametersspecificfor Se are in the lit-
erature(e.g., Sanzolone& Chao, 1987; Makita and Fujii, 1992). Automatedcon-
tinuous-flow hydride generators(e.g., Varian VGA-76; Fig. 30-1) are common-
ly used;they generallyhavefewer interferences compared to batchsystems(San-
zolone & Chao, 1987). The automatedcontinuous-flow hydride generatorsare
designedto combinethe sample,the NaBH4 reagent,and the 7 M HCl reagentto
form the H2Se(Eq. [1 D. The mixture goesthrougha gas/liquidseparator,an inert
gas, such as Ar, carriesthe H2Se into the quartz absorptioncell mountedin the
light path of the AAS where Se is ionized, and its absorbanceis measuredat a
wavelengthof 196.0 nm. Heating of the quartz absorption cell is commonly
accomplishedusing an air-acetyleneflame, but more uniform heatingand better
precision have been reported for electrically heated quartz absorption cells
(Chapman& Dale, 1979).
The absorbanceof the sampleis comparedto the standardcurve to deter-
mine the total Se concentrationin microgramsper liter. This value is multiplied
by any dilution factor resulting from dilution of the original digestion solution,
and by 0.2 to obtain the concentrationof total Se in soil in milligrams per kilo-
gram. This methodhasa practicaldetectionlimit of 0.2 mg kg-! Se.
Standardsolutionsshouldbe periodicallyanalyzedassamples(aboutevery
S-10 samples).If the value of the standarddeviatesby more than 10% from its
known value,the instrumentshouldbe recalibratedusing eithera one-pointrecal-
ibration (reslope)or the entire rangeof standardsand the recalibrationchecked
for adequacy.Any samplesanalyzed prior to recalibration that have suspect
resultsshouldbe reanalyzed.
Selenium compounds may be toxic. Use caution and handle with care.
Fluorimetry
APPARATUS
1. Fluorimeter(e.g., Perkin-ElmerModel LS-2 Filter Fluorimeter).

2. Autoanalyzermanifold (e.g., Alpkem Se analysismanifold; Fig. 30-2,
flow diagramof autoanalyzermanifold).
3. Autosampler.
4. Peristalticpump (e.g., Gilson Minipump 2, Gilson Medical Electron-
ics, Inc., Middleton, WI).
5. Data-acquisitionsystem (e.g., strip-chart recorder, microprocessor-
basedintegrator).
6. Taylor tubes,50 mL.
7. Water bath.
8. Separatoryfunnels, 1 L.
9. pH electrodeand meter.
10. Whatmanno. 41 filter paperor equivalent.
11. Large glassfunnel.
12. Dark glassbottle, 1 L.
REAGENTS
1. Ethylenediaminetetraacetic acid (EDTA), disodiumsalt, 0.016M: Pre-

paredby dissolving5.96 g of disodiumEDTA in 1 L of deionizeddis-
tilled water.
2. Sodium hydroxide, 10 M: Prepareby dissolving 100 g NaOH in 250
mL of deionizeddistilled water.
3. Hydrochloric acid, 6 M and 0.1 M: Preparedby dilution of concentrat-
ed HCI, 12M.
4. Cyclohexane,HPLC gradeor better.
5. Ammonium hydroxide (NH40H), 7 M: Prepareby dissolving 24.53 g
~OH in 100 mL of deionizeddistilled water.
6. 2,3-diaminonaphthalene dihydrochloride (0.5% w/v) in 0.1 M Hel:
Preparationof this reagentis describedin detail in the next section.
802 HUANG & FUJII
7. Seleniumstandardsolutions:Prepareat leastfive concentrationsfrom

a commerciallyavailableAAS standard(1000 mgIL-l Se) by making
serial dilutions in 0.1 M HCl to concentrationsthat fall within the lin-
ear range of the instrument. Standardsolutions should be made for
eachset of analysesand be treatedas samples.
8. Cresolred pH indicator, commerciallyavailable.
PROCEDURE. Digestedsamplesmust first be treated prior to fluorimetric
analysisfor Se (Brown & Watkinson, 1977). Interferencesfrom trace elements
are reducedby adding EDTA and adjustingthe pH of the solution to between1
and 2. To treat the solution resultingfrom the digestionprocedure,the following
procedureis recommended(R.G. Burau and A. Jacobson,1990, personalcom-
munication):(i) quantitativelytransferthe solution to 50-mL Taylor tubes(Fish-
er Scientific, Pittsburgh,PA), and add5 mL of 0.016M EDTA solution and 1.6
mL of 10 M NaOH to bring the pH to approximately1; (ii) agitatethe tube to mix
reagentsinto the dropletsadheringto the walls of the test tube; (iii) using a pH
electrodeand meter, adjust the pH to between1 and 2 by adding either 7 M
N~OH or 6 M HCl dropwise;(iv) bring the sampleto a fmal volume of 50 mL
with 0.1 M HCl, cover, and mix. As an alternativeto electrochemicalpH adjust-
ment, add four dropsof cresol red pH indicator, add 7 M N~OH dropwiseto a
bright yellow end-point,andbacktitrate with 6 M HCl to a deepgold color (about
pH 1.8).
Preparationof the 0.5% (w/v) DAN reagentin 0.1 M HCL: (i) in a 1-L
beakeror flask combine2.50 g of DAN and 250 mL of 0.1 M HCl; (ii) heat at
60°C in a water bath for 20 min while stirring occasionally;(iii) add 250 mL of
0.1 M HCI and transfer to a 1-L separatoryfunnel; (iv) while working in an
appropriatehood, add about a 5-cm layer of cyclohexaneand mix carefully; (v)
draw off and saveaqueouslayer and appropriatelydiscardthe cyclohexane;(vi)
placethe aqueouslayer in a cleanseparatoryfunnel and repeatsteps(iv) and (v)
twice more or until the aqueoussolution is clear; (vii) filter the purified DAN
solution through a Whatmanno. 41 filter paperusing a large glass funnel and
savethe filtrate in a dark glassbottle; (viii) add a 2.5-cm layer of cyclohexaneto
the top of the purified reagentand store in the refrigeratorbeing careful not to
freeze the aqueoussolution; (ix) before using the reagent,remove it from the
refrigeratorand let the cyclohexanemelt, shakethe solution, and allow separa-
tion of the two phasesfor 1 to 2 min.
The fluorimetric method couples an autoanalyzermanifold with a fluo-
rimeter to measuretotal Se in the solution resultingfrom soil digestion.Autoan-
alyzer manifolds can be constructed(e.g., Brown & Watkinson, 1977), or pur-
chasedcommercially(e.g., Fig. 30-2). Specific procedureswill vary depending
on the particular autoanalyzermanifold, fluorimeter, autosampler,and other
equipmentused,and the readeris referredto appropriateoperationalmanualsfor
detailed instructions for the particular system. Autoanalyzer manifolds are
designedto form the piazselenolcomplex betweenSe(IV) and DAN, and to
extractthe complexinto cyclohexane.The resultingsolution is pumpedto a flu-
orimeterwhere the piazselenolcomplex is irradiatedat 369 nm and its fluores-
cenceis measuredat 525 nm.
The fluorescenceof the sampleanalyzedis comparedto the standardcurve

to determinethe total Seconcentrationin microgramsper liter. This value is mul-
tiplied by any dilution factor resultingfrom dilution of the original digestedsolu-
tion and by 0.2 to obtain the concentrationof total Se in soil in milligrams per
kilogram. Calibrationfor fluorimetric analysesshouldbe doneon a daily basisor
with eachset of samplesanalyzed,whicheveris more frequent.
Comments
The methodpresentedabove to determinetotal Se in soil is comprisedof
two parts-soildecompositionand analysisof total Se in the solution resulting
from the soil digestion.The recommendeddigestionmethodis a modification of
the procedureof Welschet al. (1990), which is a modification of the methorl ini-
tially developedby Sanzoloneand Chao (1987) for determinationof Se in geo-
logic materials.The modificationsallow analysisof both Se andAs in the digest-
ed solution.
Interferencesduring the determinationof Se by HGAAS are well docu-
mentedin the literature (e.g., Pierce & Brown, 1977; Roden & Tallman, 1982;
Briggs & Crock, 1986; Sanzolone& Chao,1987).Interferencesgenerallyare due
to high concentrationsof transition metals [e.g., Fe(II,III), Cu(II), Ni(II)] and
otherelements(e.g.,As, Sb) in the samplethat cause areductionin the Se absorp-
tion signal of the AAS. For most soils, concentrationsof theseelementsgeneral-
ly are low enoughso that their interferenceis not a problem.For samplesthat
have concentrationsof transition metals high enough to cause interference,a
strong-basecationexchangeresin canbe usedto removethe interfering transition
metals(Am. Public Health Assoc., 1992).
Other interferencescan be result from incomplete oxidation of organic
materialduring the digestion,which impedesthe formation and volatilization of
H2Se (Roden& Tallman, 1982). Also, somereagent-gradeHel containsenough
el2 to causeback oxidation of Se(IV) to Se(VI) during the HCl reductionstep.
Bubbling an inert gas such as He through the concentratedHel will remove
excessCl2 prior to use (Cutter, 1986).
Brown and Watkinson(1977) testedmany elementsfor potential interfer-
ence during fluorimetric determinationof Se. With the addition of EDTA or
oxalate,they found no significant interferencefor the elementstested,exceptfor
Pd(II) and Sb(IV). They also caution about the presenceof strong oxidizing or
reducing agentsthat can react with either Se(IV) or DAN and causeinaccurate
results.
The above recommendeddigestion procedureand HGAAS were used to
analyzeSe in standardreferencesoils and geologicmaterialshaving diversemin-
eral compositionsand reported Se concentrationsranging from 0.3 to 5.6 mg
kg-I. The resultsof analysesof six samplesrelative to Se values reportedin the
literature,rangedfrom -6.7 to +4.2% relative error; resultsof one samplehad a
-19.8% relative error. The precisionof the measurements (6-25 replicateanaly-
ses)rangedfrom 2.0 to 6.3% relative standarddeviation(E.P. Welsch, 1994, per-
sonalcommunication).
804 HUANG & FUJII
Total Seleniumand SeleniumSpeciesin Soil Solutions

and WaterExtractsof Soil
The total concentrationof Se in natural waters dependson the biogeo-
chemicalprocessescontrolling the solubilities of Se species.Under equilibrium
conditions,the predominantinorganicSespeciesis a function of the redox poten-
tial and pH of the systemas describedabove. The solubility of inorganic Se
speciesin soils generally is controlled by mechanismssuch as mineral dissolu-
tion andprecipitation(Geeringet aI., 1968; Elrashadiet aI., 1987;McNeal & Bal-
istreiri, 1989), ion pair formation in solution (Elrashadiet aI., 1987; Cowan,
1988), adsorptionand desorption(Neal et aI., 1987a,b;Neal & Sposito, 1989;
Balistreiri & Chao,1990; Fio et aI., 1991),and biological uptake(Fleming, 1980;
Mikkelsen et aI., 1989). In contrast,concentrationsof organicforms of Se (e.g.,
selenomethionine,selenocysteine,methylatedSe species)in naturalwatersgen-
erally result from microbiologicalreactionssuchas mineralizationof soil organ-
ic matterby microorganisms(Wereset aI., 1989;Abramset aI., 1990)andmethy-
lation of inorganicSe speciesby fungi and other soil microorganisms(Franken-
berger& Karlson, 1989; Thompson-Eagleet aI., 1989; Karlson & Frankenberg-
er, 1990a,b).
Total Seleniumin Soil Solutionsand WaterExtractsof Soil

Principles.Total Se in soil solutionsand waterextractsof soil can be com-
posedof inorganicandorganicSespeciesof differing oxidationstates;therefore,
water samplesmust be digestedto quantitativelytransformall Se speciesto the
speciesdetectedby the analytical methodused.Both of the recommendedana-
lytical detectionmethods,HGAAS and fluorimetry, are sensitiveonly to SeelY).
The recommendeddigestionprocedure,similar to that describedby Makita and
Fujii (1992), is a modification of a proceduredescribedby Presserand Barnes
(1984) and requiresan initial oxidationof the sampleusing potassiumpersulfate
(KzSzOg) at elevatedtemperature.This oxidative digestion transformsall Se
speciesto Se(VI). Next, the sampleis digestedafter the addition of HCI to quan-
titatively reduceSe(Vl) to Se(IV) (Brimmer et aI., 1987)
[2]
Total Se,all specieshavingbeentransformedto Seely),is then detectedby either

HGAAS or fluorimetry, as describedbelow.
Apparatus
1. Acid fume hood.
2. Laboratoryhot plate.
3. Erlenmeyerflasks, 250 mL, with tuttle caps(availablefrom FisherSci-
entific).
4. All apparatusdescribedin "Apparatus"sectionfor HGAAS and in the
"Apparatus"sectionunder"Fluorimetry."
Reagents
1. Nitric acid, 0.4% (v/v): Prepareby dilution of concentratedHN03, 16
M, with deionizeddistilled water.
SELENIUM AND ARSENIC 80S
2. Oxalic acid (C2H20 4), 3.5% (w/v): Prepare by dissolving 35.0 g

C2H20 4 powder in deionizeddistilled water and diluting to 1 L with
deionizeddistilled water.
3. Potassiumpersulfate,0.2% (w/v): Prepareby dissolving 2.0 g K2S20 g
in deionizeddistilled water,and diluting to 1 L with deionizeddistilled
water.
5. All reagentsdescribedin the "Reagents"sectionfor "HGAAS" and the
"Reagents"sectionfor "Fluorimetry." The only differenceis that 0.4%
HN03 (insteadof 1.2 M HCI) is used to make serial dilutions of Se
standardsolutions for HGAAS analysis.Standardsolutionsprepared
for both HGAAS and fluorimetric methodsare treatedas samplesand
are digestedas describedbelow.
Procedure.Pipet 20 to 40 mL of filtered (0.45 11m), acidified (pH < 2 with
either HN03 or HCI) sampleinto a 250-mL Erlenmeyerflask and dilute to about
40 mL with 0.4% HN03. Add 3.3 mL of 7 M HCI and 4 mL of 0.2% K 2S20 g to
eachsample.Cover flasks with tuttle capsand boil on the hot plate for 15 to 20
min. Removesamplesfrom the hot plate and let cool. Add 4 mL of 3.5% oxalic
acid to eachsampleand boil samplescoveredwith tuttle capson the hot plate for
an additional 15 to 20 min. Removesamplesfrom the hot plate and let cool. Add
15 mL of concentratedHCI to eachsample.Boil samplescoveredwith tuttle caps
on the hot plate for 35 to 40 min, watch carefully to preventevaporationof any
sampleto dryness.Removesamplesfrom the hot plate and let cool.
Quantitatively transfer samplesto 50-mL volumetric flasks and dilute to
volume with deionizeddistilled water for analysisof Se as describedabovein the
section "Selenium Determination."Samples to be analyzed fluorimetrically
should be first treatedas describedabove in "Procedure"under the section for
"Fluorimetry." Resultsfrom the HGAAS or fluorimetric analysesare compared
to the respectivestandardcurve and multiplied by any dilution factor resulting
from dilution of the original sample to obtain the concentrationof total Se in
microgramsper liter.
Seleniteand SelenateSpeciesin Soil Solutionsand Water

Extractsof Soil
Principle. The HGAAS and fluorimetry methodsdetect only the Se(IV)
species.This specificity is usedto determineSe(IV) and Se(VI) speciesin water
by HGAAS and fluorimetry. The recommendedHGAAS methodis similar to that
reported by Presserand Barnes(1984) and Fio and Fujii (1990). The recom-
mendedfluorimetric method usesthe samesamplepreparationprocedures,but
fluorimetry is usedto detectSe in the resulting solutions.Seleniteis determined
directly using one aliquot of sample.A separatesamplealiquot is digestedwith
HCl at elevatedtemperatureto quantitativelyreduceSe(VI) to Se(IV) (Brimmer
et aI., 1987). Analysis of this aliquot representsthe concentrationof Se(IV) plus
Se(VI), and Se(VI) is calculatedfrom the differencebetweenthe two analyses.A
third samplealiquot is analyzedfor total Se, and an estimateof soluble reduced
Sespeciesis calculatedas the differencebetweentotal SeandSe(IV) plus Se(VI).
806 HUANG & FUJII
The reduced Se speciesrepresentSe present in the -II or 0 valence states.

Becausethe concentrationof reducedSe is usually much lower than Se(VI) and
Se(IV), and the errorsassociatedwith the Se(VI) and Se(IV) analysescan be rel-
atively large comparedto the reducedSe concentration,it is frequently difficult
to accuratelyquantify the concentrationof reducedSe. An analysisof organic
forms of Se also can be directly measuredand is further discussedbelow.
Apparatus
1. SO-mL borosilicateglasstest tubeswith Teflon lined caps.
2. Water bath, 90°C.
3. All apparatusdescribedin "Apparatus"for HGAAS and fluorimetry.
Reagents
1. 1000 mg L-l selenite[Se(IV)], as Se, solutions for spiking samples:
Prepareby first drying sodium selenite(Na2Se03)at lOSoC overnight
and cooling in a desiccator;dissolve 2.1902g Na2Se03in deionized
distilled water,add 8.3 mL of concentratedHCI, and dilute to 1 L with
deionizeddistilled water. Store the Se(IV) solution in the refrigerator
and preparea new solution at leastevery 6 mo.
2. 1000 mg L-l selenate[Se(VI)], as Se, solutionsfor spiking samples:
prepareby first drying sodiumselenate(Na2Se04)at lOSoC overnight
and cooling in a desiccator;dissolve 2.3928 g Na2Se04in deionized
distilled water, add 8.3 mL of concentratedHCI, and dilute to 1 L with
deionizeddistilled water. Store the Se(IV) solution in the refrigerator
and preparea new solution at leastevery 6 mo.
3. All reagentsdescribedin "Reagents"sectionfor HGAAS and "Fluo-
rimetry."
Procedure.Analyze an aliquot of the samplefor total Se as describedin
"Procedure"for both HGAAS and fluorimetric method.Analyze anotheraliquot
of samplefor concentrationof Se(IV) directly as describedin "Procedure"for
HGAAS and "Procedure"for fluorimetry. A third aliquot of sampleis digestedto
selectivelyand quantitativelyreduceSe(VI) to Se(IV) using a methodsimilar to
that describedby Brimmer et al. (1987).Pipet lO-mL aliquotsof eachsampleinto
a borosilicateglass test tube. Add an equal volume of concentratedHCI and
loosely capthe test tubeswith Teflon-linedcaps.Heat the samplesin a waterbath
at about90°C for 30 min. Removethe samplesfrom the water bath and let cool.
Add 20 mL of water toeachsample,this resultsin a samplematrix of 3 M HCl.
Analyze the digestedthird aliquot of sample as describedin "Procedure"for
HGAAS or "Procedure"for fluorimetry to obtain the concentrationof Se(IV)
plus Se(VI). The concentrationof Se(VI) is calculated from the difference
betweenthe resultsfor Se(IV) only and Se(IV) plus Se(VI), and an estimateof
reducedSe is calculatedfrom the difference betweentotal Se and Se(IV) plus
Se(VI).
Comments
The methodfor analysisof total Se in soil solutionsand water extractsof
soil using HGAAS has been used by the U.S. Geological Survey (USGS),
ResearchLaboratory in Sacramento,California, for the past severalyears. The
laboratory has participated in numerous"round-robin" and referencematerial

exchangeprogramsconductedby the USGS(V. Janzer,1989 and 1990, personal
communication)and by the U.S. Bureau of Reclamation,San Joaquin Valley
DrainageProgram(W.w. Walker, R.G. Burau,andA. Jacobsen,1988 and 1989,
personal communication).Results using this method always were acceptable,
within at least 12.5% of the certified or most probablevalue for referencesam-
ples. Repetitive analyses(n = 64) of an internal USGS referencewater sample
(agriculturaldrain water) over a 2-yr period resultedin a meanand standarddevi-
ation of 62.2 ± 2.4 ).lg L-l total Se. The reportedmost probablevalue and stan-
dard deviation for the sample is 64.2 ± 2.3 ).lg L-l total Se. The method has a
practical detectionlimit of 0.5 ).lg L-l.
This procedure has been successfullyapplied by the USGS California
ResearchLaboratoryalso using 125-mL Erlenmeyerflasks and reducingsample
aliquots and reagentsby one-half. It also has beensuccessfullyusedwith small-
er volumesof sampleand proportionaldecreasesin reagents,and with test tubes
or Taylor tubesand an aluminum heatingblock.
Soil solutionsand water extractsof soil to be analyzedfor total Se gener-
ally are filtered (0.45 ).lm) and preservedwith eitherHN03 or HCl to a pH < 2 to
preventloss of Se from solution. Aqueoussamplestakenfor total metal analysis
generally are acidified to pH < 2 with HN03 for preservation,but acidification
with HCl also is acceptablefor total Se analysis.The procedurefor total Se in
aqueoussampleswas originally developedfor samplesacidified to pH < 2 with
HN03, which is the rationalefor using 0.4% HN03 for initial dilution of samples
prior to digestion.Acidification preventspotential loss of Se dueto coprecipita-
tion of Se specieswith supersaturatedmineral phases[e.g., coprecipitationof
Se(IV) with CaC03]. Acidification also preventscoprecipitationof Se species
with precipitating Fe and Mn solid phases resultingwhen reducing water con-
taining high concentrationsof Fe(II) and Mn(I1) becomesoxidized. Methylation
of Se and subsequentvolatile loss of Se also is inhibited under acidic conditions
(Frankenberger& Karlson, 1989; Thompson-Eagleet aI., 1989).
Environmental water samples and soil extracts can contain interfering
organic (Workman & Soltanpour, 1980; Roden & Tallman, 1982; Am. Public
Health Assoc., 1992) and inorganic (Pierce & Brown, 1977; Narasaki& Ikeda,
1984; Briggs & Crock, 1986) substancesthat can cause erroneously low
absorbances during analysisof total Se and Se speciesusing HGAAS.
When initially analyzing sampleshaving a new or different matrix for
Se(IV) and Se(VI), a split of the sampleshouldbe spikedwith known concentra-
tions of both Se(IV) and Se(VI). Ideally, the massof Se(IV) and Se(VI) added
should result in an addedconcentrationcomparableto that originally presentin
the sampleso that recoveriesare easily evaluated.Also, the Se speciesconcen-
tration of the spiking solution shouldbe high enoughthat the volume of the sam-
ple is not significantly changed.If spike recoveriesare lessthan 85% (refer to the
ONOC criterion in Chapter2, Klesta & Bartz, 1996), the method of standard
additionsshouldbe usedto determineSe(IV) and Se(VI) (Friedman& Erdmann,
1982; Fishman& Friedman,1989). For recoveriesless than about75%, the stan-
dard additionsmethodmay not work (Am. Public Health Assoc., 1992) and the
interferant(s)may needto be removed(this is discussedin more detail below).
808 HUANG & FUJII
It is much more difficult to assessthe accuracyand precisionof Se species

analysesbecauseof problemsrelatedto preservingthe speciesdistribution. Cut-
ter (1986) determinedSe speciesusing HGAAS combined with a liquid Nr
cooledtrappingand preconcentrationsystem.The precisionof the methodhad a
relative standarddeviation no greater than 5.4% for a reservoir sample, the
methodhas a theoreticaldetectionlimit of 0.2 ng Se. Water samplesto be ana-
lyzed for Se speciesshouldbe filtered (0.45 Ilm) and preservedwith HCl to pH
< 2 and refrigerated(Cutter, 1986).
As discussedabove, high concentrationsof transition metals can causea
reductionin the Se absorptionsignal of the AAS causinginaccuraciesduring the
analysisof Se species.Undertheseunusualcircumstances,interfering metalscan
be removed using a strong-basecation exchange resin (Am. Public Health
Assoc.,1992).
Roden and Tallman (1982) reported interferencesby dissolved organic
compoundsduring the analysis of Se(IV) and Se(VI) speciesin groundwater
using HGAAS. Removalof hydrophobicorganiccompoundsusing XAD-8 resin
(SigmaChemicalCo., St. Louis, MO) hasbeenshownto amelioratethis problem
(Roden& Tallman, 1982; Fio & Fujii, 1990). Cutter (1986) useda Sep-PakC18
cartridge (WatersAssoc., Milford, MA) to remove organic selenidefrom solu-
tions and analyzedtotal Se in the eluatesas a direct measureof organic Se.
Cutter(1983) reportedthat concentrationsof nitrite as low as 10 Ilg L -1 can
interferesignificantly during the analysisof Sespecies.Addition of sulfanilamide
hasbeenshownto alleviatethis interference(Cutter, 1983).
The abovediscussionof Se speciesfocusedmainly on Se(IV), Se(VI), and
reducedSe species,where reducedSe representsSe presentin the -II or 0 oxi-
dation states.Under oxidizing to mildly reducingconditions,Se(VI) and Se(IV)
usually predominateand generallyare the most solubleSe species(Masscheleyn
et aI., 1989; McNeal & Balistreiri, 1989).Although usually presentin relatively
low concentrations,methylatedforms of Se have beenreportedin the literature
in both soil and water systems(Doran & Alexander, 1977; Cutter, 1978; Cooke
& Bruland, 1987; Frankenberger& Karlson, 1989; Masscheleynet aI., 1989;
Thompson-Eagleet aI., 1989; Thompson-Eagle& Frankenberger,1990; Mass-
cheleynet aI., 1991a).Also, the importanceof other organic forms of Se in the
biogeochemicalcycling of Se has beenextensivelystudied(Cutter, 1982; Cutter
& Bruland, 1984; Takayanagi& Wong, 1984; Wereset al., 1989; Abrams et aI.,
1990; Velinsky & Cutter, 1991).Therefore,it is importantto initially evaluatethe
significanceof volatile and organicforms of Se in any systemstudied.
SeleniumAvailability
The discussionsand methodsdescribed above focusedon the total concen-

tration of Se in soils and total Se and inorganic Se speciesin soil solution and
water extractsof soil. Uptake of Se by plants (e.g., Mikkelsen et aI., 1989) and
soil-test methods aimed at predicting plant uptake of Se (e.g., Soltanpour&
Workman, 1980;Jump& Sabey,1989)are reviewedin the literature.Additional-
ly, an understandingof the partitioning of Se speciesbetweensolution and solid
phasesand potential mechanismsof Se releaseassociatedwith solid phasesis
neededto assessthe short-termand long-term bioavailability and mobility of Se

in soils.
Recently,Jump and Sabey(1989) comparedSe extractedfrom 18 different
soils and mine-spoil materialsby severalsoil-test extractantswith Se uptake by
two Se-accumulatingplant species.They found that Seconcentrationsin extracts
of water-saturatedsoil pastes correlated highest with plant concentrations.
Soltanpourand Workman (1980) used an ammoniumbicarbonate-diethylenetri-
aminepentaacetic acid (DTPA) extractantand found a high degreeof correlation
betweenextractedSe and Se taken up by alfalfa (Medicago sativa L.) for five
North Dakotasoils treatedwith five levels of Se(VI) in a greenhousestudy.
The soil-testextractionsdiscussedabovewere not analyzedfor Se species.
Mikkelsen et ai. (1989) discuss the different mechanismsrelated to uptake of
Se(VI) (energydependent)and Se(IV) (energy independent)that should be con-
sideredin relation to the concentrationsof Se(VI) and Se(IV) extractedor present
in the soil solution. An addedcomplicationis the variable uptakeof Se by differ-
ent plant species(Mikkelsen et aI., 1989). Determinationof Se speciesalso may
provide insight into the geochemicalprocessescontrolling Se solubility.
Anotherapproachusedto evaluaterelatively labile forms of Se in soils uses
orthophosphate(P04) as a soil extractant.Fujii et al. (1988) used a 0.32 M P04
extract to estimateadsorbedSe in alkaline soils from the westernSan Joaquin
Valley and found relatively low concentrationsof Se extracted«4% of the total
soil Se), comprised mostly of Se(IV). The method is basedon the assumptions
that P04 exchangeswith adsorbedforms of Se and that the dominantadsorbedSe
speciesin thesesoils is Se(IV). Further refinementof this methodshowedthat a
0.1 M P04 solution adjustedto pH 8 extractedbetween89 and 103%of the
adsorbedSe(IV) which was determinedby exchangewith 75Se(IV) for three sur-
face soils (Fujii & Burau, 1989).
Other forms of Se are presentin soils as relatively insoluble solid phases
and representpotential long-term sourcesof Se. Sequentialselective-extraction
or partial-dissolutionmethodscommonly used to identify solid-phaseassocia-
tions of trace-metalcationsin sedimentsand soils (e.g.,Tessieret ai., 1979; Chao,
1984) are not readily applicable to redox-sensitivetrace elements,such as Se.
Gruebelet ai. (1988) reportedthat Se(IV) associatedwith amorphousiron oxides
is readsorbedonto other mineralsafter reductivedissolution,a methodcommon-
ly used to evaluate trace-metal associationswith iron oxides (Chao & Zhou,
1983).They also confirmedthat oxidantsusedto dissolveorganicmatter,suchas
hydrogenperoxideor sodium hypochlorite,oxidized Se(IV) adsorbedon miner-
al surfacesto Se(VI), which was subsequentlyreleasedto solution. Thus, if these
selective extraction techniquesare applied to soils and sediments,they may
underestimateSe associatedwith amorphousiron oxides and overestimateSe
associatedwith organic matter.
Sequential selective-extractionmethods recently have been developed
specifically for fractionatingSe in soils (Chao& Sanzolone,1989; Lipton, 1991).
The method developedby Chao and Sanzolone(1989) determinesSe in five
operationally defined soil fractions. General information regarding extractant,
forms extracted,mechanismsof extraction, and relative availability index for
eachfraction are listed in Table 30-1 [seeChao & Sanzolone(1989) for details].
810 HUANG & FUJII
Table 30-1. Generalinformation on the partial dissolutiontechniques.

Availability for
Formsextracted leachingand
Extractant or dissolved Mechanismsinvolved plant uptaket
O.25MKCI H20 soluble,nonspecifically Massaction, anion Highly available

adsorbedselenate exchange
Exchangeable,specifically Anion competition, Available
adsorbedselenite ligand exchange
4MHCI AssOciatedwith oxide minerals Acid dissolution, Conditionally
(Fe, Mn, and Al and amorphous acid hydrolysis available
materials),carbonates,acid-
volatile sulfides,and acid
hydrolizableorganicmatter
KCI04 + concentrated Sulfides,complex humified Oxidative acid Unavailable
HCI organicmatter dissolution
HF+HN03 Siliceousmaterials Acid destructionof Resistant
+ HCI04 mineral-lattice
structure
t Availability for leachingandplant uptakebasedwason the conceptualrelative rating betweenbeing
highly availableand being resistant.
Their resultsfor soil samplesfrom Hawaii and the westernSan JoaquinValley,

California, and four USGS geochemicalreferencematerials,show contrasting
distributionsof Sein variousfractions,generallyrelatedto the geochemicalenvi-
ronmentsfrom which the sampleswere taken.
Lipton (1991) developeda more extensivesequentialselective-extraction
methodthat determinesSe associatedwith nine operationallydefined soil frac-
tions. This methodwas developedand applied to three agricultural surfacesoils
from the westernSan JoaquinValley, California (Lipton et aI., 1988; Lipton &
Doner, 1989; Lipton, 1991).Generalinformation regardingextractantsandforms
extractedare listed in Table 30--2, along with resultsof the three soils analyzed.
Detaileddescriptionand evaluationof the methodis availablein Lipton (1991).
This method uses P04 or KOH extracts following certain extraction steps to
assuredesorptionof any adsorbedSe(IV) that originally was associatedwith the
selectivelydissolvedsolid phase.Data in Table 30--2 show that most of the Se
was associatedwith the carbonate,oxidizable,crystalline-oxide,and alkali-solu-
ble alumino-silicatephases.Theseresultsimply that substantialconcentrationsof
Table 30-2. Selectivesequentialextractionfor Se, three surfacesoils from the westernSan Joaquin
Valley.
Percentage
Extractant Formsextracted of total Set
1. 0.25MKCI Soluble 2~
2.0.1MP04 Ligand exchangeable 4
3. 1 M NaOAc, pH 5 (P04 following) Carbonates 8-19
4. NaOCl, pH 9.5 (boil) Oxidizable(organicmatter) 10-23
5. 0.1 M NH20H-HCI, pH 2 (KOH following) Easily reducibleoxides 2-7
6. 0.25 M NH20H-HCI, 0.25M HCI (KOH following) Amorphousoxides 3-8
7. 4 M HCI (boil) Crystallineoxides 20-32
8. 0.5 M NaOH (boil) Alkali solubleAl/Si 20-30
9. HN03 + HCI04 Residual 1-5
t Total Se rangedfrom 0.8 to 1.0 mg/kg.
Se are associatedwith thesesoil fractions that may be releasedover time through

weatheringprocesses.
The methodsdescribedin "Total Selenium in Soil Solutions and Water
Extractsof Soil" for analysisof total Se in soil solutionsandwaterextractsof soil
can be usedto measureSe extractedby various extractionmethods,but matrix-
matchingtechniquesand extensiveQNQC proceduresshould be usedto assure
the quality of the analysis.
ARSENIC
Introduction
Arsenic is presentin the Earth'scrust at an averageof 2 to 5 mg kg-I. The
mean concentrationsof As in sandstones,igneous rocks, shales,and volcanic
rocks are 1.0, 1.8, 13, and 20 mg kg-I, respectively(Onishi & Sandell, 1955;
Kipling, 1977; Lemmo et aI., 1983). Sedimentaryrocks, including coal, contain
0.1 to 2000 mg kg-I as As (Irgolic et aI., 1983). The most commonAs-bearing
minerals are arsenopyrite(FeAsS), enargite (Cu3AsS4),orpiment (As2S3), and
realgar(As4S4).
Arsenic is presentin soils as sulfide minerals, complex sulfides of metal
cations, and as a constituentretained by soils in the adsorbedor precipitated
forms. The role of hydrous oxides in the adsorptionof As is well established
(Walsh & Keeney, 1975; Livesey & Huang, 1981). Arsenic also is adsorbedon
other mineral colloids, bound to organic matter, and may form sparingly water-
soluble compounds with AI, Fe, Ca, and Mg in soil environments(Huang, 1987).
In general,arsenateis the most strongly adsorbedAs species(Creceliuset aI.,
1986), althoughadsorptionof arsenitealso is reportedin the literature (Ferguson
& Anderson,1974; Oscarsonet aI., 1980; Pierce& Moore, 1980; Elkhatib et aI.,
1984).
Soils overlying sulfide ore depositshave beenreportedto contain as much
as 10 000 mg kg- I As (Ganje & Rains, 1982). Arsenic in uncontaminatedsoils
rangesfrom 0.2 to 40 mg kg-I, with an averageof about5 mg kg-I, generallynot
high enoughto causephytotoxicity. Application of As herbicidesor pesticidesto
soils may increasesoil As levels to about 550 mg kg- I (Walsh & Keeney,1975;
Nati. Res. Council, 1977).
Weatheringof As-containingmineralsliberatesAs in the form of inorgan-
ic compounds,which include arsenic trioxide, arsenite,and arsenate.Microor-
ganismsincreasethe rate of As releaseby catalyzingthe oxidation of sulfide to
sulfate and ferrous to ferric Fe. Weatheringof As-containingmineralsis consid-
eredthe major naturalsourceof As (Ferguson& Gavis, 1972).The weatheredAs
compoundsmay be retainedin soils or dissolvedin water and transported, and
further redistributed(Wakao et aI., 1988). Two additional major sourcesof As
influx to soil are precipitationfrom the atmosphereand the usesof arsenicalsin
agriculture. Inorganic forms of As, such as insecticides,herbicides,fungicides,
desiccants,defoliants, and additives to animal feeds, were extensively used in
agriculture prior to 1968 (Ganje & Rains, 1982).These inorganic forms of As
havesincebeenreplacedby organoarsenicals to decreasephytotoxic responsesof
812 HUANG & FUJII
cropsand potentialhazardsto humansand animals.Many of thesearsenicalsdo

not persistin soils and are believedto be removedrapidly by volatilization and
possiblyleaching(Morrison, 1969).However,excessiveapplicationof all arseni-
cals can lead to pollution of agricultural drainageand surface run-off waters
(DubIe et aI., 1978).
Dissolved As can occur in natural waters in both inorganic and organic
forms (Anderson & Bruland, 1991). Inorganic forms of dissolved As include
arsenateand arsenite.Primary aqueousspeciesundernatural pH conditions(pH
4--8) are anionic in arsenate[As(V), H2AsO.$" and HAsOl-] underoxidizing con-
ditions, or neutralfor arsenite[As(III), As(OH)30] under reducingconditions.In
terrestrialand aquaticenvironments,Mn oxidescan serveas catalyststo oxidize
As(III) to As(V) (Oscarsonet aI., 1981a,b;Huang, 1991). Arsenateis less toxic
than As(III) (Oscarsonet aI., 1981a).
The USEPA classifiesAs as a priority pollutantwith an aquatic-lifecriteri-
on for chronic exposureto As(III) of 190 Ilg L-l (USEPA, 1986).The maximum
contaminantlevel for As in drinking water is 50 Ilg L-l and a draft advisory con-
centrationof 21lg L-l hasbeenproposedrelative to cancerrisk (USEPA, 1993).
Measurableconcentrationsof methylated As also are found in natural
waters (Anderson & Bruland, 1991). Organismsmay have developedmecha-
nisms to isolate and detoxify As by producingorganoarsenicals (Wood, 1974).
Furthermore,incorporationof As into arsoniumzwitterionssuchasarsenobetaine
and arsenocholinemay servedual purposesof detoxificationand osmoregulation
analogousto someS compounds(Yancey et aI., 1982).
Total Arsenic in Soil

Many analytical methodsare availablefor the determinationof total As in
soils. Theseinclude: (i) nondestructivemethodssuchas NAA (Salmon& Cawse,
1991) and XRF (Jones,1991); and (ii) methodsthat require sampledecomposi-
tion followed by analysisof As in the resultingsolution using methodssuch as
colorimetry(Ganje& Rains, 1982; Assoc.Official Anal. Chern.,1990),HGAAS
(Welschet aI., 1990), ETAAS (Am. Public Health Assoc., 1992), ICP-AES, and
inductively coupledplasma-massspectrometricmethods(Sharp,1991). Because
NAA requiresaccessto nuclearreactorsand XRF canonly be appliedto samples
with relatively high concentrationsof As, the most common methods used
require initial decompositionof the soil as discussedin this section.
Two methodsare recommendedfor the determinationof total As in soil and
are discussedbelow in detail in "Principles" and "Methods." Unlike the recom-
mendedmethodsfor analysisof total Sein soil ("Total Seleniumin Soil") that use
one soil-digestionprocedure andtwo methodsto analyzeSe in the resultingsolu-
tion, eachAs determinationmethodis coupledto a specific digestionprocedure
for analysisof total soil As. Analysisof As by the diethyldithiocarbamate
method
(colorimetry) is an official methodapprovedby the AOAC (1990) and the Amer-
ican Public Health Association(1992) and is used following the HN03-H2S04
digestionprocedure(Method A). However, continuous-flowHGAAS is usedto
determine As in solutions resulting from the HN03-HCI-HCI04-H2S04-HF
digestionprocedure(Method B).
The molybdenumblue method is another colorimetric method which is

commonly usedand also is approvedas an official methodby the AOAC (1990).
This method is more sensitivethan the silver diethyldithiocarbamate(Ag-DDC)
methodbut consumesmore reagentsand is not as rapid.
Determinationof As using ETAAS is subject to significant interferences
from molecularabsorption,as well as chemicaland matrix effects.Although pre-
vious indicationswere that low optimum concentrationranges(5-100 Ilg L- 1 As)
were attainable,more recentdatashow that this may not be so. Exerciseextreme
caution when applying the methodto the lower concentrationranges(Am. Pub-
lic Health Assoc., 1992). In ETAAS, furnace temperaturesettingsmust be care-
fully optimized to avoid samplespatteringor prematurevolatilization of the ele-
ment being determined.Atomization temperaturemust be selectedby determin-
ing the temperatureproviding maximumsensitivity withoutsignificantly eroding
precision.The charringtemperaturemust be high enoughto maximizevolatiliza-
tion of interfering matrix componentsyet too low to volatilize the element of
interest.When the optimum charringtemperatureis exceeded,therewill be a sig-
nificant decreasein sensitivity.
Recentapplicationsof HGAAS coupledto ICP-AES (Bakhtaret aI., 1989)
and FIA combinedwith HGAAS (Chan & Sadana,1992) to As analysisin envi-
ronmentalsamplesindicate the potential for further developmentof thesemeth-
ods for As analysisin soils and st:diments.
Principles
Digestion of Soils. Digestion of soils with acid mixtures is essentialto
decomposesoil componentsthat bind As. Oxidizing-aciddigestiondissolvespre-
cipitated,adsorbed,sulfide-bound,loosely silicate-bound,and organically bound
As. The HNOr H 2S04 digestionprocedure(Method A) (Ganje & Rains, 1982) is
the recommendedmethodfor soil digestionprior to analysisof As by the colori-
metric methoddescribedbelow in "Colorimetry-Silver Diethyldithiocarbamate
Method (for Analysis of Soil Sample Digestedusing Method A)." The second
recommendeddigestion procedure(Method B) usesHNOr HCI-HCI04-H zS04-
HF (E.P. Welsch, 1992, personalcommunication)to digest soil samplesand is
designed for analysis of As using the HGAAS method described below in
"Hydride GenerationAtomic Absorption Spectrometry(for Analysis of Soil
SampleDigestedUsing Method B)", This digestionprocedureis a modification
of the method describedby Welsch et ai. (1990) and also is recommendedfor
total Se in soil (see"Digestion of Soil"). Thus, solutionsresulting from samples
digestedusing Method B can be analyzedfor both As and Se by continuous-flow
HGAAS. For samplescontainingrelatively high concentrationsof organicmatter
(greaterthan about 40 g kg- 1 organic C), Method B requiresthat samplesstand
overnight at room temperatureafter the initial addition of HN03 and HCI (E.P.
Welsch, 1992, personal communication).
Arsenic Determination.Two commonly usedmethodsto determineAs in
solution, namely, colorimetry and HGAAS, are the recommendedmethodsand
are outlined in this section.The colorimetric method is the Ag-DDC method
(Ganje & Rains, 1982; AOAC, 1990). The HGAAS method is similar to the
814 HUANG & FUJII
methodsdescribedby Welschet al. (1990)andthe AmericanPublic HealthAsso-

ciation (1992).
Colorimetry-Silver Diethyldithiocarbamate Method. In an arsinegenera-
tor, inorganicAs is reducedto arsine,AsH3, by Zn in acid solution. The arsineis
then passedthrough a scrubbercontaining glass wool impregnatedwith lead
acetatesolution and into an absorbertube that containsAg-DDC dissolvedin
pyridine or chloroform. In the absorber,As reactswith the silver salt, forming a
solublered complex that is suitablefor spectrophotometric determination.
Hydride Generation Atomic Absorption Spectrometric Method. After initial
conversionof all inorganicand organicAs compoundsto As(V) by the oxidizing
digestion,As(V) is convertedby the sodiumborohydride reagent in acid solution
to volatile AsH3• The hydride is continuouslypurgedby Ar into a heatedquartz
cell mountedin the light path of an AAS where Asabsorptionis measured.The
dilution of the hydride by the carrier gas is minimized by rapid generationof the
hydride in an appropriatereactioncell.
Methods
Digestion of Soils
Method A: For Colorimetric Method
APPARATUS
1. Digester, with rheostat-controlledelectric heaters, comparable to
micro-Kjeldahl rotary digestionapparatus.
2. Digestionflask, micro-Kjeldal, 100 mL.
3. Glassbeads.
4. Acid fume hood.
REAGENTS
1. Nitric acid, concentrated,16 M.

2. Sulfuric acid, concentrated,18 M, high purity.
3. Ammonium oxalatemonohydratesolution, 7.5% (w/v): Dissolve 75 g
of (NH4nC204• H20 in deionizeddistilled water, and dilute to 1 L.
PROCEDURE. Place 1.000 g of the ground «0.15 mm) soil sample in the
micro-Kjeldahl flask, add three glass beads,30 mL of concentratedHN03, and
swirl to mix. Placethe flask on the micro-Kjeldahl-typedigesterin an acid fume
hood. Slowly heat the samplefor 45 min to avoid foaming or bumping. Rotate
the flask when necessaryto preventsoil caking. Graduallyincreasethe tempera-
ture to producesteady boiling until 2 to 4 mL of HN03 remains in the flask.
Removethe flask from the digesterand allow to cool. Add 5 mL concentrated
H2S04, swirl to mix, placethe flask on the digester,and increasethe temperature
to boiling. After fumes cease,removethe flask from the digesterand cool. Add
25 mL of saturatedammoniumoxalatesolution, swirl to mix, and place the flask
on the digesterand continuedigestionuntil fumescease.Upon completionof the
digestion,the contentsin flask shouldbe light gray to nearly white. About 3 hare
1- 40mm~
TI
B mm 0 .0. GAS
DISPERSION T UBE
I
..
40
COARSE FRITTED
,ro
DISC 130 mm
24
46
1CON ICAL
TES T TUBE
KJELDAHL
FLASK
Fig. 30-3. Arsine generatorand absorptiontube (Ganje & Rains, 1982).
requiredto completethe entire process.After the digestionis completed,transfer

the flask to a portablerack in an acid fume hood and allow the flask to cool prior
to As determination(seenext "Procedure"section).
Method B: For Hydride Generation Atomic Absorption Spectrometric
Method. This method(modification of Welsch et aI., 1990) is the samedigestion
method describedfor Se in "Digestion of Soil." See this section for complete
descriptionsof apparatus,reagents,and procedurefor this method.Solutions
resultingfrom this digestionprocedurecan be analyzedby HGAAS for both As
[see"Hydride GenerationAtomic AbsorptionSpectrometry(for Analysis of Soil
SampleDigestedUsing Method B)"] and Se (see "Hydride GenerationAtomic
AbsorptionSpectrometry")to determinetotal As and Se in soil.
Arsenic Determination
Colorimetry-Silver Diethyldithiocarbamate Method (for Analysis of Soil Sample
Digested using Method A)
APPARATUS
1. Arsine generatorand absorptiontube (seeFig. 30--3). Funneltrap with

coarse-fitteddisc at upperend and separatebentglassdispersiontube,
comparableto Coming no. 33680 from Coming Glass Works (Com-
ing, NY). The funnel trap is fitted with two female standard-taper
ground-glassjoints and connectsthe Kjeldahl flask to the dispersion
816 HUANG & FUJII
tube. Placeglasswool above fitted disc in upper part of funnel trap.

Saturateglass wool with Pb(C2H30 2)z by using a disposableplastic
eye dropper.Removeexcessby light suction.
2. Glasswool.
3. Plasticeye dropper,disposable.
4. Test tube, conical, 15 mL.
5. Spectrophotometer, double-beamtype.
REAGENTS

2. Potassiumiodide solution, 15% (w/v). Dissolve 15 g KI in 100 mL
deionized-distilledwater. Store in a brl)wn bottle. Keep in the dark.
Discardwhen solution turns yellow.
3. Stannouschloride (SnCI2) reagent,40% (w/v). Dissolve 40 gAs-free
SnCl2 • 2H20 in 100 mL concentratedHCI.
4. Lead acetate [Pb(C2H30 2)z] solution, 10% (w/v). Dissolve 10 g
Pb(C2H30 2)z • 3H20 in 100 mL deionized distilled water. Prepare
weekly or when solution becomescloudy.
5. Silver diethyldithiocarbamatereagent,0.5%. Dissolve 0.5 g Ag-DDC
in 100 mL of colorless,spectral-qualitypyridine. Mix and store in a
dark container.Reagentis stablefor severalmonths.
6. Zinc metal, 0.85 to 0.60 mm (20 to 30 mesh),As-free.
7. Arsenic standardsolution: (i) Stock As solution, 1.00 mg mL-l As:
Dissolve 1.320g arsenictrioxide, As20 3 in 10 mL deionizeddistilled
water containing4 g NaOH, and dilute to 1 L with deionizeddistilled
water. (ii) IntermediateAs solution, 10.0 mg L- 1 As: Dilute 5.00 mL
stockAs solution to 500 mL with deionizeddistilled water. (iii) Stan-
dard As solution, 1.00 mg L-l As: Dilute 10.00 mL intermediateAs
solution to 100 mL with deionizeddistilled water, and mix. Prepare
daily. (Note: CommerciallyavailableAs standardsolutionsalso can be
used.)
PROCEDURE.Pipette4.0 mL Ag-DDC into the conical test tube.To the flask

that contains the digestedsample (see previous "Procedure"),wash down the
neck of the flask with 40 mL deionizeddistilled water, then add 5 mL concen-
tratedHCI, 2 mL KI reagentand 10 drops(0.5 mL) SnCl2 reagent.Allow to stand
at room temperaturefor 15 min but not longer than 30 min. Add 4.5 g of granu-
lar Zn to the flask for reduction of As(V) to As(III). Immediately connectthe
flask to the funnel trap containingglasswool saturatedwith Pb(C2H30 2)2 (Fig.
30-3). Connectthe otherendof trap to the bentglassdispersiontube with the fit-
ted end submergedin the Ag-DDC solution in a conical test tube. Lubricate
ground-glassjoints with water.
Allow 50 min for completeevolution of arsine. Warm the Kjeldahl flask
slightly to insure that all arsine is released.Pour solution from conical test tube
directly into a l-cm cell and measureabsorbanceat 535 nm, using a reagentblank
as the reference.
Treat portionsof standardsolution containing0, 1.0,2.0,5.0,and 10.0 jlg
As as describedabove. Comparesample absorbancesto the standardcurve to
determinethe concentrationof total As in soil in milligrams per kilogram.
Arsenic compoundsare toxic. Use caution and handlewith care.
Hydride Generation Atomic Absorption Spectrometry (for Analysis of Soil

Sample Digested using Method B)
ApPARATUS. The apparatusrequiredfor this method are the sameas those
listed in "Apparatus"for determinationof Se by HGAAS exceptthat an As EDL
is requiredinsteadof the Se EDL.
REAGENTS
1. Arsenic standardsolutions:Preparefrom commerciallyavailableAAS
standard(1000mg L- 1 As) by makingserialdilutions to concentrations
of 2,5, 12, and 18 jlg L -I As in 1.2 M HCI. Standardsolutionsshould
be madefor eachset of analysesand be treatedas samples.
2. Reagentsdescribedin "Reagents"under"Hydride GenerationAtomic
Absorption Spectrometry,"except that the Se standardsolutions are
replacedby the As standardsolutionsdescribedin step No.1 above.
PROCEDURE. After digestion is completed(see "Method B: For Hydride
GenerationAtomic Absorption SpectrometricMethod"), a suitable aliquot of
digestedsampleis analyzedusing a continuous-flowHGAAS (Fig. 30-1). Stan-
dard analytical proceduresfor determinationof As by continuous-flowHGAAS
are available in manufacturer'soperating manuals. The reader is referred to
Chapter4 (Wright & Stuczynski,1996), which also providesdetailsfor analysis
by HGAAS. Examplesof As analysisby HGAAS are available in the literature
(e.g., Welschet aI., 1990; Am. Public Health Assoc., 1992).
The continuous-flowHGAAS functions as describedin "Procedure"under
"Hydride GenerationAtomic AbsorptionSpectrometry"for Se, except for As
analysis,the arsine(AsH3) is formed and its absorbanceis measuredat 193.7nm.
The absorbanceof the analyzedsample is comparedto the standardcurve to
determinethe total As concentrationin microgramsper liter. This vaiue is multi-
plied by any dilution factor resultingfrom dilution of the original digestionsolu-
tion and by 0.2 to obtain the concentrationof total As in soil in milligrams per
kilogram. This methodhas a practical detectionlimit of 0.4 mg kg-I As.
Standardsolutionsshouldbe analyzedperiodically as samples(aboutevery
8 to 10 samples).If the value of the standarddeviatesby more than 10% from its
known value, the instrumentshouldbe recalibratedusingeithera one-pointrecal-
ibration (reslope)or the entire range of standardsand the recalibrationchecked
for adequacy.Any samples analyzed prior to recalibration that have suspect
resultsshould be reanalyzed.
818 HUANG & FUJII
Comments.For soils that are resistantto dissolution,smaller soil particle

sizes should be used for soil digestion. Use of a mechanizedgrinder such as
Angstrom grinder (Angstrom Inc., Belleville, MD) that grinds soils to <25 Ilm
will acceleratedissolution.
Microwave dissolutionmay improve the efficiency of acid digestionof soil
samplesand should be considered(Kingston & Jassie,1988).The advantagesof
microwavedissolutioninclude fasterreactionratesthat result from the high tem-
peraturesand pressuresattained inside the sealedcontainers.Thesecontainers
generallyare madeof polymersthat do not contaminateor adsorbthe sampleand
do not absorb microwave energy. The use of closed vessels also eliminates
uncontrolledlossesof volatile molecularAs speciesthat may be presentin a sam-
ple or that are formed in the courseof a dissolution.
The Ag-DDC method is a widely acceptedcolorimetric procedureand is
recommendedas an official AOAC method (AOAC, 1990). This method has a
detectionlimit of 1 Ilg As. Certain metalsand metalloids(Cr, Co, Cu, Hg, Mo,
Ni, Pt, Ag and Se) interfere in the generationof arsine (Am. Public Health
Assoc., 1992). If Sb salts are presentin the sample,stibine will form and inter-
fere with color developmentby yielding a red color with maximum absorbance
at 510 nm. Normally the concentrationof theseconstituentspresentin soil do not
interferesignificantly. At pH 1, methylarseniccompoundsare reducedto methy-
larsines, which form colored complexeswith the absorbersolution.If methy-
larseniccompoundsare present,measurements of total As by this methodis unre-
liable.
Continuous-flowHGAAS offers the advantagesof simplicity in operation,
excellentreproducibility, low detectionlimits, and high-samplevolume through-
put for As analysis.The detectionlimit of this methodfor aqueoussamplesis 2
Ilg L-l, and the optimum concentrationrange is 2 to 20 Ilg L-l (Am. Public
Health Assoc., 1992). Interferencesare minimized becausethe As hydrides are
removed from the solution containing most potential interfering substances.
Slight responsevariationsoccur when acid matricesare varied. Thesevariations
can be controlled by treating standardsand samplesin the samemanner.Con-
centrationsof Se in excessof 20 mg L-l in the samples,although unlikely, can
suppressthe responseof As hydride (Ganje & Rains, 1982). Low concentrations
of noble metals (approximately100 Ilg L-l of Ag, Au, Pt, Pd, etc.), concentra-
tions of Cu, Pb, and Ni at or greaterthan 1 mg L-t, and concentrationsbetween
0.1 and 1 mg L- 1 of otherhydride-formingelements(Bi, Sb, Sn, andTe) also may
suppressthe responseof As hydride (Am. Public Health Assoc.,1992). Reduced
nitrogen oxides resulting from HN03 digestion and nitrite also suppressinstru-
mental responsefor As.
The above recommendeddigestion procedure(Method B) and HGAAS
were used to analyzeAs in standardreferencesoils and geologic materialshav-
ing diverse mineral compositionsand reportedAs concentrations,ranging from
3.1 to 17.7 mg kg-1 As. The resultsof analysesof threestandardsamplesrelative
to As valuesreportedin the literature rangedfrom 0 to +3.7% relative error. The
precisionof the measurements (10-25 replicates)rangedfrom 1.7 to 5.2 relative
standarddeviation(E.P. Welsch, 1994, personalcommunication).
Total Arsenic andArsenic Speciesin Soil Solutions

and WaterExtractsof Soil
Total Arsenic in Soil Solution and WaterExtractsof Soil

Principles.Total As in soil solutionsandwaterextractsof soil is composed
of inorganicand organicAs speciesof variousoxidation states.Therefore,all As
speciesmust be quantitatively transformed to the speciesdetectedby the method
used.The recommendeddigestionprocedureis the sameproceduredescribedfor
total Se in soil solutionsand water extractsof soil (see"Total Seleniumin Soil
Solutionsand Water Extractsof Soil"), similar to that describedby Makita and
Fujii (1992). This method requires an initial oxidation of the sample using
K 2S20 S at elevatedtemperature,which transformsall As speciesto As(V). The
sampleis then digestedafter the addition of Hel. Samplesresulting from this
digestion procedureare suitable for analysisof both total As and Se. The Hel
digestion reducesSe(VI) to Se(IV) (Eq. [2]), but it is not reducing enough to
reduceAs(V) to As(III). Under theseacidic conditions,total As is determinedby
continuous-flow HGAAS as describedabove in "Hydride GenerationAtomic
AbsorptionSpectrometricMethod" and"Hydride GenerationAtomic Absorption
Spectrometry(for Analysis of Soil SampleDigestedUsing Method B)".
Apparatus
1. Apparatus1 to 3 describedin "Apparatus"under "Total Seleniumin
Soil Solutionsand Water Extractsof Soil."
2. All apparatusdescribedin "Apparatus" under "Hydride Generation
Atomic AbsorptionSpectrometry(for Analysisof Soil SampleDigest-
ed Using Method B)."
Reagents
1. Reagents1-4 describedin "Reagents"under "Total Seleniumin Soil
Solutionsand Water Extractsof Soil."
2. All reagents described in "Reagents" under "Hydride Generation
Atomic AbsorptionSpectrometry(for Analysis of Soil SampleDigest-
ed Using Method B)"; the only differenceis that 0.4% HN03 (instead
of 1.2 M Hel) is usedto makeserialdilutions of As standardsolutions,
andstandardsolutionsare treatedassamplesand digestedas described
below.
Procedure.The digestionprocedureis the sameas that describedin "Pro-
cedure"under"Total Seleniumin Soil Solutionsand Water Extractsof Soil" for
total Se in soil solutionsandwaterextractsof soil. Quantitativelytransferdigest-
ed samplesto 50-mL volumetric flasks and dilute to volume with deionizeddis-
tilled water. Analyzetotal As in resultingsolutionsasdescribedin "Hydride Gen-
erationAtomic Absorption Spectrometry(for Analysis of Soil SampleDigested
Using Method B." Resultsof analysesare comparedto the standardcurve and
multiplied by any dilution factor resultingfrom dilution of the original sample,to
obtain the concentrationof total As in microgramsper liter. This methodhas a
practicaldetectionlimit of 2 Jlg L-1.
820 HUANG & FUJII
Comments.This digestionmethod has beenusedby the U.S. Geological

Survey (USGS), California District Laboratory to analyzetotal As since 1992.
Repetitiveanalysesof two internal USGS referencewater samples(agricultural
drainwater)resultedin meansand standarddeviationsof 4.36 ± 0.221lg L-l As
(n = 23) and 8.09 ± 0.40 Ilg L-1 (n = 35). The reportedmost probablevalue and
standarddeviationsfor the samplesare 4.20 ± 0.57 f.1g L-l and 8.09 ± l.11f.1g L- 1
As, respectively.
Determinationof Arseniteand Arsenate

The two recommendedmethodsto determineAs(I1I) and As (V) species
utilize colorimetricandcontinuous-flowHGAAS techniques and are discussedin
detail below.
Another method that combineshydride generation,a liquid-N trap, and
AAS (Creceliuset aI., 1986; Masscheleynet aI., 1991b) also has been used to
determineAs(III) and As(V). This is one of the more reliable methods,especial-
ly for complex matrices,and has the capability to quantify monomethylarsonic
acid (MMAA) and dimethylarsinicacid (DMAA), and possiblyothervolatile As
species.
AqueousAs(III) and As(V) speciescan be separatedby a strong anion-
exchangeresin (Ficklin, 1983, 1990). Elution of the resin providestwo separate
portions of solution, eachcontaininga different inorganicAs species. Each por-
tion is analyzedby ETAAS to determinethe respectiveinorganicAs species.The
detectionlimit for eachspeciesis 1 Ilg L-1 As. One of the primary advantagesof
this methodis that the separationcan be done in the field immediatelyafter sam-
pling, thereby eliminating the problems related to interconversionsbetween
As(III) and As(V) speciesdiscussedbelow (see"Comments").In addition to the
disadvantagesof ETAAS (see "Total Arsenic in Soil"), the strong anion-
exchangeresins may be adverselyaffectedby saline samplematricesand high
temperatures.
Arsenic also is present in natural waters as monoethyl, dimethyl, and
trimethyl arsine,MMAA, and DMAA, and trimethylarsineoxides(Takamatsuet
aI., 1982; Van Loon, 1985; Anderson& Bruland, 1991).TheseAs speciescan be
determinedby selectivevolatilization followed by AAS (Andrae, 1977;Ander-
son & Bruland, 1991). High-pressureliquid chromatographycombined with
AAS also has beenapplied to the determinationof As(III), As(V), methylarson-
ic acid, and dimethylarsinicacid in soil extracts(Brinckmanet aI., 1980).
Principles. Both As(I1I) and As(V) can be presentin soil solutions and
waterextractsof soil. Arsenateforms ablue complexwith molybdate,but As(III)
does not (Johnson& Pilson, 1972). Phosphate,like As(V), also forms a blue
complexwith molybdate.Oscarsonet ai. (1980) developeda colorimetricmethod
to determineAs(III) and As(V) and to minimize phosphateinterference.Analy-
sis of one samplealiquot using the molybdenumblue methodprovidesthe con-
centrationof As(V) plus P in the sample.By addingan oxidizing agentto a sec-
ond samplealiquot, As(III) is oxidizedto As(V), and analysisof this treatedsolu-
tion providesthe concentrationof As(V) plus As(III) plus P. Addition of a reduc-
ing agentto a third samplealiquot convertsAs(V) to As(III) and analysisof this
reducedsolution gives the concentrationof P. The concentrationof As(V) and

As(III) can be calculatedfrom the differencebetweenSamples1 and 3 and that
betweenSamples1 and 2, respectively.Unless one knows with certainty that
there is no P in the sample,Treatment3 must be included. To accuratelydeter-
mine As by this method, the sampleshould have a high AslP molar ratio-the
higher the better.This is a limitation of the method.
Aqueous As(III) can be determined using automated continuous-flow
HGAAS (Glaubig & Goldberg,1988).Total inorganicAs(III) plus As(V) can be
determinedby reducingAs(V) to As(III) using KI, then generatingthe hydride in
6 M HCl. Arsenite can be determinedby generatingthe hydride at pH 4.0 to 4.5
using an oxalatebuffer. Arsenateis calculatedby difference.Detectionlimits for
As(III) and As(III) plus As(V) are about 21lg L- 1 range.
Colorimetry
APPARATUS
1. Volumetric flasks, 25, 250, and 500 mL.

2. Pipettes,2 mL.
3. Spectrophotometer, for use at 840 nm.
4. Plasticbottles,500 mL.
5. Glassbottles,250, 500, and 1000 mL.
REAGENTS
1. Iodine solution,0.02M. Dissolve2.54 gof I and 8.00 g of KI in 25 mL

of deionizeddistilled water. When dissolutionis complete,dilute to 1
L with deionizeddistilled water in a volumetric flask and store in a
dark bottle.
2. Iodine solution, 0.002M. Preparedas neededby dilution of the 0.02M
I solution with deionizeddistilled water.
3. Special color reagent(Strickland & Parsons,1968). (i) Ammonium
molybdate solution, 3% (w/v): Dissolve 15 g of analytical reagent-
quality ammonium paramolybdate[(NH4)6M07024 • 4H20] (prefer-
ably fine crystalline) in 500 mL of deionizeddistilled water. Store in a
plastic bottle out of direct sunlight. The solution is stableindefinitely.
(ii) Sulfuric acid solution: Add 140 mL of concentratedH2S04, 18 M,
to 900 mL of deionizeddistilled water. Allow to cool and store in a
glass bottle. (iii) Ascorbic acid (C6Hs0 6), 5.4% (w/v): Dissolve 27 g
C6Hs0 6 in 500 mL of deionizeddistilled water. Store the solution in a
plastic bottle frozen solid in a freezer. Thaw for use and refreezeat
once.The solution is then stablefor monthsbut shouldnot be kept for
more than a week at laboratory temperatures.(iv) Potassiumanti-
monyl-tartratesolution,0.136%(w/v): Dissolve0.34 g potassiumanti-
monyl-tartrate(tartar emetic) in 250 mL of deionizeddistilled water,
warming if necessary.Store in a glassor plastic bottle. The solution is
stable for many months. (v) Mixed reagent: Mix together 100 mL
ammoniummolybdate,250 mL H2S04, 100 mL C6Hs0 6, and 50 mL
potassiumantimonyl-tartratesolutions.Preparethis reagentfor imme-
822 HUANG & FUJII
diate use,and discardany excess.Do not storefor longer than about6

h. This quantity is suitablefor about50 samples.
4. Sodium metabisulfitesolution, 14% (w/w) NaZSZ05 in deionizeddi-
stilled water. Preparefresh for use.
5. Sodiumthiosulfate solution,1.4% (w/w) NaZSZ03 • 5HzO in deionized
distilled water.
6. Reducingagent(Johnson,1971). Mix 20 mL of 1.75 M HzS04 solu-
tion into 40 mL of the sodium metabisulfitesolution. This should be
done slowly to avoid excessivebubbling causedby the liberation of
SOz. Finally, mix in 40 mL of the sodium thiosulfate solution. This
procedurewill provide enoughreagentfor about20 aliquots.If refrig-
erated,the reducingreagentis stablefor up to 24 h.
7. Arsenic standard solutions. Prepare As standard solutions with
NaAsO z in deionizeddistilled water over the concentrationrange of
0.1 to 10 mg L-1. Standardsolutionsshould be made for eachset of
analysesand be treatedas samples.
Procedure. For eachsample,transferan aliquot of the soil solutionor water
extract of soil to each of three 25-mL volumetric flasks. Leave the first flask
untreated,add 2 mL of the 0.002 M I solution (oxidizing agent) to the second
flask and add 2 mL of a solution of sodium metabisulfite,sodium thiosulfate
(NaZS203)'and sulfuric acid (reducingagent)(Johnson,1971) to the third flask.
After 15 min, add 2 mL of the specialcolor reagent(mixed reagent)to all flasks;
dilute the solutionsto volume with deionizeddistilled waterand mix thoroughly.
After 90 min measurethe absorbanceof the samplesand standardsspectropho-
tometrically at 840 nm. The absorbanceof the analyzedsampleis comparedto
the standardcurve to determinethe concentrationof As(V) plus P (first flask),
As(III) plus As(V) plus P (secondflask) and P (third flask). This value is multi-
plied by any dilution factor resulting from dilution of the original solution to
obtain the concentrationin the original solution. The concentrationof As(V) is
then obtainedfrom the differencebetweenthe first flask and the third flask and
that of As(III) from the differencebetweenthe first flask and the secondflask.
Hydride Generation Atomic Absorption Spectrometry
APPARATUS. The apparatusrequired for this method are the sameas those
listed in "Apparatus"under"Hydride GenerationAtomic Absorption Spectrom-
etry (for Analysis of Soil SampleDigestedUsing Method B)."
REAGENTS
1. Stock As(II1) solution, 10.0 mg L-1 As(III): Dissolve 1.320 g arsenic

trioxide, AsZ0 3, in deionizeddistilled water containing4.00 g NaOH,
and dilute to 1 L to obtain a 1 g L-1 As(III) stock solution. Further
dilute this solution by 100 times with deionized distilled water to
obtain the 10 mg L -1 As(III) stock solution.
2. StandardAs(III) solutions:StandardAs(III) solutions of 10.0, 25.0,
50.0, and 75.0 Ilg L-1 As(III) are prepareddaily by diluting the 10.0
mg L-l As(III) stock solution.
3. Sodium borohydride solution, 0.35% NaBHJ0.30%NaOH in H20:

Dissolve0.35 g NaBI4 and 0.30g NaOH in 100 mL of deionizeddis-
tilled water.
4. Sodiumoxalatebuffer solution, 1.15%H2C20JO.60%sodiumoxalate
(Na2Cz04)in H20: Dissolve1.15g H2C20 4 and0.60g Na2Cz04in 100
mL deionizeddistilled water.
6. Urea [(NH2)2CO] solution, 80% (w/v) in H20: Dissolve 80 g
(NH2)2CO in 100 mL deionizeddistilled water.
7. Potassiumiodide solution,40% (w/v) in H20: Dissolve40 g KI in 100
mL deionizeddistilled water.
8. Arsenic standard solutions:Prepared from commercially available
AAS standard(1000 mg L-1 As) by makingserial dilutions to concen-
trationsof 2, 5, 12, and 18 J..lg L-1 As in 1.2 M HCI. Standardsolutions
shouldbe madefor eachset of analysesand be treatedas samples.
Procedure. For more information see Glaubig and Goldberg(1988). The
HGAAS analysisusesa continuous-flowhydride generator(e.g., Varian VGA-
76; Fig. 30-1) coupledto an AAS (e.g., Perkin-ElmerModeI3030B).The AAS
conditionsare asfollows: wavelengthof 193.7nm, slit width of 0.7 nm, andlamp
power supply of 7 W using a reducingair/acetyleneflame or electrical heating
device. GaseousAs hydride, AsH3, is formed by pumping2.5 mL min-1 sample
(or standard),1.0 mL min-1 acid reagent,and 1.0 mL min-1 NaBI4 reagent
througha reactioncoil and into a gas-liquidseparator.Argon is usedas the carri-
er gas to strip the hydride from the liquid and carry it into an open-ended,air-
acetyleneor electrically heatedquartz absorptioncell. Manufacturer'soperation
manualsspecific to the hydride generatorand AAS usedalso shouldbe consult-
ed for recommendedsettingsand procedures.
For analysisof As(III) plus As(y), solutionscontaining10.0 mL of sample
(or standard),10 mL of concentratedHCl, and 0.5 mL of the (NH2)2CO reagent
are reducedby adding0.5 mL of the KI reagent30 to 60 min prior to analysis.
The As hydride is generatedusing concentratedHCI and the NaBHJNaOH
reagentin deionizeddistilled water. Sample absorbance is comparedto the stan-
dard curve to calculateAs(III) plus As(V) contentsof soil solutions and water
extractsof soils.
For analysisof As(III) only, no reductionof standardsor samplesis need-
ed prior to analysis.The As hydride is generatedusing the sodiumoxalatebuffer
reagentin deionizeddistilled water and the NaBI4fNaOH reagent.The reagent
concentrationsshouldbuffer the samplesto a final pH between4.0 and 4.5.Sam-
ple absorbanceis comparedto the standardcurve to calculatethe As(III) contents
of soil solutionsandwaterextractsof soil. From the determinationof As(III) plus
As(V) and As(III) only, the concentrationof As(V) is calculated.
Comments.Using continuous-flow HGAAS for analysis of As(V) and
As(III) significantly increasesthe rateof samplesanalysisover traditional batch
hydride generators.Recoveriesof As(V) and As(III) spikes (10-40 J..lg L-l of
eachspecies)addedto a soil extract, a sampleof synthesizedriver water, and a
sampleof synthesizedagricultural drain water ranged from 96.0 to 105% for
824 HUANG & FUJII
As(V) and from 102 to 114% for As(II1) (Glaubig & Goldberg,1988). Standard
deviationsof triplicate analysesfor this study were 0.4)lg L-l As or less.For the
determinationof As(III) species,samplesshouldbe spikedwith As(III). For the
determinationof both As(III) and As(V) species,samplesshouldbe spikedwith
both As(III) and As(V). Refer to Chapter2 (Klesta & Bartz, 1996) of this book
for ONOC purposes.
Dissolved organic compoundshave been reported to interfere with the
analysisof Se(IV) and Se(VI) speciesin groundwaterusing HGAAS (Roden &
Tallman, 1982).Use of XAD-8 resin to removehydrophobicorganiccompounds
has beenshown to amelioratethis interference(Roden& Tallman, 1982; Fio &
Fujii, 1990). Similar interferenceto As speciesdeterminationmight occur and
shouldbe investigated.
Rateof transformationbetweenAs(III) and As(V) are of concernfor accu-
rate determinationof thesespecies.Oxidation of As(II1) to As(V) has beensug-
gestedto occur both spontaneously(Creceliuset aI., 1986) and to take at least3
wk (Tallman & Shaikh, 1980). Glaubig andGoldberg(1988) report oxidation of
As(III) to As(V) in about 3 h. Creceliuset ai. (1986) also report spontaneous
reduction of As(V) to As(II1) in filtered (O.4-)lm) river water. These findings
emphasizethe need for sampling and preservationtechniquesthat assurethe
quality of As(V) and As(II1) determinations.For example,Glaubig and Goldberg
(1988) recommendacidification to pH 2 and refrigerationnear O°C to preserve
microgramsper liter levels of As(III) for up to 6 wk. In contrast,Crecilius et ai.
(1986) found considerableand variable transformation between As(V) and
As(III) speciesusing various preservationtechniques,including acidification
(0.1% HCI plus 0.01% C6Hs06 at room temperature).They recommendedthat
samplesbe quick-frozento -196°Cin liquid N andstoredat -80°C or below until
analysis. Preservationof microgramsper liter levels of As(III) for 6 wk was
achievedin interstitial water samplesby acidification to pH 2 and refrigeration
nearO°C (Aggett & Kriegman, 1987).
The HGAAS methodto determineAs(III) buffers the pH of the solution to
between4.0 and 4.5. Masscheleynet al. (1991b)showedthat generationof AsH3
from As(III) is essentiallyindependentof pH below a pH of about7. Arsinesgen-
eratedfrom As(V), MMAA, and DMAA were negligiblebetweenpH 6 and8, but
increasedwith decreasingpH below a pH of about 6. Thus, for solutions con-
taining As(III), As(V), MMAA, and DMAA, quantitativegenerationof arsines
solely from As(III) only occurredbetweenpH 6 and 7. For thesereasons,Mass-
cheleyn et ai. (1991b) prefer to buffer solutionsat pH 6.0 for As(III) analysis.
This modification should be consideredfor As(III) analysis,especiallyfor sam-
ples containingsignificant concentrationsof As(V), MMAA, or DMAA.
Becauseof the potentialrapid interconversionsbetweenAs(V) andAs(II1),
ONOC measuresare extremelyimportant. Samplesshouldbe analyzedas soon
as possibleafter sampling.For the determinationof As(III) and As(V) species,
duplicate samplesshould be spiked with As(III) and As(V) at the time of sam-
pling and in the laboratory.Becausethere is no consensuson a single preserva-
tion techniquefor As(V)/As(III) analysis,for each sample matrix it is recom-
mendedthat preservationstudies be done to confirm reliability of whichever
preservationtechniquewill be used.
Availability Indicesof Soil Arsenic

Arsenic is not consideredto be essentialfor the growth of higher plants.
However,As will contaminatethe food chain when it is taken up by plants
(Woolson,1973). Furthermore,As hasbeenshownto be phytotoxicwhen soil As
concentrationsare elevated(Ganje & Rains, 1982). The availability of soil As is
not necessarilyrelatedto the total As contentof soils but dependson many fac-
tors, including plant species,soil properties,geographicareas,climactic condi-
tions, farming practices,and other anthropogenicactivities.
Plant speciesvary substantiallyin their ability to take up As (Liebig, 1966;
Woolson, 1973). The influenceof As specieson the mechanismof As uptakeby
plants also merits attention. Moreover, As accumulationin plant parts within a
speciesshow a consistenttrend of the following order: plant roots > vegetable
tops> edible seedsand fruits (Fleming et aI., 1943; McLean et aI., 1944; Jones
& Hatch, 1945; Ganje & Rains, 1982).
Soil propertiesknown to influenceAs availability includeconcentrationsof
hydrousoxidesof Fe, AI, and Mn,organicmatter,soil pH, soil texture,and P lev-
els of soil solutions (Walsh & Keeney, 1975; Huang, 1987). Arsenite(lII) and
As(V) are adsorbedby hydrous oxides of AI, Fe, and Mn (Livesey & Huang,
1981; Huang, 1987) and may form compoundsof varying degreesof solubility
witli elementssuchas AI, Fe, Ca, and Mg (Walsh & Keeney,1975). Arsenitecan
be convertedto As(V) through abiotic catalysisby Mn oxides (Oscarsonet aI.,
1981a).Arsenateand As(III) are quite immobile in fine-texturedsoils. However,
if soils are low in hydrousoxides of AI and Fe, As may leach from coarse-tex-
tured soils and contaminategroundwater.Recentimprovementsin samplecol-
lection and analyticaltechniqueshavesuggestedthat As(lII) is more prevalentin
groundwaterthan previously believed(Korte & Fernando,1991).
The concentrationof P in soil solutions also affects As behavior in soils.
Plant-availableAs and phytotoxicity of As to corn (Zea mays L.) are alteredby P
additions to both sandy and clay-type soils (Woolson et aI., 1973). Under more
restrictedleachingconditions,As phytotoxicity is enhancedby the addition of P
and is more prevalentand acutein coarse-textured soils then in fine-texturedsoil.
Phytoxicity of As to com is decreasedin the coarse-texturedsoils when the soils
are leachedwith P. Therefore,bioavailability and phytotoxicity of As in soils
should be affectedby farming practicessuch as phosphatefertilization and irri-
gation.
Despite the complex chemical behavior of soil As, many methods have
beenproposedto measurethe bioavailability of soil As. Theseinclude the use of
separatemedia extraction or the fractionation approachcommon to soil P. The
oxyanionsof As and P have similar chemical propertiesand, therefore,extrac-
tants commonly used to measureP bioavailability are used for As (Ganje &
Rains, 1982). Johnstonand Barnard (1979)classified14 extractingsolutionsinto
three groupsaccordingto reactionswith soils and their effectivenessin extract-
ing As from agriculturalsoils:
1. Neutral extractants:Theseextractantsare deionizeddistilled water, 1
M NH4Cl, 0.5 M NH4C2H30 2, and 0.5 M NH4N03• Generally these
extractantsremovedless than 0.1 mg of As per kilogram of soil.
826 HUANG & FUJII
2. Basic extractants:Arsenic removed by these reactantsfollows the

order of 0.5 M NH4F (pH 8.2) < 0.5 M NaHC03 (pH 8.5) < 0.5 M
(NH4hC03 < 0.1 M NaOH. Basic extractantsare more effective than
the neutralextractantin removingAs from soils.
3. Acid extractants:Arsenic removalby acid extractantsfollows the order
0.5 M HCI + 0.0125M H2S04 - 0.5 M HCI < 0.5 M KH 2P04 < 0.5 M
H 2S04.
For detailed description of procedureand application, refer to the refer-
encescited by Johnstonand Barnard (1979) and Ganje and Rains (1982). The
methods described in "Arsenic Determination" may be used to measureAs
extractedby various extraction methods,but matrix-matchingtechniquesand
extensiveQNQC proceduresshouldbe usedto assurethe quality of the analysis.
The ammonium bicarbonate-diethylenetriaminepentaacetic acid (AB-
DTPA) soil test was developedby Soltanpourand Schwab(1977) to simultane-
ously extract labile nutrientsfrom neutral and calcareoussoils. It was later used
for extractionof potentially toxic elementsfrom soils and mine spoils and soils
treatedwith sewagesludge.Bioavailability of As may be measuredwith the AB-
DTPA soil test (Soltanpour,1991). It is likely that HC03" will desorbor solubi-
lize As(V) and As(III). Also, 0.5 M NaHC03 has beenshown to extract plant-
available As (Woolson et aI., 1971).The relationshipbetweenAs extractedby
AB-DTPA and plant uptakeof As still remainsto be evaluated.
No one extractantmay serve as a primary index of all bioavailableAs in
soils. The extractantchosenshould be specific for certainAs forms in the soils.
Therefore,eachsoil type or soil areashould be treatedindependentlyin testing
for its bioavailableAs. Furthermore,an understandingof the equilibrium rela-
tions of As speciesbetweensolution and solid phasesand the kinetics and mech-
anismsof the releaseof As associatedwith solid phasesis essentialto evaluate
the short-termand long-termmobility and bioavailability of As in soils.
ACKNOWLEDGMENTS
Contribution from the Department of Soil Science, University of

Saskatchewan,Saskatoon,SK, CanadaS7N 5A8 and the United StatesDepart-
ment of the Interior, GeologicalSurvey,Sacramento,CA 95825.This work is in
part supportedby the Natural Sciencesand EngineeringResearchCouncil of
CanadaGrant GP2383.
REFERENCES
Abrams, M.M., R.G. Burau, and R.J. Zasoski. 1990. Organicseleniumdistribution in selectedCali-
fornia soils. Soil Sci. Soc. Am. J. 54:979-982.
Aggett, J., and M.R. Kriegman. 1987. Preservationof arsenic(III) and arsenic(V) in samplesof sed-
iment in interstitial water. Analyst (London) 112:153-157.
American Public Health Association. 1992. Standardmethodsfor the examiantionof water and
Anderson,L.C.D., and K.W. Bruland. 1991.Biogeochemistryof arsenicin naturalwaters:The impor-
tanceof methylatedspecies.Environ. Sci. Technol. 25:420-427.
Andrae,M.O. 1977. Determinationof arsenicspeciesin naturalwaters.Anal. Chern.49:82~30.

Associationof Official Analytical Chemists.1990. Official methodsof analysis.15th ed. Assoc.Offi-
cial Anal. Chern.,Arlington, VA
Bakhtar, D., G.R. Bradford, and L.I. Lund. 1989. Dissolution of soils and geologic materials for
simultaneouselementalanalysisby inductively coupledplasmaoptical emissionspectrometry
and atomic absorptionspectrometry.Analyst 114:901-909.
Balistrieri, L.S., and T.T. Chao. 1990.Adsorption of seleniumby amorphousiron oxyhydroxideand
manganesedioxide. Geochim.Cosmochim.Acta 54:739-75l.
Beath, O.A., 1.H. Draize, H.F. Eppson,e.s. Gilbert, and O.e. McCreary. 1934. Certain poisonous
plants of Wyoming activatedby seleniumand their associationwith respectto soil types. J.
Am. Pharm.Assoc.23:94-97.
Berrow, M.L., andAM. Ure. 1989. Geologicmaterialsandsoils. p. 213-242.In M. Ihnat (ed.) Occur-
renceand distribution of selenium.CRC Press,Boca Raton,FL.
Briggs, P.H., and 1.G. Crock. 1986. Automateddeterminationof total seleniumin rocks, soils, and
plants.U.S. Geol. Surv. Open-FileRep. 86-46. U.S. Geol. Surv., Denver,CO.
Brimmer, S.P.,W.R. Fawcett,and K.A Kulhavy. 1987.Quantitativereductionof selenateion to selen-
ite in aqueoussamples.Anal. Chern. 59:1470--147l.
Brinckman, F.E., K.L. Jewett, W.P. Iverson, KJ. Irgolic, K.e. Ehrhardt, and R.A. Stockton. 1980.
Graphitefurnaceatomic absorptionspectrophotometers as automatedelement-specificdetec-
tors for high-pressureliquid chromatography.The determinationof arsenite,arsenate,methy-
larsonicacid, and dimethylarsenicacid. 1. Chtomatogr.191:31-46.
Brown, M.W., and 1.H. Watkinson.1977.An automatedfluorimetric methodfor the determinationof
nanogramquantitiesof selenium.Anal. Chim. Acta 89:29-35.
Chan, C.C.Y., and R.S. Sadana.1992. Determinationof arsenicand seleniumin environmentalsam-
pleesby flow-injection hydride generationatomic absorptionspectrometry.Anal. Chim. Acta
270:231-238.
Chao, T.T. 1984. Use of partial dissolution techniquesin geochemicalexploration. J. Geochem.
Explor.20:101-135.
Chao, T.T., and L. Zhou. 1983. Extraction techniquesfor selective dissolution of amorphousiron
oxidesfrom soils and sediments.Soil Sci. Soc. Am. J. 47:225-232.
Chao,T.T., and R.F. Sanzolone.1989. Fractionationof soil seleniumby sequentialpartial dissolution.
Soil Sci. Soc. Am. J. 53:385-392.
Chapman,1.F., and L.S. Dale. 1979.Atomic absorptionspectrometricdeterminationof someelements
forming volatile hydrideswith a heatedcell atomizerand gas handling system.Anal. Chim.
Acta 111:137-144.
Cooke,T.D., and K.W. Bruland. 1987. Aquatic chemistryof selenium:Evidenceof biomethylation,
Cowan, C.E. 1988. Review of selenium thermodynamicdata. Electric Power Res. Inst. Rep. EA-
5655. EPRI, Palo Alto, CA
Crecelius,E.A, N.S. Bloom, e.E. Cowan,and E.A Jenne.1986. Speciationof seleniumand arsenic
in natural waters and sediments.Vol. 2. EA-464l. Battelle Pacific Northwest Lab., Sequim,
WA
Cutter, G.A 1978. Speciesdeterminationof seleniumin natural waters.Anal. Chim. Acta 98:59-66.
Cutter, G.A 1982. Seleniumin reducingwaters.Science(Washington,DC) 271:829-83l.
Cutter,G.A 1983.Elimination of nitrite interferencein the determinationof seleniumby hydride gen-
eration.Anal. Chim. Acta 149:391-394.
Cutter, GA. 1986. Speciationof seleniumand arsenicin natural watersand sediments.Vol. L Elec-
tric PowerRes. Inst. Rep. EA-464l. EPRI, Palo Alto, CA
Cutter, G.A, and K.W. Bruland. 1984. The marine biogeochemistryof selenium:Are-evaluation.
Limnol. Oceanogr.29:1172-1192.
Doran, 1.W., and M. Alexander. 1977. Microbial transformationof selenium.Appl. Environ. Micro-
bioI. 3:31-37.
Dubie, R.L., J.e. Thomas,and K.w. Brown. 1978. Arsenic pollution from underdrainageand runoff
from golf greens.Agron. 1. 70:71-74.
Elkhatib, E.A, O.L. Bennet,and R.J. Wright. 1984.Arsenitesorptionand desorptionin sols. Soil Sci.
Soc. Am. 1. 48:1025-1030.
Elrashadi,M.A., D.e. Adriano, S.M. Workman,and w.L. Lindsay. 1987. Chemicalequilibria of sele-
nium in soils: A Theoreticaldevelopment.Soil Sci. 144:141-152.
Ferguson,J.F., and M.A. Anderson. 1974. Chemical forms of arsenic in water supplies and their
removal. In AJ. Rubin (ed.) Chemistry of water supply, treatment,and distribution. Ann
Arbor Sci. Publ., Ann Arbor, ML
828 HUANG & FUJII
Ferguson,J.F., and J. Gavis. 1972. A review of the arseniccycle in natural waters. Water Res.
6:1259-1274.
Ficklin, W.H. 1983. Separationof arsenic(In) and arsenic(V) in ground watersby ion-exchange.
Talanta30:371-373.
Ficklin, W.H. 1990.Extractionandspeciationof arsenicin lacustrinesediments.Talanta37:831-834.
Fio, J.L., and R. Fujii. 1990.Seleniumspeciationmethodsand applicationto soil saturationextracts
from SanJoaquinValley, California. Soil Sci. Soc.Am. J. 54:363--369.
Fio, J.L, R. Fujii, and S.I. Deverel.1991. Seleniummobility and distribution in irrigated and nonir-
rigatedalluvial soils. Soil Sci. Soc.Am. J. 55:1313--1320.
Fishman,M.J., andLC. Friedman.1989.Methodsfor determinationof inorganic substances in water
and fluvial sediments.u.S. Geol. Surv. Tech.Water-Res.Invest.
Fleming, GA 1980.Essentialmicronutrients.Vol. 2. p. 199-234.In B.E. Davis (ed.) Applied soil
traceelements.JohnWiley & Sons,New York.
Fleming, W.P., F.E. Baker, and L Koblitsky. 1943. Effect of lead arsenatein soil on vegetables.J.
Econ.Entomol. 36:231-233.
Frankenberger, W.T., Jr., andU. Karlson. 1989.Environmentalfactorsaffectingmicrobial production
of dimethylselenide in an selenium-contaminatedsediment.Soil Sci. Soc. Am. J.
53:1435-1442.
Friedman,L.C., and D.E. Erdmann.1982. Quality assurancepracticesfor the chemicaland biologi-
cal analysesof waterand fluvial sediments.U.S. Geol. Surv. Tech. Water-Res.Invest.
Fujii, R., D.B. Hatfield, andS.J.Deverel.1988.Distribution of seleniumin soils of agriculturalfields,
westernSanJoaquinValley, California. Soil Sci. Soc.Am. J. 52:1274-1283.
Fujii, R., and R.G. Burau. 1989.Estimationof adsorbedSe(lV) in soils, westernSanJoaquinValley,
California. p. 200. In Agronomy abstracts.ASA, Madison,WI.
Ganje,T.I., and D.W. Rains. 1982. Arsenic. p. 385-403.In A.L Pageet al. (ed.) Methodsof soil
analysis.Part2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Geering,H.R., E.E. Cary, L.H.P. Jones,andW.H. Allaway. 1968.Solubility and redoxcriteria for the
possibleforms of seleniumin soils. Soil Sci. Soc.Am. Proc. 32:35-40.
Giaubig, RA, and S. Goldberg.1988. Determinationof inorganicarsenic(III) and arsenic(III plus
V) usingautomatedhydride-generationatomic-absorptionspectrometry.Soil Sci. Soc.Am. J.
52:536-537.
Goyal,S.S.,A Hafez,andD.W. Rains.1991.Simultaneousdeterminationof arsenite,arsenate,selen-
ite, and selenatein watersusing suppressedion chromatographywith ultraviolet absorbance
detection.J. Chromatogr.537:269-276.
Gruebel,K.A, J.A Davis, and J.O. Leckie. 1988.The feasibility of usingsequentialextractiontech-
niquesfor arsenicand seleniumin soils and sediments.Soil Sc. Soc.Am. J. 52:390--397.
Helmke,P.A. 1982.Neutronactivationanalysis.In AL Pageet al. (ed.) Methodsof soil analysis.Part
2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Huang,P.M. 1987.Agricultural soils: Fateof toxic substances. p. 143--148.In M. Singh(ed.) Systems
andcontrol encyclopedia.PergamonPress,Oxford, England.
Huang,P.M. 1991. Kinetics of redox reactionson surfacesof Mn oxidesand its impact on environ-
mentalquality. p. 191-230.In D.L Sparksand D.L Suarez(ed.) Rateof chemicalprocesses
in soils. SSSASpecialPbu\. 27. SSSA,Madison,WI.
Irgolic, K.J., R.A Stockton,andD. Charkaborti.1983.Determinationof arseniccompoundsin water
supplies.p. 282-305.In W.H. Ledererand R.I. Fensterheim(ed.) Arsenic: Industrial, bio-
medical,and environmentalperspectives.Van Nostrand,New York.
Johnson,D.L 1971. Simultaneousdeterminationof arsenateand phosphatein natural waters.Envi-
ron. Sci. Technol.5:411-414.
Johnson,D.L, and M.E. Q. Pilson. 1972.Spectrophotometric determinationof arsenite,arsenate,and
phosphatein naturalwaters.Anal. Chim. Acta 58:289-299.
Johnston,LR., andW.M. Barnard.1979.Comparativeeffectivenessof fourteensolutionsfor extract-
ing arsenicfrom four New York soils. Soil Sci. Soc.Am. Proc. 43:304-308.
Jones,A.A. 1991.X-ray fluorescenceanalysis.p. 287-324.In K.A Smith (ed.) Soil analysis.2nd ed.
Marcel Dekker,New York.
Jones,J.S.,andM.B. Hatch. 1945.Sprayresiduesandcrop assimilationof arsenicand lead.Soil Sci.
60:277-288.
Jump,R.K., andB.R. Sabey.1989.Soil testextractantsfor predictingseleniumin plants.In Jacobset
al. (ed.) Seleniumin agricultureandthe environment.SSSASpec.Pub\. 23. ASA and SSSA,
Madison,WI.
Karlson,U., andW.T. Frankenberger, Jr. 1986a.Determinationof selenateby single-columnion chro-
matography.J. Chromatogr.368:153--161.
Karlson,U., andW.T. Frankenberger, Jr. 1986b.Single-columnion chromatographyof selenitein soil

extracts.Anal. Chern.58:2704-2708.
Karlson, U., and W.T. Frankenberger.1990a.Volatilization of seleniumfrom agriculturalevaporation
pond sediments.Sci. Total Environ. 92:41-54.
Karlson, U., and W.T. Frankenberger,Jr. 1990b.Alkylselenideproductionin salinizedsoils. Soil Sci.
149:56--61.
Kingston, H.M., and L.B. Jassie(ed.) 1988. Introduction of microwave sample preparation.Am.
Chem.Soc.,Washington,DC.
Kipling, M.C. 1977.Arsenic: p. 93-120.In J. LenihanandW.W. Fletcher(ed.) Environmentandman.
Vol. 6. Acad. Press,Inc., New York.
Klesta, EJ., and J.K. Bartz. 1996. Quality assuranceand quality control. p. 19-48.In D.L. Sparkset
al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSA Book Ser. 5. SSSA and
AsA, Madison,WI.
Korte, N.E., and Q. Fernando.1991. A review of arsenic(III) in groundwater.Crit. Rev. Environ.
Control 21:1-39.
Lakin, H.W. 1973. Seleniunin our environment.Adv. Chem. Ser. 123:96-111.
Leckie, J.O., M.M. Benjamin, K. Hayes,G. Kaufman,and S. Altmann. 1980.Adsorption/coprecipi-
tation of trace elementsfrom water with iron oxyhydroxide.Electric Power Res. Inst. Rep.
CS-1513.StanfordUniv., Stanford,CA.
Lemmo, N.V., S.D. Faust,T. Belton, and R. Tucker. 1983.Assessmentof the chemicaland biological
significanceof arsenicalcompoundsin a heavily contaminatedwatershed.Part 1. The fate and
speciationof arsenicalcompoundsin aquaticenvironments-aliterature review. J. Environ.
Sci. Health AI8:335-387.
Liebig, G.F., 1966.Arsenic. p. 12-23.In H.D. Chapman(ed.) Diagnosticcriteria for plantsand soils.
Univ. California, Riverside.
Lipton, D.S. 1991.Associationof seleniumin inorganicand organicconstituentsof soils from a semi-
arid region. Ph.D. diss. Univ. California, Berkeley,CA.
Lipton, D.S., and H.E. Doner. 1989. Labile pools of seleniumin semi-aridsoils: Carbonatesand soil
organicmatter.p. 202. In Agronomy abstracts.ASA, Madison,WI.
Lipton, D.S., J.A. Davis, R. Fujii, and H.E. Doner. 1988. Seleniumassociatedwith hydrousoxides
and organicmatterin surfacesoils from the SanJoaquinValley, California. Trans.Am. Geo-
phys. Union 69:1181-1182.
Livesey, N.T., and P.M. Huang. 1981. Adsorption of arsenateby soils and its relation to selected
chemicalpropertiesand anions.Soil Sci. 131:88-94.
Makita, S.N., and R. Fujii. 1992. Quality assurancepracticesof the U.S. GeologicalSurvey laborato-
ry in Sacramento,California. U.S. Geol. Surv. OpenFile Rep. 91-522.U.S. Geol. Surv., Den-
ver, co.
Martell, A.E., and R.M. Smith. 1982. Critical stability constants.Vol. 5. PlenumPress,New York.
Masscheleyn,P.o., R.D. Delaune,and W.H. Patrick, Jr.. 1989. Transformationof seleniumas affect-
ed by sedimentoxidation-reductionpotentialand pH. Environ. Sci. Technol.24:91-96.
Masscheleyn,P.H., R.D. Delaune,and W.H. Patrick, Jr. 1991a.Arsenic and seleniumchemistry as
affectedby sedimentredox potential and pH. J. Environ. Qual. 20:522-527.
Masscheleyn,P.H., R.D. Delaune,and W.H. Patrick,Jr. 1991b.A hydride generation/separation tech-
nique for arsenicspeciation.J. Environ. Qual. 20:96-100.
McLean, H.C., A.L. Weber,and J.S. Ioffe. 1944. Arsenic contentof vegetablesgrown in soils treat-
ed with lead arsenate.J. Econ. Entomol. 37:315-316.
McNeal, I.M., and L.S. Balistrieri. 1989. Geochemistryand occurrenceof selenium:An overview. p.
1-13. In L.W. Jacobset al. (ed.) Seleniumin agriculture and the environment.SSSA Spec.
Publ. 23. ASA and SSSA,Madison,WI.
Mikkelsen, R.L. D.C. Adriano, and W.L. Lindsay. 1989. Factorsaffecting seleniumaccumulationby
agricultural crops. p. 65-94. In L.W. Jacobs etal. (ed.) Seleniumin agricultureand the envi-
ronment.SSSASpec.Pubol. 23. ASA and SSSA,Madison,WI.
Morrison, J.L. 1969. Distribution of arsenicfrom poultry litter in broiler chickens,soils, and crops.I.
Agric. Food Chem. 17:1288-1290.
Muth, O.H., I.E. Remmert,and I.R. Schubert.1958. Effects of seleniumand vitamine E on white
muscledisease.Science(Washington,DC) 128:1090.
Narasaki, H., and M. Ikeda. 1984. Automated determinationof arsenic and selenium by atomic
absorptionspectrometrywith hydride generation.Anal. Chern. 56:2059-2063.
National ResearchCouncil. 1977. Committeeof medicaland biological effectsof environmentalpol-
lutants:Arsenic. Natl. Acad. Sci., Washington,DC.
Neal, R.H., and G. Sposito. 1989. Selenateadsorption of alluvial soils. Soil Sci. Soc. Am. J.
53:70--74.
830 HUANG & FUJII
Neal, R.H., G. Sposito,K.M. Holtclaw, and S.J.Traina. 1987a.Seleniteadsorptionon alluvial soils:

I. Soil Compositionand pH effects.Soil Sci. Soc.Am. J. 51:1161-1165.
Neal, R.H., G. Sposito,K.M. Holtclaw, and S.J.Traina. 1987b.Seleniteadsorptionon alluvial soils:
II. Solutioncompositioneffects.Soil Sci. Soc. Am. J. 51:1165-1169.
Ohlendorf, H.M., R.L. Hothem, C.M. Bunck, T.W. Aldrich, and J.F. Moore. 1986. Relationships
betweenselenium concentrationsand avian reproduction.p. 330-342. In Trans. N. Am.
Wildlife Nat. Resour.51st Conf., Reno,NV. March 1986.Wildlife Manage.Inst.
Onishi, H., and E.B. Sandell.1955.Geochemistryof arsenic.Geochim.Cosmochim.Acta 7:1-33.
Oscarson,D.W., P.M. Huang,and W.K. Liaw. 1980.The oxidation of arseniteby aquaticsediments.
J. Environ. Qual. 9:700-703.
Oscarson,D.W., P.M. Huang,C. Defosse,andA Herbillion. 1981a.The oxidative powerof Mn(IV)
andFe(lll) oxideswith respectto As(lII) in terrestrialandaquaticenvironments.Nature(Lon-
don) 291:50-51.
Oscarson,D.W., P.M. Huang,andw.K. Liaw. 1981b.The role of manganese in the oxidationof arsen-
ite by freshwaterlake sediments.Clays Clay Miner. 29:219-225.
Pearlman,I., and F. Assaro.1971. Pottery analysisby neutronactivation. p. 182-195.In R.H. Brill
(ed.) Scienceand archaeology.4th Symp. Archaeol. Chern., Atlantic City, NJ. 1968. MIT
Press,Cambridge,MA.
Pierce,F.D., and H.R. Brown. 1977.Comparisonof inorganicinterferentsin atomicabsorptionspec-
trometric determinationof arsenicand selenium.Anal. Chern.49:1417-1422.
Pierce,M.L., and C.B. Moore. 1980.Adsorptionon amorphousiron hydroxidefrom dilute aqueous
solution.Environ. Sci. Technol. 14:214-216.
Presser,T.S., and I. Barnes.1984. Seleniumconcentrationsin waterstributary to and in the vicinity
of KestersonNational Wildlife Refuge,Fresnoand Merced Counties,California. U.S. Geol.
Surv. Water-Res.Invest. Rep. 84-4122.
Presser,T.S.,andH.M. Ohlendorf.1987.Biogeochemicalcycling of seleniumin the SanJoaquinVal-
ley, California. Environ. Manage.11:805-821.
Robberecht,H., and R. Van Grieken.1982.Seleniumin environmentalwaters:Determination,speci-
ation and concentrationlevels.Talanta29:823-844.
Roden,D.R., and D.E. Tallman. 1982.Determinationof inorganicseleniumspeciesin groundwaters
containing organic interferencesby ion chromatographyand hydride/atomicspectrometry.
Anal. Chern.54:307-309.
Rosenfeld,I., and O.A Beath. 1964. Selenium:Geobotany,biochemistry,toxicity, and nutrition.
Salmon, L., and P.A Cawse.1991. Instrumentalneutronactivation analysis.p. 377-432.In K.A
Smith (ed.) Soil analysis.2nd ed. Marcel Dekker, New York.
Sanzolone,R.F., andT.T. Chao.1987.Determinationof seleniumin thirty-two geochemicalreference
materialsby continuous-flowhydride generationatomic absorptionspectrophotometry.Geo-
stand.Newslett.11:81-S5.
Sharma,S., and R. Singh. 1983.Seleniumin soil, plant, andanimal systems.p. 23-50.In C.P. Straub
(ed.) Crit. Rev. Environ. Control. Vol. 13.
Sharp, B.L. 1991. Inductively coupled plasmaspectrometry.p. 63-109. In K.A. Smith (ed.) Soil
analysis.2nd ed. Marcel Dekker,New York.
Siu, K. W.M., and S.S.Berman.1984.Determinationof selenium(IV) in seawaterby gaschromatog-
raphy after coprecipitationwith hydrousiron (III) oxide. Anal. Chern.56:1806-1808.
Soltanpour,P.N. 1991.Determinationof nutrientavailability andelementaltoxicity by AB-DTPA soil
test and ICPS. Adv. Soil Sci. 16:165-190.
Soltanpour,P.N., and A.P. Schwab.1977.A new soil test for simultaneousextractionof macro-and
micro-nutrientsin alkaline soils. Commun.Soil Sci. Plant Anal. 8:195-207.
Soltanpour,P.N., and S.M. Workman. 1980.Use of the N14HC03-DTPA soil test to assessthe avail-
ability and toxicity of selenium to alfalfa plants. Commun.Soil Sci. Plant Anal.
11:1147-1156.
Strickland,J.D.H., and T.R. Parsons.1968. A practical handbookof seawateranalysis.p. 50. Fish.
Res. Board Can. Bull. 167. Queen'sPrinter, Ottawa,Ontario.
Takamatsu,T., H. Aoki, and T. Yoshida. 1982. Determinationof arsenate,arsenite,monomethylar-
sonateand dimethylarsinatein soil pollutedwith arsenic.Soil Sci. 133:239-246.
Takayanagi,K., andG.T.F. Wong. 1984.Organic andcolloidal seleniumin southernChesapeake Bay
and adjacentwaters.Mar. Chern. 14:141-148.
Tallman, D.E., and AU. Shaikh. 1980. Redox stability of inorganicarsenic(III) and arsenic(V) in
aqueoussolution.Anal. Chern.52:199-201.
Tessier,A, P.G.C.Campbell,andM. Bisson.1979.Sequentialextractionprocedurefor the speciation
of particulatetracemetals.Anal. Chern.51:844-851.
Thompson-Eagle,E.T., and w.T. Frankenberger,Jr. 1990. Volatilization of seleniumfrom agricultur-

al evaporationpond water. 1. Environ. Qual. 19:125-131.
Thompson-Eagle,E.T., W.T. Frankenberger,Jr., and U. Karlson. 1989. Volatilization of seleniumby
alternaria alternata. Appl. Environ. Microbiol. 55:1406-1413.
Tidball, R.R., WD. Grundy, and K.L. Sawatzky.1986. Kriging techniquesappliedto elementdistrib-
ution in soils of the SanJoaquinValley, California. p. 992-1009.In Proc. HAZTECH INTER-
NATIONAL, Hazardous Materials ManagementExhibition & Conference, Denver, CO.
13-15 August 1986. Pollut. Eng. Magazine,Chicago,IL.
Tyson, J.F. 1991. Flow injection atomic spectrometry.Spectrochim.Acta Rev. 14:169-233.
Ure, A.M., and M.L. Berrow. 1982.The elementalcompositionof soils. In H.1.M. Bowen (ed.) Envi-
ronmentalchemistry.Vol. 2. Royal Soc. Chern.,London.
U.S. EnvironmentalProtection Agency. 1986. Quality control for water. EPA 440/5-86-001.U.S.
Gov. Print. Office, Washington,De.
U.S. EnvironmentalProtectionAgency. 1987. Ambient water quality criteria for selenium-1987.
EPA 440/5-87-006.U.S. Gov. Print. Office, Washington,DC.
U.S. Environmental Protection Agency. 1988. Water quality criteria documents. Fed. Reg.
53(2):177-179.
U.S. Environmental Protection Agency. 1993. Drinking water regulations and health advisories.
Office of Water, USEPA, Washington,DC.
Van Loon, J.e. 1985. Selectedmethodsof trace metal analysis:Biological and environmentalsam-
ples. JohnWiley & Sons,New York.
Velinsky, D.1., and G.A. Cutter. 1991. Geochemistryof seleniumin a coastalsalt marsh.Geochim.
Wakao, N., H. Koyatsu, Y. Komai, H. Shimokawara,Y. Sakurai,and H. Shiota. 1988. Microbialoxi-
dation of arseniteand occurrenceof arsenite-oxidizing bacteria in acid mine water from a sul-
fur-pyrite mine. Geomicrobiol.J. 6:11-24.
Walsh, L.M., and D.R. Keeney. 1975. Behaviorand phytotoxicity of inorganic arsenicalsin soils. p.
35-52. In EA Woolson (ed.) Arsenical pesticides.ACS Symp. Ser. 7. Am. Chern. Soc.,
Washington,DC
Welsch,E.P.,1.G. Crock, and R. Sanzolone.1990. Trace-leveldeterminationof arsenicand selenium
using continuous-flowhydride generationatomic absorptionspectrophotometry(HG-AAS).
p. 38-45. In B.F. Arbogast(ed.) Quality assurancemanual for the Branch of Geochemistry,
U.S. Geological Survey. U.S. Geol. Surv. Open-File Rep. 90-668. U.S. Geol. Surv., Denver,
CO.
Weres, 0., A.R. Jaouni,and L. Tsao. 1989. The distribution, speciationand geochemicalcycling of
selenium in a sedimentary environment, Kesterson Reservoir,California, U.S.A. Appl.
Geochem.4:543-563.
Wood, J.M. 1974. Biological cycles for toxic elementsin the environment.Science(Washington,
DC). 183:1049-1052.
Woolson,E.A. 1973.Arsenic phytotoxicity and uptakein six vegetablecrops.WeedSci. 21:524-527.
Woolson, E.A., J.H. Axley, and P.e. Kearney. 1971. Correlationbetweenavailablesoil arsenicesti-
mated by six methods and responseof com (Zea mays L.). Soil Sci. Soc. Am. Proc.
35:101-105.
Woolson, E.A., I.H. Axley, and P.e. Kearney. 1973. The chemistry and phytotoxicity of arsenicin
soils: II: Effects of time and phosphorus.Soil Sci. Soc. Am. Proc. 37:254-259.
Workman, S.M., and P.N. Soltanpour.1980. Importanceof prereducingselenium(VI) to selenium
(IV) and decomposingorganicmatterin soil extractsprior to determinationof seleniumusing
hydride generation.Soil Sci. Soc. Am. 1. 44:1331-1332.
Wright, R.J., and T. Stuczynski.1996.Atomic absorptionand flameemissionspectrometry.p. 65-90.
In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSA Book
Ser. 5. SSSAand ASA, Madison,WI.
Yancey,P.H., M.E. Clark, S.e. Hand, R.D. Bowlus, and G.N. Somero.1982.Living with water stress:
Evolution of osmolytesystems.Science(Washington,DC) 217:1214-1222.
Zhaolun, F., S. Xu, X. Wang, and S. Zhang. 1986. Combinationof flow-injection techniqueswith
atomic spectrometry in agricultural and environmental analysis. Anal. Chim. Acta
179:325-340.
Published 1996
Chapter31
Bromine, Chlorine, & Fluorine
w. T. FRANKENBERGER,JR., University of California, Riverside,

California
D. C. ADRIANO, Savannah River Ecology Laboratory, Aiken, South Carolina
H. E. DONER, University of California, Berkeley, California
This chapteris a revisedversion of the comprehensivetreatmenton CI, Br, and

F by Adriano andDoner(1982),which wasa revisionof earlierchaptersby Stout
and Johnson(1965) and Brewer(1965a).Becausethe methodshavechangedlit-
tle, if any, over the past20 yr, they are essentiallythe sameas thosedescribedin
the previousedition.
The halidesBr, CI, and FI are ubiquitousin natureand are found in agri-
cultural lands from a variety of sources.Soil is adulteratedwith Br from fumi-
gants,CI from irrigation water, animal wastes,fertilizers, and rainwater,and F
from fertilizers, insecticides,and rainwater.Bromide and Cl- are consideredrel-
atively solubleand can be readily leachedin soils. Becauseof this property, Br-
and Cl- are usedas tracersof nitrate,salt, andwatermovementin soils. Chlorine
is an essentialplant micronutrient,and F is essentialfor animal nutrition in trace
amounts.Their occurrencein soils is quite variable,rangingfrom almostnoneto
as high as severalhundred milligrams per kilogram. Therefore,the analytical
methodsshould have the sensitivity, accuracy,precision,and speedfor the mea-
surementof Br, CI, and F in the milligrams per kilogram range.A variety of ana-
lytical proceduresthat will allow a greatdeal of flexibility for the analystsis out-
lined in this chapter.Limitations and advantagesare discussedwhere appropri-
ate.
SegoeRd., Madison,WI 53711,USA. Methods o/Soi/Analysis. Part 3. Chemical Methods-SSSA
Book Seriesno. 5.
833
834 FRANKENBERGER ET AL.
BROMINE
Introduction
Bromide contentsof mineral soils rangefrom 0.3 to 40 mg kg-l (average

about 6 mg kg-I), and in peatsrangefrom 12 to 70 mg kg-l (averageabout 30
mg kg-I) (Vinogradov, 1959). Organic matter, sedimentaryrocks and brine
waters are commonly enrichedwith Be Volcanic gasesmay often contain Br.
Bromargyrite (AgBr) and embolite (AgBrx Clx- l ) are the main insoluble inor-
ganic forms of Be Like Cl-, Be is consideredsoluble in soil and is readily
leached.However,Vinogradov(1959) points outthat soil organicmatter retains
Br- preferentiallyto Cl- and hypothesizesa bromo-organicinteraction.High cor-
relation (r = 0.77) betweenC and Br contentsin volcanic ashsoils in Japanhave
been reported(Yamada,1968), with Br concentratedin the humic acid of the
soils. It was suggestedthat Br in thesesoils was derivedfrom seawater,brought
by rain over a very long period.
Nematocidefumigants, such as methyl bromide and ethylenedibromide,
are sourcesof Br- in soils. Methyl bromide is widely used and is known to
decomposein soil to produceinorganic Br- (Hoffman & Malkomes, 1974). A
temporaryincreasein Br- of nearly lO-fold following fumigation was reported.
The amountof Br- residuedependson soil conditionssuch as moisturecontent,
temperature,and drainage.Someplant species,such as citrus (Citrus spp.), may
be sensitiveto elevatedlevels of Be following soil fumigation (Martin et aI.,
1956).
Two methodsof Br- determinationare presentedhere.One is the Conway
microdiffusion procedure(Stout & Johnson,1965), and the other is the use of a
Br- specific ion electrode(SIE).
Direct PotentiometricMethod!
Principles
The Be SIE usesa Ag-AgBr or AgBr-AgzS solid pellet as a sensor(Ross,
1969). When the electrodeis placedin a test solution, equilibrium is established
accordingto the reaction
AgBr = Ag+ + Br-.
The millivolt outputof the sensordependson the activity of Ag+. Thus, an

increasein Br- shifts the reactionto the left, resulting in a decreasedAg+ activi-
ty and subsequentdecreasedmillivolt output. Principal ions that causeinterfer-
enceswith the millivolt output are those that react with Ag+. Ammonia com-
plexeswith Ag+ resultingin positiveerror. Adjustmentof solution pH to 7 or less
will eliminate NH3 interference.Becauseboth Cl- and 1- form insoluble Ag
compounds,they will interferewith Be determination.The extentof this inter-
1 Modified from Orion Research,Inc. (1976),Saffignaet al. (1976),andAbdalla andLear(1975).

BROMINE, CHLORINE, & FLUORINE 835
ference dependsdirectly on the solubility of thesehalides with Ag. Thus CI-,

forming a more soluble salt AgCl than AgBr, will not interfere as much as 1-,
which forms a much more insolublesalt, Agl. According to one supplierof Br-
electrodes(E & K Scientific, Saratoga,California) when Cl- concentrationis 300
times greaterthan Br- and 1-, its interferenceis 0.00004times Br-. Fortunately,
watersoluble1- in soil is normally extremelylow in concentration.Cyanidealso
is commonly listed as an interfering ion. Again, CN- would be extremelyrare in
soils. The use of an 1- SIB to determinethe concentrationsof 1-, Br-, and Cl- by
titration with Ag has been proposed(Csakvari & Meszaros,as cited by Durst,
1969) but has not beenapplied to soil analysis.
SpecialApparatus
1. Bromide SIB, preferably a solid-statemodel, with suitable reference
electrode such as a double junction referenceelectrode with 10%
KN03 solution in the outer chamber.
2. Electronicvoltmeteror pH/millivolt meterwith expandedscaleor spe-
cific ion meter.
3. Magneticstirrer with Teflon-coatedstirring bar: stirrer should be well
insulatedto preventsampleheating.
4. Microburettes.
5. Graphpaper,semilogarithmicpaperfor preparingcalibrationcurves.
Reagents
1. StandardBr- stock solution, 0.01 M Be Dissolve 1.029g of NaBr in
1 L of deionizedwater to give 0.010M NaBr.
2. Total ionic strengthadjustmentbuffer (TISAB) solution,5 M: Dissolve
210 g NaND3 (previouslyheatedat lOSoCfor severalhoursandcooled
to room temperaturein a dessicator)in 500 mL of deionizedwater.
Procedure
By making proper dilutions of 0.010M NaBr standardstock solution, the
analystcan make a seriesof dilute standardscoveringthe expectedBr- rangein
the samples,probably0.01 to 1.0 meqL-l (0.8-80mg L-l). Startingwith the low-
est concentration,place 100 mL of standardin a 250-mL beaker,and add2 mL
of the TISAB buffer solution to maintainconstantionic strength.Gently stir with
the magneticstirrer with electrodesin the solution. After the readinghas stabi-
lized or after a given period of time (usually <5 min), record the potentiometer
(millivolt) reading.Avoid stirring the solution before immersingthe electrodes,
becauseentrappedair around the crystal can produceerratic results or fluctua-
tions in the potentiometerreading.Rinsethe electrodeswith deionizedwater, and
blot dry betweenreadings.Repeatthis for the remainderof the standardsamples,
and plot the resultson semilog graph paperwith concentrationon the log axis.
Using the samealiquot sizes,repeatthe measuresfor the samplesolutions.Bring
the standard andsamplesolutionsto the sametemperature,preferablyroom tem-
perature,before taking any reading. Determinethe Be concentrationby com-

paring millivolt readingsto concentrationfrom the graph.
The above procedurecan be used when the soil extract composition is
known, andstandardscanbe madeto duplicateit. Soil extractcomposition,how-
ever, may be complex and can contain interfering ions, such as Cl-, at various
concentrations.To circumvent this problem, the method of "known additions"
may be used.A standardcurve is made as describedabove. From the standard
curve, slopeS can be determined,if one realizesthat S is equalto the difference
in potentialsdivided by the difference inlog concentrations.Potentialof the sam-
ple solution is measured.A known amountof standardis added,and the solution
potential is measured.The concentrationof Be in the samplesolution, Cx , can
be calculatedby the equation
whereCs and Vs are the concentrationand volume, respectively,of the standard

addedto a known volume, Vx , a samplesolution, and AE = E z - El is the differ-
encebetweenthe potentialof the unknownmeasured before Eb and after, Ez, the
additionof standardsolution(Durst, 1969).This assumesthat S remainsconstant
and that the volume of Vs is negligible comparedwith Vs. If Vs is not negligible,
then allowancefor volume changecan be madeusing the equation
Another methodusedto determineion concentrationsby SIE in complex

solutionsis that of Gran (1952).The procedureinvolves running a seriesof stan-
dardsand standardsaddedto samples.The resultsare plotted on specialantilog
paper, which is often supplied by the electrodemanufacturer.This method is
time-consumingbut doesimprove precisionand accuracy.Additional discussion
can be found in Reevesand Brooks (1978).
Comments
The TISAB solution is addedto all standardsand samplesso that the back-
ground ionic strengthis high and constantrelative to variable concentrationsof
Br-. Sodium nitrate is recommendedfor the Br- electrodes.Other solutionscan
be used as long as they do not contain ions that would interfere with electrode
responseto Be. Thus, if the backgroundionic strengthis high and constantrel-
ative to the sensedion concentration,the activity coefficient is constant,and
activity is directly proportionalto concentration.
No attempthas beenmadeto discussmethodsof extractingBr- from soil
for analysis.Vinogradov (1959) reportedtotal Br- analysisby a NazC03 fusion
method.Water-solubleBr- in 1:1 soiVwater extractsalso has beenused(Onken
et aI., 1975). Abdalla and Lear (1975) reportedsuccessfulBe determinationby
direct measurementby the SIE of a solution extractedfrom two parts of 0.2 M
NaN03 to one part soil. Saffignaet al. (1976) extractedBe from soil using 0.05
M K ZS04 (2:1 soiVsolutionratio, 24-h shakingperiod) and determinedBr- with
direct potentiometry.
MicroditTusion Method
Principles
The following discussionon Be determinationby the Conway microdif-
fusion methodis basedon the procedureoutlined by Stout and Johnson(1965).
Bromide in soil extractscan be determinedby oxidation of Be in an acid envi-
ronment.The resultinggaseousBr2, is collectedin a KI trap causingoxidationof
the 1-. The amount of 1 formed by this reaction is measuredby either titration
with standardK 2S20 3 or by spectrometricmeasurement of the blue-purplestarch
1--1 color. The following equationsillustrate the reaction
6Be + Cr20?- + 14 W --t 2Cr3+ + 7H20 + 3Br2
Br2 + 31- --t 13 + 2Bc.
Iodide, if present,will be quantitativelyoxidized with Br- and interferewith Br-

determinations.Normally, 1- is too low in concentrationto causeany error. Con-
way (1950, p. 182-207)describesa simple methodfor removing1- interference.
Chloride doesnot interfereup to amountsof 1 to 2% in the samplesolution. The
methoddescribedis adaptedfrom thosereportedby Stout and Johnson,1965.
SpecialApparatus
1. Microdiffusion units and accessories,
standardConway-typeunits, 70-
mm in diam. and 18 mm in depth, with a glasslid: To provide a gas-
tight seal betweenlids and units, preparean adhesivefixative as fol-
lows. Melt together30 g of heavyliquid petrolatumand 13 g of paraf-
fin (mp = 50-55°C), or changethe ratio as neededto provide a mix-
ture with suitableconsistencyfor the operatingtemperatureemployed.
Preparean aspirationdevice as shown in Fig. 31-1 to transfer the I
solution from the microdiffusion unit with a 5-mL volumetric flask.
2. Shaker:To hastenthe diffusion of Br2 gasfrom the water to the inner
chamberin the diffusion units, shakethe units gently in a reciprocat-
ing shaker.A shakermaking a 3-cm strokeat the rate of 60 cycles/min
is satisfactory.Mount the units on the shakerfirmly as shown in Fig.
31-1.
Reagents
1. StandardBe stock solution: Dissolve 119 mg of KBr in deionized
water, and dilute the solution to 1000 mL with deionizedwater. One
milliliter of this solutioncontains80 /lg of Br-. Make appropriatedilu-
tions of the stock for use in preparinga standardcurve in the rangeof
approximately0 to 30 /lg of Be.
2. Sulfuric acid solution: Add 800 mL of cone H2S04 to 200 mL of
deionized water, cool the solution, and dilute it to 1000 mL with
deionizedwater.
Fig. 31-1. Tray, Conway vessels,lid supports,and aspirationdevice for transfer of solution from
inner chamberof Conway vesselto volumetric flask.
3. Potassiumchromate(KZCr04) solution: Dissolve 400 g of KZCr04 in

deionizedwater, and dilute the solution to a volume of 1000 mL with
deionizedwater.
4. Starchsolution: Grind 4 g of soluble starchand 10 mg of mercuricio-
dide (HgI2) with water to a smoothpaste.Pour the pastewith stirring
into 1 L of boiling deionizedwater.
5. Starch-iodidesolution: Preparethe solutionfresh daily by dissolving3
g of KI in 100 mL of the starchsolution.
Procedure
Place3 mL of the starch-iodidesolution in the centercompartmentof the
Conway unit.
Place2 mL of the sample,standardor blank, in the outer compartmentof
the Conwayvessel.Add 1 mL of K 2Cr04 solution and 1 mL of the H2S04. Place
the cover lid on the unit quickly, and shakethe units for 1 to 1.5 h. At the end of
this time, all the Br- will havediffused into the centerwell, which will now show
the usualblue-purplestarch1--1 color.
Draw the contentsof the centerwell into a 5-mL volumetric flask by aspi-
ration. Wash the center well with several small portions of deionized water,
drawing each washing into the volumetric flask. Make the solution to volume
with deionizedwater, and mix it thoroughly. Transfer it to a spectrometercell,
and determinethe absorbanceat 540 nm. Determinethe Be contentof the sam-
ple from the standardcurve of 0 to 30 Ilg of Br-.
Comments
No interference fromamountsof Cl- up to 1 to 2% in the samplesolution
is observed(Conway, 1950, p. 182-207). Iodide interferesquantitatively but is
rarely, if ever, found in soils and plant samplesin amountslarge enough to be
detectablein this method.
The H2S04 concentrationin the reaction is critical and should be con-
trolled within the limits of 19 to 24%.
The precipitate of K2Cr207 that forms in the reaction vessel when the
H 2S04 is addedcausesno harm.
Ion Chromatography
For information aboutthe use of ion chromatographyfor determinationof

Be in soil solution, seeTabatabaiand Frankenberger(1996, seeChapter8).
CHLORINE
Introduction
Chlorine, as the CI ion, is ubiquitousin the soil environment.It is addedto

the soil from irrigation waters, animal manures,fertilizers, plant residues,rain-
water, dust, and others. Measurementof Cl- in soil extractsis a routine task in
laboratoriesinvolved with salinity problems. In arid and semiarid areasof the
USA, Cl- is used as an important parameterin irrigation management.For
example,Cl- is used in estimatingleachingfractions and salt balance.A mea-
surementof the cation exchangecapacitiesof soils is basedon the determination
of Cl-, along with NUt, in soil extracts(Greenhill & Peverill, 1977; Smartet aI.,
1974; McLeod & Zarcinas,1976). Chloridein soil extractsmay rangefrom a few
parts milligrams per kilogram to severalhundredmilligrams per kilogram.
An essentialplant micronutrient(Broyer et aI., 1954), Cl- can rangefrom
as little as 35 mg kg- I in severelydeficient plant tissuesto as much as several
percentagepoints. Ozanne(1958) reportedthat soils containing from 30 to 50
/lmol kg- I of CI-IO.1 kg (approximately1-2 mg kg-I) failed to supportgrowth of
subterraneanclover (Trifolium subterraneum L.). Becauseof the importanceof
Cl- in plant nutrition, soil analysis,and environmentalchemistry,many labora-
tories include Cl- in their routine analysis.Therefore,the methodsshould have
the sensitivity, accuracy,precision,and speedfor the measurementof Cl- in the
milligrams per kilogram range.
Principles
Of the various methodsemployedfor Cl- determination,i.e., gravimetric,

volumetric, and electrometric,the latter two appearto be widely used(Chapman
& Pratt, 1961; Carlson& Keeney, 1971). However,with the adventof autoana-
lyzers, severalcolorimetric procedureswere adapted.One methodwidely usedis
the Hg(II) thiocyanatemethod that dependson the displacementof thiocyanate

by the Cl-. In the presenceof Fe3+, a highly colored ferric thiocyanatecomplex
is formed
2CI- + Hg(SCN)z+ 2Fe3+ -7 HgClz + 2Fe(SCN)z+,
the color of which is stable and proportional to the original ion concentration.
This method was earlier describedby Bergmannand Sanik (1957) and Rowe
(1965) and was subsequentlyutilized and modified by Canelli (1976), Fuge
(1976), and Huang and Johns(1967).
Severalelectrometricmethodsare being usedin both plant and soil analy-
ses. The three most commonly used are: (i) potentiometrictitration, (ii) direct-
readingpotentiometry(SIE), and (iii) coulometric-amperometric automatictitra-
tion. Potentiometrictitration of CI- with AgN03 has beenwidely usedfor many
yearsfor the determinationof Cl- in plant tissuesand soil samples.The end-point
can be determinedfrom a plot of electrodepotential againstthe volume of the
titrant. The rapid changein the activity of the ion in questionin the region of the
end-point generateda correspondinglylarge changein electrodepotential per
incrementof titrant. Silver-silver chloride electrodeswere commonly used for
end-pointdetectionbefore the developmentof solid-stateCl- electrodes.Solid-
stateelectrodescan be usedin the titration with the advantageof lessinterference
from oxidizing and reducing agents.Liquid membraneCl- electrodesalso are
availablebut are subjectto interferencefrom certainions commonlyfound in soil
extracts,namely,OH-, NO), HCO), and SO~-. Solid-stateCl- electrodesare vir-
tually free of interferencesfrom theseions.
Chloride in soil extractswas determinedby Hipp and Langdale(1971)
using threemethods:(i) direct-readingwith the solid-stateCl- electrode,(ii) titra-
tion with AgN03 using the solid-stateCl- electrodeas the end-point indicator,
and (iii) titration with AgN03 using a Ag+ electrodeas the end-pointindicator.
The solid-stateelectrodegave results very similar to those obtainedwith a Ag+
electrode(r = 0.999) when used as an end-point indicator. However, the solid-
stateelectrodegave higher valueswhen usedon a direct-readingelectrodethan
by titration with a Ag+ electrode.La Croix et al. (1970) also found that the solid-
stateCl- electrodewas unsatisfactoryfor direct readingmeasurements of Cl- in
plant extracts. Magalhaesand Page (1978, personal communication)working
with tropical soils, obtainedslightly higher valueswith the solid-stateelectrode
usedfor direct readingthan with the use of coulometric-amperometric automatic
titrator (r = 0.997). Adriano et al. (1973) obtainedvery similar resultswith the
use of the automatic Cl- titrator and potentiometrictitration of plant extracts.
Similarly, Gillian (1971) obtained satisfactoryCl- results with the automatic
titrator.
In direct potentiometry,the potential or voltage producedis related to the
activity of the specific ion accordingthe Nemst equation.The activity of the ion
of interestis not identical to the concentrationof the ion. Rather,the activity for
a specific type of ion in solution is relatedto the concentrationof the ion by an
activity coefficient. At low concentrationsof total ions in solution, the coefficient
approachesunity. It can be calculatedthat the error in activity resulting from an
electromotive force (EMF) measurementerror correspondsto approximately
4.0%/mV. This value representsthe most seriouslimitation of theseelectrodesfor

direct potentiometry,since at the presentstateof the art, EMF measurements are
seldommore accuratethan :to.1 mY, which even under the bestconditions,lim-
its the accuracyof ion-selectiveelectrodesto about0.5% (Ross,1969). This lim-
itation is easilyovercome,however,by using theseelectrodesas end-pointdetec-
tors in titrations wherean accuracyof betterthan 1 part per 1000can be achieved.
Specificion electrodesare becomingwidely acceptedfor the following rea-
sons:
1. They senseionic activity, which in many casesis more physiochemical-
Iy meaningfulthan concentration.However, theseelectrodesalso can
be madeto measureconcentrationwhen usedin titration.
2. Electrode measurementis extremely rapid, sometimesin a matter of
seconds.
3. Electrodesare virtually nondestructiveof the sample.
4. Pretreatmentof the samplesolution in many casesis not necessary.
5. The apparatusrequired is quite simple and relatively inexpensiveand
can be madeportable.
The automaticCI- titrator was conceivedand developedby Cotlove et al.
(1958) and has beenmodified since. It was designedoriginally for the rapid and
accuratedeterminationof solute CI- concentrationsin biological fluids, by titra-
tion with Ag ions. This processis basedon the well-establishedprinciples of
coulometric generationof reagentand of amperometricindication of the end-
point. A constantdirect current is passedbetweena pair of Ag generatorelec-
trodes in the generator(coulometric)circuit, causingreleaseof Ag ions into the
titration solution at a constantrate. The end-point is indicated after all CI- has
beenprecipitatedby the increasingconcentrationof free Ag ions, which causesa
rising currentto flow through a pair of Ag indicator electrodesand a meter relay
in the indicator(amperometric)circuit. At a presetincrementof indicatorcurrent,
the relay is actuated,stoppinga timer that runs concurrentlywith the generation
of Ag ions. Sincethe rate of generationof Ag ions is constant,the amountof CI-
precipitatedis proportionalto the elapsedtime. Therefore,CI- can be determined
by three electrometricand one coulometricmethodsoutlined in this chapter.In-
herent advantagesof the automaticCI- titrator are numerous,and results have
proved superiorto thoseobtainedby colorimetric or other electrometricmeans.
The microdiffusion methodfor CI- presentedby Stout and Johnson(1965) was
deleted,it is not commonly used.
PotentiometricTitration Method2
SpecialApparatus
1. Electronic voltmeter or pH meter with expanded scale
or digital read-
out for measuringthe potential difference betweenthe electrodes.(A
pH metercan be employedby using the appropriateelectrode.)
2 Modified from Stout and Johnson(1965) and Am. Public Health Assoc. (1980).
842 FRANKENBERGER ET AL
2. Glassand Ag-AgCl electrodes:Preparein the laboratory(seeproce-

dureoutlinedby StoutandJohnson,1965)or purchasea Ag+ electrode
coatedwith AgCl for usewith specialapparatus.Follow the manufac-
turer'sinstructionson the useand careof the electrodes.An alternate
referenceelectrodementionedin "Special Apparatus"under "Direct
PotentiometricMethod" will do.
3. Magnetic stirrer, with variable speedsetting, and Teflon-coatedstir-
ring bar.
4. Microburettes,1- to 5-mL capacity.
Reagents
1. Standardpotassiumchloridestocksolution,0.01 M: Dissolve0.746g
of pure KCl in deionizedwater, and makeup the solution to 1 L in a
volumetricflask. This solution is 0.01 M with respectto Cr and is to
be usedin standardizingthe silver nitrate (AgN03) solution.
2. StockAgN03 solution,0.1 M: Dissolve8.5 g of AgN03 in deionized
water,dilute the solution to a volume of 500 mL, and storein a dark-
coloredbottle.
3. Silver nitrate solution, 0.01 M: Dilute exactly 10 mL of the stock 0.1
M AgN03, solution to 100 mL with deionizedwater. Determinethe
correctnormality of the dilute AgN03solutionby potentiometrictitra-
tion of standardKCl solutionbeforeuse.Preparethe dilute AgN03 at
frequentintervals,and storein a dark bottle
Normality of AgN03 =milliliters of standardKCI x O.Ol/mL AgN03.
4. Concentrated,redistilled nitric acid (HN03).
Procedure
Standardization.Make the necessaryinstrumentadjustmentby following
the manufacturer'sinstructions.After allowing sufficient time for warm-up,bal-
ancethe intern-al electricalcomponentsto give an instrumentsettingof 0 mV, or
if a pH meteris used,a pH readingof 7.0. Place10 mL of standardKCl solution
into a-sufficiently largetitration vessel,dilute to about25 mL, andadd2 mL con-
centratedHN03. Immersethe electrodesin the solutionandplacethe stirring bar.
Placethe titration vesselon the stirrer. Setthe instrumentto the desiredrangeof
millivolts or pH units. Startthe stirrer.
Add standardAgN03 titrant recordingthe scalereadingafter eachaddi-
tion. At the start, add large incrementsof AgN03; then, as the end-pointis ap-
proached,add smallerand equalincrements(0.1 or 0.2 mL) very slowly, so that
the exact end-pointcan be determined.Determinethe volume of titrant usedat
end-pointat which there is the greatestchangein instrumentreadingper unit
addition of titrant. Plot a differential titration curve if the exactend-pointcannot
be determinedby visual inspectionof the data.
SampleAnalysis. Preparea soil extract using the desiredsoiVwater ratio,

or use an aliquot of an extract intendedfor such purposesas salinity measure-
ment, NO]" analysis,Cl- adsorption,etc.
Transferan aliquot of the extract preferablycontaining<0.1 mmol of Cl-
to the titration vessel.Add deionizedwater to make a volume of about 25 mL,
and addconcentratedAgN03 dropwiseuntil acidic to litmus paper,then 2 mL in
excess.Cool, immersethe electrodes,placea stirring bar in the solution,and start
the stirrer. Follow the rest of the stepsin "Standardization"under"Potentiomet-
ric Titration Method." If an end-pointreading has beenestablishedfrom previ-
ous analysesfor similar samplesand conditions, use this predeterminedend-
point. For accurateresults,makea blank titration by carryingthe deionizedwater
and other reagentsthroughthe procedure.Recordthe volume of AgN03 solution
required,and calculatethe Cl- concentrationin the aliquot, or on soil solution, or
soil dry-weight basis.
Comments
The methodoutlined is essentiallythat of Stout and Johnson (1965)except
that a supportingelectrolytesolution was not used. Iodide and bromide also are
titrated as Cl-. Ferricyanidecauseshigh resultsand must be removed.Chromate
and dichromateinterfereand shouldbe reducedto the chromicstateor removed.
Ferric ions interfere if presentin excessof that of the Cl ions. Chromic, ferrous,
and phosphateions do not interfere.
In the presenceof organiccompounds,sulfite and/orlarge amountsof fer-
ric ions, cyanide,or sulfite, the sampleshould be pretreatedbeforetitration. The
sampleshouldbe acidified with H2S04 to litmus paper.Boil for 5 min to remove
volatile compounds.Add more H2S04 if necessary,to keep the solution acidic.
Add 3 mL of H 20 2, and boil for 15 min, addingdeionizedwater to keepthe vol-
ume at about 25 mL. Dilute to 25 mL, add NaOH solution dropwise until alka-
line to litmus, then add 10 drops in excess.Boil for 5 min, filter into the titration
vessel,and washthe precipitateand filter paperseveraltimes with hot deionized
water.
The titration vesselsshould be protectedfrom direct sunlight during the
titration to preventslow drift of end-point.Rapid stirring also may causedrift of
end-point.
Direct PotentiometricMethod3
SpecialApparatus
1. Chloride SIE, preferably a solid-statemodel, with suitable reference
electrodeas indicatedin "SpecialApparatus"under"Direct Potentio-
metric Method."
2. ExpandomaticpH/millivolt meteror a specific ion meter.
3. Magneticstirrer with Teflon-coatedstirring bar: Stirrer shouldbe well
insulatedfrom heating.
3 Modified from Orion Research,Inc. (1976), and Hipp and Langdale(1971).

4. Microburettes
Reagents
1. StandardNaCl stock solution, 1000 mg L- 1 of Cl-: Dissolve 1.65 g of
reagent-gradeNaCI in a 1-L volumetric flask. Working standardscan
be preparedfrom this solution.
2. Total ionic strengthadjustmentbuffer solution for maintaininga con-
stantbackgroundionic strength (see"Reagents"under"Direct Poten-
tiometric Methods"): For sampleswith a total ionic strength<0.1 M
(see manufacturer'sinstruction manual on how to calculate ionic
strength), prepare 5 M NaN03 dissolving 42.5 g of reagent-grade
NaN03 in 100 mL of deionizedwater. Sufficient TISAB solution is
addedto the standardor samplesolutionsto bring the backgroundto
0.1 M. For sampleswith over 0.1 M ionic strength,preparestandard
solutionssimilar to the samplecomposition.
Procedure
Check the electrodeoperation following the manufacturer'sinstruction
manual.Chloride can be measuredin units of molesper liter, milligram per liter,
or any other convenientconcentrationunit.
Preparea calibrationcurve using from 1 mg L-I to about100 mg L-I of Cl-
standardsby serial dilution of the 1000 mg L- 1 stock solution. Add 2 mL of the
TISAB buffer solution to every 100 mL of the standard.Use a semilogarithmic
paperfor preparingthe calibration curve.
Transfera 50- to 100-mL aliquot of the sample(or extract) to a 250-mL
beaker.Add 2 mL of TISAB buffer solution per 100 mL of sample.Rinse elec-
trodes,blot dry, and place in sample.Stir thoroughly. Recordthe millivolt read-
ing. Determinethe Cl- concentrationfrom the calibrationcurve.
Restandardizeafter about 1 to 2 h of use. If the ambienttemperaturehas
changed,recalibratethe standardcurve.
Comments
The analystis referredto the instructionmanualissuedby the manufactur-
er of the electrodesfor detailed information about the electrodes,ion meter, or
both. Severalmodelsof the ion meterare now available,somewith digital out-
put.
The presenceof OH-, Br-, 1-, S2-, CN-, NH3 and S20~- in the solutionsto
be measuredinterferedirectly with the results(Van Loon, 1968a).
Automatic Titration Method
SpecialApparatus
1. Cotlove automaticCl- titrator.
2. Pipettes,0.5- to 2.0-mL capacity.
3. Specialtitration vials suppliedby the manufacturerof the titrator.
Reagents
1. Standardsodium chloride stock solution, 1 mmol L-': Dissolve 58.44
mg of dried, reagent-gradeNaCI in a 1-L volumetric flask.
2. Nitric acid-aceticacid reagent,0.1 M HN03 plus 10% glacial acetic
acid: Placeabout500 mL of deionizedwater in a 1-L volumetricflask.
Add 6.2 mL of concentratedHN03 and then add 100 mL glacial acetic
acid, and make up to volume.
3. Gelatin reagent:Mix Knox unflavoredgelatin no. 14 thymol blue, and
thymol at a ratio of 60:1: 1 by weight, and dissolvein 1 L of hot deion-
ized water. Store in refrigeratorwhen not in use. Immersegel in hot
water to liquefy when ready to use.
Procedure
Transfera 1- to 2-mL aliquot of the samplesolution into the specialtitra-
tion vial, add 3 mL of the HN03-acetic acid reagent,and add four drops of the
gelatin reagent.Titrate the samplewith the automatictitrator. The concentration
of Cl- in the extract is computedby referring the titration time of the sampleto
the titration time of 1 or 2 mmollL of Cl-. Subtractthe titration time of the rea-
gentsfrom the sampleand the standard.The methoddescribedis adaptedfrom
thosereportedby Cotlove et ai. (1958) and Adriano et ai. (1973).
Comments
The gelatin reagentis addedto the sampleto preventflocculation of AgCl
during titration and servesto smoothout the end-point.
The analystis referredto the manufacturer'sinstructionmanual fordetailed
information. AmericanInstrumentCo. (1969) hasseveralmodelsavailableon the
market, including a solid-stateanalytical Cl- titrator.
The Cl- titrator hasbeenusedwidely in severallaboratories(Adriano et aI.,
1972; Endelmanet aI., 1974) for Cl- determinationand also can be usedfor Be
and 1- determinations(Saffigna et aI., 1976).
Mercury(II) ThiocyanateMethod
SpecialApparatus
1. Spectrometer.
2. Volumetric flasks.
3. Pipettes.
4. Dilutor.
Reagents
1. StandardNaCl stocksolution: Use any of the previousNaCI standards
(see "Reagents"under "PotentiometricTitration Method" or "Rea-
gents" under"Direct PotentiometricMethod").
4 C.B. Knox Gelatin Co., Inc., Johnstown,NJ, or J.T. Baker Co., Phillipsburg, NJ.
2. Saturatedmercury(II) thiocyanate[Hg(SCN)21 solution, 0.075%:Add

about 0.75 g of Hg(SCN)2 to 1 L of deionizedwater, and stir over-
night. Filter througha Whatmanno. 40 paper.It is importantthat this
solution be saturated,becauseit may then be storedfor long periods
of time.
3. Ferric(III) nitrate monohydrate[Fe(N03)3 • 9H20 solution: Dissolve
20.2g of Fe(N03)3.9H20in about500 mL of deionizedwater,and add
concentratedHN03 until the solution is almostcolorless.Make up to
1 L with deionizedwater. ExcessHN03 is not important as long as
there isenoughto preventdarkeningof the solutionon prolongedstor-
age.
Procedure
Transfera known volume of the extract or samplesolution into a 25-mL
volumetric flask. Add 2 mL of the Fe(N03)3 • 9H20 solution and 2 mL of
Hg(SCN)2solution. Mix the solution,make upthe volume with deionizedwater,
and mix again.After about 10 min, measurethe absorbanceor transmittanceof
the sampleat 460 nm. Treat a blank, containingonly the reagents,in the same
manner.
Determine the Cl- concentrationfrom a standardcalibration graph pre-
paredfrom standardsolutionscontaining0 to about5 mg L-I of CI-. The method
describedis adaptedfrom those reportedby Canelli (1976), Fuge (1976), and
USEPA Staff (1974).
Comments
Severalinvestigatorshave substitutedferric perchlorate[Fe(CI04hl (Zall
et aI., 1956) and ferric ammoniumsulfate [FeNH4(S04h• 12H201 (Huang &
Johns,1967; Fuge,1976) for ferric(III) nitrate nonahydrate.Nitrite, sulfide, cya-
nide, thiocyanate,and the halides Br and I can causeinterference(Huang &
Johns,1967) when presentin sufficient quantities.This methodhas been used
with autoanalyzerin the analysisof soil and rock extracts(McLeod & Zarcinas,
1976; Selmer-Olsen& 0ien, 1973; Fuge, 1976).
Ion Chromatography
For information aboutthe useof ion chromatographyfor determinationof

Cl- in soil solution, seeTabatabaiand Frankenberger(1996, seeChapter8).
FLUORINE
Introduction
Fluorine is commonin the terrestrialenvironmentand is alwayspresentin

plants,soils, and phosphaticfertilizers. As a rule of thumb, the F concentrations
in thesematerialsare on the order of 3 x 10°,3 x 102, and 3 x 104 mg kg-I for
plants, soils, and phosphatic fertilizers, respectively (Larsen & Widdowson,

1971).Fluorineis a commonconstituentof rocks and soils. The continentalrocks
of the earth's crust contained, on average, 650 mg kg-1 of F (Fleischer &
Robinson,1963).Fleischerand Robinson(1963) gavethe following meanvalues
in milligrams F per kilogram for various geological materials: basalts, 360;
andesites,210; rhyolites, 480; phonolites,930; gabbrosand diabases,420; gran-
ites and granodiorites,810; alkalic rocks, toOO; limestones,220; dolomites,260;
sandstonesand graywackes,180; shales,800; oceanicsediments,730 and soils,
285. It occursmainly in the silicate mineralsof the earth'scrust and is an essen-
tial elementin the following minerals:fluorite, apatite,cryolite, topaz, phlogo-
phite, lepidolite,and other less importantminerals.
Very common soil minerals, such as biotite, muscovite,and hornblende,
may contain as much as several percentagepoints of F and, therefore,would
seemto be the main sourceof F in soils. However, Steinkoenig(1919) believed
that micas,apatite,andtourmalinein the parentmaterialswere the original source
of F in soils. It appears,therefore,that the F contentof soils is largely dependent
on the mineralogicalcompositionof the soil's inorganic fraction. Since the soil
environmentis often adulteratedwith various kinds of phosphaticfertilizers and
varioustypesof industrial pollutants,soil F can be very variable.Phosphaticfer-
tilizers, especially the superphosphates, are perhapsthe single most important
~ource of F in agriculturallands. Repeatedapplicationof rock phosphates, which
containedseveralpercentagepoints of F, significantly elevatedthe F contentsof
soils (Omueti & Jones,1977b).Other possiblesourcesthat may affect soil Fare
precipitation,fallout of combustionproductsof coal and other industrially pol-
luted air, and F-bearinginsecticides.It is reasonableto expectthat the soil con-
tent of F rangesfrom a trace to severalhundredmilligrams of total F per kilo-
gram.
Fluorine is not an essentialplant nutrient but is essentialfor animals.How-
ever, continuousingestionby animalsof excessiveamountsof F can lead to flu-
orosis, and suboptimallevels in the diet can have an equally damagingeffect.
Therefore,the plant contentof F is of interestto livestock producers.It hasbeen
conclusivelyshown that plant uptakeof F from soil is not usually relatedto the
F contentofthe soil but ratheris largely dependenton the soil type, Ca and P con-
tents,and soil pH (Brewer, 1965b).
Fluorineis found in soils asthe singly chargedF""", or occasionallyasa com-
ponentof suchcomplexanionsas(BF4t, (AlF6)3-, and (SiF6)2- (Hopkins, 1977).
Principles
Until recently, the classicalWillard-Winter distillation method(Willard &

Winter, 1933)(and its modifications and subsequenttitration or colorimetric
determinations)has beenusedextensivelyfor F determinationin soil and rock
samples.This well-known proceduredependson: (i) the releaseof F from the
sampleby meansof a suitabledecompositiontechnique,(ii) separationof F from
interfering ions by steamdistillation, and (iii) determinationof F by a suitable
analytical method. Although quite accurateand reliable, the distillation step is
time-consuming,potentially hazardous,and requirescareful control to obtain re-
liable results. Recovery of F largely dependson the design of the distillation

apparatus,the concentrationand type of acid, temperature,distillation rate, and
distillate volume collected. Modem techniquesbasedon the original Willard-
Winter methodhaveF recoveriesof 95 to 100% (Brewer, 1965a).Becauseof the
time-consumingdistillation and the cleaningprocedureafterward,this procedure
is becomingless and less widely used and probably is not practical in analysis
involving large numbersof samples.
In titrimetric procedures,prior eliminationof interferingions suchas SO~-,
P~-, and CI- is necessary.This can be accomplishedby steamdistillation in
HCI04 but requiresthe addition of Ag2S04 to the distillation flask to precipitate
the Cl- (Brewer, 1965a).The F in the final distillate can be determinedby the tra-
ditional titrimetric methodusing Th(N03)4 in the presenceof Alizarin red S or
other suitablecolor indicator. The titration is performedaccordingto the follow-
ing equation(Dahle et aI., 1938):
When all the P-- is bound, the first excessof Th(N03)4 reacts with the indi-
cator alizarin sulfonate by forming a peach-blossompink (red) lake
(C14H404S03H)4 Th (Nommik, 1952). This techniqueis sensitivebut is not reli-
able, and end-point detection is probably the largest single sourceof variation
between analysts (Hardin, 1952). However, spectrometricmethods are more
reproduciblebut lesssensitive(Brewer, 1965a).
Becauseof the inherent disadvantagesof the classicaldistillation method
and its companiontitrimetric techniques,two alternatemethodsare presented.
The first method involves the use of P-- SIE, which can tolerate a relatively
greateramountof interferingions andprovidesa muchsimplerandfastermethod
of analysis.The second methodis the spectrometric sodium 2-(parasulfophenyl-
azo)-1,8-dihydroxy-3,6-naphthalene disulfonate [also called 4,5-dihydroxy-3-
(parasulfophenylazo)-2,7-naphthalenedisulfonic acid trisodiumsalt, or SPADNS]
method(see"Fluorine DeterminationUsing Sodium2-(parasulfophenylazo)-1,8-
dihydroxy-3,6-Naphthalene disulfonateMethod"), which can be interphasedto
an autoanalyzer.Both are standardmethodsadaptedfor the American Public
Health Association(1980). The SIE procedureis becominga popularand wide-
ly acceptedmethod.
Total Fluorine
Decompositionof Fluorine.ContainingSamples
As previously mentioned,it is important to completelydecomposeall F-
bearingcompoundsto allow completedissolutionof F before any distillation or
analytical step is attempted.A very reliable, but time-consuming,procedurefor
decomposition of F-containing samples involves the high temperature
(16~165°C) treatmentwith H2S04 (Brewer, 1965a).The F, togetherwith vari-
ous volatile compounds,is removedfrom the hot H2S04 by steamdistillation.
After a quantitativetransferof F from the distillation step, the distillate is made
alkaline with NaOH, reducedto smaller volume by evaporation,and redistilled

from H2S04 at lower temperature.Prior distillation from H2S04 at 165°C was
superiorto prior incinerationwith CaO at 800°C for determiningF in soils resis-
tant to decompositionby HCI04 (Maclntire et al., 1951). Many soils that do not
contain large amountsof micas and topaz may be satisfactorilydecomposedby
incinerationwith CaO or Ca(OHhwhich is less time-consumingthan the H2S04
treatment(Brewer, 1965a).
Since the introduction of the r SIE in 1966 (Frant & Ross,1966) several
modificationsof traditional methodsor new decompositionprocedureshavebeen
developed.Most of thesemethodsdo not require distillation prior to analysisby
SIE.
Ficklin (1970, p. 186--188)developeda simple methodwhere the rock or
soil samplewas sinteredwith a mixture of Na2C03 and KN0 3. The sinter was
dissolvedwith deionizedwater and the flux neutralizedwith citric acid. The pH
of the solution was between5.5 and 6.5. This methodhas been adaptedand/or
modified by Pluger and Friedrich (1973), Farrel (1974), Crenshawand Ward
(1975), and Troll et al. (1977) and was found to be satisfactoryfor most purpos-
es. However,at high F concentrations(> 19 000 mg L -1), the SIE did not conform
to the Nernstian responseand consequentlyresulted in poor precision (Farrel,
1974). Crenshawand Ward (1975) useda TISAB buffer solution in conjunction
with the SIE use.
Another decompositionmethod was developed by Shergold and Selfe
(1974) and subsequentlymodified by Hopkins (1977) for the SIE method of
analysisfor rocks and soils. He fused the samplewith a Na2COrK2C03mixture.
Citric acid and sodium citrate were usedto dissolvethe fused sampleand adjust
the pH, maintain high ionic strength,and inhibit any metal-ionr complexes.
This methodgave F values in good agreementwith certified, accepted,and re-
portedvaluesfor severalstandardrock samples.
Thomasand Gluskoter(1974) usedthe Na2C03fusion methodfor F analy-
sis of coal samples.It involves two fusion-temperaturesteps,first at 475°C and
then at 1000°C.The fused samplewas then extractedwith hot water and the solu-
tion pH adjustedto 5.0 to 5.2 with H2S04. A citrate buffer solution was added
before F determinationby SIE. This methodgave satisfactorytotal F resultsfor
Illinois soils, where a coefficient of variation of about3% was obtained(Omueti
& Jones,1977a).Determinationof F in standardrock samples, using this method
agreedwithin 2% of the reportedvalues.
Another fusion procedureadaptedfor the SIE methodis the NaOH fusion
method (Van Loon, 1968b). The fused sample was dissolved with deionized
water and the solution pH adjustedto 7.0 to 8.0 with HCI solution. It is not very
accurate,giving errors up to 5% of the calculatedF contents.A simple modifica-
tion of the NaOH fusion SIE methodwas developedby Evanset al. (1970) for the
analysisof phosphaterock by using dilute HCl04 solution to further dissolvethe
resultantsolid of the fuseate.The resultscomparedvery favorably with a distil-
lation-SIE method. Pluger and Friedrich (1973) further modified Van Loon's
NaOH fusion method by adding strong buffer (pH 6) after the samplefusion to
avoid F sorption by precipitated hydroxides. They obtained favorable results
when compared with Ficklin's fusion technique (Na2COrKN03). Hopkins
(1977)also modified the NaOH fusion methodby usingcitric acid to dissolvethe

fuseateand addingsodiumcitrate buffer (pH 6) beforeSIE determination.
McQuakerand Gurney(1977) modified the NaOH fusion-SIEmethodthat
was originally proposedby Baker(1972) for plant sampleanalysis,which can be
employedfor determiningF in both soil and plant samples.This methodis pre-
sentedbecauseit provedto be relatively preciseand accurate,with recoveriesof
addedF in the neighborhoodof 95 to 100%.
Separationof Fluorine from Interfering Compounds

It is important to separateF from interfering ions before the use of titri-
metric methodswith Th(N03)4 and, in somecases,with spectrometricSPADNS
method.
Steamdistillation of soil in the presenceof H2S04 at 165°C releasesmany
other ions other than F-. Some of these,particularly PO~- and SOi-, interfere
with the Th(N03)4 determinationof F. It 'is necessary,therefore,to separatethe
F from the interfering substances.The most common method of separation
involves volatilization of the F as HF or H2SiF6 from H2S04 or HCI04 by steam
distillation at 135°C(Brewer, 1965a;Willard & Winter, 1933). Silver sulfate or
perchlorate is usually added to prevent simultaneousvolatilization of HCl.
McClure (1939) found Ag2S04 powderbetterthan Ag2S04 solution for precipi-
tating Cl- in H2S04, Frant and Ross(1966) have demonstratedthat the presence
of quartzor glassbeadsas a sourceof silica is not requiredfor volatilization of
F, becausethe F is distilled as a hydrate of HF rather than as fluosilicic acid
(H2SiF6), assomeworkershavepostulated(Willard & Winter, 1933).The useof
glassbeadsto preventbumping before the admissionof steamis highly desir-
able, however,when distillate or other solutionsare being processed.No beads
are necessarywhen F is being distilled from a calcinedsoil sample.
Steamdistillation from 50 to 60% HCI04 at 135°Cis oneof the mosteffec-
tive methodsand probably is the most popularmethodemployedfor separating
F in soils and other materials from interfering substances(Brewer, 1965a).
Double vacuum-distilled70% HCI04, which is especiallylow in F content, is
commerciallyavailable.Perchloricacid is preferredto otheracids,becausenear-
ly all perchloratesare easily soluble, and HCI04, which may distill over, has no
interfering effect (unlike H2S04) in the colorimetric procedure.Willard (1912)
has pointed out that HCI04 at the above concentrationsis not explosive. It is
unstableonly when anhydrousand does not becomea strong oxidizing agent
until it is heatedabove200°C. At lower temperatures,it is effective in releasing
F from its various compounds.The evolved HF is carried away from the distil-
lation flask in a streamof steam.
McClenahenand Schulz(1976) usedonly a single distillation to determine
total soil F with the SIE. However,the resultsthey obtainedwereslightly but sig-
nificantly higher comparedwith a double distillation-SIE method.
SodiumHydroxide FusionMethod for Total Fluorine

Special Apparatus
1. Nickel crucibles,130-mL capacity.
BROMINE, CHWRINE, & FLUORINE 851
2. Electric drying oven.

3. Muffle furnace.
4. Micropipettesor Eppendorfpipettes.
Reagents
1. StandardF- stocksolution, 1000mg L -1 of F-: Dissolve2.21 g of pre-
dried reagent-gradeNaF in deionizedwater, and dilute to 1 L.
2. Total ionic strengthadjustmentbuffer solution: Add 58 mL of glacial
aceticacid and 12 g of sodiumcitrate to 300 mL of deionizedwater,
stir to dissolve,and adjustthe pH to 5.2 using 6 N (6 M) NaOH. Cool
and dilute to 1L.
3. Sodium hydroxide solution, 17 M: Dissolve 680 g of reagent-grade
NaOH pellets in deionizedwater, and dilute to 1 L. Store in a poly-
ethylenecontainer.
4. Hydrochloric acid, concentrated.
Procedure.Homogenizethe soil sample,passthrough a 100-mesh(150-
Ilm) sieve,and dry to constantweight at 105°C.Accuratelyweigh approximate-
ly 0.5 g of the sample,and transferto the Ni crucible. Next, slightly moistenthe
samplewith deionizedwater,and add 6.0 mL of the NaOH solution.Tap the cru-
cible lightly to uniformly dispersethe samplein the NaOH solution. Place the
samplein an oven set at 150°C for about 1h, then remove after the NaOH has
solidified. Transferthe crucible to a muffle furnaceset at 300°C, gradually raise
the temperatureto 600°C, and fuse the samplefor 30 min. Removethe crucible
from the furnace,allow it to cool, and add10 mL of deionizedwater to the sam-
ple. Heat the sampleslightly to facilitate dissolutionof the NaOH fusion cake.
Then,slowly add about8 mL of concentratedHCI, with stirring to adjustthe pH
to 8 to 9 with an aid of a pH paperindicator. Cool the acidified sample,transfer
to a 100-mL volumetric flask, dilute to volume, and filter through Whatmanno.
40 paper.
Carry a reagentblank through the same procedure.Before making the
potentialmeasurement following the procedureoutlined in "Fluorine Determin-
ation Using Direct PotentiometricMethod," adjust the pH of the preparedsam-
ple and blank to 5.2 by adding 25 mL of the filtrate and 25 mL of the TISAB
buffer solution to a 150-mL polyethylenebeaker.Determinethe F valuesfrom
the standardcurve following the proceduresoutlined in "Procedure" under
"Fluorine Determination Using Direct PotentiometricMethod." The method
describedis adaptedfrom thosereportedby McOuaker andGurney(1977).
Comments.McOuakerand Gurney (1977) found that interfering cations,
suchas AI3+ and Fe3+, were removedas insolubleoxideswhen the sampleswere
filtered underthe slightly alkaline condition of pH 8 to 9. It is well documented
that some cations are capableof complexingF, depressingthe results. Bellack
(1972) found that at a level of 1 mg F L-1, a suppressionof 10% occurredif
either2 mg L-1 of AI or 200 mg L-l of Fe was present.An equivalentreduction
occurredin the presenceof 2% Cl-. The addition of citrate to buffer (TISAB)
solution will preferentially complex up to 2 mg L -1 of AI (Am. Public Health
Fig. 31-2. All-glass P-distillation unit wrappedwith asbestoscord

to minimize heat loss. The principal parts are (A) distillation
flask; (B) 0 to 200°Cthermometerin the thermometerwell; (C)
combinationstill head,steaminlet tube, aerosolbaffle, and 300-
mL Liebig condenser;(D) steamcondensatetrap; and (E) elec-
tric heatingmantle.
Assoc., 1980). Only trace quantities of these interfering ions were found by
McQuaker and Gurney (1977) in the soil sample filtrates. Silica was largely
removedasinsolublesilicateswhenfiltered underthis slightly alkalinecondition
and averagedonly 1.9 mg L-I of Si in the filtrates. Crosby(1969) indicatedthat
at Si levels <15 mg L-I (or 50 mg L-l of SiOf), formation of undissociatedSiFg
is negligible. Precisionof 4.1% was obtainedby McQuakerand Gurney (1977)
for soil samplesin the rangeof 30 to 700 mg kg-1 of F and 4.3% for plant sam-
ples in the rangeof 50 to 600 mg kg-1 of F. With a 0.5-g sample,the detection
limit was approximately3 mg kg-1 of F.
Sulfuric Acid Method for Total F1uorines

Special Apparatus
1. Steamdistillation apparatus(Fig. 31-2).
2. Soil grinder or mortar and pestle.
Reagents
1. Sulfuric acid, concentrated,reagentgrade.
2. Sodium hydroxide solution, 10%: Dissolve 100 g of reagent-grade
NaOH pelletsin 900 mL of deionizedwater.
Procedure.Pulverize sufficient air-dried, coarsely screenedsample
with a mortar and pestleor soil grinder until the entire samplepassesthrough a
200-mesh(75-J..lm) sieve. Dry the ground sampleto constantweight at 1OS°e.
Weigh accuratelysufficient sampleto yield 10 to 200 J..lg of F (1.0 g of sampleif
the F contentis not known). Transferthe samplequantitatively to a distillation
flask, add 25 mL of deionizedwater and SO mL of concentratedH2S04, Install a
o to 200°C thermometerso that at least 2.S cm of the tip is immersedin the
S Willard and Winter (1933), Dahle and Wickman (1936, 1937), MacIntire et al. (1951), and
Hardin (1952), as adaptedfrom Brewer(1965a).
H2S04, Attach the steamdistillation head,and placea 500-mL Erlenmeyerflask

underthe condenser.Slowly apply heat to the distillation flask with a gas burner
or electric heating mantle. When the temperaturereaches150°C, admit steam,
and continuethe heatinguntil a temperatureof 165°C is attained.Adjust the rate
of steamentranceand externalheatingto achievethe delivery of 6 to 8 mL of dis-
tillate per minute to the Erlenmeyerflask while maintainingthe solution in the
distilling flask at a temperatureof 165 ± 2°C. When approximately500 mL of
distillate has beenobtained,the next step dependson the methodusedfor mea-
suring F. If the direct potentiometricmethodas outlined in "Fluorine Determina-
tion Using Direct Potentiometric Method" is used, no further distillation is
required.The distillate is madeto volume in a 500-mL volumetricflask and saved
for F determination(McClenahen& Schulz, 1976).
If one of the titration methodsis used,add 10 mL of 10% NaOH solution
to the distillate, and evaporatethe distillate almostto drynesson a warm plate or
steambath.Add deionizedwater to give a volume of approximately30 mL, and
swirl the solution to dissolvethe salt residueon the walls. Then proceedwith the
seconddistillation as directed in "SteamDistillation for Separationof Fluorine
from Sulfuric Acid."
Calcium Oxide IncinerationMethod for Total Fluorine6

Special Apparatus
1. Nickel or platinum crucibles,50-mL capacity.
2. Electric muffle furnace.
Reagent
1. Calcium oxide (CaD), reagent-grade,low in F.
Procedure.Grind sufficient subsampleof air-dried soil until it passes
through a 200-mesh(75-llm) sieve, and dry to constantweight at 105°e. Ac-
curately weigh the sampleinto a Nickel or Platinum cruciblesufficient to yield
10 to 200 Ilg of F. Otherwiseuse a 1.0-g sample.Add 5 g of CaD powder, and
mix it thoroughlywith the soil. Heat the crucible at 500°C in a muffle furnacefor
at least5 h, then increasethe temperatureto 800°C for 2 h. Allow the crucible to
cool, and then proceedwith the steamdistillation of the F as describedin "Steam
Distillation Method for Separationof Fluorine from PerchloricAcid."
SteamDistillation Method for Separationof Fluorine

from PerchloricAcid'
SpecialApparatus.The most popular P- distillation apparatus,and per-
haps the easiestto fabricate, consistsof a modified 250-mL Claisen flask into
which is fed steamfrom a separatelyheatedFlorenceflask. This type of appara-
tus hasbeendescribedin detail by Churchill (1945) and Rowley et al. (1953) and
6 Willard and Winter (1933), MacIntire (1945), MacIntire et al. (1951), and Hardin (1952), as
adaptedfrom Brewer (1965a).
7 Willard (1912), Willard and Winter (1933), Dahle and Wickman (1936, 1937), and Fox and
Jackson(1959), as adaptedfrom Brewer (1965a).
854 FRANKENBERGERET AL.
is recommendedfor determinationof F in soils by the Associationof Official

Analytical Chemists(1980). This basicdesigncan be improved considerably by
replacing the rubber stopperswith ground-glassjoints and adding a Kjeldahl
connectingbulb aerosolbaffle to the side arm, thereby extendingthe distance
betweenthe distilling solutionandthe top of the condenserby at least10 em. The
extradistanceand the baffle reducecontaminationof the distillate by sprayfrom
the distillation flask. A combinationdistillation flask, steamgenerator,and con-
denserin a single piece of glasswarehas beendescribedby Mavrodineanuand
Gwirtsman(1954). Huckabayet al. (1947) have describeda unit that combines
the distillation and condenserunits and that usesrefluxed sym-tetrachloroethane
to maintaina constanttemperatureof 146°Cin the distillation chamber.This unit
is suitablefor useonly with H2S04, becauseaccidentalbreakageof the glasswall
separatingthe acid and the refluxing organicconstantboiling solution would be
disastrouswith HCl04 in the distillation flask. Fox and Jackson(1959) have
describedan elaboratethermostaticallycontrolledunit for usewith ore samples.
With slight modification (increasingthe size of all components,especiallythe
distillation flask), this unit would be suitablefor determinationof F in soils.
Brewer (1965a) strongly favors the still design illustrated in Fig. 31-2.
This design,describedin more detail elsewhere(Brewer & Liebig, 1960), is all
glass,relatively easyto constructfrom componentparts, efficient, dependable,
and convenientto operate.It consistsof a 300-mL, three-necked,standard-taper
distilling flask (A) attachedto a combinationsteaminlet tube,distilling head,and
Liebig condenser(C). A steamcondensatetrap (D) is usedbetweenthe still and
the steamsource.A modified domesticpressurecookermakesan excellentsteam
generator.A standard-taper thermometerwell (B), containinga 0 to 200°C ther-
mometerimmersedin a few milliliters of concentratedH2S04, is placedin one
of the side necksof the distilling flask. A gasburneror electricalheatingmantle
is used to heat the distilling flask. Gas burnersare less expensivebut require
more watching and more frequent adjustmentto maintain the desired distil-
lation temperature.For routine work, a bank of six or more stills is usually
constructed.
Reagents
1. Perchloricacid, 70% double vacuumdistilled, low in F (commercial-
ly availablefrom many chemicalmanufacturers).
2. Silver perchlorate·(AgCl04), 17.5% solution: Dissolve 17.5 g of
reagent-grade AgCl04 in a 100-mLvolumetricflask, anddilute to vol-
ume.
3. p-nitrophenolindicator, 0.5% solution: Dissolve 0.5 g of p-nitrophe-
nol in a 100-mL volumetric flask, and dilute to volume.
4. Sodium hydroxide, 50% solution: Dissolve 50 g of reagent-grade
NaOH in 50 mL of deionizedwater.
Procedure. Quantitatively transfer the condensed distillate from
"Procedure"under"Sulfuric Acid Methodfor Total Fluorine" or the calcinedsoil
samplefrom "Procedure"under"Calcium Oxide IncinerationMethodfrom Total
Fluorine" to a distillation flask with 50 mL of 70% HCI04 and 25 mL of deion-
ized water. Add 10 drops of 17.5% AgCI04 solution. If the solution turns milky
white, indicating the presenceof a large amount of Cl-, add an additional 10
drops to assurean excessof Ag+. If the sampleis a distillate from "Procedure"
under"Sulfuric Acid Method for Total Fluorine," add 8 to 10 glassbeadsto pre-
vent bumping before the steamis turned on. Place a 500-mL Erlenmeyerflask
under the Liebig condenser.Add one drop of p-nitrophenol indicator and one
drop of 50% NaOH to the receiverflask. Heat the distilling flask to 128°C, intro-
duce steam,and raise the temperatureto 135°C. Adjust the steamflow to main-
tain a constantlevel of liquid in the distillation flask and to produce5 to 6 mL of
distillate per minute. Continue the steamdistillation until 400 mL of distillate
havebeencollected.If samplesare extremelyhigh in Si, AI, or both, reducethe
rate of distillation to 3 to 4 mL min-l. To verify that all the F has beenremoved
from solution, distill an additional100 mL into a separateflask. Determinethe F
content of both samplesas describedin "Fluorine DeterminationUsing Direct
Potentiometric Method", "Fluorine Determination Using Sodium 2-(parasul-
fophenylazo)-1,8-dihydroxy-3,6-Naphthalenedisulfate Method," "Titrimetric
Fluorine Determinationwith Thorium Nitrate in Presenceof Chrome Azurol
Sulfur," or "Titrimetric FluorineDeterminationwith Thorium Nitrate in Presence
of Alizarin Red Sulfur". If the 100-mLsampleis found to contain>5% as much
F as the initial 400 mL, continuetaking 100-mL samplesof distillate until com-
plete recovery is indicatedby lack of measurableamountsof F. In practice,the
amountof F releasedafter 400 mL is collectedwill seldomexceed1 or 2% of
the total F containedin the sample.
SteamDistillation for Separationof Fluorine from Sulfuric AcidS

Special Apparatus
1. Steamdistillation apparatus(see "Special Apparatus" under "Steam
Distillation Methodfor Separationof Fluorinefrom PerchloricAcid").
Reagents
1. Sulfuric acid, 70%: Add slowly 1 L of reagent-gradeH2S04 to 790 mL
of deionizedwater.
2. Silver sulfate(Ag2S04), reagent-grade.
3. p-nitrophenolindicator: Dissolve 0.5 g of p-nitrophenolin 100 mL of
deionizedwater.
4. Sodium hydroxide,50%: Dissolve50 g of reagent-gradeNaOH in 50
mL of deionizedwater.
Procedure. Transfer the condenseddistillate from "Procedure"under
"Sulfuric Acid Method for Total Fluorine" to a distillation flask, rinse the Erlen-
meyerflask twice with 25 mL of 70% H2S04, and add 0.1 g of Ag2S04 (a small
measuringscoopcan be madeto containapproximatelythis amount)and 8 to 10
glassbeads.When the temperaturereaches130°C,introducesteamand continue
8 Hoskins and Ferris (1936), Armstrong (1936), Hillebrand and Lundell (1953, p. 737-748),as
adaptedfrom Brewer (1965a).
heatinguntil the temperaturereaches13SoC.Adjust the heatand steamto give S

to 6 mL of distillate per minute at a distillation temperatureof 13S ± 2°e. Distill
approximately37S mL into a SOO-mLErlenmeyerflask calibratedat 400 mL and
containingone drop of p-nitrophenoland one drop of SO% NaOH. If the yellow
color disappears,add additional NaOH to maintain the distillate alkaline.
Comments.Under certainconditions,particularly when the stills are new,
mild to violent bumpingof the acid solutionmay be experiencedbetween11S to
130°e. To avoid such an occurrence,a trickle of steam should be bubbled
through the acid solution beginningat about lOSoC. Only a minimum of steam
should be admittedbelow the prescribedtemperatureof 130°C, however.
SolubleFluorine
Principles
Soil type, pH, Ca, P, and AI contentsappearto be the predominantfactors
controlling F availability to plants (Brewer, 1965b). Prince et al. (1949) and
Hurd-Karrer (19S0) showedthat only under very special conditions (e.g., very
acid, sandysoils low in P and Ca) was plant uptakeof F from soil relatedto total
soil F. Water-solubleF is expectedto be the most readily available form for
plants. However, uptake by plants will not be controlled completely by F con-
centration in water extracts. Fluoride is adsorbedon soil surfacesand forms
insoluble compoundsmuch like phosphate.
For water-solubleF, Brewer (196Sa)recommendeda 1:1 Vol. soil extract.
Distillation of the extractfrom HCI04 (see"SteamDistillation Method for Sep-
aration of Fluorine from Perchloric Acid") or H2S04 (see "Steam Distillation
Method for Separationof Fluorine from Sulfuric Acid"), followed by titration
with Th(N03)4, was recommendedfor accurateresults.A colorimetricprocedure
was outlined when high precisionwas not necessary(Brewer, 1965a).Use of the
SIE offersanotherrapid, accuratemethodfor determiningwater solution F (see
"Fluorine DeterminationUsing Direct PotentiometricMethod").
Larsenand Widdowson(1971) useda CaCl2 solution to obtain water sol-
uble and weakly adsorbedF. They recommendedusing an anion exchangeresin
for labile F. Oncethe extractis obtained,as before,selectionof an analytical pro-
cedurefor F dependson the precisionneededand equipmentavailability.
Water-SolubleFluorine
Procedure.Mix SOO mL of deionizedwater with SOO mL of weighed,SO-
mesh(300-/lm), air-dried soil, and allow the mixture to stand2 to 3 h. Transfer
the soil suspensionto a IS-cm Buchnerfunnel fitted with Whatmanno. 1 filter
paper,and apply vacuum.Fluorine in the extractcan be determinedby a poten-
tiometric method (see "Fluorine Determination Using Direct Potentiometric
Method"), by a volumetricmethod(see"Titrimetric FluorineDeterminationwith
Thorium Nitrate in Presenceof ChromeAzurol Sulfur" or "Titrimetric Fluorine
Determinationwith Thorium Nitrate in Presenceof Alizarin Red Sulfur"), after
distillation (see"SteamDistillation Method for Separationof Fluorinefrom Per-
BROMINE, CHWRINE, & FLUORINE 857
chloric Acid" or "Steam Distillation for Separationof Fluorine from Sulfuric

Acid"), or colorimetrically (see"Fluorine DeterminationUsing Sodium2-(para-
sulfophenylazo)-1,8-dihydroxy-3,6-Naphthalene disulfateMethod"). The meth-
od describedis adaptedfrom thosereportedby Brewer (1965a).
0.01 M Calcium Chloride-ExtractableFluorine

Special Apparatus
1. End-over-endshakeror a suitablereciprocatingshaker.
Reagents
1. Calciumchloridesolution,0.01 M: Weigh 1.47g of CaCl2• 2H20, and
dilute to 1 L with deionizedwater.
Procedure.To 16 g of air-dried andsieved«0.42mm) soil, add50 mL of
0.01 M CaCl2 solution. Using an end-over-endshaker,shakethe mixture for 16
h at 25°C. Analyze the solution for F by direct SIE measurement
after filtration.
Fluorine can be analyzedby another method as outlined in "Fluorine Deter-
mination Using Sodium2-(parasulfophenylazo)-1,8-dihydroxy-3,6-Naphthalene
disulfate Method." The method describedis adaptedfrom those reported by
Larsenand Widdowson(1971).
Comments.The possibility of fluoride (CaF2) controlling the solubility of
F shouldbe acknowledged.However,Larsenand Widdowson(1971) found that
F concentrationswere too low to allow CaF2 formation. In soils they tested,F
was commonly in the 0- to 0.2 mg kg-1 rangewith the 0.01 M CaCl2 extractant.
By an anion exchangemethod,they found F in the 20-mg kg-1 range.
Measurementof Fluorine in Solution
Fluorine DeterminationUsing Direct PotentiometricMethod

Principles.The direct potentiometricmethodfor F determinationby solid-
stateSIE hasreceivedwide application.This electrode,whoseuniqueproperties
arisefrom a Eu-dopedLaF3 crystal, is sensitiveto P-- andvirtually no otheranion
or cation.It is one of the few truly specific ion-selectiveelectrodes.It gives a
Nernstianresponseto P-- from >1 M to <10-5 M (0.2 mg L-l) (Butler, 1969).
Measurementsof concentrationsto 10-6 M are possiblewith careful standard-
ization. Hydroxyl seemsto be the only ion to interfere with P-- directly. This
interferenceis eliminatedby properpH control. Since the electroderespondsto
P-- activity, if concentrationis desired,considerationof activity coefficientsor
using a constantionic strengthsolution in all measurements must be made.For-
mation of complexeswith Fe or AI will reducethe activity of P--. This also is
controlledby addition of an Fe and AI organiccomplexingagentto reducethis
interference.Whenvery low P-- concentrationsare measured,the presenceof La-
complexingagents,such as citrate, ethylenediaminetetraacetic acid (EDTA), or
cyclohexanediaminetetraacetic acid (CDTA), will increasethe solubility of LaF3,
and consequentlyP--, giving anomalouslyhigh values(Ross,1969).The method
describedis adaptedfrom thosereportedby McQuakerand Gurney(1977).
Special Apparatus
1. Fluoride SIE, preferably a solid-statemodel, and with a single-junc-
tion referenceelectrode.
2. ExpandomaticpH/millivolt meteror a specific ion meter.
3. Magnetic stirringunit, preferablya water-driventype to preventsam-
ple warming, or a well-insulatedelectric unit.
4. Microburettes.
5. Graph paper,semilogarithmicpaperfor preparingcalibrationcurves.
Reagents
1. StandardP- stock solution, 1000 mg L -1: Dissolve 2.21 g of predried
reagent-gradeNaF in deionizedwater, and dilute to 1 L.
2. Total ionic strengthadjustmentbuffer solution for maintaininga con-
stantbackgroundionic strength(see"Procedure"under "Microdiffu-
sion Method"): To 500 mL of deionizedwater, add 58 g of NaCl, 57
mL of glacial aceticacid, and 4 g of CDTA. Stir to dissolve,and then
adjust pH to 5.2 using 6 M NaOH. Cooland dilute to 1 L with deion-
ized water.
Procedure.Placean aliquot of a solution from a soil extract in a polyeth-
ylene beaker(150-mL beaker is a convenientsize). Add an equal volume of
TISAB to' the aliquot, and stir the solution.The pH of the resulting solution
shouldbe between5.0 and 5.5. While stirring gently on a magneticstirrer, place
the electrodesin the solution. After stabilizationof the potentiometer,take the
millivolt reading. By preparingstandardP- solutions from the standardstock
solution containingP- over the expected range, preparea standardcurve by plot-
ting millivolt (linear axis) againstP- concentration(log axis). As with the soil
extract solution, it is necessaryto add one volume of TISAB to one volume of
the standardsolution. Determinethe P- concentrationby comparingthe millivolt
reading from the soil solution extract with the standardcurve (see "Reagents"
under "Microdiffusion Method" for other methodsof relating potential reading
to concentration).
Insteadof making a standardcurvefor determiningP- concentrationin soil
extract solutions,usethe methodof "known addition" asdescribedin "Reagents"
under"Microdiffusion Method."
Comments.Someresearchersuse 12 g of sodiumcitrate dihydrate in the
TISAB buffer solution in placeof CDTA (McQuaker& Gurney, 1977), andoth-
ers have used a combination of 1 g of CDTA with 0.3 g of sodi'Jm citrate
(McClenahen& Schulz,1976).Soil extractsobtainedfrom alkali fusion methods
shouldbe acidified with HCI to pH 8 to 9 and filtered through Whatmanno. 40
filter paper to remove much of the AI and Fe before analysis by the P- SIE
(McQuaker& Gurney, 1977).
Commonly,25- to 50-mL aliquotsof soil extractsare usedin the analysis.
As long as analystsare consistentand the electrodeswith magneticstirring bar
are coveredwith solution, there is no other restriction on aliquot volume other
than sampleavailability.
Crosby et aI. (1968) comparedthe SIE methodwith severalwidely used

colorimetricmethodsand found it superiorin regardto speed,accuracy,and con-
veniencefor determinationof P- in watersamples.They useda modified TISAB
buffer solution that containedphosphate,citrate, and EDTA. They found virtual-
ly no interferencefrom some commonwater constituents,such as FeS04,Cl-,
POa-, alkalinity, and Caz+, except that AI markedly interfered at the 0.5-mg
L-1level.
The TISAB buffer largely minimizes the interferingeffectsof ions present
in the solution and largely eliminateserrors from pH changesand complexing
agentssuch as AI.
Fluorine DeterminationUsing Sodium2-(parasulfophenylazo)-1,8-

dihydroxy-3,6-Naphthalenedisulfate Method
Special Apparatus
1. Suitablespectrometer.
Reagents
1. StandardP- stock solution, 1000 mg L -1: Prepareas outlined in "Rea-
gents" under"Sodium Hydroxide Fusion Method for Total Fluorine."
2. Sodium2-(parasulfophenylazo )-1,8-dihydroxy-3,6-naphthalene disul-
fonate [also called 4,5-dihydroxy-3-(parasulfophenylazo )-2,7-naph-
thalenedisulfonicacid trisodium salt, or SPADNS] solution: Dissolve
958 mg of SPADNS in deionizedwater, and dilute to 500 mL. Store
in dark-coloredcontainer.This solution is stable indefinitely if pro-
tectedfrom direct sunlight.
3. Zirconyl acid solution: Dissolve 133 mg of zirconyl chloride octahy-
drate (ZrOClz • 8HzO) in about 25 mL of deionizedwater. Add 350
mL of concentratedHel, and dilute with deionizedwater to 500 mL.
4. Acid zirconyl-SPADNSreagent:Mix equal volumesof zirconyl acid
solution and SPADNSreagent.The mixed reagentis stablefor at least
2 yr.
5. Sodium arsenite(NaAsOz) solution: Dissolve 5.0 g of NaAsOz, and
Procedure.Transfer sufficient volume of the distillate from "Steam
Distillation Method for Separationof Fluorine from PerchloricAcid" or "Steam
Distillation for Separationof Fluorine from Sulfuric Acid," or extracts from
"Sodium Hydroxide Fusion Method for Total Fluorine" or "Water-Soluble
Fluorine," to a 50-mL volumetric flask. Add 10 mL of the acid zirconyl-
SPADNS reagentmixture, dilute to volume, mix well, and read the absorbance
as soon as possibleat 570-nm wavelength.Use an appropriateblank for refer-
ence readingof the absorbance.Determinethe P- of the samplefrom the stan-
dard calibrationcurve.
If the sample contains residual Cl- as from "Water-Soluble Fluorine,"
removeit by addingone drop of NaAsOz solution/0.1mg of Cl- and mix. Sodi-
um arseniteconcentrationsof 1300 mg L- 1 producean error of 0.1 mg L- t at 1.0
mg P- L-l. The methoddescribedis adaptedfrom thosereportedby the Ameri-

can Public Health Association(1980) and Peckand Smith (1964).
Comments.Peck and Smith (1964) sinteredrock sampleswith Na2C03
flux containingZnO and MgC03. Fluorine was separatedfrom the acidified fil-
trate by steamdistillation and determinedspectrometricallywith a modified zir-
conium-SPADNS method. Theyobservedthat some F was adsorbedby the
glassware.They suggestedusing Na2C03 solutionto removethe sorbedF, which
they saidwasnot cumulativeandwastoo small to affect the results.Sulfuric acid
containedin the distillate did not interferein the colorimetry,sincetraceamounts
of H 2S04 can distill dependingon the type of distillation unit (peck & Smith,
1964). Phosphate(Phosphoricacid) doesnot distill in sufficient quantity to inter-
fere. Crosby et ale (1968) observedthat the optical density using this method
changedfrom 0.215 to 0.207after 60 min but only slightly changedfurther after
6 h (0.202). They further observ~d that this methodwas comparablewith other
colorimetricmethodsin regardto reproducibility and sensitivityand had a detec-
tion limit of about 0.02 mg L-l of F. Of the interfering ions and compounds
investigated,Crosby et al. (1968) found that Al3+ and metaphosphate produced
the most noticeableeffects, whereasSO~-, Cl-, and Ca2+ hardnessgave rise to
smallerinaccuracies.
TItrimetric Fluorine Determinationwith Thorium Nitrate in Presence

of ChromeAmrol SulfurS
Special Apparatus
1. pH meter,sensitiveto 0.1 pH unit.
2. TItrating enclosure,30 cm deepby 60 em wide, with nonglossy white
walls and white opal glassbottom; lighted by two 15-W fluorescent
bulbs in a white reflector at the top rear of the enclosure.
3. Nesslertubes,50-mL, low form, calibratedat 50 mL.
4. Microburettewith lO-mL capacity and0.05-mL graduations.
Reagents
1. ChromeAzurol S indicator: Dissolve 16 mg of ChromeAzurol S in
100 mL of deionizedwater.
2. Thorium nitrate tetrahydrate[Th(N03)4 • 4H20] stock solution, 0.1 N
(0.025 M): Dissolve 13.8 g of Th(N03)4 • 4H20 in deionizedwater,
and dilute the solution to a volume of 1 L.
3. Thorium nitrate (Th(N03)4] titrating solution A, 0.01 N (0.0025 M):
Dilute 100 mL of 0.1 N (0.025 M) Th(N03)4 stock solution with de-
ionized water to a volume of 1 L.
4. Thorium nitrate titrating solution B, 0.02N (0.0005M): Dilute 20 mL
of 0.1 N (0.025M) Th(N03)4 stocksolution with deionizedwater to a
volume of 1 L.
9 Milton (1949) and Willard and Horton (1952). as adaptedfrom Brewer(1965a).

5. Sodiumhydroxide,1 M: Dissolve4 g of NaOH in water,anddilute the

solution to a volume of 100 mL.
6. Perchloricacid, 1 M: Dilute 8.63 mL of 70% HCI04 to 100 mL with
deionizedwater.
7. pH 3.3 water: Adjust 3 L of deionizedwater to pH 3.3 with approxi-
mately 1.5 mL of 1 M HCI04.
8. StandardF- stocksolution, 1000 mg L-l: Dissolveexactly 2.211 g of
reagent-gradeNaF (dried to constantweight at 100°C) in 800 mL of
deionizedwater in a 1-L volumetric flask. Dilute the solution to vol-
ume, and transferit to a polyethylenebottle for storage.
9. StandardF- stock solution, 10 mg L -1: Dilute 10 mL of 1000-mgF-
L-1 stock solution to a volume of 1 L with deionizedwater. Storethe
solution in a polyethylenebottle.
10. StandardF- solution, 1 mg L-1: Dilute 1 mL of 1000-mgF- L-1 stock
solution to 1 L with deionizedwater. Storethe solution in a polyeth-
ylene bottle.
Procedure.Add 1 M HCI04, drop by drop, to the distillate containedin the
Erlenmeyerflasksfrom "Procedure"under"SteamDistillation Methodfor Sepa-
ration of Fluorine from PerchloricAcid" or "Procedure"under"SteamDistilla-
tion for Separationof Fluorinefrom Sulfuric Acid" until the yellow color disap-
pears.Dilute the solution to the 4oo-mL mark with deionizedwater, and mix it
thoroughly.Pour approximately150 mL into a beaker,and adjustthe pH of the
solution to 3.3 using 0.1 M HCI04 and 0.1 M NaOH and checkingthe results
with a pH meter. Pour duplicate50-mL portionsof the distillate adjustedto pH
3.3 into Nesslercolor-comparisontubescalibratedat 50 mL. Fill two similar
tubeswith pH 3.3 water.Preparea color blankby adding1 mL of ChromeAzurol
S and 0.1 mL of Th(N03)4 titrating solution B (0.0005M or 0.002N) to one of
the tubescontainingpH 3.3 water. Add indicatorsolution(1.0 mL) to oneof the
tubescontainingan unknown amountof F. Slowly add Th(N03)4 titrating solu-
tion B from a microburettewith thoroughagitation (a long stirring rod with a
small loop at the baseis very effective for stirring) until the color matchesthe
color blank. If the titration requires>2.0 mL of Th(N03)4solution B, titrate the
duplicatesamplewith Th(N03)4 solution A using the samecolor blank as with
solution B, and multiply the titration value by 5. Make up a new color blank
every 15 min. Preparetwo 20-llg F standardsby pipettingexactly 2.0 mL of 10
mg L-1 of NaF solution into Nesslertubesand making the solution up to 50 mL
with pH 3.3 water. Add 1.0 mL of indicator, and titrate each standardwith
Th(N03)4 solution B until the color matchesthe color blank. Subtract0.1 mL
(color blank) from the volumesof Th(N03)4 required,and divide by 20 to obtain
the titer. From the titration valuesfor individual soil samples,subtractthe vol-
umeof Th(N03)4 solutionB requiredto titrate a reagentblank. For each12 deter-
minations,carry at leastone blank (reagentsonly-no soil samples)throughthe
completedecompositionand separationprocesses.Multiply the correctedtitra-
tion valueby the titer to obtain the milligrams of F in the 50 mL that were titrat-
ed. Multiply this value by eight to obtainthe total milligrams of F in the original
sample.To obtainthe F concentrationin milligrams perkilogram, divide the total
milligrams of F found by the weight of the soil sampleexpressedin grams.
Titrimetric Fluorine Determinationwith Thorium Nitrate in Presence

of Alizarin Red SulfurlO
Reagents
1. Sodiumalizarin sulfonate:Dissolve0.5 g of sodiumalizarin sulfonate
in 1 L of deionizedwater.
2. Hydrochloric acid, approximately1 M solution.
3. Chloroacetatebuffer: Dissolve9.45 g of monochloroaceticacid and 2
g of NaOH in 100 mL of deionizedwater.
4. Reagents2, 3, 4, 5, 8, 9, and 10 described in "Reagents"under
"Titrimetric Fluorine Determinationwith Thorium Nitrate in Pres-
enceof ChromeAzurol Sulfur."
Procedure.Pipette 100 mL of distillate from "Procedure"under "Steam

Distillation Method for Separationof Fluorine from PerchloricAcid" or "Proce-
dure" under "SteamDistillation for Separationof Fluorine from Sulfuric Acid"
into a beaker.Add 2 mL of sodium alizarin sulfonate. Neutralize the solution
with 1 M NaOH addeddropwiseuntil a pink color appears,and then add 2.0 mL
of buffer solution. Checkthe solution with a pH meterto makesurethat the pH
is between2.9 and 3.1. Divide the buffereddistillate evenly betweentwo 50-mL
Nesslertubes,and titrate eachtube with 0.002N (0.0005M) Th(N03)4 titrating
solution until a pink color that matchesthat of a color blank is obtained.Prepare
the color blank by adding 1 mL of indicator, 1 mL of buffer, and 0.1 mL of
Th(N03)4 Solution B to 50 mL of deionizedwater in a Nesslertube. The color
blank shouldbe a light peach-blossompink.
For distillatescontaining>10 Ilg of F/50 mL of solution,usetitrating Solu-
tion A, and multiply the titration by 5 to obtain the correspondingvolume of
Solution B. A color blank is not necessaryat higher concentrations unlessvery
high accuracyis desired.
To find the F equivalenceof the Th(N03)4, add 2 mL of 10 mg L-1 of NaF
solution to eachof two Nesslertubes.Add 1 mL of indicator and 1 mL of buffer
solution to eachtube,and titrate the solutionswith Th(N03)4 SolutionB until the
color matchesthe color blank. Subtract0.1 mL of Th(N03)4 from eachtitration,
and divide the remainderby 20 to determinethe milligrams of F equivalentto 1
mL of Th(N03)4 Solution B.
Subtractthe distillation blank (indicator of F in water, acid, and reagents)
from the titration value for eachunknown sample,and calculatethe milligrams
of F in the 50 mL of distillate. Averagethe valuesfrom at leasttwo titrations,and
multiply the result by eight to calculatethe total F contentof the original sample.
Divide the total microgramsof F in the sampleby the weight of the soil sample
in gramsto calculatethe F concentrationin milligrams per kilogram.
Comments.McClure (1939) statesthat the precision of the Th titration

end-point is equivalentto approximately±0.25 Ilg of F. The precision can be
\0 Hoskins and Ferris (1936), Annstrong (1936), MacIntire et al. (1951), and McHenry and
Charles(1960),as adaptedfrom Brewer (1965a).
improved considerablyby titrating to a specific end-point, i.e., matching the

color of the titrated solution with a previously preparedcolor blank as indicated
above.For very low F concentrations,the back titration proceduredevelopedby
Dahle et aI. (1938) offers someadvantages.
Ion ChromatographicMethods
For information aboutthe use of ion chromatographyfor determinationof
P- in soil solution, seeTabatabaiand Frankenberger(1996, seeChapter8).
OTHER AVAILABLE METHODS FOR DETERMINATION

OF BROMINE, CHLORINE, AND FLUORINE
The methodsjust describedare simple andinexpensiveand do not require

large facilities. There are other methodsthat can be usedfor sensitiveand accu-
rate analysisof Br, CI, and F in soil and other agriculturalsamples.Bromine can
be determinedwith x-ray fluorescence(Getzendaner et aI., 1968; Van Cauwen-
berge & Gordts, 1977; Reuter et aI., 1976; Yamada, 1968), neutron activation
(Castro & Schmitt, 1962; Guinn & Potter, 1962; Yamada, 1968; Nadkami &
Ehmann, 1969;Kline & Brar, 1969; Morrison et aI., 1969; Lag & Steinnes,1973;
Van Wambeke, 1974; Koons & Helmke, 1978), and stripping voltammetry
(Denniset aI., 1976).Chlorine can be determinedwith neutronactivation(Yama-
da, 1968; Morrison et aI., 1969; Lag & Steinnes,1973; Castro& Schmitt, 1962)
and stripping voltammetry (Dennis et aI., 1976), and F can be determinedwith
neutronactivation(Damset aI., 1975) and fluorometry (Guyon et aI., 1968; Wil-
lard & Horton, 1952). Very comparableBr resultswere obtainedwith x-ray flu-
orescenceand neutronactivation analysisof soil samples(Yamada,1968).
The paperby MacDonald(1976) summarizedthe various most commonly
used methodsfor P- provided by 13 laboratoriesthroughout the world. It out-
lined the various decompositionproceduresand analytical determinations and
included various titrimetric, spectrometric,and potentiometrictechniques.The
Th(N03)4 titration outlined in this chapterwas included, and direct potentiome-
try with SIE appearsto be popular, as is with Be and CI-.
REFERENCES
Abdalla, N.A., and B. Lear. 1975. Determinationof inorganicbromide in soils and plant tissueswith
a bromide selective-ionelectrode.Soil Sci. Plant Anal. 6:489-494.
Adriano, D.C., and H.E. Doner. 1982. Bromine, chlorine, and fluorine. p. 449-483.In A.L. Pageet
al. (ed.) Methods of soil analysis. Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,
Madison,WI.
Adriano, D.C., P.E Pratt, and K.M. Holtclaw. 1973. Comparisonof two simple methodsof chlorine
analysisin plant materials.Agron. J. 65:130--134.
Adriano, D.C., EH. Takatori, P.E Pratt, and O.A. Lorenz. 1972. Soil nitrogen balancein selected
row-crop sites in southernCalifornia. 1. Environ. Qual. 1:279-283.
American InstrumentCo. 1969. Chloride determination-automatic titrator method(instruction man-
ual). Am. Instrum. Co., Silver Springs,MD.
Armstrong,W.D. 1936. Microdeterminationof fluorine. Ind. Eng. Chern.Anal. Ed. 8:384-387.

Associationof Official Analytical Chemists.1980.Official methodsof analysis.13th ed. Assoc.Off.
Anal. Chern.,Washington,DC.
Baker, R.L. 1972. Determinationof fluoride in vegetationusing the specific ion electrode.Anal.
Chern.44:1326-1327.
Bellack, E. 1972. Methodsand materialsfor fluoride analysis.J. Am. Water Works Assoc.64:62-66.
Bergmann,J.G., and I. Sanik,Jr. 1957.Determinationof traceamountsof chlorine in naphtha.Anal.
Chern. 29:241-243.
Brewer, R.E 1965a.Fluorine. p. 1135-1148.In C.A. Black et al. (ed.) Methodsof soil analysis.Part
2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Brewer, R.E 1965b.Fluorine. p. 180-196.In H.D. Chapman(ed.) Diagnosticcriteria for plants and
soils. Qual. Print. Co., Inc., Abilene, TX.
Brewer, R.E, and G.E Liebig. 1960. Improved multiple all-glassdistillation apparatusfor determi-
nation of fluorine in plant samples.Anal. Chern. 32:1373.
Broyer,T.C., A.B. Carlton,C.M. Johnson,and P.R. Stout. 1954.Chlorine-amicronutrientelementfor
higher plants. Plant Physiol. 29:526-532.
Butler, IN. 1969.Thermodynamicstudies.p. 143-189.In R.A. Durst (ed.) Ion-selectiveelectrodes.
Natl. Bur. Stand.,Spec.Publ. 314. U.S. Gov. Print. Office, Washington,DC.
Canelli, E. 1976. Simultaneousautomateddeterminationof chloride, nitrite, nitrate, and ammoniain
water and wastewater.WaterAir Soil Pollut. 5:339-348.
Carlson,R.M., and D.R. Keeney. 1971.Specific ion electrodes: Techniques and usesin soil, plant,
and water analysis.p. 39-65. In L.M. Walsh (ed.) Instrumentalmethodsfor analysisof soils
and plant tissue.SSSA, Madison,WI.
Castro,C.E., and R.A. Schmitt. 1962. Direct elementalanalysisof citrus cropsby instrumentalneu-
tron activation:A rapid methodfor total bromide,chloride, manganese,sodium and potassi-
um residues.J. Agric. Food Chern. 10:236-239.
Chapman,H.D., and P.E Pratt. 1961. Methodsof analysisfor soils, plantsand water. Div. Agric. Sci.,
Univ. California, Riverside.
Churchill, H. V. 1945. A multiple still for use in the Willard-Winter separationof fluorine. Ind. Eng.
Chern.Anal. Ed. 17:720-721.
Conway,E.J. 1950. Microdiffusion analysisand volumetricerror. 3rd ed. D. Van NostrandCo., New
York.
Cotlove, E., H.V. Trantham,and R.L. Bowman. 1958. An instrumentand method for automatic,
rapid, accurate,and sensitivetitration of chloride in biological samples.I. Lab. Clin. Med.
51:461-468.
Crenshaw,G.L., and EN. Ward. 1975. Determinationof fluorine in soils and rocks by known-incre-
ment addition and selective-ionelectrode.U.S. Geol. Surv. Bull. 1408:77-84.
Crosby,N.T. 1969. Equilibria of fluorosilicate solutionswith specialreferenceto the fluoridation of
public water supplies.J. Appl. Chern. 19:100-102.
Crosby,N.T., A.L. Dennis,and J.G. Stevens.1968.An evaluationof somemethodsfor the determi-
nation of fluoride in potable waters and other aqueous solutions. Analyst (London)
93:643-652.
Dahle, D., R.U. Bonnar,and H.I. Wickman. 1938.Titration of small quantitiesof fluorine with thor-
ium nitrate. I. Assoc. Off. Agric. Chern. 21:459-474.
Dahle,D., and HJ. Wickman. 1936.A quantitativestudy of fluoride distillation. J. Assoc.Off. Agric.
Chern. 19:313-320.
Dahle, D., and HJ. Wickman. 1937. Further studieson fluorine distillation. J. Assoc. Off. Agric.
Chern.20:297-303.
Dams, R., J. Billiet, and I. Hoste. 1975. Neutron activation analysisof F, Sc, Se,Ag and Hf in
aerosolsusing short-lived isotopes.Int. I. Environ. Anal. Chern.4:141-153.
Dennis,B.L., G.S. Wilson, and J.L. Moyers. 1976.The determinationof bromide,chloride and lead
in airborneparticulatematterby stripping voltammetry.Anal. Chim. Acta 86:27-34.
Durst, R.A. 1969. Analytical techniquesand applicationof ion-selectiveelectrodes.p. 375-414.In
R.A. Durst (ed.) Ion-selectiveelectrodes.Natl. Bur. Stand.Spec.Publ. 314. Washington,DC.
Endelman,FJ., D.R. Keeney,J.T. Gilmour, and P.G. Saffigna. 1974. Nitrate and chloride movement
in the Plainfield loamy sandunderintensiveirrigation. J. Environ. Qual. 3:295-298.
Evans,L., R.D. Hoyle, and J.B. Macaskill. 1970. An accurateand rapid methodof analysisfor fluo-
rine in phosphaterocks. N.Z. I. Sci. 13:143-148.
Farrel,B.L. 1974. Fluorine,a direct indicatorof fluorite mineralizationin local and regionalsoil geo-
chemicalsurveys.J. Geochem.Explor. 3:227-244.
Ficklin, W.H. 1970.A rapid methodfor the determinationof fluoride in rocks and soils, using an ion-
selectiveelectrode.U.S. Geol. Surv. Prof. Pap. 700-e.
Fleischer,M., and W.O. Robinson. 1963. Some problemsof the geochemistryof fluorine. R. Soc.
Can. Spec.Publ. 6:58-75.
Frant, M.S., and 1.w. Ross,Jr. 1966. Electrodefor sensingfluoride ion activity in solution. Science
(Washington,DC) 154:1553-1554.
Fuge,R. 1976.The automatedcolorimetricdeterminationof fluorine and chlorine in geologicalsam-
ples. Chern. Geol. 17:37-43.
Getzendaner,M.E., A.E. Doty, E.L. Mclaughlin, and D.L. Lindgren. 1968. Bromide residuesfrom
methyl bromide fumigation of food commodities.1. Agric. Food Chern. 16:265-271.
Gilliam, 1.w. 1971. Rapid measurementof chlorine in plant materials. Soil Sci. Soc. Am. Proc.
35:512-513.
Gran, G. 1952. Determination of the equivalence point in potentiometric titrations. Part 2.
Analyst(London)77:661-671.
Greenhill, N.B., and K.I. Peverill. 1977. Determinationof cation exchangecapacity of soils using
ammoniaand chloride electrodes.Soil Sci. Plant Anal. 8:579-589.
Guinn, V.P., and J.e. Potter. 1962. Determinationof total bromine residuesin agricultural crops by
instrumentalneutronactivation analysis.1. Agric. Food Chern. 10:232-236.
Guyon,J.e.,B.E. Jones,and D.A Britton. 1968.A f1uorometricmethodfor determiningtrace quan-
tities of fluoride. Mikrochim. Acta 1:1180-1184.
Hardin, LJ. 1952. Report on the determinationof fluorine-contentof soils. J. Assoc. Off. Agric.
Chern.35:621-633.
Hillebrand, W.F., and G.E.F. Lundell. 1953. Applied inorganicanalysis.2nd ed. JohnWiley & Sons,
Inc., New York.
Hipp, B.w., and G.w. Langdale.1971. Use of a solid-statechloride electrodefor chloride detemina-
tions in soil extracts.Soil Sci. Plant Anal. 2:237-240.
Hoffman, G.M., and H.P. Malkomes.1974. Bromide residuesin vegetablecropsafter soil fumigation
with methyl bromide.Agric. Environ. 1:321-328.
Hopkins, D.M. 1977.An improvedion-selectiveelectrodemethod forthe rapid determinationof flu-
orine in rocks and soils. J. Res. U.S. Geol. Surv. 5:589-593.
Hoskins,W.M., and e.A Ferris. 1936.A methodof analysisfor fluoride. Ind. Eng. Chern.Anal. Ed.
8:6-9.
Huang,W.H., and W.D. Johns.1967. Simultaneousdeterminationof fluorine and chlorine in silicate
rocks by a rapid spectrophotometricmethod.Anal. Chim. Acta 37:508-515.
Huckabay,W.B., E.T. Welch, and A.V. Metler. 1947. Constanttemperaturesteamdistillation appara-
tus for isolation of fluorine. Anal. Chern. 19:154-156.
Hurd-Karrer,AM. 1950. Comparativefluorine uptakeby plants in limed and unlimed soil. Soil Sci.
70:153-160.
Kline, J.R.,and S.S. Brar. 1969. Instrumentalanalysisof neutronirradicatedsoils. Soil Sci. Soc. Am.
Proc. 33:234-238.
Koons, R.D., and P.A Helmke. 1978. Neutron activation analysisof standardsoils. Soil Sci. Soc.
Am. J. 42:237-240.
laCroix, R.L., D.R. Keeney,and L.M. Walsh. 1970. Potentiometrictitration of chloride in plant tis-
sueextractsusing the chloride ion electrode.Soil Sci. Plant Anal. 1:1-6.
Lag, J., and E. Steinnes.1973. Distribution of chlorine,bromineand iodine in Norwegianforest soils
studiedby neutronactivation analysis.p. 383-395.In Proc. of Symp. on the Use of Isotopes
and Radiation in Researchon Soil-Plant Relationshipsincluding Application in Forestry.
Vienna.
Larsen,S., and AE. Widdowson.1971. Soil fluorine. J. Soil Sci. 22:210-211.
MacDonald,AM.G. 1976. The presentstate of methodsfor the microdeterminationof fluorine in
organiccompounds.Pure Appl. Chern.45:31-37.
MacIntire, W.H. 1945. Soil contentof fluorine and its determination.Soil Sci. 59:105-109.
MacIntire, W.H., LJ. Hardin, and L.S. Jones.1951. Report on fluorine-the direct double distillation
for the determinationof fluorine contentof soils. J. Assoc. Off. Agric. Chern. 34:597-603.
Martin, J.P.,G.K. Helmkamp,andJ.O. Ervin. 1956. Effect of bromidefrom a soil fumigant and from
CaBr on growth and chemical composition of citrus plants. Soil Sci. Soc. Am. Proc.
20:209-212.
Mavrodineanu,R., and J. Gwirtsman. 1954. Improved apparatusfor the distillation of fluorine as
hydrofluosilicic acid. Contrib. Boyce ThompsonInst. 17:489-494.
McClenahen,J.R., and E.R. Schulz. 1976. Total soil fluoride determinationby a single distillation
selectiveion electrodeprocedure.Soil Sci. 122:267-270.
FRANKENBERGER ET AL.
McClure, FJ. 1939. Microdeterminationof fluorine by thorium nitrate titration. Ind. Eng. Chern.
Anal. Ed. 11:171-173.
McHenry, C.R., and H. Charles.1960.Monitoring fluoride contentof air, waterand vegetation.Farm
Chern. 123:58-62.
Mcleod,S., andB. Zarcinas.1976.The determinationof ammoniumandchloride by an autoanalyser
for the measurementof cation exchangecapacityof soils. Soil Sci. Plant Anal. 7:743-750.
McQuaker,N.R., and M. Gurney. 1977. Determinationof total fluoride in soil and vegetationusing
an alkali fusion-selectiveion electrodetechnique.Anal. Chern.49:53-56.
Milton, R.F. 1949. Titrimetricestimationof fluorine. Analyst (London) 74:54.
Morrison, G.H., J.T. Gerard, A. Travesi, R.L. Currie, S.F. Peterson,and N.M. Potter. 1969.
Multielement neutronactivation analysisof rock using chemicalgroup separationsand high
resolutiongammaspectrometry.Anal. Chern.41:1633-1637.
Nadkarni,R.A., andW.D. Ehmann.1%9. Determinationof traceelementsin biological standardkale
by neutronactivation analysis.1 Radioanal.Chern. 3:175-185.
Nommik, H. 1952.Fluorine in Swedishagriculturalproducts,soil and drinking water.Acta Polytech.
Incl. MetaJl. Ser. 105:1-121.
Omueti,J.A.!., and R.L. Jones.1977a.Regionaldistribution of fluorine in Illinois soils. Soil Sci. Soc.
Am. J. 41:771-774.
Omueti, lA.I., and R.L. Jones.1977b.Fluorine contentof soil from Morrow plots over a period of
67 years.Soil Sci. Soc. Am. J. 41:1023-1024.
Onken,A.B., R.S. Hargrove,c.w. Wendt, and O.C. Wilke. 1975.The useof a specific ion electrode
for determinationof bromide in soils. Soil Sci. Soc. Am. Proc. 39:1223-1225.
Orion Research,Inc. 1976. Orion ionalyzerand electrodes. OrionRes. Inc., Cambridge,MA.
Ozanne,P.G. 1958. Chlorine deficiency in soils. Nature(London) 182:1172-1172.
Peck, L.c., and V.C. Smith. 1964. Spectrophotometricdeterminationof fluorine in silicate rocks.
Talanta11:1343-1347.
Pluger,W.L., and G.H. Friedrich. 1973. Determinationof total and cold-extractablefluoride in soils
andstreamsedimentswith an ion-sensitivefluoride electrode.p. 421-427.In MJ. Jones(ed.)
Geochemicalexploration.V.K. Inst. Min. MetaJl., London.
Prince, A.L., F.E. Bear, E.G. Brennan,I.A. Leone, and R.H. Daines. 1949. Fluorine: Its toxicity to
plantsand its control in soils. Soil Sci. 67:269--277.
Reeves,R.D., and R.R. Brooks. 1978.Traceelementanalysisof geologicalmaterials.p. 304-309.In
Chemicalanalysis.Vol. 51. JohnWiley & Sons,Inc., New York.
Reuter,F.W., G.E. Secor,and M. Friedman.1976. A methodfor brominedeterminationin wool fab-
ric by X-ray fluorescencespectrometry.Text. Res.J. 46:463-465.
Ross, J.W., Jr. 1969. Solid-stateand liquid membraneion-selectiveelectrodes.p. 57-88. In R.A.
Durst (ed.) Ion-selectiveelectrodes.Natl. Bur. Stand. Publ. 314. U.S. Gov. Print. Office,
Washington,DC.
Rowe, R.D. 1965. Wickbold combustion and spectrophotometricanalysis procedure for trace
amountsof organicchlorine in viscouspolybutenepolymers.Anal. Chern.37:368-370.
Rowley, R.I., J.G. Grier, and R.L. Parsons.1953. Determinationof fluoride in vegetation.Anal.
Chern. 25:1061-1065.
Saffigna, P.G., D.R. Keeney,and L.L. Hendrickson.1976. Halide analysisin soils with a chloride
titrator and a bromideelectrode.Soil Sci. Plant Anal. 7:691~99.
Selmer-Olsen,A.R., and A. 0ien. 1973. Determinationof chloride in aqueoussoil extractsand water
samplesby meansof a chloride-selectiveelectrode.Analyst (London) 98:412-415.
Shergold,H.L., and F.L. Selfe. 1974. Determinationof fluorine content of ores with fluoride-ion
selectiveelectrode.Inst. Min. MetaJl. Trans. 83:256-257.
Smart,R.St.C.,A.D. Thomas,and D.P. Drover. 1974. Selectiveion electrode measurements of chlo-
ride concentrationsin the determinationof cation exchangecapacitiesof soils. Soil Sci. Plant
Anal. 5:1-11.
Steinkoenig,L.A. 1919. The relation of fluorine in soils, plants and animals. J. Indus. Eng. Chern.
11:463-465.
Stout, P.R., and C.M. Johnson.1965. Chlorine and bromine.p. 1124-1134.In C.A. Black et al. (ed.)
Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Tabatabai,M.A., and W.T. Frankenberger,Jr. 1996. Liquid chromatography.p. 225-245. In D.L.
Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemical methods.SSSABook Ser. 5.
Thomas,1, Jr., and HJ. Gluskoter. 1974. Determinationof fluoride in coal with the fluoride ion-
selectiveelectrode.Anal. Chern.46:1321-1323.
Troll, G., A. Farzaneh,and K. Cammann.1977.Rapid determinationof fluoride in mineral and rock

samplesusingan ion-selectiveelectrode.Chern.Geol. 20:295-305.
u.s. EnvironmentalProtectionAgency Staff. 1974. Methods for chemical analysisof water and
wastes.EPA 625/6-74-003.USEPA, Office Technol.Transfer,Washington,DC.
Van Cauwenberge,P.P., and L.A. Gordts. 1977. X-ray fluorescencespectroscopicdeterminationof
Br residuesin crops after soil treatment with methyl bromide. J. Agric. Food Chern.
25:1000-1002.
Van Loon, J.C. 1968a.Determinationof chloride in chloride-containingmaterialswith a chloride
membraneelectrode.Analyst (London)93:788-791.
Van Loon, J.C. 1968b.The rapid determinationof fluoride in mineral fluorides using a specific ion
electrode.Anal. Lett. 1:393-398.
Van Wambeke,E. 1974. Bromide residuesin lettuceafter soil fumigation with methyl bromide,and
somefactorsinvolved. Agric. Environ. 1:277-282.
Vinogradov,A.P. 1959. The geochemistryof rare and dispersedchemical elementsin soils. 2 ed.
ConsultantsBur., New York.
Willard, H.H. 1912.The preparationof perchloricacid. J. Am. Chern.Soc. 34:1480-1485.
Willard, H.H., and C.A. Horton. 1952. Fluorometric determinationsof traces of fluoride. Anal.
Chern.24:862-865.
Willard, H.H., and O.B. Winter. 1933. Volumetric methodfor determinationof fluorine. Ind. Eng.
Chern.Anal. Ed. 5:7-10.
Yamada,Y. 1968.Occurrenceof brominein plantsand soils. Talanta15:1135-1141.
Zall, D.M., D. Fisher,andM.O. Garner.1956.Photometricdeterminationof chloridesin water.Anal.
Chern.28:1665-1668.
Published 1996
Chapter 32
Phosphorus
s. KUO, Washington State University, Pullman, Washington
The total P concentrationin soils is generally in the rangefrom 200 to 5000 mg

P kg-I with an averageof 600 mg P kg-I (Lindsay, 1979). Physicochemicaland
biological reactionsin soils and sedimentsact In concertto regulateP solubility
which in turn affectsboth agronomicproductionas well as eutrophicationof sur-
face water.
Phosphorusexists in soil as organic and inorganic P forms. Oxidation of
organicconstituentsand acid dissolutionof mineralsare necessaryfor total P de-
termination.This is generally accomplishedby Na2C03 (sodium carbonate)fu-
sion, aciddigestion,H20 2 (hydrogenperoxide)or NaOBr (sodium hypobromite)
oxidation.
The soil organic P fraction may be derived from plant residuesand from
soil flora and fauna tissueand residuesthat resist rapid hydrolysis. While a large
proportion of soil organic P remains uncharacterized,inositol phosphate,phos-
pholipids, nucleic acids and their derivatives, and polyphosphateshave been
identified. Chromatographicand nuclear magnetic resonancetechniqueshave
beenusedto quantify various organic P fractions. Quantificationof organic P is
necessaryto better understandthe mineralization-immobilizationturnover of P
under particular environmentsand cropping systemsin soils. Dalal (1977), An-
derson(1967, 1980) and Stevenson(1982) have provided detaileddiscussionof
organic P and its transformationsin soil.
To understandthe inorganic P statusand availability in soil, diverse frac-
tionation schemesand soil testshave beendeveloped.The P fractions have been
usedto study the transformationof applied P fertilizer in soils and interpretation
of P soil test values.The fractionation schemeproposedby Changand Jackson
(1957), which is intendedto separateCa-P, AI-P, and Fe-P fractions, has been
modified to considersoil type effects.
When the P concentrationin soil solution, or P intensity, is diminishedby
P removal, it is replenishedby labile P which in turn is replenishedmuch more
slowly by nonlabile P.
Book Seriesno. 5.
869
870 KUO
Nonlabile soil P ... labile soil P ... soil solution P [1]
The quantity of labile P, the concentrationof P in soil solution, as well as

the P buffering capacitythat governsthe distribution of P betweenthe solution
and solid phases,are the primary factors characterizingsoil P availability. The
basic function underlying P soil tests is to determinethe quantity and intensity
factorsof soil P. The soil test P levels, however,are dependenton both test meth-
ods and soil characteristics.Thus, in many respects,P soil test levels are opera-
tionally defined unlessthe relatedP buffering capacityof the soil also is includ-
ed in the interpretationof soil test results(Kuo, 1991).
A wide variety of soil test schemeshave evolvedover the years,reflecting
regional preference,considerationof soil types and efficiency of operation.The
newer methods that include chelates [e.g., ethylenediaminetetraacetic acid
(EDTA) or diethylenetriaminepeI1taacetic acid (DTPA)] are designedto test, in
addition to P, other anionsas well as cations.The recentlydevelopediron oxide-
impregnatedfilter papermethod,which removesP in the samemanneras anion
exchangeresin does,can overcomethe tediousnessof separatingresin from soils
requiredby the latter method.This methodwill deservegreaterconsiderationand
acceptanceif a uniform quality of oxide-impregnatedpaperbecomesavailable.
Phosphorusin soil extractsand plant ashcan be determinedby instrumen-
tation or by wet chemistry.Both inductively coupledplasma spectrometry (ICPS)
and ion chromatography (IC)are convenientparticularly for multielementanaly-
sis. For wet chemistry,four colorimetric methodsfor P determinationare includ-
ed in this chaptervarying in sensitivity, color stability, and toleranceto interfer-
ing cations,anions and labile organic P. Users have the choice of selectingone
that bestsuits the conditionsof their test solution and sensitivity desired.
TOTAL PHOSPHORUS
Principles
Four methodshavebeenusedto determinetotal P in soils: sodiumcarbon-

ate fusion (Jackson,1958), perchloric acid (HCI04) digestion (Jackson,1958),
H2S04 (sulfuric acid)-H20z-HF (hydrogenfluoride) digestion(Bowman, 1988),
andsodiumhypobromiteoxidationfollowed by dissolutionin dilute H2S04 (Dick
& Tabatabai,1977a).All four methodsconvertorganicP to inorganicP to facil-
itate total P determination.
The HCI04 and NaOBr digestions,however,do not readily dissolveP im-
bedded in the matrix of silicate minerals (e.g., quartz). Thus, the HCI04 or
NaOBr digestionsmay underestimatetotal P in proportionto the quantity of the
P imbedded(Syers et aI., 1967; Sommers& Nelson, 1972). Sodium carbonate
fusion extractsmore P than HCI04 or NaOBr, providedthat the melt is extracted
with dilute H2S04 (Sherrell & Saunders,1966). Differencesbetweenthe total P
determinedby fusion and that by HCI04 digestionbecomewider as the propor-
tion of fme and coarsesandsin the soil increases(Mattingly, 1970).
PHOSPHORUS 871
SpecialApparatus
1. Platinumcrucibles,30-mL capacity,with lids.
2. l00-mL Teflon beakerswith lids.
3. Stainless steelperchloricacid fume hood.
4. Hot plate.
Methods
SodiumCarbonateFusion
Reagents
1. Sodiumcarbonate,anhydrous.
2. Sulfuric acid, 4.5 M: Slowly add 250 mL of concentratedH2S04 (18 M)
to 500 mL of deionizedwater in a l-L volumetric flask. Cool the solu-
tion to room temperatureand dilute to 1 L.
3. Sulfuric acid, 1 M: Add 56 mL of concentratedH2S04 to 800 mL of
deionizedwater in a l-L volumetric flask. Cool the solution to room
temperatureand dilute to 1 L.
4. Sodiumhydroxide (NaOH),5 M: Dissolve200 g of NaOH in deionized
5. p-nitrophenol(C6HsN03), 0.25%: Dissolve 0.25 g of p-nitrophenolin
100 mL of deionizedwater.
Procedure.Mix 1.0 g (or 0.5 g of soil containinghigh Fe) of air-dried soil
«0.15 mm) and 4 g of reagentgrade anhydrousNa2C03 in a 30-mL platinum
crucible. Placean additional 1 g of Na2C03on top of the mixture. Cover the cru-
cible with a lid but leave 10% opento allow gasesto escape.Place a Meeker
burnerwith a low flame underone side oppositeto the side with the openingfor
10 min to remove moistureand fuse the massgently. Continuethe heatingwith a
full flame of the Meekerburnerfor 10 or more min to allow the massto fuse com-
pletely and form a liquid melt. Adjust the lid to provide a larger openingduring
the heating. Incline the crucible to insure the complete fusion of soil particles
sticking on the uppersidesof the crucible.
When the bubblesof gas ceaseto come off, tum off the flame, and rotate
the crucible to spreadthe contentsover the sides of the crucible to expedite
removalof the melt. Quantitativelytransferthe melt with 30 mL of 4.S M H 2S04
with care to avoid loss by effervescence,to a 2S0-mL volumetric flask. Boil the
crucible and the lid in 25 mL of 1 M H2S04 in a beaker.Removethe crucible and
lid, and transferthe solution to the 250-mL volumetric flask. Add five drops of
0.2S%p-nitrophenolindicator and adjust thesolution pH with S M NaOH until
the color of the indicator just changesfrom colorlessto yellow.
Determine the P concentrationby the ascorbic acid method outlined in
"Ascorbic Acid Method" or by any othermethodin "PhosphorusDetermination."
The quantity of the total P presentin the soil is calculatedas follows.
SO 2S0
Total P =P Concentration(llglmL) • - • . [2]
VI g soIl used
where VI =samplevolume usedfor the determinationof P concentration.

872 KUO
Digestionwith PerchloricAcid
Reagents
2. Nitric acid (HN03), concentrated,15.8M.
3. Sodium hydroxide,SM: Dissolve 200 g of NaOH in deionizedwater
and dilute to 1 L.
4. p-nitrophenol,0.25%: Dissolve 0.25 g of p-nitrophenolin 100 mL of
deionizedwater.
Procedure.Add 2.0 g of air-dried soil «0.5 mm) and 30 mL of 70%

HCI04 in a 250-mL volumetric flask or Erlenmeyerflask that is coveredwith a
pyrex funnel to ensurethe reflux of the HCI04. Digest the mixture in a preheat-
ed sandbathon a hot plate at 130°Cin a well-ventilated,stainlesssteelhood until
the dark color due to organicmatterdisappears.Continueheatingat 203°Cfor 20
min. Heavy white fumes of HCI04 appearwhen the digestionis completed,and
the silica becomeswhite. Add 1 or 2 mL of HCI04 to washdown any black par-
ticles sticking to the sidesof the flask.
If the soil containsa high organic matter content, add 20 mL of concen-
tratedHN03 and heatto oxidize the sample(or alternativelyallow the oxidation
to continue overnight at room temperature)in a well-ventilated hood prior to
digestionwith HCI04 as previously described.
When the digestion is complete,remove the flask and cool the mixture.
Dilute with deionizedwater to 250 mL and mix well. Allow the solid materialto
settle.
Transferan aliquot containing2 to 40 )lg P to a 50-mL volumetric flask.
Add five dropsof 0.25%p-nitrophenolindicator, and adjustthe solution pH with
dropwiseadditionof 5 M NaOH until the color of the indicatorjust changesfrom
colorlessto yellow. Determinethe P concentrationby the ascorbicacid method
outlined in "Ascorbic Acid Method."
Use Eq. [2] to calculatethe total P concentrationin the soil.
Digestionwith Sulfuric Acid-HydrogenPeroxide-HydrofluoricAcid
Reagents
3. Hydrogenfluoride, concentrated,24 M.
and dilute to 1 L.
deionizedwater.
Procedure.Add 0.5 g of air-dried soil «0.15 mm), or 0.25 g for soils that
are sandyor have a high organicC content,to a 100-mL fluoropolymer(Teflon)
beaker.Add 5 mL of concentratedH2S04 and swirl to suspendthe soil particles
PHOSPHORUS 873
adheringto the bottom of the beaker.Placethe beakerin a well-ventilatedhood

and slowly add 3 mL of 30% H20 2, 0.5 mL at a time. Mix well to facilitate the
oxidation following each addition of H20 2• Care should be taken to avoid the
overflow of foam, particularly for soil with high organicmattercontent.After the
reaction subsides,add 1 mL of concentratedHF, 0.5 mL at a time, using a
polypropylenepipette,mix well, and placethe beakeron a preheatedhot plate at
150°C for 10 to 12 min to eliminate excessH20 2• Removeand cool the beaker.
Quantitatively transferthe contentsto a 50-mL volumetric flask with deionized
water and dilute to volume.
Transferan aliquot containing2 to 40 flg P to a 50-mL volumetric flask.
Add five drops of p-nitrophenolindicator, and adjust the solution pH with 5 M
NaOH until the indicatorcolor just changesfrom colorlessto yellow. Add 15 mL
of 0.8 M H3B03 (boric acid) to eliminate the interferenceof r if necessary.
Determinethe P concentrationby the ascorbicacid methodoutlined in "Ascorbic
Acid Method."
Calculatethe total P concentrationas follows.
50 50
Total P = P Concentration(flg/mL)· - . [3]
VI g soil used
where VI = samplevolume usedfor the determinationof P concentration.
SodiumHypobromiteOxidation
Reagents
and dilute to 1 L.
2. Sodiumhypobromite:Add 3 mL of Br2 slowly (about0.5 mL per min)
to 100 mL of 2 M NaOH with constantstirring. Preparethis reagent
immediatelybefore use.
3. Formic acid (HCOOH), 90%.
4. Sulfuric acid, 0.5 M: Dilute 27.8 mL of concentratedH2S04 to 1 L with
deionizedwater.
Procedure.Add 0.2 or less of air-dried soil «0.15 mm) to a 50-mL boil-
ing flask. Add 3 mL of NaOBr solution and swirl to mix. Allow the suspension
to standfor 5 min and mix again. Placethe flask in a preheatedsandbath at 260
to 280°C on a hot plate situatedin a well-ventilatedhood.
Heat the flask until its contentsare evaporatedto dryness,which takes 10
to 15 min, and continueto heat for an additional 30 min. Removeand cool the
flask. Add 4 mL of deionizedwater and 1 mL of HCOOH. Swirl to mix and add
25 mL of 0.5 M H2S04. Mix well and quantitativelytransferthe mixture to a 50-
mL volumetric flask with about 15 mL of deionizedwater and dilute to volume.
Filter the contentsor allow the solid material to settleto the bottom of the volu-
metric flask.
Transferan aliquot containing2 to 40 flg to a 50-mL volumetric flask. Add
five drops of p-nitrophenolindicator and adjust the solution pH with 5 M NaOH
874 KUO
until the indicator color just changesfrom colorlessto yellow. Determinethe P

concentrationby the ascorbicacid methodoutlinedin "AscorbicAcid Method."
Calculatethe total P concentrationin the soil usingEq. [3].
Comments
The amountsof P determinedby the NaOBr andHCl04 methodsare about

the same(Dick & Tabatabai,1977a)but less than that by the Na2C03 fusion
(Sherrell & Saunders,1966; Syderset aI., 1968) or H2S04-H20 r HF method
(Bowman,1988).This is becausethe HCl04 or 0.5 M H2S04 addedafter NaOBr
oxidation does not dissolve silicate mineralscontaining imbeddedP minerals
(e.g.,apatite).
Sommersand Nelson(1972) modified the HCl04 digestionprocedureso
that digestioncould be carriedout in an aluminum digestionblock to facilitate
routine analysisof large numbersof samples.The NaOBr methodis useful for
routine analysisas well.
Heatedmixture of HCl04 andorganicmattermay explodeviolently. Avoid
adding HCl04 to heatedsampleswith high organic matter contents.For such
samples,initiate HN03 pretreatmentfirst. The digestionwith HCl04 shouldbe
done using a specially constructedstainlesssteel hood and the hood shouldbe
washedafter eachuseto removeperchlorates.
TOTAL ORGANIC PHOSPHORUS
Principles
The total organic P (Po) is generallydeterminedby the ignition method

(Saunders& Williams, 1955; Walker & Adams, 1958) or by extraction using
concentratedHCl (hydrogenchloride)(Mehtaet al., 1954),concentratedH2S04
(Bowman,1988)or acetylacetone (CH3COCH2COCH3; 2,4-pentanedione) (Hal-
steadet aI., 1966)as a primary extractant.In the ignition method,Po is convert-
ed to inorganicP (Pi) by high temperatureoxidation. The Po is then determined
by the differencebetweenthe amountsof H2S04-extractablePi for the ignited
and unignited soils. However, high temperaturecan alter the solubility of Pi in
soils, affecting the accuracyof Po measurements. Strongmineral acidsdissolve
iron and aluminumoxides,and removeAI, Fe and other polyvalentcationsthat
precipitatewith Po. This, coupledwith heatfrom the addition of water to con-
centratedH2S04, facilitates Po extraction.However, the acids can hydrolyze
somePo [e.g., glucose-I-phosphate, ribonucleicacid (RNA), and deoxyribonu-
cleic acid (DNA)] and lower the estimateof Po.
Acetylacetone(PH 8.0) is used as a complexing agent for polyvalent
cationsto facilitate extractionof organicmatteror Po. Although this extractionis
lesseffectivethanthe ignition or acid extractionmethod,it is lessdestructiveof
the organicstructureandovercomesthe problemof the hydrolysisof Po inherent
in the acid extractions.Pretreatmentof soil with dilute mineral acid, however,is
neededto removeCa that precipitateswith Po underalkalineconditions.
PHOSPHORUS 875
The oxidation of Po in alkaline extractsby HCI04 digestion requiresthe

additionof magnesiumchloride(MgCI2) to preventP lossthroughthe conversion
of P to pyrophosphate(Na2H2P207)and then to metaphosphate(Na30309) at
temperaturesgreaterthan 240°C (Brookes& Powlson,1981).
SpecialApparatus
1. Muffle furnace.
2. Centrifuge.
3. Aluminum blockfor digestion.
4. Hot plate.
6. pH meterequippedwith a combinationelectrode.
7. Stainlesssteelperchloricacid fume hood.
9. Ultrasonicdisperser.
10. Hot water bath.
Methods
Ignition Method
Reagents
1. Sulfuric acid, 0.5 M: Dilute 27.8 mL of concentratedH2S04 (18 M) to
1 L with deionizedwater.
2. Sodiumhydroxide,5 M: Dissolve200 g NaOH in deionizedwater and
dilute to 1 L.
deionizedwater.
Procedure.Add 2.0 g of soil «2 mm) in a porcelaincrucible and placethe
crucible in a cool muffle furnace. Increasethe temperatureof muffle furnace to
550°C and maintainit for 1 h. Allow the crucible to cool and transferthe ignited
soil to a lOO-mL centrifugetube. Place2.0 g of unignitedsoil into another100-
mL centrifuge tube.
Add 50 mL of 0.5 M H2S04 to eachcentrifuge tubeand shakethe tubesfor
16 h. Centrifuge or filter through a 0.45-~m membranefilter to obtain a clear
solution. Transferan aliquot containing 2to 40 ~g of P into a 50-mL volumetric
flask, add five dropsof 0.25%p-nitrophenolindicatorand adjustthe solution pH
with 5 M NaOH until the indicator color just changesfrom colorlessto yellow.
Determinethe P concentrationusingthe ascorbicacid methodoutlinedin "Ascor-
bic Acid Method."
The total P extracted(Pex) for the ignited or unignitedsoil is calculatedas
follows.
50 50
Total Pex =P concentration(~glmL)· -VI . g soil used
[4]
876 KUO
where VI =samplevolume usedfor the determinationof P concentration.

Po =total Pignited- total Punignited. [5]
Extractionwith Dilute SodiumHydroxide, ConcentratedHydrochloric

Acid, and Dilute SodiumHydroxide
Reagents
and dilute to 1 L.
2. Sodiumhydroxide,0.5 M: Dilute 100 mL of 5 M NaOH solution to 1 L
3. Sodium hydroxide,0.3 M: Dilute 60 mL of 5 M NaOH solution to 1 L
5. Hydrochloric acid, 2 M: Dilute 167 mL of concentratedHCI (12 M) to
6. Sulfuric acid, concentrated18 M.
8. Magnesiumchloride (MgCI 2) saturated:Suspend600 g of MgCl2 in 1
L of deionizedwater.
9. p-nitrophenol,0.25%: Dissolve 0.25 g of p-nitrophenol in 100 mL of
deionizedwater.
Procedure.Weigh 1 g of soil «0.15 mm) in a 100-mL polypropylenecen-
trifuge tube. Add 70 mL of 0.3 M NaOH. Shakethe suspensionfor 16 h and cen-
trifuge. Decantthe supernatantliquid into a 100-mL volumetric flask. Add five
dropsof p-nitrophenolindicatorand adjustthe pH with dropwiseaddition of con-
centratedH2S04 with constantstirring until the color just changesfrom yellow to
colorless.Bring up to 100 mL with deionizedwater.
Add 10 mL of concentratedHCI to the soil residueand heat at 70°C in a
water bath for 10 min. Removethe tube and addan additional 10 mL of concen-
tratedHCl. Swirl to mix and allow the suspensionto cool at room temperaturefor
1 h. Add 50 mL of deionizedwater, and mix well. Centrifuge, and decantthe
supernatantliquid into a 200-mL volumetric flask.
Add 30 mL of 0.5 M NaOH to the centrifugetube, mix well, and allow to
standat room temperaturefor 1 h. Centrifuge,and decantthe supernatantinto the
200-mL volumetricflask containingthe acid extract.Add 60 mL of 0.5 M NaOH
to the tube and mix thoroughly. Cover the tube loosely with a beaker andheatin
an oven at 90°C for 8 h. Cool the tube, centrifuge,and decantthe supernatantliq-
uid into the 200-mL volumetricflask. Bring up to 200-mL volumewith deionized
water, and mix well.
Transferaliquotscontaining2 to 40 )lg P of the baseextractin the 100-mL
volumetric flask and the acid and baseextractsin the 200-mL volumetric flask to
Folin-Wu (or comparable)digestiontubesfor total P determination.Add 0.5 mL
of saturatedMgCl 2 solution, one drop of concentratedH2S04 and 1 mL of 70%
HCI04 to each tube and digest in an AI digestion block at 205°C for 30 min.
Removethe tube, and let cool. Quantitatively transfer the digest with about 30
PHOSPHORUS 877
mL of deionizedwater to a 50-mL volumetric flask for P determination.Adjust

the solution pH in a similar manneras describedbelow, prior to P determination
by the ascorbicacid methodoutlined in "Ascorbic Acid Method."
Allow the suspendedmaterials in the 100-mL and 200-mL volumetric
flasks to flocculate. Pipettean aliquot containing2 to 40 J..lg P to a 50-mL volu-
metric flask for inorganicP determination.For the acid and baseextractadd five
dropsof p-nitrophenolindicatorand adjustthe solution pH with 5 M NaOH until
the indicator color just changesfrom colorlessto yellow. Determinethe P con-
centrationby the ascorbicacid methodoutlinedin "Ascorbic Acid Method" or by
the modified ascorbicacid methodin "Modified Ascorbic Acid Method" for the
determinationof inorganic P in the first base extract if sufficient acid-labile
organicP is present.A blank containingall reagentsusedin the extractionshould
be included.
InorganicP in the first baseextract(Pf) or the acid plus baseextract(pY)
is calculatedas follows:
50 V2
Pf or pY = P concentration(J..lglmL)· - •. [6]
Vl g soli used
where Vl =volume of extractsusedfor the determinationof inorganicP concen-

tration; V2 =volume of extracts.Total P in the first baseextract(TP") or the acid
plus baseextract(Tpb) is calculatedusing the following equation.
50 V2
Tpa or Tpb =P concentration(J..lglmL)·-·. [7]
v3 g soli used
where vl = volume of baseor acid plus baseextractusedin the digestion.

The total organic P concentration(Po) is calculatedusing the following
equation
[8]
Extractionwith ConcentratedSulfuric Acid and Dilute

Sodium Hydroxide
Reagents
2. Sulfuric acid, 5.5 M: Slowly add 305 mL of concentratedH2S04 to 500
mL of deionizedwater in a l-L volumetric flask. Cool the solution to
room temperatureand dilute to 1 L.
3. Hydrochloric acid,SM: Dilute 417 mL of concentratedHCI to 1 L.
4. Sodiumhydroxide, 10 M: Dissolve 400 g of NaOH in deionizedwater
and dilute to 1 L.
5. Sodium hydroxide, 2 M: Dilute 200 mL of 10 M NaOH to 1 L with
deionizedwater.
878 KUO
6. Sodiumhydroxide,0.5 M: Dilute SO mL of 10 M NaOH to 1 L.

7. Potassiumpersulfate,K 2S20 g•
8. p-nitrophenol,0.2S%:Dissolve 0.25 g of p-nitrophenolin 100 mL of
deionizedwater.
Procedure.Weigh 2.0 g of air-dried soil «0.18 rom) into a SO-mL volu-

metric flask and add3 mL (or 4 mL if the soil is calcareous)of concentrated
H2S04• After mixing, add 4 mL of deionizedwater in I-mL incrementswhile
mixing vigorously for S to 10 s after eachaddition. Rinse the interior side of the
flask with S to 10 mL of deionizedwater, andmix the suspensionvigorously. Add
about20 mL of deionizedwater and, after coolingto room temperature,filter the
suspension through Whatmanno. 1 filter paper.Savethe filtrate in a SO-mL vol-
umetricflask for P determination.Quantitativelytransferthe remainingsoil in the
flask to the filter paperwith aboutS mL of deionizedwater or more if necessary.
Transfer the filtrate to the SO-mL volumetric flask and dilute to volume with
deionizedwater. Savethe filter papercontainingthe soil residuefor baseextrac-
tion.
Placethe soil residueand filter paperin a 2S0-mL Erlenmeyerflask. Add
98 mL of 0.5 M NaOH, shakefor 2 h, and filter through Whatmanno. 1 filter
paper.
For total P (TP) determination,pipette an aliquot containing2 to 40 Ilg P
from the acid or baseextractinto a SO-mL volumetric flask. Add 1 g of K 2S20 g
with a calibratedscoopand 2 mL of S.5 M H2S04• Digest the sampleon a hot
plateat about IS0aC for 20 to 30 min or until the vigorousboiling subsides.Cool,
addfive dropsof p-nitrophenoland adjustthe pH with 10M NaOH until the color
just changesto yellow. Determine the P concentrationusing the ascorbicacid
methoddescribedin "AscorbicAcid Method." Calculatethe TP in the acid (TP")
and base(Tpb) extractsusing Eq. [7].
For inorganicP (Pi) determination,transferan aliquot from the acid or base
extractcontaining2 to 40 Ilg P to a 50-mL volumetricflask. Add five dropsof p-
nitrophenoland adjustthe pH of the acid extractwith 2 M NaOH and of the base
extractwith 5 M HCl until the indicatorcolor just changes.Determinethe P con-
centrationusingthe ascorbicacid methoddescribedin "AscorbicAcid Methods."
Calculatethe quantitiesof Pi in the acid (Pf) or base(P~) extractsusing Eq. [6].
The total organicP (Po) fraction in the initial soil sampleis calculatedusing Eq.
[8].
AcetylacetoneExtraction
Reagents
1. Hydrochloric acid, S M: Dilute 417 mL of concentratedHCl (12 M) to
2. Hydrochloric acid, 0.1 M: Dilute 20 mL of 5 M HCl to 1 L with deion-
ized water.
3. Acetylacetone(CH3COCH2COCH3), 0.2 M: Add 20 mL of acetylace-
tone (>99%) to 950 mL of deionized water,adjust pH to 8.0 with 3 M
NaOH and dilute to 1 L.
PHOSPHORUS 879
4. Sodium hydroxide, 3 M: Dissolve 120 g of NaOH in deionizedwater

and dilute to 1 L.
5. Sodium hydroxide,0.05 M: Dilute 16.7 mL of 3 M NaOH to 1 L with
deionizedwater.
6. Ethyl ether(CH3CH20CH2CH3).
8. Magnesiumchloride, saturated:Suspend600 g of MgCl 2 in 1 L of
deionizedwater.
Procedure. Add 2.0 g of soil «0.15 mm) to 40 mL of 0.1 M HCI in a 100-
mL centrifuge tube. Shake the suspensionmechanicallyfor 30 min, and cen-
trifuge. Discardthe supernatantliquid and repeatthe washingwith another40 mL
of 0.1 M HCI. Quantitativelytransferthe soil residueto a 50-mL beakerwith 30
mL of 0.2 M acetylacetone(pH 8.0) and give a 2-h ultrasonictreatment.Transfer
the suspensionto a centrifugebottle with 10 mL of acetylacetoneand adjust the
pH to 8.0 with 3 M NaOH. Shakethe suspensionfor 16 h and centrifuge.Decant
the supernatantliquid in a 500-mL Erlenmeyerflask. Repeatthe extractionfour
more times with 40 mL of 0.2 M acetylacetone(pH 8.0) and a 24-h shakingperi-
od, with an ultrasonic treatment for the final extraction. Add diethyl ether
(C2HsOC2HS)and shake well to extract the acetylacetoneand its complexes.
Removethe ether using a liquid-liquid separatoryfunnel. Centrifuge the ether-
extractedsolution at high speedto clarify the solution.
Transferan aliquot containing2 to 40 f.lg P to a Folin-Wu digestion tube
(Kimble Glass,Vineland, NJ). Add 0.5 mL of saturatedMgCl 2 and 1 mL of 70%
HCI04 and digest in an aluminum digestionblock. Slowly raise the temperature
to 205°C. When the white fumes appear,cover the tubeswith funnels or beakers,
and digest at 205°C for an additional 30 min. Remove the tube, and let cool.
Quantitativelytransferthe digestto a 50-mL volumetricflask for P determination
by the ascorbicacid methodas outlined in "Ascorbic Acid Method."
The digestionalso can be done in a beakeron a hot plate. Cover the beaker
with a watch glasswhen white fumes appear.
Calculatethe Po as follows.
50 50
Po = P Concentration(f.lglmL) • - • - - - - [9]
Vt g soil used
where Vt = samplevolume usedfor the P determination.
Comments
Condronet al. (1990) showedthat the ignition methodand the acid-alkali

extractionmethodspresentedaboveare about equally effective in extractingPo.
The concentrated H 2S04 extractionmethodextractsslightly higher Po (Bowman,
1989; Condronet aI., 1990) but less total soil P (Po + Pi). Efficiency in operation
and requirementsfor further fractionation of Po are important considerationsin
deciding which methodto choose.The ignition method,while simple and suit-
able for routine soil testingof a large numberof soil samples,destroysthe struc-
880 KUO
ture of Po and is unsuitableif further fraction of Po is needed.The method of

Mehta et al. (1954), for instance,was adoptedby Caldwell and Black (1958a),
and Thomasand Lynch (1960) for determiningthe inositol hexaphosphate using
column chromatography.Fractionationof soil Po is presentedin more detail in
"Fractionationof OrganicPhosphorus."
The ignition methodis subjectto severalpotentialerrors.Ignition at 550°C
canenhancethe P solubility particularlyfor the strongly weatheredsoils that con-
tain primarily iron- and aluminum-phosphates, leading in some casesto a con-
siderableoverestimateof Po (Williams & Walker, 1967; Williams et ai., 1970).
The Pi associatedwith organic matter through bridge bonding of Fe or Ai can
contribute to the error in Po estimatebecauseit is taken as Po after the organic
matteris oxidized throughignition (Hong & Yamane,1980). In addition, incom-
plete oxidation of Po (e.g., RNA and phytin) (Dormaar & Webster, 1964) can
result in small errorsin estimatesof Po.
The strengthof H2S04 and the length of extractioJ)time usedfor the igni-
tion method vary with investigators.The H2S04 concentrationoriginally pro-
posed by Saundersand Williams (1955) is 0.1 M. The concentrationwas
increasedto 0.5 M by Walker and Adams(1958) and further to 1 M by Anderson
(1960), HanceandAnderson(1962), Bowman(1989),and Condronet ai. (1990).
The extractionperiod usedis either2 or 16 h by the aboveauthors.Saundersand
Williams (1955) extractedsimilar amountsof Po with 0.1 M H2S04 using 2- or
16-h extractionperiod. Their choice of a 16 h or overnight extractionperiod is
simply a matterof convenience.
Concentratedmineral acid (e.g., HCI) hydrolyzes native Po (Halstead&
Anderson,1970). Anderson(1960) found the similar resultswith inositol hexa-
phosphate (C6HlS024P6), glycerophosphate(C3H706PNa2), sugar phosphate
(C6H l1 0 9PNa), and nucleic acid, and proposeda pretreatmentwith dilute alkali
(0.3 M NaOH) to remove the majority of Po prior to acid extraction.The pre-
treatmentremovedthe majority of Po prior to acid extraction.The pretreatment
removedclose to 60% of Po (Condron et aI., 1990), including all of the glyc-
erophosphateand the majority of phytate (Martin, 1964). Ribonucleic acid
(RNA), for instance,is not solublein 0.3 M NaOH (Martin, 1964)but hydrolyzed
extensivelyin strongacid (Anderson,1960). The hydrolysiscontributeserrorsin
Po estimatesproportionateto the quantity of RNA presentin soils.
The concernover the hydrolysis of Po in strong acid solution led to some
modificationof the Mehtaet ai. (1954)procedure.Kaila and Virtanen(1955)pro-
posedto use 2 M H2S04 insteadof concentratedHCI, whereasAnderson(1960)
introduceda pretreatmentwith dilute alkaline solution as a remedial measure.
However,the modification by Anderson(1960),while improving the accuracyof
Po extraction,increasesthe sequentialextractionsfrom threeto four steps.In con-
trast, Bowman(1989)observedonly a slight hydrolysisof inositol hexaphosphate
among several phosphomonoesters and phosphodiestersadded to concentrated
H2S04 and developeda two-sequentialextractionprocedurethat is equally effec-
tive as the Anderson-Mehtamethod.The Bowman methodmerits consideration
if efficiency is vital. However,Martin (1964) showedthat threeserial extractions
with 5 M H2S04 heatedat 100°C for the first extractioncan extract as much as
85% of Po, a level generally achievedby the Bowman method. Several other
PHOSPHORUS 881
methodsalso havebeendevelopedfor determiningPo over the years.An impor-

tant considerationin their developmentis to facilitate the fractionationof Po. Pre-
treatmentwith 0.1 M HCl to removeCa and Mg is usually required.Thesemeth-
ods include three serial extractionswith 0.3 M KOH at room temperature(Mar-
tin, 1964)-thecombinationof exchangeresin with alkaline solution (De Serra
& Schnitzer,1972)or with acetylacetone (pH 8.3) (Hong & Yamane,1980)-and
0.3 M NaOH extraction sonified to dispersesoil particles (Soltanpour et aI.,
1987). Dependingon the method,between78 and 99% of Po can be extracted.
IncreasedPo accumulationin the fulvic acid fraction occursin the extrac-
tion of Po by the Anderson-Mehtamethod but not by the resin-acetylacetone
method(Hong & Yamane,1980).
Organic P associatedwith fulvic acid is consideredto be relatively more
labile than humic acid associatedP in soils (Bowman & Cole, 1978). When ful-
vic acid is used to index P transformationand bioavailability in soils, mild and
less destructiveextractantssuch as acetylacetonemay be preferredover strong
mineral acids. For Pi determination,the "Modified Ascorbic Acid Method"
developedby Dick and Tabatabai(1977b) should be employed to avoid the
potentialinterferencefrom acid-labile Po, if presentin a sufficient amount.
FRACTIONATION OF SOIL PHOSPHORUS
Fractionationof InorganicPhosphorus
Principles
Inorganic P can reactwith Ca, Fe, or AI to yield discretephosphatessuch
as hydroxyapatite[Cas(P04hOH],octacalciumphosphate[Ca4H(P04h2.5 H20]
and variscite (AlP04 • 8 H20) (Lindsay, 1979; Lindsay & Vlek, 1977). Their
identification in soil, however,is done mostly by solubility equilibria. Scanning
transmissionelectron microscopy in conjunction with energy dispersive x-ray
analysis of density separatesyields P-rich particles containing various cations
including AI, Si, Ca, and Fe (Pierzynskiet aI., 1990).
Fractionationschemesutilize the complexingability of r from ammoni-
um fluoride (NH4F) to separateAI-P from Fe-P, followed by removal of Fe-P
with NaOH and of the reductant-solubleP with sodium citrate (Na3C6Hs07•
2H20)-sodium dithionite (Na2S204)-sodiumbicarbonate (NaHC03) (CDB)
extractions.The calcium-phosphate, which is insoluble in CDB (Williams et aI.,
1980), is extractedwith H2S04 or HCI. However, NH4F reactswith CaC03 to
form calcium fluoride (CaF2) in calcareoussoils (Smillie & Syers,1972), which
in tum forms secondaryprecipitateswith solubilizedP and reducesthe effective-
nessof NH4F to extract P. As a result, the NH4F extractionis not recommended
for calcareoussoils (Williams et aI., 1971a).
SpecialApparatus
1. pH meterequippedwith a single combinationelectrode.
882 KUO
3. Hot waterbath.
4. Centrifuge.
Methods
Fractionation for Noncalcareous Soils
Reagents
1. Ammonium chloride(NH4Cl), 1 M: Dissolve 53.3g of NH4CI in deion-
ized water and dilute to 1 L.
2. Ammonium fluoride (NH4F), 0.5 M (pH 8.2): Dissolve 18.5 g of NH4F
in deionizedwater and dilute to 1 L. Adjust pH to 8.2 with 4 M ammo-
nium hydroxide (NH40H).
3. Sodiumhydroxide,2 M: Dissolve80 g of NaOH in deionizedwaterand
dilute to 1 L.
4. Sodium hydroxide,0.1 M: Dissolve 4.0 g of NaOH in deionizedwater
and dilute to 1 L.
5. Sodiumhydroxide,0.1 M, + sodiumchloride (NaCI), 1 M: Dissolve4.0
g of NaOH and 58.5 g of NaCI in deionizedwater and dilute to 1 L.
6. Sulfuric acid, 0.25 M: Dilute 14 mL of concentratedH 2S04 (18 M) to 1
L with deionizedwater.
8. Sodiumcitrate, 0.3 M: Dissolve 88.2g of Na3CJls07• 2 H20 in about
900 mL of deionizedwater and dilute to 1 L.
9. Sodiumchloride, 1 M: Dissolve58.5 g of NaCI in deionizedwater and
dilute to 1 L.
10. Sodium chloride, saturated:Add 400 g of NaCI to 1 L of deionized
water.
11. Sodiumbicarbonate,1 M: Dissolve 84g of NaHC03 in deionizedwater
and dilute to 1 L.
12. Sodiumdithionite reagentgrade.
13. Boric acid, 0.8 M: Dissolve50 g of H3B03in deionizedwateranddilute
to 1 L.
deionizedwater.
Procedure. Add 1.0 g «2 mm) and50 mL of 1 M NH4Cl to a 100-mL cen-
trifuge tube and shakefor 30 min to extractthe solubleand loosely boundP. Cen-
trifuge and decantthe supernatantinto a 50-mL volumetricflask and bring to vol-
ume with deionizedwater (ExtractA). Add 50 mL of 0.5 M NH4F (pH 8.2) to the
residueand shakethe suspensionfor 1 h to extractaluminumphosphate(AlP04).
Centrifuge and decantthe supernatantinto a 100-mL volumetric flask (Extract
B). Wash the soil sampletwice with 25-mL portionsof saturatedNaCI and cen-
trifuge. Combinethe washingswith Extract B and bring to volume. Add 50 mL
of 0.1 M NaOH to the soil residuesand shakefor 17 h to extract iron phosphate
(FeP04). Centrifugeand decantthe supernatantsolution into a 100-mL volumet-
ric flask (Extract C). Wash the soil twice with 25-mL portionsof saturatedNaCl
PHOSPHORUS 883
and centrifuge.Combinethe washingswith extractC and bring to volume. Add

40 mL of 0.3 M Na3C6H507and 5 mL of 1 M NaHC03 to the residueand heat
the suspensionin a water bath at 85°C. Add 1.0 g of Na2S204(sodiumdithion-
ate) and stir rapidly to extract reductant-solubleP. Continueto heat for 15 min
and centrifuge.Decantthe supernatantsolution into a 100-mL volumetric flask
(Extract D). Washthe soil twice with 25-mL portionsof saturatedNaCl and cen-
trifuge. Combinethe washingswith Extract D, and dilute D to volume. Expose
Extract D to air to oxidize Na2S204'
Add 50 mL of 0.25 M H2S04 to the soil residueand shakefor 1 h. Cen-
trifuge the suspensionfor 10 min and decantthe supernatantinto a 100-mL vol-
umetric flask (Extract E). Wash the soil twice with 25-mL portionsof saturated
NaCl, andcentrifuge.Combinethe washingswith the ExtractE and dilute to vol-
ume.
Transferan aliquot containing2 to 40 Ilg P from eachof ExtractsA, B, C,
D, and E to a 50-mL volumetric flask. Add somedeionizedwater and five drops
of p-nitrophenolindicatorto the volumetricflask containingExtractsC and E and
adjustthe pH with 2 M HCl or 2 M NaOH until the indicator color just changes.
Add 15 mL 0.8M H3B03to the volumetricflask containingExtractB. Determine
P concentrationusing the ascorbicacid method as outlined in "Ascorbic Acid
Method." PrepareP standardsthat containthe samevolumeof extractingsolution
as in the extracts.
The calculationof variousP fractions can be madeas follows.
50 V2
Pextract =P concentration(llglmL) 0
-0
[10]
VI g soil used
where VI = samplevolume from ExtractA, B, C, D, or E usedfor P determina-

tion.
Vz = the volume of Extract A, B, C, D, or E.
Fractionation of Calcareous Soils

Reagents. Preparethe reagentsas describedin the previous"Reagents"sec-
tion.
Procedure. Add 1.0 g «2 mm) of soil and 50 mL of 0.1 M NaOH + 1 M
NaCl, shakefor 17 h and centrifuge.Decantthe supernatantinto a 100-mL volu-
metric flask (Extract A). Wash the soil twice with 25-mL portionsof 1 M NaCl.
Combinethe washingswith Extract A and bring to volume.
Add 40 mL of 0.3 M Na3C6H507and 5 mL of 1 M NaHC03 to the soil
residueand heatthe suspensionin a waterbath at 85°C. Add 1 g of NaZSZ04and
stir rapidly. Continueto heatfor 15 min. Centrifugeand decantthe supernatant
solution into a 100-mLvolumetricflask (ExtractB). Washtwice with 25-mL por-
tions of saturatedNaCI andcentrifuge.Combinethe washingswith ExtractBand
dilute to volume. Exposethe solution to air to oxidize Na2S204'Add 50 mL of
0.5 M HCI to the soil andshakefor 1 h. Centrifugethe suspensionanddecantthe
supernatantsolutioninto a lOO-mL volumetricflask (ExtractC). Washtwice with
884 KUO
25-mL portions of saturatedNaCl, combine the washingswith Extract C, and

dilute to volume.
Transferan aliquot containing2 to 40 Ilg P from eachextract to a 50-mL
volumetricflask. Add somedeionizedwater and five dropsof p-nitrophenolindi-
cator to the volumetric flasks containingExtractsA and C, followed by adjust-
ment of pH with 2 M HCI or 2 M NaOH. The indicator color changesfrom yel-
low to colorlessfor Extract A and from colorlessto yellow for Extract C. Deter-
mine the P concentrationusing the ascorbicacid methodas outlined in "Ascor-
bic Acid Method." PrepareP standardscontainingthe samevolume of extracting
solution as in the extract.
The calculationof various P fractions can be madeusing Eq. [10].
Comments
Therehavebeenseveralmodificationsof the fractionationschemepresent-
ed by ChangandJackson(1957). Petersonand Corey(1966) alteredthe pH of the
NH4F extractingsolutionaswell as the sequenceof the extraction.Increasingthe
pH from neutral,as originally proposedby Changand Jackson(1957) for NH4F,
to 8.2 increasesP extractabilityby NH4F (Fife, 1962). Williams et al. (1967) pro-
posed a second NaOH extraction for calcareoussoil. Williams et al. (1971a)
includedsodiumcitrate-sodiumbicarbonateto minimize the resorptionof the dis-
solvedP by CaC03 in 0.1 M NaOH solution and a secondHCI extractionfor 4 h
to increasethe extraction of occluded apatite in the matrix of minerals (e.g.,
quartz)(Syerset aI., 1967).
The inclusion of NaHC03 is consideredto be necessaryto buffer against
decreasesin pH during the extractionof reduntant-solubleP to preventthe dis-
solution of apatite(Williams et aI., 1967). Thus, the recommendedfractionation
schemeby Petersonand Corey (1966) was slightly modified to include NaHC03
in the extractionof the reductant-solubleP fraction.
Loosely boundP generallyrepresentsa very small fraction of the total P in
soils or sediments.While only trace amountsare found in noncalcareoussoils,
Ca-boundP can constitutea large proportionof the P in calcareoussoils (Sharp-
ley & Smith, 1985).The majority of Pin noncalcareous soils or sedimentsis pre-
sent as aluminum phosphateand iron phosphatethat are extractablewith NH4F
and NaOH (Williams et aI., 1971b).
Fractionationof OrganicPhosphorus
Principles
The phosphatemonoesters,which include inositol phosphates,are more
resistant to degradationby soil microorganismsthan the phosphatediesters,
which include phospholipidsand nucleic acids (Condron et aI., 1990). Of the
50% or lessof soil Po that hasbeenaccountedfor in known compounds,inositol
phosphatespredominate(Anderson,1967).The quantitiesof lower phosphatesof
inositol are much smallerthan thoseof higher phosphates(penta-and hexaphos-
phate).The higherphosphatesof inositol (CJI 1206),which canbe synthesizedby
PHOSPHORUS 885
soil microorganisms (Caldwell & Black, 1958b),are more inaccessibleto micro-

bial or enzymedegradation(Greaves& Webley, 1969)due to their higher charge
densitiesand high binding strengthon the surfacesof soil minerals(Andersonet
aI., 1974; Stewart & Tiessen,1987). Stereoisomersof myo-, chiro-, neo- and
scyllo-phosphateestershavebeencharacterizedin soils.
Inositol phosphatesin soils can be extractedwith alkaline solution. Basi-
cally, the organicP is precipitatedas Fe or Ba salts at appropriatepH levels fol-
lowing hypobromitetreatment.The precipitatedFe- or Ba-phytatesare dissolved
againin acid solutoin or with H+-resin and fractionatedby anion-exchangechro-
matography.Separationof Po is basedon the strength of interactionsbetween
organicP componentsin the mobile phaseand the exchangesiteson the station-
ary phase(resin).
Phospholipids,a small fraction of total Po «5%) (Stevenson,1982), are
definedasphosphateesterssolublein fat solvents such as ether(C2Hs)O, benzene
(C6H6), or chloroform (CHCI3). However,soil mineralsinterferewith the extrac-
tion of phospholipidsby the solvents,and pretreatmentof soil with a mixture of
HCI and HF is necessary toimprove the effectivenessof solventextraction.Phos-
phatidyl choline (1,2-Diacyl-sn-glycero-3-phospho-choline), phosphatidyl
ethanolamine (1,2-Diacyl-sn-glycero-3-phospho-ethanolamine), and phos-
phatidyl serine(1,2-Diacyl-sn-3-phospho-L-serine) are the predominantforms of
phospholipidsin soils. .
Like phospholipids,nucleic acids and their derivatives representa small
fraction of Po in soil (Anderson,1975; Stevenson,1982). Ribonucleic acid and
deoxyribonucleicacid (DNA), which consist of chains of nucleotides,can be
adsorbedby soil particles(Goring & Bartholomew,1952). The methodsusedto
characterizethis Po fraction havebeenreviewedby Anderson(1967) and are not
included in this chapter.
Microbial biomass P representsa small fraction of soil P held in soil
microorganisms(Tate, 1984). It contains,in addition to Pj, organicP compounds
such as RNA, DNA, polyphosphates,and inositol phosphatesand turns over
rapidly to supply Pi for plant use.The microbial biomassP is measuredbiologi-
cally in fresh soil by CHCl3 fumigation, followed by chemicalextractionto deter-
mine the increaseof Pi. The procedurefor its measurementis not includedin this
chapter. It is describedin detail by Brookes et al. (1982), Hedley & Stewart
(1982), and Walbridge and Vitousek (1987).
Special Apparatus
2. Centrifuge.
3. Hot water bath.
4. Refrigerator.
5. Fractioncollectors.
6. Aluminum block for tube digestion.
7. Stainlesssteelperchloricacid fume hood.
8. pH meterequippedwith a single combinationelectrode.
886 KUO
Methods
Inositol Phosphates
Reagents
1. Hydrochloric acid, concentrated(12 M).
2. Hydrochloric acid, 1.5 M: Dilute 125 mL of concentratedHCI (12 M)
3. Hydrochloricacid, 0.1 M: Dilute 67 mL of 1.5 M HCI to 1 L with deion-
ized water.
4. Sodiumhydroxide, 10 M: Dissolve400 g of NaOH in deionizedwater
and dilute to 1 L.
deionizedwater.
deionizedwater.
7. Ferric chloride (FeCI3), 60% w/v: Dissolve 1000g of FeCl3 • 6 H20 in
deionizedwater and dilute to 1 L.
8. Bromine.
10. Magnesium chloride saturated: Suspend600 g of MgCl2 to 1 L of
deionizedwater.
11. Anion exchangeresin, Dowex 1-X8 (0.038--0.075mm) (Dow Chemi-
cal, Midland, MI).
12. Cation exchangeresin, Dowex-50W (W).
13. p-nitrophenol,0.25%: dissolve 0.25 g of p-nitrophenolin 100 mL of
deionizedwater.
Procedure. Add 5.0 g of soil «0.18mm) and50 mL of 0.1 MHCI to a 100-

mL centrifugetube and shakethe mixture for 10 min. Centrifugethe mixture to
obtain a clear supernatantsolution. Decantthe supernatantliquid and determine
the Ca concentration.Repeatthe processuntil the supernatantis free of Ca. Add
50 mL of 1 M NaOH to the soil residueandshakethe mixture for 16 h. Centrifuge
and decantthe supernatantliquid to a 250-mL Erlenmeyerflask. Add an addi-
tional 50 mL of 1 M NaOH to the residualsoil and heat the mixture in a water
bath at60°C for 4 h with occasionalstirring. Centrifugeand combinethe super-
natant liquid with the first extract. Wash the residualsoil twice with "50 mL of
H20, centrifuge,and add the supernatantto the two alkali extracts.
Adjust the pH of the combinedextractsand washingswith concentrated
HCI to pH 0.5. Mter 30 min, centrifugethe mixture and decantthe supernatant
liquid, which containsfulvic acid, to a 5OO-mL Erlenmeyerflask. Redissolvethe
residualhumic acid by twice washingthe residuewith 25 mL of 1 M NaOH. Pre-
cipitate the humic acid again with concentratedHCI each time as described
above.Centrifugeand combinethe washingswith the fulvic acid fraction in an
Erlenmeyerflask. Cool the combinedfulvic acid solution to 5°C and add 20 mL
of Br2 in 2-mL incrementswith constantmixing. Keep the mixture at 5°C for 18
h, after which heatthe solution at 60°C for 1 h andremovethe excessBr2 by low
temperaturedistillation in vacuo or alternativelyby extractionwith 50-mL por-
PHOSPHORUS 887
tions of ether (C2HS)O four times. Transfer an aliquot to a Folin-Wu digestion

tube, add 0.5 mL of saturatedMgCl2 solution, 1 mL of 70% HCI04 and deter-
mine the P concentrationas describedin "Ascorbic Acid Method."
Adjust the solution pH to 2.0 with 10 M NaOH solution. Add 60% FeCl3
(w/v) to give a FelP (W/W) ratio of about two and heat the solution in a steam
bath for 20 min to coagulatethe precipitate.Centrifugethe solution after it has
cooled and decantthe liquid. Dissolve the precipitatein 2 M NaOH and repre-
cipitate at pH 2 using enoughFeCl3 to obtain a FelP ratio of about 1. Centrifuge
and decantthe liquid. Dissolve the precipitatein 2 M NaOH and neutralizethe
excessalkali by addition of H+ -resin until the pH is about seven.Filter to sepa-
rate the solution from the resin and reducethe volume of the solution to 5 to 10
mL in vacuo.
Ion-Exchange Chromatography. Preparea resin column (10- by 1.2-cm
diam.) with the Cl- form of Dowex 1 x 8 anionexchangeresin (0.038-0.075mm)
and convert the resin to the formate form by washing with 6 M formic acid
(HCOOH) and then with H20. Add an aliquot containing5 to 8 mg of organic P
to the column and elute the column with increasingconcentrationof HCI up to
0.7 M by an apparatusthat consistsof two connectedreservoirsof the type
describedby Parr (1954): one containing 1550 mL H20 (ReservoirA) and the
other 900 mL 1.5 M HCI (Reservoir B). Stir the solution in ReservoirA con-
stantly and keepthe reservoirsat identical levels. Keep the flow rate from Reser-
voir A at 1 mL per 3 min. Collect successivelO-mL samplesof the eluateup to
50 in an automaticfraction collector.
Transferan aliquot containing2 to 40 l!g P to a Folin-Wu digestion tube.
Add 0.5 mL of saturatedMgCI2, 1 mL of concentratedH2S04 and 1 mL of 70%
HCI04 and digest at 205°C for 30 min as describedin "Procedure"for "Acety-
lacetoneExtraction."Transferthe digestto a 50-mL volumetricflask with 15 mL
of deionizedwater. Add five dropsof 0.25%p-nitrophenolindicator and neutral-
ize the acidity with 10 M NaOH in dropwise additions until the color just turns
yellow. Determinethe P concentrationusing the ascorbicacid methoddescribed
in "Ascorbic Acid Method."
Comments. Treatmentof fulvic acid with NaOBr following the removalof
humic substancesby centrifugationis necessaryto destroy extraneousorganic
matter that can interfere with the adsorptionand elution of inositol phosphates.
The excessBr2 shouldbe removedby low temperaturedistillation as describedin
the procedure,by boiling under a fume hood (Wrenshall & Dyer, 1941) or by
extraction with ether to facilitate the formation of ferric phytate (Cosgrove,
1963).The brominationof fulvic acid hasminimal effect on the stability of phytin
presentin the fulvic acid (Wrenshall& Dyer, 1941).
Anderson(1963) observedthat when the Fe concentrationis high and inos-
itol phosphateconcentrationis low, quantitativeyields may not be achieveddue
to the formation of solublecomplexesof sodium-phytatewith FeCI3. The desired
weight ratio of FelP is about 0.81 (Wrenshall & Dyer, 1941).The ratio of FelP
calledfor by the Omotosoand Wild's (1970a)methodis between1:1and 2:1. Fer-
ric chloride applicationbasedon theserations is more than adequateto precipi-
tate the phytin as well as avoid high concentrationsof Fe.
888 KUO
The elution diagramrevealssevenfractions for soil inositol phosphatesas

opposedto six for the commercialphytin (Omotoso& Wild, 1970a; Cosgrove,
1963). The first fraction yields Pi> inositol mono-, di-, and triphosphates,plus an
unknown inositol phosphate.The lower estersconstituteonly a minor portion of
the total Po (Anderson,1956; Martin & Wicken, 1966; Dormaar,1967). Howev-
er, Cosgrove(1963), using a Cl--form of Dowex 1 x 8 (0.038-0.075mm), failed
to identify the presenceof lower estersand the free inositol from the hydrolysis
of the Po in this fraction. Fractions 2 and 3 containedtetrainositol phosphate
(C6H 1601SP4), and Fractions 4 and 5 contained pentainositol phosphate
(C6H 170 21Ps). Fractions6 and 7 containedthe inositol hexaphosphate and scyl-
loinositol hexaphosphatethat was referred to as isomers of inositol hexaphos-
phateby Smith and Clark (1951) and later identified by Cosgrove(1963) as scyl-
loinositol hexaphosphatebased on paper chromatographyand comparisonof
infrared spectra.
A two-stepfractionatioll of Po by chromatographyin alkaline soil extracts
without acidification to separatethe humic acid from fulvic acid fractions has
been developed(Martin & Wicken, 1966; Omotoso& Wild, 1970b; Steward&
Tate, 1971).The Po is separatedfrom Pi by eluting the extractthrougha Sephadex
G-25 gel (PharmaciaBiotech, Alameda,CA) column and is further fractionated
by eluting through a secondSephadexcolumn following the destructionof extra-
neousorganics(Omotoso& Wild, 1970b)or by isolation asbariumsaltsand con-
version to the free acids (Steward& Tate, 1971). The two-step approachavoids
the use of FeCl3 to precipitatephytin, which has a relatively low reactivity with
lower esters(Anderson,1975), possibly due to a relatively low chargedensity.
Severalother fractionationschemes,differing mainly in the type and form
of resinsand elutantsused,also havebeenproposed.Caldwell and Black (1958a)
obtainedPo in the fulvic fraction using the procedureof Mehta et al. (1954). The
Po was separatedin a stepwiseelution with increasingconcentrationof HCI using
weak base polyamine resin. The method, designedprimarily for isolating the
polyphosphates (inositol pentaphosphate andhexaphosphate), haspoor resolution
for the lower phosphateesters (Dormaar, 1967). McKercher and Anderson
(1968a)extractedsoils with 3 M NaOH in boiling water and successfullyeluted
the lower and polyinositol phosphatesfrom a column of Dowex-l (format form)
with increasingconcentrationsof NH4-formate. Penta-and hexaphosphates of
inositol and their isomerscan be separated when the eluatescontainingthem are
further eluted with a gradientof HCI from 0 to 1.5 M (McKercher & Anderson,
1968b). Myoinositol hexaphosphateis generally the major componentof the
polyphosphateester.
Polyphosphateis the major componentof soil Po, although its content
varies widely amongsoils. Its concentrationrangesfrom 11 to 30% of total Po
(averaging24%) for someEnglandand Nigerian soils (Omotoso& Wild, 1970b),
2 to 30% for some Canadiansoils (Dormaar, 1967; McKercher & Anderson,
1968a,b;Thomas & Lynch, 1960), 8 to 25% for some U.S. soils (Caldwell &
Black, 1958c),and 12 to 16% for someAustrian soils(Cosgrove,1963).
Other isolates of Po that have been identified include acid-labile sugar
phosphate[e.g., glucose-I-phosphate (C6H 130 9P), Omotoso& Wild, 1970b] and
pyrophosphate(Anderson& Russell, 1969). In addition, phosphonates of bacte-
PHOSPHORUS 889
ria origin have been identified by nuclear magnetic resonance(NMR) spec-

troscopy(Tate & Newman,1982; Hawkeset aI., 1984).
Another method of characterizingPo is basedon successiveextra~tions
with various extractants(Bowman & Cole, 1978; Tiessenet aI., 1983). These
investigatorsdivided the total Po into labile (NaHC03-extractable),moderately
labile (dilute NaOH and dilute H2S04-extractable),moderatelyresistant(associ-
atedwith fulvic acid), highly resistant(associatedwith humic acid), and residual
fractions. This fractionationschemeaccountsfor 98% of total Po.
Phospholipids
Reagents
1. 1.25% HCI + 1.25% HF: Dilute approximate34 mL of concentrated
HCI (12 M) and 26 mL of concentratedHF (24 M) to 1 L with deion-
ized water in a l-L polypropylenebottle.
2. Acetone(CH3COCH3).
3. Ether.
4. Chloroform (CHCI3).
5. Light petroleum.
6. Benzene(C6H6)'
7. Methanol (CH30H).
Procedure. Add 1.0 g of soil «0.15 mm) to 10 mL of 1.25%HCI + 1.25%
HF solution in a 100-mL polyethylenecentrifuge tube. Shake the suspension
overnightand centrifuge.Discardthe supernatantliquid and repeatthe extraction.
Wash the soil residuewith 10-mL portions of deionizedH20 until acid free and
discardthe washings.Add 50 mL of acetone,cover the tube with a polyethylene
sheetheld in placewith a rubberband.Allow the suspensionto standfor 4 h with
thoroughshakingevery 30 min. Centrifugeand transferthe supernatantliquid to
a 500-mL bottle with a ground-glassstopper.Add 50 mL of light petroleum(boil-
ing point == 40-60°C) and allow it to standfor 4 h with occasionalshaking.Cen-
trifuge and combinethe supernatantwith the acetoneextract. Repeatthe extrac-
tion with 50 mL of a 1:4 (v/v) mixture of ethanollbenzeneand then with 50 mL
of a 1:1 (v/v) mixture of ethanol(C2HsOH):chloroform. Combinetheseextracts
with the previous extracts and evaporatethem to dryness at low temperature
(30°C) and pressureunder a fume hood. To purify the P-containinglipid, re-
extract the residuewith the following sequence:20 mL of a cold 1:1 (v/v) mix-
ture of ether/light petroleumfor 2 min; 20 mL of cold chloroform for 2 min; 20
mL of cold 1:1 (v/v) ether/lightpetroleumbroughtto boiling in a water bath; and
finally 20 mL of cold chloroform, heatedto boiling. Combinetheseextractsand
evaporateto drynessin a beakerin a water bath and determinethe TP concentra-
tion by digestionin HCI04 as outlined in "Procedure"for "AcetylacetoneExtrac-
tion."
Comments. Soil drying greatly affects the extractability of phospholipids,
and for air-dried soils, at least two pretreatmentswith the mixture HCl and HF
are necessaryto increasethe efficiency of extraction(Hance& Anderson,1963a).
Clay is known to decreasethe extraction of lipid P in bacteriacellular extracts
(Goring & Bartholomew,1949).
890 KUO
There have beenseveralmodificationsto simply the Hanceand Anderson

(1963a)method.Theseinclude two successiveextractionswith hexane-acetone
(Kowalenko & McKercher, 1970), extractionwith ethanol-benzene followed by
extraction with methanol (CH30H)-chloroform (Baker, 1975), and extraction
with methanol-chloroform(Chae & Tabatabai,1981). High-performanceliquid
chromatographycan be used in combinationwith methanol-chloroformextrac-
tion for identifying phospholipidsin soils (Stott & Tabatabai,1985).
The concentrationof phospholipidsrangesfrom 0.1 to 13 mg kg-I, repre-
senting0 to 14% of the total Po of somesoils (Kowalenko & McKercher, 1970;
Hance & Anderson, 1963a; Dormaar, 1970; Chae & Tabatabai,1981; Stott &
Tabatabai, 1985). Alkaline hydrolysis of soil lipids yields glycerophosphate,
whereas acid hydrolysis produces choline (2-hydroxy-N,N,N-trimethylethan-
ammoniumhydroxide) and ethanolamine(2-aminoethanol)(Hance& Anderson,
1963b).
AVAIlABILITY INDICES
General Principles
To evaluateP availability in soils, numeroussoil testshavebeendeveloped
that extractvarying amountsof P, dependingon the typesof extractantsused.The
extractantscan be generallyclassifiedinto severalcategories:
1. Water or unbufferedsalt solutions(e.g., CaCI2).
2. Dilute concentrationsof weakacids(e.g.,lactate,acetate)with or with-
out a complexingagent(P- or EDTA).
3. Dilute concentrationsof strongacids(e.g., HCI, H 2S04) with or with-
out a complexingagent(e.g., P-, lactate,EDTA).
4. Buffered alkaline solutions [e.g., NaHC03, NH4HC03 (ammonium
bicarbonate)]with or without a complexingagent(DTPA).
5. Anion exchangeresin or iron oxide-impregnatedfilter paperstrips.
6. isotopic exchangewith 32p.
The dilute strongacid solutionssolubilizeCa-P,Al-P, and to a lesserextent,
Fe-P(Nelson et aI., 1953). Fluoride is includedwith dilute strong acid solutions
(Bray & Kurtz, 1945) to complexAl and preventreadsorptionof P by Fe oxides.
Salts and EDTA are further included with P- and dilute strong acid (Mehlich,
1984) to form multielementteststhat simultaneouslyextract macro-and micro-
elements. Like EDTA, DTPA was added to the buffered alkaline solutions
(Soltanpour& Schwab, 1977) to facilitate the extractionof microelementsthat
can be simultaneouslydeterminedby inductively coupled argon plasma spec-
troscopy(Soltanpouret aI., 1979).
Anion exchangeresin, iron-oxide impregnatedfilter paperstrips, and iso-
topic exchangemethodsare nondestructiveon soil constituentsin contrastto the
chemicalextractants.Iron-oxide impregnatedfilter paperor anionexchangeresin
functionsas a sink that simulatesthe actionof plant roots by continuouslyremov-
ing dissolved P from the soil solution. The quantity of P that is isotopically
exchangeablewithin a specifiedtime interval gives an estimateof labile surface
P.
PHOSPHORUS 891
Methodsfor a numberof soil testsare presentedalongwith their basicprin-

ciples. For the soil teststo adequatelyreflect soil P availability, the P testsshould
respondto soil characteristicsin a similar manneras plants. The relationships
amongsoil tests and betweensoil test and plant yield or P uptakeare discussed
in the commentssections.
Methods
Extractionwith Wateror Dilute Salt Solution

Principles.Soil solution P representsthe portion of soil P that is in equi-
librium with the solid phaseP under the conditionsprevailing in the soil. It is a
very small fraction of availablesoil P, but readily accessibleto plant roots.
Centrifugation and displacementare the two most commonly used tech-
niques for obtaining soil solution. However, these methodsare impractical for
routine soil testing, consideringthe time and the volume of soil solution for P
determination.A wider ratio of water to soil is generally recommendedin the
interestof efficient operation,recognizingthat the resultingchangesof the chem-
ical characteristicsof the soil solution, such as ionic strength,can influence soil
P solubility.
Dilute CaCl2 (0.01 M) is usedin placeof waterfor obtaininga clear filtrate
(Aslyng, 1964). The amountof P solublein CaCl2 solution is'smallerthan that in
water due in part to the enhancementof Ca2+ on P sorption by soils.
Reagents
1. Calcium chloride, 0.01 M: Dissolve 1.47 g of CaCl2 • 2H20 in deion-
ized water and dilute to 1 L.
Procedure.Add 5 g of air-dried soil «2 mm) and 50 mL of deionized
water (or 0.01 M CaCI2) to a flask. Shakethe suspensionfor 1 h and centrifuge
to obtain a clear supernatantliquid. If the solution is not free of suspendedsoil
particles,filter through a membranepaper(0.45 Ilm) or repeatedlyfilter through
the sameWhatmanno. 42 filter paperuntil clear.
Pipette an aliquot containing 1 to 20 Ilg P into a 25-mL volumetric flask
and determinethe P concentrationby the "Ascorbic Acid Method" if significant
amountsof acid-labile Po are expected.PreparestandardP solutionsand a blank
that contain the samevolume of extracting solution. Calculatethe amount of P
extracted(Pex) as follows
25 V2
Pex (mg kg-I) = P concentration(llglmL)· - • [12]
VI g soil used
whereVI =volume of extractusedfor P determination,

V2 =volume of extract.
Comments.The ratios of soil weight to the volume of H 20 that have been
used to measurethe water-solubleP vary widely, including 1:1.25 (Olsen &
Watanabe,1970), 1:10 (Olsen & Sommers,1982), 1:60 (van der Paauw, 1971),
892 KUO
and 1:100(van Diest, 1963). The reactiontimes usedfor the extractioninclude 5

min (Kuo & Jellum, 1987; Olsen & Sommers,1982), 1 h (Olsen & Watanabe,
1970; Mackay et aI., 1984)or 15 h (van Diest, 1963).The water-solubleP should
be a function of the quantity of P sorbedand the P sorptioncapacityor, alterna-
tively, the degreeof P surfacesaturation(Murrmann& Peech,1969). It also may
be related to the concentrationof physically sorbed P (Ryden & Syers, 1977).
However,due to the unbufferedcharacteristicof water, the quantity of water-sol-
uble P, or the P concentrationin solution, is highly dependenton pH (Kuo & Jel-
lum, 1987; Guptaet aI., 1990),ionic strength(Bolan et aI., 1986; Bar-Yosefet aI.,
1988) and type and valenceof exchangeablecation (Smillie et aI., 1987; Stoop,
1983; Sharpleyet aI., 1988). For example,Na saturationof soil increasesP solu-
bility in water, whereasCa saturationgenerallydoesthe opposite.
Water-solubleP correlatedwell with P uptakeby plants (van Diest, 1963;
Thompsonet aI., 1960; Tran et aI., 1988; Keramidas& Polyzopoulos,1983), al-
though results to the contraty also have been shown (Fried & Shapiro, 1956).
Underlong-termgrowth conditions,especiallywith plantshaving high P require-
ments,the water-solubleP may be inadequateto accuratelyreflect the overall soil
P availability (Ballard & Pritchett, 1975). The replenishmentof solution P in-
volves the P quantity factor as well,which becomesincreasinglycritical as more
P is withdrawn by plants.
At comparablesolution P concentrations,soils with higher P buffering ca-
pacitiesor clay contentsshouldcontain more labile P to replenishthe solution P
removedby root absorption(Hedley et aI., 1982; Jungk, 1987). Thus, the critical
solution P concentrationor the critical quantity of water-solubleP required for
optimum plant growth is lower for fine-textured soils than for coarse-textured
soils (Kamprath& Watson,1980).
Unlike water extraction, the dilute CaCl2 (0.01 M) extraction does not
require a high-speedcentrifugationto obtain a clear extract (Schofield, 1955).
The ratios of the weight of soil to the volume of CaCl2 solution as well as the
lengths of time that have been used for the extraction vary considerably.The
ratios range from 1:1.25 (Soltanpouret aI., 1974) to 1:10 (White & Beckett,
1964); and the reaction times range from less than 1 h (Baker & Hall, 1967;
Aslyng, 1964) to 7 d (Dalal & Hallsworth, 1977).
The amount of CaClz-extractableP correlatedwell with the eqilibrium P
concentration(Moody et aI., 1983), NaHCOF or NH4HC03-DTPA-extractable
P (Labhsetwar& Soltanpour,1985). At the same level of CaClz-extractableP,
more P is availablein soils with high clay contents(Olsenet aI., 1983). Thus, the
CaCl2-s01ubleP level critical for optimum plant growth is lower for silty clay
loam soil than for very fine sandyloam (Fox & Kamprath,1970).
The phosphatepotential(1/2 pCa + pH2P04") in 0.01 M CaCl2 extractswas
consideredto be a viable parameterfor describingP availability or P solubility in
soils (Aslyng, 1964; Schofield, 1955), as the potential is dependenton the P
buffering capacityof soil (Barrow, 1967). However, for the phosphatepotential
to adequatelydescribeP uptakeby plants,the solution activity of Ca2+ shouldbe
maintainedconstantor at least should vary much less than that of phosphate
PHOSPHORUS 893
(White & Beckett,1964;Wild, 1964;Olsen& Khasawneh,1980).In this manner,

the potentialvaries in proportionto the P activity.
Extractionwith Dilute Concentrationof StrongAcids

Reagents
1. Extracting solution (0.05 M HCI + 0.0125M H2S04): Add 14 mL of
concentratedH2S04 (18 M) and 83.3 mL of concentratedHCI (12 M)
to 18 L of deionizedH20 and dilute to 20 L.
Procedure.Add 5.0 g of air-dried soil «2 mm) and about200 mg of char-
coal (Darco G60, J.T. Baker, Phillipburg, NJ) to a 50-mL Erlenmeyerflask. Add
20 mL of extractingsolution and shakethe suspensionfor 5 min. Filter the sus-
pensionthrougha membranefilter «0.45mm) or Whatmanno. 42 filter paper(if
Whatmanno. 42 filter paper is used and the filter is not clear, pour the filtrate
back through the samefilter paper).Transferan aliquot containing1 to 20 I!g of
P to a 25-mL volumetric flask and dilute to volume. Determinethe P concentra-
tion by the "Ascorbic Acid Method." PreparestandardP solutions and a blank
which containthe samevolume of extractingsolution.
Calculate the amount of P extracted using Eq. [12] in "Procedure"for
"Extraction with Water or Dilute Salt Solution."
Comments.The dilute acid extractionprocedureof Nelson et ai. (1953),
also known as the North Carolina or Mehlich-l P test (Kamprath & Watson,
1980),extractslarge amountsof nonlabileP in soils that have pHgreaterthan 6.0
(Holford, 1980). It is unreliable for calcareousor alkaline soils (Thomas &
Peaslee,1973), soils that have recently received rock phosphateapplications
(Yost et aI., 1982),or soils with high cationexchangecapacity(CEC) or high base
saturation(Thomas& Peaslee,1973). Thesetypes of soils tend to neutralizethe
acid, thereby reducing the capability of the dilute acid to extract P. Clay or
hydrousaluminum and iron oxides also can reducethe quantity of P extractable
by the acids(Nelsonet aI., 1953; Lins & Cox, 1989; Lins et aI., 1985).
The dilute acid, or Mehlich-l, soil test generally correlateswell with the
Bray-1 test (Welch et aI., 1957; Shumanet aI., 1988),but not aswell with Olsen's
NaHC03 test (Shumanet aI., 1988) amongdiversesoils. However,for the same
soil type, they are all highly correlated(Welch et aI., 1957). The amount of P
extractedor the recoveryof a known amountof addedP by the Mehlich-1 test is
generallygreaterthan that by Olsentest, particularly for soils that have received
rock phosphateapplicationsrecently(Menon et aI., 1988). It is slightly less than
that by the dilute acid fluoride test (Bray-I) (Ballard & Pritchett, 1975; McLean
et aI., 1982; Menon et aI., 1988).The Mehlich-l test has the advantageof simul-
tmeouslyextractingCa and Mg as well (Nelson et aI., 1953).
A P level of 20 to 30 mg P kg-1 soil for the Melich-l test is generallycon-
sideredin the high category(Thomas& Peaslee,1973),althoughadjustmentsfor
different soil types are necessary.Kamprathand Watson(1980) indicatedthat a
Mehlich-l P level of 20 to 25 mg P kg-1 soil is consideredadequatefor plant
894 KUO
growth in sandy soils but only 10 ppm P is required for fine-textured soils.
Increasingclay contentgenerallydecreasesthe critical P level requiredfor opti-
mum plant growth (Lins & Cox, 1989).
Extractionwith Dilute Acid Fluoride
Principles. For acid soils, F"" promotesP desorption by decreasingAI
activity through the formation of AI and F complexes.Fluoride also is effective
in suppressingthe readsorptionof solubilized P by soil colloids. However, the
Bray-1 test (Bray & Kurtz, 1945) performsunsatisfactorilyin highly calcareous
soils due to the neutralizationof the acid by calcium carbonate(CaC03) and for-
mation of CaF2, that reactswith dissolvedP to form secondaryprecipitates.
Reagents
1. Ammonium fluoride, 1 M: Dissolve 37 g of NH4F in deionizedwater
and dilute to 1 L. Storethe solution in a polypropylenebottle.
2. Hydrochloric acid, 0.5 M: Dilute 20.8 mL of concentratedHCI (12 M)
to 500 mL with deionizedwater.
3. Extracting solution: Add 15 mL of 1.0 M NH4F and 25 mL of 0.5 M
HCI to 460 mL of deionizedwater to obtain a solution containing0.03
M NH4F and 0.025M HCI.
Procedure.Add 1.0 g of air-dried soil «2 mm) to a flat-bottomedglass
vial or bottle. Add 7 mL of the extractingsolution. Shakethe suspensionvigor-
ously for 1 min and filter through a membranefilter (0.45 flm) or Whatmanno.
42 filter paper(if Whatmanno. 42 filter paperis usedand the filtrate is not clear,
pour the filtrate back through the samefilter). Transferan aliquot containing1 to
20 flg of P to a 25-mL volumetric flask and determinethe P concentrationby the
"Ascorbic Acid Method" or by the "Modified Ascorbic Acid Method" if suffi-
cient amountsof labile Po are present.PrepareP standardsolutionsand a blank
which include the samevolume of extractingsolution.
Calculate the amount of P extracted using Eq. [12] in "Procedure"for
"Extraction with Water or Dilute Salt Solution."
Comments.The soil/extracting solution ratios and shaking times vary
amongusers(Jackson,1958; Mackay et aI., 1984; Chien et aI., 1980; Olsen &
Sommers,1982). For soil with a high CEC or a CaC03 equivalentthat exceeds
7% base saturation,the Bray-1 test is not suitable (Thomas & Peaslee,1973;
Baker & Hall, 1967; Nesseet aI., 1988) due to neutralizationof the acids, unless
the ratio of extractantto soil is increasedconsiderably(Randall & Grava, 1971).
Maintaininga pH level <2.9 during the extractionis necessaryfor betterP extrac-
tion (Mehlich, 1978). The formation of CaF2 resultingfrom the reactionof Ca2+
and F"", which rapidly immobilizes P, is anotherconcernregardingthe effective-
nessof the test for calcareoussoils. Van Lierop (1988) replacesHCI in the Bray-
1 test with 0.25 M acetic acid to form the Kelowna test that is comparablewith
the Bray-1 test for acid soils.
Fluoride, arsenic, or ferric ions may interfere with color formation. The
addition of boric acid (Bray & Kurtz, 1945) to form noninterferringfluoroborate,
and reduceAs5+ to As3+ and Fe3+ to Fe2+ by adding sodium bisulfite (NaHS03)
(Jackson,1958) may be requiredto preventsuch interference.
PHOSPHORUS 895
Aluminum phosphateis the primary soil P fraction extractableby the

Bray-l test (Susukiet aI., 1963).The formation of stableAlF4 complexesreleas-
esP from AI-P (Chang& Jackson,1957).However,the solubilizedP canbe read-
sorbedby iron oxide (Bruce & Bruce, 1972) as substantiatedby the decreaseof
the amountof P releasedfrom AI-bound P with the addition of iron oxide (Brom-
field, 1967). Holford (1980), McLean et ai. (1982), and Kuo et ai. (1988) also
showedthat the Bray-l test is sensitiveto the soil P buffering capacityor P sorp-
tion capacity,and the fraction of addedP it recoversvariesconsiderablyamong
diversesoils. The degreeof P saturationon soil surfacesis the main soil parame-
ter that regulatesthe level of P extractableby the Bray-l test.
The Bray-l test can dissolveunreactedphosphaterock appliedto acid soils
(Chien, 1978; Mackay et aI., 1984).Thus,for soils that havereceivedrock phos-
phate application recently, the Bray-l test could overestimatethe P availability
until all of the rock phosphateis fully dissolved(Mackayet aI., 1984).The recov-
ery of addedsoluble fertilizer P to soils by the l3ray-l test varies dependingon
the P buffering capacityor P sorptioncapacity,averagingfrom 28 to 39% (Baker
& Hall, 1967; McLean et aI., 1982; Kuo et aI., 1988).
Phosphorustest valuesexceeding30 mg P kg-1 soil are consideredin the
high category(Thomas& Peaslee, 1973; Kamprath & Watson, 1980). For very
strongly buffered soils, the required P test valuesfor maximum crop yields are
lower (Holford, 1980). Increasingclay content decreasesthe critical P level
requiredfor optimum plant growth (Lins & Dox, 1989).
Extractionwith ButTeredAlkaline Solution

Principles.The OW and C01- in the NaHC03 solution decreasethe con-
centrationor activity of Ca2+ andAl3+, resultingin increasedP solubility in soils.
The extractantis useful for both acid and calcareoussoils. In calcareoussoils,
increasedcalcium phosphatesolubility resultsfrom the decreasedCa concentra-
tion by the high concentrationof C01-and the precipitationof CaC03. In acid or
neutral soils, the solubility of aluminum and iron phosphatesincreasesas
increasedOH- concentrationsdecreasesthe concentrationsof Al3+ by aluminate
complexformation and of Fe3+ by precipitationas the oxide. The increasedsur-
face negativechargesand/or decreasednumberof sorption sites on Fe and AI
oxide surfacesat high pH levelscould be responsiblefor the desorptionof sorbed
P as well.
Reagents
1. Sulfuric acid 2.5 M: Add 139 mL of concentratedH2S04 (18 M) to 800
mL of deionizedH20. Cool the solution and dilute to 1 L with deion-
ized water.
2. Sodium hydroxide, 1 M: Dissolve 40 g of NaOH in deionizedwater
and dilute to 1 L.
3. Sodium bicarbonatesolution, 0.5 M: Dissolve 42.0 g of NaHC03 in
about950 mL of deionizedH20. Adjust the pH with 1 M NaOH to pH
8.5 and dilute the solution to 1 L with deionizedwater. Add a layer of
mineral oil to prevent the direct exposureof the solution to atmos-
phere.
896 KUO
4. Carbonblack.
5. p-nitrophenol,0.25%. Dissolve 0.25 g of p-nitrophenolin 100 mL of
deionizedwater.
Procedure.Add 2.0 g of soil «2 mm) and 40 mL of the extractingsolu-

tion in a 125-mL Erlenmeyerflask. Shakethe suspensionfor 30 min and filter
througha membrane(0.45-~m) filter or Whatmanno. 42 filter paper(if Whatman
no. 42 filter paper is used and the filtrate is not clear, pour the filtrate back
throughthe samefilter paper).Add half a teaspoonof carbonblack to the filtrate
and filter if necessaryto removethe dissolvedorganics.(This treatmentof the fil-
trate is not necessaryfor most soils when the ascorbicacid methodis usedfor P
determination).
Transferan aliquot of the extractcontaining1 to 20 ~g P to a 25-mL volu-
metric flask. Add five drops of p-nitrophenolindicatort.Adjust the solution pH
by slowly adding2.5 M H2S04 down the side of the flask to preventrapid evo-
lution of CO2 and loss of solution. Adjust the pH until the solution becomescol-
orless.Determinethe P concentrationby the "Ascorbic Acid Method" or by the
"Modified AscorbicAcid Method" if a sufficient quantity of acid-labilePo is pre-
sent.
Preparestandardsand a blank using the same volumes of the NaHC03
solution and acid. Calculatethe amountof P extractedusing Eq. [12].
Comments.The NaHC03 test (Olsenet ai., 1954)generallyextractslessP

than the Mehlich-1 or Bray-1 test (Ballard & Pritchett, 1975; Shumanet ai.,
1988) exceptin highly bufferedsoils (Holford, 1980). The quantity of P extract-
ed is well correlatedwith resin-extractableP (Adepoju et ai., 1982; Matar et ai.,
1988; Nesseet ai., 1988; Reith et ai., 1987) and the Al-P contentin soil (Kam-
prath & Watson,1980). The Olsen methodwas modified by Cowell (1963) who
increasedthe soil/solution ratio from 1:20 to 1:50 and the reactiontime from 30
min to 16 h to decreasethe influence of the soil P buffering on P extractability
particularly for highly weatheredsoils. The Cowell test extractsa higher propor-
tion of availablesoil P than doesthe Olsentest (Adepoju et ai., 1982).The aver-
age recovery of P fertilizer by the Olsen test was 21% for some diverse acidic
soils (Kuo et ai., 1988).
The dependenceof the NaHC03 .extractionon soil clay content (Pratt &
Garber,1964),citrate-dithionite-bicarbonate Fe (Ryan et ai., 1985),and P buffer-
ing capacity (Holford, 1980; Barrow & Shaw, 1976) is well known. Kuo et ai.
(1988) and Kuo (1990) illustrate that the NaHC03 test levels are a function of the
fraction of P coverageon the surfacesof soil particles, reflecting P intensity
rather than P quantity. To properly interpretthe NaHC03 soil test valuesamong
diverse soils, the soil P sorption capacitiesshould be considered.The NaHC03
solution extracts Pi as well as a small amount of Po (Bowman & Cole, 1978;
Tiessenet ai., 1983).If the dissolvedPo is not expectedto hydrolyze in the acid-
ified molybdatesolution, the "Ascorbic Acid Method" or the isobutanolextrac-
tion technique(Jayachandran et ai., 1992)could be usedfor P determination;oth-
erwise, the "Modified Ascorbic Acid Method" should be used to prevent inter-
ferencefrom the dissolvedPo on P determination.
PHOSPHORUS 897
A NaHC03 test level of 10 mg P kg-1 soil is consideredin the high cate-

gory (Thomas & Peaslee,1973). The need to adjust the critical P test value
requiredfor optimumplant growth with respectto P buffering capacityof the soil
is unclear.However,Holford (1980) suggestedthat the correctionis unnecessary
evenfor strongly bufferedsoils.
Extractionwith Ammonium Bicarbonate-Diethylenetriaminepentaacetic

Acid
Principles.The test (Soltanpour& Schwab, 1977)was developedto simul-
taneouslyextractP, K, Zn, Fe, Cu, and Mn. The extractingsolution consistsof 1
M NH4HC03 and 0.005 M DTPA. Whereasthe DTPA reacts withthe trace ele-
ments(Zn, Fe, Cu, and Mn), N~+ exchangeswith K+. The HC03" and OH- facil-
itate P desorption.
Reagents
1. Ammonium bicarbonate,1 M, and DTPA, 0.005 M: Add 1.97 g of
DTPA to 800 mL of deionizedwater. Add 79.06 g of NH4HC03, and
stir the solution gently until it dissolves.Adjust the pH to 7.6 with
NH40H or HCI and dilute to 1 L. Use immediatelyor add a 3-cm layer
of mineral oil.
2. p-nitrophenol,0.25%: Dissolve0.25 g in 100 mL of deionizedwater.
3. Sulfuric acid, 2.5 M: Add 139 mL of concentratedH2S04 (18 M) to
800 mL of deionizedwater and dilute to 1 L.
Procedure.Place 10 g of air-dried soil «2 mm) in a 250-mL Erlenmeyer
flask or bottle. Add 20 mL of extractingsolution and shakethe suspensionon a
reciprocatingshakerat 180 cycles per min for 15 min with the flask kept open.
Filter through Whatmanno. 42 or other medium-speedfilter paper. Pipette an
aliquot containing 1 to 20 ~g P into a 25-mL volumetric flask and add some
deionizedwater. Add five dropsof p-nitrophenolindicatorand adjustthe solution
pH by adding 2.5 M H2S04 slowly down the side of the flask until the indicator
just changesfrom yellow to colorless. Determine the P concentrationby the
"AscorbicAcid Method," or by the "Modified AscorbicAcid Method" if the con-
centrationof acid-labile organic P is high, or by an inductively-coupledplasma
(ICP) spectroscopy(see"Other Methods").PrepareP standardsand a blank con-
taining the samevolume of extractingsolution.
Calculatethe amountof P extractedusing Eq. [12].
Comments.Completeanalysisof P, K, Cu, Zn, Fe, and Mn can be made
by ICPS(Soltanpouret aI., 1979; Soltanpour,1991).The amountof P determined
by ICPS is highly correlatedwith that by colorimetry (Soltanpouret aI., 1979).
However,colorimetry may be more desirablewhen the P concentrationis low or
the solution containsa high organic P level. High plasmatemperaturescan oxi-
dize Po, causingan overestimateof the extractablePi.
The quantity of P extractableby NH4HC03-DTPA correlateswell with P
determinedby NaHC03, 0.01 M CaCl2 (Labhsetwar& Soltanpour, 1985), or
resin extraction(Nesse etaI., 1988),but not with the Mehlich-3 (Mehlich, 1984),
898 KUO
which contains a mixture of extractants(0.2 M CH3COOH-0.25M NH4N03-

0.015 M NH4F-0.013M HN03-O.oo1 M EDTA) for multielementextraction.
Extractionwith Anion ExchangeResin

Principles.Anion exchangeresin in aqueoussuspensionwith soil simulates
plant roots by removing the dissolvedphosphatefrom the soil solution via sur-
face adsorption.The solution P concentration,quantity of sorbedP, and temper-
ature all affect the quantity of P extractedby the resin. The rate of P adsorption
by resin is controlled by diffusion, which is a function of solution P concentra-
tion (Amer et aI., 1955; Viadyanathan& Talibundeen,1970).The resin maintains
the solution P concentrationat a low level to facilitate continuedP desorption
from the soil. The quantity and rate of P adsorptionby the resin corresopndto the
quantity and the rateof P desorptionfrom the soil (Amer et aI., 1955).
Reagents
1. Strong-baseanion exchangeresin (Dowex-l x 8, saturatedwith ap-
proximately25% Cl- and 75% HCOj"), particle size >0.4 mm and <0.8
mm. To transformthe Cl--form anion exchangeresin to one saturated
with 25% Cl- and 75% HCOj", place 1 g of Cl-- form resin in a poly-
ester bag «O.4-mm opening). Transfer the bag to a 250-mL Erlen-
meyer flaskcontaining100 mL of 0.5 M NaHC03. Shakethe suspen-
sion occasionallyfor 30 min. Repeatthe processin another100 mL of
0.5 M NaHCo3 solution, followed by two washingswith deionized
water. Keep the bag in water until use.
2. Hydrochloricacid, 0.5 M: Dilute 42 mL of concentratedHel (12 M) to
3. Sodiumbicarbonate,0.5 M: Dissolve 42.0 g of NaHC03 in deionized
Procedure.Add 4.0 g of soil «0.30mm) and a resin bag containing1 g of
anion exchangeresin saturatedwith 25% Cl- and 75% HCOj" to 100 mL of
deionizedwater in a 250-mL flask or bottle. Shakethe suspensionat an oscilla-
tion rate of about 30 cyclesper minute at constanttemperature(24°C) for 17 h.
Removeand thoroughly rinse the resin bag with deionizedwater to removesoil
particles.
Transferthe resin bag to a 250-mL Erlenmeyerflask and add 80 mL of 0.5
M HCI to extract the adsorbedP. When the releaseof CO2 has subsided,shake
the suspensionfor 30 min. Transferthe solution to a 100-mL volumetric flask.
Wash the resin bag twice with lO-mL portions of 0.5 M HCI and transfer the
washingsto the 100-mLvolumetricflask. Dilute to volume with deionizedwater.
Transfer an aliquot containing 1 to 20 Ilg P to a 25-L volumetric flask.
Determinethe P concentrationby the "Ascorbic Acid Method."
Calculate the amount of P extractedusing Eq. [12]. The volume of the
extractis 100 mL.
Comments.The amountof P extractableby anion exchangeresin is com-
parablewith isotopically exchangeableP in soils with mediumand high P levels
(Amer et aI., 1955; Wolf et aI., 1985), and NaHCOrextractableP for someacid
PHOSPHORUS 899
soils (Sharpley et aI., 1989; Wolf et aI., 1985), and NaHCOrextractableP for
some alkaline, caicareous,and acid soils (Nesseet aI., 1988; Sharpley et aI.,
1989; Wolf et aI., 1985).The recoveryof addedP by anion exchangeresin ranges
from 100%for a calcareoussoil (Bowman& Olsen,1979)to about60% for some
acid soils (Chen & Barber, 1990). Resin-extractableP is closely related to P
uptakeby plants (Amer et aI., 1955; Olsenet aI., 1983; Chen& Barber, 1990).
The quantity of P extractableby the Cl--form of the resin is somewhat
related to the solution/soil ratio which affects the salt concentrationin the bulk
solution (Sibbesen,1978).A wide ratio (1000:1)hasbeenrecommendedfor soils
with high salt contentsin order to reduce the salt effect on P extraction by the
resin (Cooke & Hislop, 1963).
The amountsof P extractableby the Cl--saturatedand the HCO) -saturat-
ed resins are similar, and both correlatewell with P uptake by wheat (Triticum
aestivum L.) seedlings(Lin et aI., 1991). However,in the HCO)-form (Sibbesen,
1978), the resin can better maintain a constant solutionpH, extractsa P quantity
that is independentof resin type and soil-waterratio, and doesnot inducethe dis-
solution of rock phosphate.However, in highly weatheredsoils, the HCO)-form
of the resin can increasethe pH to a high level that enhancesP extractability
(Sibbesen,1978; Tiessen,1989; van Raij et aI., 1986).
Anion exchangeresin also can be addeddirectly to soil suspensionswith-
out the polyesterbag. In this case,the resin beadsare separatedfrom the soil at
the completionof extractionby washingwith a streamof waterover a sieve«0.4
mm). However, this is time consumingand unsuitedfor a large-scaleoperation.
Addition of cation exchangeresin to anion exchangeresin decreasesthe solution
concentrationsof cations such as Ca and AI with a concomitantincreaseof P
extractionby the anionexchangeresin (Vaidyanathan& Talibudeen,1970; Curtin
et aI., 1987). A modified resin extraction that places both cation and anion
exchangeresinsin a bag hasbeenintroducedfor multi-elementtests(van Raij et
aI., 1986; Skogley et aI., 1990; Yang et aI., 1990, 1991).
Extraction with Iron Oxide-ImpregnatedFilter Paper

Principles. Iron oxide impregnatedon filter paperstrips provides sites to
specifically sorb P from soil solutions.In the presenceof a sufficient quantity of
impregnatediron oxide, the P concentrationin solution can be kept negligibly
low to facilitate continuedP desorptionfrom the soil (van der Zee et aI., 1987).
The rate of P desorption is first-order and is controlled by the quantity of
reversibly sorbedP. The amountof P sorbedon the iron oxide impregnatedstrips
over an extendedperiod of time is usedto index the P availability in soils.
Reagents
1. Ferric chloride, 0.6 M: Dissolve 162 g of FeCl3 • 6 H20 in deionized
2. Ammonium hydroxide,2.7 M: Dilute 365 mL of concentratedNH40H
(7.5 M) to 1 L with deionizedwater.
3. Sulfuric acid, 0.2 M: Dilute 11 mL of concentratedH2S04 (18 M) to 1
900 KUO
4. Calcium chloride, 0.005 M: Dissolve 0.735 g of CaCl2 • 2 H20 in

deionizedwater and dilute to 1 L.
Procedure
A. Preparationof iron-oxide impregnated fIlterpaperstrips.
Soak hardened,low ash filter papers(e.g., Whatmanno. 50) in 0.6 M
FeCl3 for 2 h. Removethe saturatedpaperand allow the excesssolu-
tion to drain from the treatedfilters for 5 min at room temperatureon
an inclined glasssheet.Immersefor 45 s in 2.7 M NH40H and with-
draw the paperswith constantmovementto preventuneveniron oxide
deposition.Rinsethe paperswell with deionizedwater and dry at room
temperature.Cut the filters into 2-cm wide by to-cm long strips.
B. Extractionof phosphorus
Add 1.0 g of air-dry soil «2 mm) to 40 mL of 0.005M CaCl2 solution
in a 125-mL Erlenmeyerflask. Add five iron oxide-impregnatedstrips
(2 by 10 em), and shakegently for 24 h. Removethe strips and rinse
thoroughlywith deionizedwater to removeany adhering soilparticles.
Shakethe strips with 40 mL of 0.2 M H2S04 for 4 h to dissolvethe P-
containingiron oxide. The strips can be air dried for later analysisat a
more convenienttime if desired.Transferthe solution to a tOO-mL vol-
umetric flask. Wash the strips twice with 20-mL portions of 0.2 M
H2S04 and combinethe washingswith the extractin the l00-mL volu-
metric flask. Dilute to volume with deionizedwater.
Transferan aliquot which contains1 to 20 Ilg P to a 25-mL volumetric
flask and determine the P concentration by the "Ascorbic Acid
Method." Calculatethe amountof P extractedusing Eq. [12]. The vol-
ume of extractis 100 mL.
Comments.Lin et al. (1991)showedthat a numberof different typesof fIl-

ter paperscanbe impregnatedwith iron oxide. However,the contentof iron oxide
and resulting PlFe molar ratios on the strips can vary considerably.The authors
recommendedWhatman no. 50 since the fibers of nonhardenedfilter papers
could be tom by soil particlesduring long shakingperiods.Avoid oven-dryingthe
strips, as it decreasesthe capability of the impregnatedoxide to retain P (Lin et
aI., 1991),presumablydue to increasedcrystallinity of the iron oxide. The effect
of long-termstorageon the effectivenessof the strips to retain P is unknown.
The quantity of P desorbedfrom soil onto the impregnatediron oxide is re-
lated to the numberof strips used(Lin et aI., 1991; van der Zee et aI., 1987).The
molar ratiosofPlFe rangefrom 0.07 to 0.15 dependingon the typesof fIlter paper
used,contentof impregnatedFe oxide, as well asthe concentration of desorbable
P in soils. Generally,four strips are sufficient to bring solution P concentrationto
as low as5 x to-7 mol L-l to facilitate continuedP desorptionfrom soils (van der
Zee et aI., 1987).As the transferof soil P to the oxide on the strips is sensitiveto
temperature,maintainingconstanttemperaturefor all extractionsis important.
The ioron oxide-impregnatedfilter paperP test correlatesvery well with
the anion exchangeresin test (Lin et aI., 1991) sincethe basicprinciple for both
PHOSPHORUS 901
P extractionsis similar. The filter paperP test also correlateswell with P uptake
by variousplant speciesunderglasshouseconditions(Lin et aI., 1991; Menon et
aI., 1988).
Isotopic Dilution of Phosphorus-32

Principles.The exchangeof 32p in solutionC2Pl) with solid-phasesoil 31P s
leads to adsorptionof 32p and releaseof 31p into soil solution (Olsen & Dean,
1965).
[13]
The equilibrium constant equals 1, which implies the following relationship

(Russellet aI., 1954; Olsen, 1952; Bowman& Olsen, 1979)
[14]
31p
s
= 31p • v. ( __
I 32p
1
32p.1 - - 1
[15]
I· V
where 32Pj =
initial activity (cpm/g),
32Pt= final activity (cpmlL),
31Pt = final 31p concentrationin solution (mg PIL),
31PS =
isotopically exchangeablesurfaceP (mg Pig),
=
v solution/soil ratio (L/g).
In soil systemsthat havevery low concentrationsof 31p(, 31p is often added

as a carrierwith 32p and adsorptionof the addedphosphateoccursalongwith iso-
topic exchange.Underthesecircumstances,the labile 31pS is calculatedusing Eq.
[16].
[16]
wherex = concentrationof carrier added(mg PIL),

Xi = initial P concentrationin solution (mg PIL),
Si (initial specific activity) =e2p;/x • v) (cpm/mg P)
e
St (final specific activity) = 2P1P1P1 • v) (cpm/mg P)
If the added31p (x) is much greaterthan the amountof 31p (Xi) in the original soil
solution (White, 1976), (x» Xi), Equation[16] reducesto
[17]
902 KUO
The carrier-freetechniqueis suitedfor soils with mediumto high P levels, while

the 31P-carriertechniqueis recommendedfor soils with low to medium P buffer-
ing capacities.The isotopic exchangemethod is unsuitedfor soils with high P
sorptioncapabilitiesand low solution P concentrations.
Special Apparatus
1. Geiger-MUeller(or scintillation) counter.
Reagents
1. Potassiumdihydrogen phosphate(KH 2P04), 2 x 10-5 M: Dissolve
0.136 g of KH 2P04 in deionizedwater and dilute to 1 L. Mix well.
Transfer a 20-mL aliquot of the P solution to a 1-L volumetric flask
and dilute to 1 L.
2. Phosphorus-32:Dilute a portion of 32p with 2 x 10-5 M potassium
dihydrogenphosphate(KH 2P04) to obtain a suitablespecific activity.
3. Toluene(C6H5CH3) or chloroform.
Procedure
A. Soils with moderateto high concentrationsof water-solubleP.
Add 5.0 g of soil «2 mm) and 99 mL of deionizedwater in a 250-mL
Erlenmeyerflask and shakethe suspensionat constanttemperaturefor
1 h. Add a 1-mL aliquot of carrier-free32p solution and add two drops
of C6HSCH3or CHCl3 to suppressthe microbial activity. Shakethe sus-
pensionfor 24 h and centrifugea portion of the suspension.Filter the
supernatantliquid through a O.22-llm membranefilter.
Transferan aliquot containing1 to 20 flg P to a 25-mL volumetricflask
and determinethe concentrationof P by the "Ascorbic Acid Method."
Determinethe 32p by liquid scintillation or countingthe activity with a
Geiger-MUellerend-window tube following evaporationof an aliquot
on an AI planchet.Calculatethe 31PS using Eq. [15].
B. Soils with low concentrationsof water-solubleP and low to medium P
buffering capacities
Add 5.0 g of soil and 100 mL of a 2 x 10-5 M KH 2P04 solution con-
taining a suitableamountof 32p to a 250-mL Erlenmeyerflask. Add two
dropsof C6H5CH3 or CHCI3. Shakethe suspensionfor 24 h, centrifuge,
and"determine32p and 31p as describedin the precedingparagraph.Cal~
culate 31PS using Eq. [16] or [17] if 31p addedis much higher than 31p
in the original solution.
Comments.The isotopicdilution techniquehas long beenusedto estimate
the labile P pool which includes31ps and the amountof 31p in solution. Various
investigatorsuse different ratios of solution to soil, backgroundelectrolytes,and
equilibraiton times. Solution/soil ratios which have beenusedinclude 10 (Mur-
rmann& Peech,1969; Salcedoet aI., 1991;Tran et aI., 1988),20(Olsen& Som-
mers, 1982), 25 (Wolf et aI., 1986), 50 (White & Taylor, 1977), and 200 (Vaid-
yanathan& Talibudeen,1968), with dilute potassiumchloride (KCl) (0.02 M)
(Viadyanathan& Talibudeen, 1968) or CaCl2 (0.01 M) (Murrmann & Peech,
PHOSPHORUS 903
1969) as backgroundelectrolytes. Wider solution/soil ratios (>20), however,

decreasethe quantity of labile P measured(Bowman & Olsen, 1979). Although
dilute CaCl2 solutionsdo not havea markedeffect on the estimateof the labile P
pool (Bowman & Olsen, 1979), it is consideredto be unsuitedparticularly for
acid soils with both highP buffering capacitiesand low solutionP concentrations.
IncreasedCa concentrationscan further lower P concentrationsin solution,there-
by aggravatingthe uncertaintyregardingthe accuracyof 31p, determination.
Equilibration times greaterthan 24 h may be requiredfor the 32p, and 31pS
exchangeto attain equilibrium. Much shorter exchangetimes have been used,
which include 1 min to index P sorption capacity (Salcedoet aI., 1991; Tran et
aI., 1988) and 30 min (Amer et aI., 1969) that are unlikely to be long enoughfor
equilibrium to be achieved.The measurementof labile P with a short equilibra-
tion time should specify the equilibration time period (Olsen & Khasawneh,
1980).
Measurements of labile P by isotopicexchangefor soils with high P buffer-
ing capacitiesand low solution P concentrationshave been a major challenge.
Direct 32p adsorptionby hydrousaluminumand iron oxideswould result in a sub-
stantialoverestimationof the labile P pool (Amer et aI., 1969; Wolf et aI., 1985).
The addition of 31p, in principle, should reduceor eliminate such a problem by
increasingthe soluble P concentrationto facilitate the isotopic exchange.How-
ever, for high P buffering and low P concentrationsoils, Wolf et al. (1986) found
that labile P fractions were still poorly estimatedeven with the addition of 1 x
10-4 M in 31P. The failure was attributedto the carriereffect on the kineticsof iso-
topic exchange,the difficulty of measuringsolution P and the formation of solu-
ble aluminophosphate complexes(White & Taylor, 1977). Wolfet al. (1986) rec-
ommendedusing the anion exchangeresin techniquefor this type of soil.
BUFFERING INDICES
Principles
The P buffering capacity,an inherentsoil characteristic,is closely associ-

atd with P diffusion in soil. It is definedas the ratio of the changeof the quanti-
ty of P sorbed(ap surface)to the changeof P solution concentration(ap solu-
tion), which reflectethe ability of the soil to moderatechangesin solution P con-
centrationwhen P is addedto or withdrawn by plants from soil systems.The P
buffering capacity varies with solution P concentration.It attains a maximum
value that is the productof P soprtioncapacityand the P sorptionconstantrelat-
ed to bonding energy at the solution P concentrationat which no P is sorbedor
desorbed.The buffering capacity at the natural equilibrium P concentrationis
termed the equilibrium buffering capacity, which approximatesthe maximum
buffering capacity(Holford, 1979; Keramidas& Polyzopoulos,1983).
The equilibrium P buffering capacitycan be approximatedby isotopicdilu-
tion with 32p or by resin extraction, provided that the surface P measuredby
either techniquecontrolsthe P concentrationin solution. The ratio of the isotopi-
e
cally exchangeableP C1PS ) to solution P concentration 1P l) representsan ap-
proximate measureof the equilibrium buffering capacity. The P extraction by
904 KUO
anion exchangeresin will result in a reductionof solution P concentrationto the

extentdependingon the soil type and labile P content.The equilibrium buffering
capacityby the anion exchangeresin techniqueis determinedby the ratio of the
quantity of P extractedto the changeof solution P concentrationbeforeand after
P extractions.
Methods
Equilibrium PhosphorusButTering Capacityby Isotopic Exchange

Procedure.Determinationof P concentrationin solution and the amountof
isotopically exchangeableP are outlined in the previous"Procedure"sectionfor
the carrier-freetechnique.For the carrier technique,determine31pS as outlined in
the previous "Procedure"section. The initial solution P concentrationis deter-
minedby adding5.0 g of soil and 100 mL of deionizedwater to a 2S0-mLErlen-
meyer flask. Add two drops of C6HsC1I3 or CHCl3 to suppressthe microbial
activity. Shakethe suspension for 24 h and centrifugea portion of the suspension.
Filter the supernatantliquid througha 0.22-~m membranefilter. Determinethe P
concentrationby the "Ascorbic Acid Method."
The (EBC) (mL g-l) is the ratio ofthe amountof isotopically exchangeable
e e
P 1Ps ) (~g g-l) to the P concentrationin solution 1Pt) (~g P mL- 1).
[18]
Equilibrium PhosphorusBuffering Capacityby Anion ExchangeResin

Procedure.Determinethe amountof P extractableby anoinexchange resin
accordingto the procedureoutlined in "Extraction with Anion ExchangeResin."
Savethe soil suspension.
To determinethe solution P concentrationand pH, shakethe soil suspen-
sion for 48 h, followed by determinationof the pH of the suspensionand cen-
trifugation to obtain a clear supernatantliquid. Determinethe P concentrationby
the "Ascorbic Acid Method."
To determinethe P concentrationin solution for the original soil, add 4.0 g
of soil «0.30mm) and 100 mL of deionizedwaterto a 2S0-mLErlenmeyerflask.
Mix thoroughly and adjustthe pH to a level comparablewith that of the soil sus-
pensionafter extractionwith anion exchangeresin. Shakethe suspensionfor 30
min and centrifugeto obtain a clearsupernatantliquid for P determinationby the
"Ascorbic Acid Method."
The EBC (mL g-l) is determinedby the ratio of the quantity of resin-ex-
tractableP (~g P g-l) (Ps) to the differencebetweenthe solution P concentration
(J.Lg P mL- 1) for the original soil and the solution P concentration (~g P mL- 1)
after extractionwith the resin (I!J>[).
Ps
EBC = LlPt [19]
PHOSPHORUS 905
Comments
Phosphorusbuffering capacity has been related to the amount of P

extractableby the commonly used soil tests (Holford & Mattingly, 1979; Hol-
ford, 1980; Barrow & Shaw, 1976; Mendoza& Barrow, 1987),the determination
of P fertilizer requirements(McLean et aI., 1982; Fitter, 1974; Lins & Cox, 1989,
Ozanne& Shaw,1968; Nunozawa& Tanaka,1984),and plant P uptake(Holford
& Mattingly, 1976; Nair & Mentel, 1984). It servesas a partition coefficient to
relate the distribution of P betweenthe solid and liquid phases,providing little
quantitative information relative to P availability. As such, it cannot be used
aloneto index P availability particularly amongdiversesoils (Keramidas& Poly-
zopoulos,1983; Dalal & Hallsworth, 1976; Holford, 1979; Moody et aI., 1983).
It is generallycombinedwith P quantity to reflect P intensity or with P intensity
to reflect P quantity (Moody et aI., 1988; Kuo et aI., 1988; Kuo, 1990).
The EBC determinedby isotopic dilution of 32p or by anion exchangeresin
differs from that determinedby the slopeof the tangentto the Langmuir isotherm
at the natural equilibrium P concentration(Holford, 1979). Even though P sorp-
tion by soils resemblesP desorption by plant removal (Bowman & Olson,
1985b),it is complicatedby the requirementof a P sorption isothermthat is too
time consumingto be carried out in routine soil testing. For the measurementof
the EBC by the isotopic or anion exchangetechnique,an assumptionwas made
that the labile P controls the solution P concentration.The P buffering capacity
measuredby the anion exchangetechniqueis highly correlatedwith that by the
sorption techniqueat low solution P concentrations(Moody et aI., 1988).
The P sorption isothermis rarely linear over a wide range of P concentra-
tions. A variety of P buffering capacitytests(e.g., the buffering index and sorp-
tion tests)havebeenproposedand evaluatedfor their effectivenessto accountfor
more of the variability in yield and P uptakeby plantsand to improve the deter-
mination of P fertilizer requirements(Holford, 1979; Moody et aI., 1988). The
equilibrium buffering capacityis an effective and consistentindex to increasethe
variancein P uptakeaccountedfor.
Isotopicdilution of 32p and anion exchangeare relatively simple techniques
for estimatingsoil P buffering capacity.The P buffering capacity can be deter-
mined in conjunction with P availability tests by the isotopic dilution or anion
exchangeresin technique.However, the isotopic dilution technique,even with
31p as a carrier, is not suitable for high P-fixing soils (e.g., Ultisols). For those
soils, the assumptionthat the soil reactionsare confinedto exchangebetween31p
and 32p may 110t be valid as in addition to exchangeof 32p with 31p, sorption of
32p by soil constituentscan occur as well (Amer et aI., 1969; Dalal & Hallsworth,
1977; Wolf et aI., 1986). Extraction of P by anion exchangeresin gives a better
measureof labile P comparedwith 32p isotopic dilution for high P fixing soils
(Wolf et aI., 1986), and is recommendedfor suchsoils. In addition, P sorptionor
desorptionisotherms,which is useful on a broad rangeof P buffering capacity,
can be utilized if necessary(Bowman & Olsen, 1985b; Kovar & Barber, 1988;
Moody et aI., 1988).
906 KUO
PHOSPHORUS DETERMINATION
Principles
The reaction of phosphoricacid with molybdate ions forms a heteropoly

molybdophosphate complex as follows:
The complexis yellow in color, and at high P concentrationsforms ayellow pre-

cipitate. In the presenceof V, the yellow color is intensified,which providesthe
basisfor the Vanado-molybdophosphoric acid colorimetric method.
In the presenceof reducing agents,the Mo in the molybdateor its com-
plexesis partially reducedfrom 6+ to 3+ and/or5+, which resultsin a character-
istic blue color. Severalmolybdenumblue color methodshave b~en developed,
that vary mainly in the kind of reducingagentused.They are the chlorostannous
acid-reducedmethod, the molybdenum-reducedmethod, the 1,2,4-aminonaph-
tholsulfonic acid-reducedmethod, and ascorbicacid-reducedmethod. Jackson
(1958) and Murphy and Riley (1962) give more detailed discussionon these
methods.WatanabeandOlsen(1965) modified the Murphy and Riley singlesolu-
tion methodfor soil systems.Dick and Tabatabai(1977b) modified the Murphy
and Riley methodby introducinga citrate and arsenitereagentto complexexcess
molybdateions. For solutionsthat containpolyphosphatesand/oracid-labile or-
ganicP, this methodpreventsfurther formation of blue color as thesecomponents
hydrolyzeand improvesthe accuracyof orthophosphatedetermination.
The sensitivity required, stability, and freedom from interference are
amongthe important considerationsin the selectionof a method. Although the
vanadomolybdophosphoric acid method is least sensitive, its yellow color re-
mainsstablefor a long period and the acid concentrationin the solution is not as
critical for the color development.While more sensitivethan the vanadomolyb-
dophosphroicacid method, the chlorostannousacid method gives a blue color
that is stablefor only 15 to 20 min. The ascorbicacid methodis equally sensitive
(150 Ilg L- 1 with l-cm light path) as the chlorostannousacid method, andhas a
blue color that is stable for up to 24 h. Consequently,the ascorbicacid method
hasbeenwidely usedin soil research.
Motomizu et al. (1983) developeda new spectrophotometric methodbased
on an ion associationof molybdophosphatewith Malachite Green in an acid
mediumto further increasethe sensitivity. Although the methodis suitablein the
nanogramper milliliter concentrationranges,it is unsuitedfor soil solutions in
part due to interferencefrom dissolvedsilica. Extractionof the ion associatewith
solvents{benzene-isobutylmethyl ketone[(CH3)zCHCH2COCH3]) is necessary
to overcomethe interference(Motomizu & Oshima,1987).
Special Apparatus
PHOSPHORUS 907
Colorimetric Methods
Vanadomolybdophosphoric
Acid Method
Reagents
1. Ammonium molybdate[(NH4)6M07024] (solution A): Dissolve 25 g of
reagentgrade[(NH4)6M07024• 4H20] in 400 mL of deionizedwater.
2. Ammonium vanadate (NH4V0 3) (Solution B): Dissolve 1.25 g of
N~ V03 in 300 mL of boiling water. Allow to cool to room tempera-
ture.
3. Mixed reagent:Transfersolution B to a l-L volumetric flask and slow-
ly add 250 mL of concentratedHN03 (15.8 M) with rapid stirring.
Allow the solution to cool to room temperatureand addSolution A.
Dilute the mixture to volume.
4. Sulfuric acid, 3.5 M: Slowly add 194 mL of concentratedH2S04 (18 M)
to deionized water and dilute 1 L with deionized water after it has
cooledto room temperature.
5. Working phosphatesolution, 50 mg P L- 1: Dissolve 0.2197g of oven-
dried (40°C) KH 2P04 in deionizedwater. Add 25 mL of 3.5 M H2S04,
and dilute to 1 L.
Procedure.Transferan aliquot of solution that contains100 to 900 I!g of
P to a 50-mL volumetricflask. Add 10 mL of mixed reagent.Dilute to 50 mL and
mix well. Measurethe absorbanceat a wavelengthbetween400 and 470 nm,
dependingon the P concentrationrange.
Prange Wave length

mgJL nm
1.0-5.0 400
2.0-10 420
4.0-18 470
Preparea blank that containsall reagentsexceptP solution.

Commeuts.Despite low sensitivity, the vanadomolybdophosphoric acid
methodis free from interferencesfrom a wide rangeof ionic speciesin concen-
trationsup to 1000mg L-l. The interferenceby arseniccan be eliminatedby pre-
treatingthe test solution with HBr to removeAs as AsBr3 (Jackson,1958).
Even though the method is more sensitive at 400 than at 470 nm, Fe3+
causes more interferenceat 400 than at 470 nm. If Fe3+ interferenceoccurs,use
a wave length of 470 nm that accommodatesa wider P concentrationrange
(American Public Health Association,American Water Works Association,and
Water Pollution Control Federation,1989).
ChlorostannousAcid Method
Reagents
908 KUO
2. Sulfuric acid, 3.5 M: Slowly add 194 mL of concentratedsulfuric acid

(18 M) to deionizedwater. Dilute to 1 L after it hascooledto room tem-
perature.
3. Ammonium molybdate, 1.5%: Dissolve 15 g of reagent grade
[(NH4)6M07024• 4H20] in 300 mL of deionizedwater. Slowly add 350
mL of 10.0M HCI slowlyl to the molybdatesolutionwith rapid stirring.
After the solution hascooledto room temperature,dilute to 1 L andmix
well. Preparea fresh solution every 8 wk.
4. Stannous chloride (SnCI2), 50% stock solution: Dissolve 25 g of
reagentgrade SnCl2 • 2H20 in 50 mL of concentratedHCl. Keep the
solution in a black, glass-stopperedbottle. Preparea fresh solution
every 6 wk.
5. Stannouschloride, dilute solution: Transfer1 mL of SnCl2 stock solu-
tion to 132 mL of deionizedwater and mix well. Preparea fresh solu-
tion every 2 h as neededdue to the rapid oxidation of Sn2+ in air.
6. Phosphatestocksolution,50 mg P L-1: Dissolve0.2197g of oven-dried
(40°C) KH 2P04 in deionizedwater. Add 25 mL of 3.5 M H2S04 and
dilute to 1 L. Mix the solution well.
7. Working phosphatestandardsolution, 5 mg P L- 1: Dilute 10 mL of the
50 mg P L- 1 stock solution to 100 mL with deionizedwater.
Procedure.Transferan aliquot of sampleor P standardsolution that con-
tains 2 to 40 Ilg P to a 50-mL volumetricflask. Add 10 mL of ammoniummolyb-
date solution and 1 mL of dilute SnCl2 solution. Bringto volume with deionized
water and mix well. Measurethe absorbanceat 660 nm after 5 to 6 min and be-
fore 10 min. Preparea blank that containsall of the reagentsexceptthe phosphate
solution.
Comments.The chlorostannousacid methodis not as sensitivein the hy-
drochloric acid systemas in the sulfuric acid system(Jackson,1958). However,
in the hydrochloricacid system,the methodis more tolerantof high Fe3+ (15 mg
of Fe L -1) and Cl-1 (2000 mg CI L -1) concentrations.
Interferencefrom high P- concentrations(>5 mg F L- 1) in the color forma-
tion can be eliminatedby the addition of 15 mL of 0.8 M H3B03 to form fluoro-
borate (BF,n since neither boric acid nor fluoroborate interferes with the Mo
complexformation. Extractionwith benzene-isobutanol to transferthe phospho-
molybdatecomplexinto the solventphase(AmericanPublic Health Association,
American Water Works Association, and Water Pollution Control Federation,
1989) also is an effective meansto overcomethe interference.
Ascorbic Acid Method

Reagents
1. Sulfuric acid, 2.5 M: Dilute 70 mL of concentratedH2S04 (18 M) to
500 mL with deionizedwater.
2. Ammonium molybdate:Dissolve 20 g of [(NH4)6M070z4 • 4HzO] in
500 mL of deionizedwater. Storethe solution in a glass-stoppered
bot-
tle.
PHOSPHORUS 909
3. Antimony potassiumtartrate [K(SbO) • C4H40 6 • 1/2 HzO] (1 mg Sb

mL-1): Dissolve0.2728g of K(SbO) • C4H40 6 • 1/2 HzO in 100 mL of
deionizedwater.
4. Ascorbic acid, 0.1 M: Dissolve 1.76 g of C6Hg06 in 100 mL of deion-
ized water. Preparethe solution on the day it is required.
5. Mixed reagent:Mix thoroughly 50 mL of 2.5 M HZS04, 15 mL of
ammoniummolybdatesolution, 30 mL of ascorbicacid solution and 5
mL of antimony potassiumtartrate solution. Preparea fresh mixed
reagentdaily.
6. Phosphatestocksolution,50 mg P L- 1: Dissolve0.2197g of oven-dried
(40°C) KH ZP04 in deionizedwater. Add 25 mL of 3.5 M HZS04, and
7. Working phosphatestandardsolution,Smg P L-1: Dilute 10 mL of the
50 mg P L-1 stock solution to 100 mL with deionizedwater.
Procedure.Transferan aliquot of sampleor P standardsolution that con-
tains 2 to 40 Ilg P to a 50-mL volumetric flask. Dilute with deionizedwater to
about 25 mL, and add 8 mL of mixed reagent.Dilute the solution to volume and
mix well. Measurethe absorbanceat 880 nm after 10 min. Preparea blank that
containsall reagentsexceptthe P solution.
Comments.The advantagesof the ascorbicacid method(Murphy & Riley,
1962) over the stannouschloride methodare the longer stability of the molybde-
num blue color and toleranceof high salt and Fe3+ (up to 2.5 mg L-1) concentra-
tions. Polyphosphatesand the organic P compoundswhich hydrolyze in acid
solution (Dick & Tabatabai,1977b) and arsenateconcentrationsexceeding0.05
mg L -1 all inter!'erewith P determination.The interferencefrom arsenatecan be
avoided by reducing AsO~- to AsO:~- with NaHS03 becauseAsOj- does not
develop the sameblue color as phosphate.For solutions containing acid-labile
organic P, the inorganic P determinationcan be made using the modified ascor-
bic acid method (Dick & Tabatabai,1977b) to minimize the interference.The
ascrobicmethodhas beenused extensivelyfor determiningP concentrationsin
extracts from various soil tests, acid (HCl04 or HF) digestion, and NazC03
fusion.
Watanabeand Olsen(1965) madea slight modification of the Murphy and
Riley method by introducing two mixed reagents.ReagentA contains dilute
HZS04 and the recommendedamountsof ammoniummolybdateand antimony
potassiumtartrate,which shouldbe storedin a darkand cool place. ReagentB is
preparedon the day it is neededby dissolving ascorbicacid in ReagentA.
Modified Ascorbic Acid Method

Reagents
1. ReagentA-ascorbicacid (0.1 M)-trichloroaceticacid (CzHCl 30 Z) (0.5
M): Dissolve 8.8 g of ascorbicacid and 40.9 g of CzHCl 30 z in about
400 mL of deionizedwater and dilute to 500 mL with deionizedwater.
2. ReagentB-ammoniummolybdate(0.01 M): Dissolve 6.2 g of ammo-
nium molybdatein about 400 mL of deionizedwater and dilute to 500
mL with deionizedwater.
910 KUO
3. Reagent C-sodium citrate (Na3C6Hs07) (0.1 M)-sodium arsenite

(NaAs02) (0.2 M)-acetic acid (5%): Dissolve 29.4 g of Na3C6Hs07•
2H20 and 26.0 g of NaAs02 in about 800 mL of deionizedwater, add
50 mL of glacial acetic acid (99.9%) and dilute to 1 L with deionized
water.
4. Phosphatestocksolution,50 mg P L-l: Dissolve0.2197g of oven-dried
(40°C) KH 2P04 in deionizedwater and dilute to 1 L with deionized
water.
5. Working phosphatestandardsolution, 5 mg P L-l: Dilute 10 mL of the
50 mg P L-l stock solution to 100 mL with deionizedwater.
Procedure.Add 10 mL of ReagentA to a 25-mL volumetric flask and
transferan aliquot of sampleor P standardsolution containing2 to 25 Ilg of P to
the flask. Add 2 mL of ReagentBand 5 mL of ReagentC to the flask immedi-
ately and mix the contentswell. Dilute to volume with deionizedwater and mix
well. After 10 min, measurethe absorbanceat 700 nm. Preparea blank contain-
ing all reagentsbut the P solution.
Comments.The citrate-arsenitereagent is added to remove theexcess
molybdateions. In this manner,the orthophosphatederived from the hydrolysis
of polyphosphatesand/or some acid-labile organic P can not form complexes
with molybdateions to intensify the blue color. In this regard,it is important ot
add ReagentBand C immediatelyfollowing the additionof the samplealiquot as
the interferencefrom acid labile organic P and polyphosphatehydrolysis to
orthophosphateincreasesin proportionto the delay of the addition of ReagentB
and C (Dick & Tabatabai,1977b).
Asher (1980) developedan automatedmethod for the determinationof
orthophosphatein the presenceof polyphosphates.However, replacementof
trichloroaceticacid with HCI is necessaryto preventthe formation of blue pre-
cipitate possibly due to the reaction of trichloroacetic acid with the standard
polyvinyl chloride tubing usedin the system.
Other Methods
Inductively CoupledPlasmaSpectrometry
The ICPS is commonly used for simultaneouselementalanalysisof soil
extractsand plant tissue ash (Jones,1977). The basic principles and analytical
capabilitieshavebeendescribedin detail by Soltanpouret al. (1979) and Soltan-
pour (1991). In general, the excited atoms or ions under high temperature
(5000-8000K) generatedby the interactionof the induced magneticfield with
argon plasmaproducetheir characteristicradiation lines with intensitiesin pro-
portion to their concentrations.The high temperatureand longer residencetime
allow completesamplevaporizationand atomization,therebygreatly improving
the sensitivity and detectionlimits.
The ICP equippedwith scanningmonochromatorspermits simultaneous
multielement analysis. Soltanpouret al. (1979) used ICP in conjunction with
ammoniumbicarbonate(AB)-DTPA to analyzeP and a numberof otherelements
to improve the efficiency of this soil test. The ICP presumablycan be used in
PHOSPHORUS 911
combinationwith other multielementsoil testsas well (Mehlich, 1984). Howev-

er, the P determinedby ICP spectrometryincludesboth organicand inorganicP.
If the amountof organicP that AB-DTPA extractsis high, the colorimetric meth-
ods would be more appropriatefor the determinationof inorganicP.
31-Phosphoms-Nuclear
MagneticResonanceSpectroscopy
Nuclearmagneticresonanceis a powerful tool for the nondestructivedeter-
minationof organicstructure,requiringvery small samples.Phosphoruspossess-
es a nuclearspin that can be observedby the NMR spectrometer.The magnetic
field at the nucleuschangeswith respectto the degreeof electronshieldingand
the magneticfield strengthis neededfor the resonanceto shift higher(upfield) or
lower (downfield). The chemicalshift (0, ppm) representsthe ratio of the shift
from an externalreferenceH3P04 (85%) to the field strengthof the NMR spec-
trometer.Solution and solid stateNMR have beenusedto quantify variousdis-
solvedP speciesin alkaline soil extracts(Newman& Tate, 1980; Hawkeset aI.,
1984; Condronet aI., 1985; Condronet aI., 1990) and the solid P speciationin
fish waste composts,sludges,and sludge-amendedsoils (Hinedi et aI., 1989;
Hinedi & Chang,1989; Prestonet aI., 1986).
Using NMR spectroscopy,Newmanandtate(1980) identified in the NaOH
extracts the presenceof inorganic P, orthophosphatemonoestersand diesters,
phosphonates,pyrophosphate,and polyphosphates.Inorganic P, mainly ortho-
phosphate,constitutes78 to 100% of the dissolvedP. Phosphonate,while con-
sideredto be of microbial origin, is presentonly in minutequantitiesin somesoils
at high altitudesin high precipitationzones(Hawkeset aI., 1984).
A major limitation for the applicationof solid state31p NMR to identify the
solid phaseP in soils stemsfrom the presenceof paramagneticions (e.g., Fe),
which broadenthe lines and increasethe intensity of sidebands.Hinedi et al.
(1989) and Hinedi and Chang(1989) removedthe paramagnetics from a soil that
had been enrichedwith P from continuedsludge applications,using the DeB
extraction(Jackson,1958).The treatmentsucceededin reducingthe intensity of
sidebandsand the noise level. They identified calcium phosphate,aluminum
phosphate(AlP04), andpyrophosphatein onesoil sample,andcarbonatedapatite
and pyrophosphatein anothersoil sample.
Ion Chromatography
Ion chromatography(IC) can be used for simultaneousdeterminationof
soluble ions including H2P04, N03", Cl-, and SO~- (Nieto & Frankenberger,
1985; Tabatabai& Dick, 1983). The suppressedion chromatographyis a con-
ductimetricchromatographicsystemconsistingof a precolumnto removepartic-
ulatesandpotentiallypoisonoussubstances, a separatorcolumnpackedwith low-
capacityanion agglomeratedresin in HC03" form, a suppressorcolumn packed
with a high density cation exchangeresin, and a conductivity detector.The IC
methodis compatiblewith the ascorbicacid methodof Murphy and Riley (1962)
(see"Ascorbic Acid Method") in the P concentrationrangefrom 0.02to 1.16 mg
L-l (Tabatabai& Dick, 1983) and is not subjectto interferencefrom arsenate.
Karlson and Frankenberger (1987) usedthe singlecolumn IC, which elim-
inates the suppressorcolumn to reducethe postcolumndead volume and im-
912 KUO
provesthe resolutionand efficiency of analysis.Phthalicacid (1,2-benzenedicar-

boxylic acid; CSIft;04) adjustedto pH 2.7 with formic acid replacesNaHC03and
Na2C03as the eluantin this procedure.The detectionlimit was as low as 0.3 Ilg
L -1 (Karlson & Frankenberger,1987).
REFERENCES
Adepoju,A.Y., P.F. Pratt,and S.Y. Mattigod. 1982.Availability andextractabilityof phosphorusfrom
soils having high residualphosphorus.Soil Sci. Soc. Am. J. 46:583-588.
Amer, F., D.R. Bouldin, C.A. Black, and F.R. Duke. 1955. Characterizationof soil phosphorusby
anion exchangeresin adsorptionand p32-equilibration. Plant Soil 6:391-408.
Amer, F., S. Mahdi, andA. Alradi. 1969.Limitations in isotopic measurements of labile phosphatein
soils. J. Soil Sci. 20:91-100.
AmericanPublic Health Association,AmericanWater Works Association,and Water Pollution Con-
trol Federation.1989.Phosphorus.In L.S. Clesceriet al. (ed.) Standardmethodsfor the exam-
ination of water and wastewater.17th ed. Am. Public Health Assoc.,Washington,DC.
Anderson,G. 1956. Identification and estimation of soil inositol phosphates.J. Sci. Food Agric.
7:437-444.
Anderson,G. 1960. Factorsaffecting the estimationof phosphateestersin soil. J. Sci. Food Agric.
11:497-503.
Anderson,G. 1963.Effect of iron/phosphorusratio andacid concentrationon the precipitationof fer-
ric inositol hexaphosphate. J. Sci. Food Agric. 14:352-359.
Anderson,G. 1967. Nucleic acids,derivatives,and organic phosphates.p. 67-90. In A.D. Mclaren
and G.H. Peterson(ed.) Soil biochemistry.Marcel Dekker,New York.
Anderson,G. 1975. Other organic phosphoruscompounds.p. 305-331.In J.E. Gieseking (ed.)Soil
components.Vol. 1. SpringerVerlag, New York.
Anderson,G. 1980.Assessingorganicphosphorusin soils. p. 411-431.In F.E. Khasawnehet al. (ed.)
The role of phosphorusin agriculture.ASA, CSSA, andSSSA,Madison,WI.
Anderson,G., and 1.0. Russell. 1969. Identification of inorganicpyrophosphatein alkaline extracts
of soil. J. Sci. Food Agric. 20:78-81.
Anderson,G., E.G. Williams, and J.O. Moir. 1974. A comparisonof the sorption on inorganic
orthophosphateand inositol hexaphosphate by six acid soils. J. Soil Sci. 25:51-62.
Asher, LE. 1980. An automatedmethodfor the determinationof orthophosphatein the presenceof
labile polyphosphates. Soil Sci. Soc.Am. J. 44:173-175.
Aslyng, H.C. 1964.Phosphatepotentialandphosphatestatusofsoils. Acta Agric. Scand.14:261-285.
Baker, D.E., andJ.K. Hall. 1967. Measurementsof phosphorusavailability in acid soils of Pennsylo-
vania. Soil Sci. Soc. Am. Proc. 31:662-667.
Baker, R.T. 1975. A new method for estimating the phospholipid content of soils. J. Soil Sci.
26:432-436.
Ballard, R., and W.L Pritchett. 1975. Evaluationof soil testing methodsfor predicting growth and
responseof Pinus elliottii to phosphorusfertilization. Soil Sci. Soc. Am. Proc. 39:132-136.
Barrow, N.J. 1967. Relationshipbetweenuptakeof phosphorusby plants and the phosphoruspoten-
tial and buffering capacity of the soil-an attempt to test Schofield'shypothesis.Soil Sci.
104:99-106.
Barrow, N.J., and T.C. Shaw. 1976. Sodium bicarbonateas an extractantfor soil phosphate.III.
Effectsof the buffering capacityof a soil for phosphate.Geoderma16:273-283.
Bar-Yosef, B., U. Kafkafi, R. Rosenberg,and G. Sposito.1988. Phosphotusadsorptionby kaolinite
and montmorillonite:I. effect of time, ionic strength, and pH. Soil Sci. Soc. Am. J.
52:1580-1585.
Bolan, N.S., J.K. Syers,and R.w. Tillman. 1986.Ionic strengtheffectson surfacechargeand adsorp-
tion of phosphateand sulfateby soils. J. Soil Sci. 37:379-388.
Bowman,R.A. 1988. A rapid method to determinetotal phosphorusin soils. Soil Sci. Soc. Am. J.
52:1301-1304.
Bowman, R.A. 1989. A sequentialextraction procedurewith concentratedsulfuric acid and dilute
basefor soil organicphosphorus.Soil Sci. Soc. Am. J. 53:362-366.
Bowman,R.A., and C.V. Cole. 1978.An exploratorymethodfor fractionationof organicphosphorus
from grasslandsoils. Soil Sci. 125:95-101.
PHOSPHORUS 913
Bowman, R.A, and S.R. Olsen. 1979. A reevaluationof phosphorus-32and resin methodsin a cal-
careoussoil. Soil Sci. Soc. Am. J. 43:121-124.
Bowman, R.A, and S.R. Olsen. 1985a. Assessmentof phosphate-bufferingcapacity: 1. Laboratory
methods.Soil Sci. 140:287-29l.
Bowman, RA, and S.R Olsen. 1985b. Assessmentof phosphatebuffering capacity:2. Greenhouse
methods.Soil Sci. 140:387-392.
Bray, R.H., and L.T. Kurtz. 1945. Determinationof total, organic,and availableforms of phosphorus
in soils. Soil Sci. 59:39-45.
Bromfield, S.M. 1967. An examinationof the useof ammoniumfluoride as a selectiveextractantfor
aluminum-boundphosphatein partially phosphatesystems.Aus!. J. Soil Res. 5:225-234.
Brookes,P.e., and D.S. Powlson. 1981. Preventingphosphoruslossesduring perchloric acid diges-
tion of sodium bicarbonatesoil extracts.J. Sci. Food Agric. 32:671-{i74.
Brookes, P.e., D.S. Powlson,and D.S. Jenkinson.1982. Measurementof microbial biomassphos-
phorusin soil. Soil BioI. Biochem. 14:319-329.
Bruce, R.e., and 1.1. Bruce. 1972.The correlationof soil phosphorusanalyseswith responseof trop-
ical pasturesto superphosphate on some north Queenslandsoils. Aus!. J. Exp. Agric. Anim.
Husb. 12:188-194.
Caldwell, AG., and e.A Black. 1958a. Inositol hexaphosphate: l. Quantitative determination in
extractsof soils and manures.Soil Sci. Soc. Am. Proc. 22:290-293.
Caldwell, AG., and C.A. Black. 1958b. Inositol hexaphosphate:II. Synthesisby soil microorgan-
isms. Soil Sci. Soc. Am. Proc. 22:293-296.
Caldwell, AG., and e.A Black. 1958c. Inositol hexaphosphate: Ill. Contentin soils. Soil Sci. Soc.
Am. Proc. 22:296-298.
Chae, Y.M., and M.A Tabatabai.1981. Sulfolipid and phospholipidin soils and sewagesludgesin
Iowa. Soil Sci. Soc. Am. 1. 45:20-25.
Chang,S.e., and M.L. Jackson.1957. Fractionationof soil phosphorus.Soil Sci. 84:133-144.
Chen, J.H., and S.A Barber. 1990. Effect of liming and adding phosphateon predictedphosphorus
uptakeby maize on acid soils of three orders.Soil Sci. 150:844-849.
Chien, S.H. 1978. Interpretationof Bray-l extractablephosphorusfrom acid soil treatedwith phos-
phaterocks. Soil Sci. 126:34-39.
Chien, S.H., L.A Leon, and H.R. Tejeda. 1980. Dissolutionof North Carolinaphosphaterock in acid
Colombiansoils as relatedto soil properties.Soil Sci. Soc. Am. J. 44:1267-1271.
Chien, S.H., L.A Leon, and H.R Tejeda. 1980. Dissolutionof North Carolinaphosphaterock in acid
Colombiansoils as relatedto soil properties.Soil Sci. Soc. Am. 1. 44:1267-1271.
Condron, L.M., K.M. Goh, and R.H. newman. 1985. Nature and distribution of soil phosphorusas
revealedby a sequentialextractionmethodfollowed by 31 P nuclearmagneticresonanceanaly-
sis. 1. Soil Sci. 36:199-207.
Condron,L.M., E. Frossard,H. Tiessen,R.H. Newman,and 1.W. B. Stewart. 1990. Chemicalnature
of organic phosphorusin cultivated and uncultivatedsoils underdifferent environmentalcon-
ditions.l. Soil Sci. 41:41-50.
Condron,L.M., 1.0. Moir, H. Tiessen,and 1.W.B. Stewart. 1990. Critical evaluationof methodsfor
determiningtotal organicphosphorusin tropical soils. Soil Sci. Soc. Am. 1. 54:1261-1266.
Cooke, I.J., and 1. Hislop. 1963. Use of anion-exchangeresin for the assessmentof available soil
phosphate.Soil Sci. 96:308-312.
Cosgrove,DJ. 1963. The chemicalnatureof soil organicphosphorus.1. Inositol phosphates.Aus!. 1.
Soil Res. 1:203-214.
Cowell, 1.D. 1963. The estimationof phosphorusfertilizer requirementsof wheat in southernNew
South Wales by soil analysis.Aus!. 1. Exp. Agric. Anim. Husb. 3:190-197.
Curtin, D., 1.K. Syers,and G.W. Smillie. 1987. The importanceof exchangeablecationsand resin-
sink characteristicsin the releaseof soil phosphorus.1. Soil Sci. 38:711-716.
Dalal, RC. 1977. Soil organic phosphorus.Adv. Agron. 29:83-117.
Dalal, Re., and E.G. Hallsworth. 1976. Evaluationof the parametersof soil phosphorusavailability
factors in predictingyield responseand phosphorusuptake.Soil Sci. Soc. Am. 1. 40:541-546.
Dalal, R.e., and E.G. Hallsworth. 1977. Measurementof isotopic exchangeablesoil phosphorusand
interrelationshipamong parametersof quantity, intensity, and capacity factors. Soil Sci. Soc.
Am. 1. 41:81-86.
De Serra,M.D., and M. Schnitzer.1972. Extractionof humic acid by alkali and chelatingresin. Can.
1. Soil Sci. 52:365-374.
Dick, W.A, and M.A Tabatabai.1977a. An alkaline oxidation method for determinationof total
phosphorusin soils. Soil Sci. Soc. Am. 1. 41:511-514.
914 KUO
Dick, W.A, and M.A Tabatabai.1977b.Determinationof orthophosphatein aqueoussolutionscon-

taining labile organic and inorganicphosphoruscompounds.J. Environ. Qual. 6:82-85.
Dormaar,J.F. 1967. Distribution of inositol phosphatesin somechernozemicsoils of southernAlber-
ta. Soil Sci. 104:17-24.
Dormaar,J.F. 1970. Phospholipidsin chernozemicsoils of southernAlberta. Soil Sci. 110:136-139.
Dormaar,J.F., and G.R Webster.1964. Lossesinherentin ignition proceduresfor determiningtotal
organicphosphorus.Can. J. Soil Sci. 44:l--Q.
Fife, C.v. 1962. An evaluationof ammoniumfluoride as a selectiveextractantfor aluminum-bound
soil phosphate:IV. Detailedstudieson soils. Soil Sci. 96:112-120.
Fitter, AH. 1974.A relationshipbetweenphosphorusrequirement,the immobilizationof addedphos-
phate,and the phosphatebuffering capacityof Colliery shales.J. Soil Sci. 25:41-50.
Fox, R.L., and E.I. Kamprath. 1970. Phosphatesorption isotherms for evaluating the phosphate
requirementsof soils. Soil Sci. Soc. Am. Proc. 34:902-907.
Fried, M., and R.E. Shapiro.1956. Phosphatesupply patternof varioussoils. Soil Sci. Soc.Am. Proc.
20:471-475.
Goring, C.AI., and w.v. Bartholomew. 1949. Microbial productsand soil organic matter: 2. The
effect of clay on the decompositionand separationof the phosphoruscompoundsin microor-
ganisms.Soil Sci. Soc. Am. Proc. 14:152-156.
Goring, C.AI., and W.v. Bartholomew. 1952. Adsorption of mononucleotides,nucleic acids, and
nUcleoproteinsby clays. Soil Sci. 74:149-164.
Greaves, M.P., and D.M. Webley. 1969. The hydrolysis of myoinositol hexaphosphateby soil
microorganisms.Soil BioI. Biochem. 1:37-43.
Gupta, RK, R.R. Singh, and KK Tanji. 1990. Phosphorusreleasein sodium ion dominatedsoils.
Soil Sci. SOc. Am. J. 54:1254-1260.
Halstead,R.L., and G. Anderson. 1970. Chromatographicfractionation of organic phosphatesfrom
alkali, acid, and aqueousacetylacetoneextractsof soils. Can. J. Soil Sci. 50:111-119.
Halstead,R.L., G. Anderson,and N.M. Scott. 1966.Extractionof organicmatterfrom soils by means
of ultrasonicdispersionin aqueousacetylacetone.Nature (London) 211:1430-1431.
Hance,R.J.,and G. Anderson.1962.A comparativestudy of methodsof estimatingsoil organicphos-
phorus.J. Soil Sci. 13:225-230.
Hance, R.I., and G. Anderson. 1963a. Extraction and estimation of soil phospholipids.Soil Sci.
96:94-98.
Hance,R.I., and G. Anderson.1963b.Identificationof hydrolysisproductsof soil phospholipids.Soil
Sci. 96:157-161.
Hawkes,G.E., D.S. Powlson,E.W. Randall, and KR Tate. 1984. A 31p nuclearmagneticresonance
study of the phosphorusspeciesin alkali extractsof soil from long-term field experiments.J.
Soil Sci. 35:35-45.
Hedley, M.J., and J.W.B. Stewart. 1982. Method to measuremicrobial phosphatein soils. Soil BioI.
Biochem. 14:377-385.
Hedley, M.I., R.E. White, and P.H. Nye. 1982. Plant-inducedchangesin the rhizosphereof rape
(Brassica Napus var. Emerald) seedlings.III. Changesin L-value, soil phosphorusfractiona-
tions, and phosphataseactivity. New Phytol. 91:45-56.
Hinedi, Z.R., and AC. Chang. 1989. Solubility and phosphorus-31magic angle spinning nuclear
magnetic resonance of phosphorus in sludge-amendedsoils. Soil Sci. Soc. Am. J.
53:1057-1061.
Hinedi, Z.R., AC. Chang,and J.P. Yesinowski. 1989. Phosphorus-31magic angle spinning nuclear
magneticresonanceof wastewatersludgesand sludge-amendedsoil. Soil Sci. Soc. Am. J.
53:1053-1056.
Holford, I.C.R 1979. Evaluationof soil phosphatebuffering indices.Aus!. J. Soil Res. 17:495-504.
Holford, I.C.R. 1980. Greenhouseevaluationof four phosphorussoil tests in relation to phosphate
buffering and labile phosphatein soils. Soil Sci. Soc. Am. J. 44:555-559.
Holford, I.C.R, and G.E.G. Mattingly. 1976. Phosphateadsorptionand plant availability of phos-
phate.Plant Soil 44:377-389.
Holford, I.C.R,and G.E.G. Mattingly. 1979.Effectsof phosphatebuffering on the extractionof labile
phosphateby plantsand by soil tests.Aus!. J. Soil Res. 17:511-514.
Hong, J.K, and I. Yamane.1980. Proposalfor a more suitablemethodto extract soil organic phos-
phorus.Soil Sci. Plant Nutr. 26:383-390.
Jayachandran,K, AP. Schwab, and B.AD. Hetrick. 1992. Partitioning dissolved inorganic and
organic phosphorus using acidified molybdate and isobutanol. Soili Sci. Soc. Am. J.
56:762-765.
PHOSPHORUS 915
Jones,1.B., Jr. 1977.Elemental analysis of soil extractsand plant tissueash by plasmaemissionspec-

troscopy.Commun.Soil Sci. Plant Anal. 8:349-365.
Jungk,A. 1987. Soil-root interactionsin the rhizosphereaffecting plant availability of phosphorus.1.
Plant Nutr. 10:1197-1204.
Kaila, A., and O. Virtanen. 1955.Detenninationof organicphosphorusin samplesof peatsoils. 1. Sci.
Agric. Soc. Finland 27:104-115.
Kamprath, E.J., and M.E. Watson. 1980. Conventionalsoil and tissue tests for assessingthe phos-
phorusstatusof soils. p. 433-469.In EE. Khasawnehet al. (ed.) The role of phosphorusin
agriculture.ASA, CSSA, and SSSA,Madison,WI.
Karlson, U., and W.T. Frankenberger,Jr. 1987. Single column ion chromatography:III. Detennina-
tion of orthophosphatein soils. Soil Sci. Soc. Am. 1. 51:72-74.
Keramidas,V.Z., and N.A. Polyzopoulos.1983. Phosphorusintensity, quantity, and capacity factors
of representativeAlfisols of Greece.Soil Sci. Soc. Am. J. 47:232-236.
Kovar, J.L., and S.A. Barber. 1988. Phosphorussupply characteristicsof 33 soils as influenced by
sevenratesof phosphorusaddition. Soil Sci. Soc. Am. J. 52:160-165.
Kowalenko,C.G., and RB. McKercher. 1970.An examinationof methodsfor extractionof soil phos-
pholipids. Soil BioI. Biochem. 2:269-273.
Kuo, S. 1990. Phosphatesorption implications on phosphatesoil testsand uptake by com. Soil Sci.
Soc. Am. J. 54:131-135.
Kuo, S. 1991. Phosphatebuffering and availability in soils. TrendsSoil Sci. 1:203--214.
Kuo, S., and E.J. JelJum.1987. Influenceof soil characteristics and environmentalconditionson sea-
sonal variation of water-solublephosphatein soils. Soil Sci. 143:257-263.
Kuo, S., E.J. JelJum,and W.L. Pan. 1988. Influenceof phosphatesorption parametersof soils on the
desorptionof phosphateby variousextractants.Soil Sci. Soc. Am. J. 52:974-979.
Labhsetwar,V.K., and P.N. Soltanpour.. 1985. A comparisonof NH4HCOr DTPA, NaHC03, CaCI2,
and Na2-EDTA soil testsfor phosphorus.Soil Sci. Soc. Am. J. 49:1437-1440.
Lin, T.H., S.B. Ho, and K.H. Houng. 1991. The use of iron oxide-impregnatedfilter paper for the
extractionof availablephosphorusfrom Taiwan soils. Plant Soil 133:219-226.
Lindsay, w.L. 1979. Chemicalequilibria in soils. JohnWiley & Sons,New York.
Lindsay, W.L., and P.L.G. Vlek. 1977. Phosphateminerals.p. 639-672.In 1.B. Dixon and S.B. Weed
(ed.) Minerals in soil environments.SSSABook Ser. 1. SSSA,Madison,WI.
Lins, I.D.G., and ER Cox. 1989. Effect of extractantand selectedsoil propertieson predicting the
optimum phosphorusfertilizer rate for growing soybeansunder field conditions. Commun.
Lins, I.D.G., ER. Cox, and 1.1. Nicholaides,III. 1985. Optimizing phosphorusfertilization ratesfor
soybeansgrown on Oxisols and associatedEntisols. Soil Sci. Soc. Am. 1. 49:1457-1460.
Mackay,AD., 1.K. Syers,P.E.H. Gregg,and R.W. Tillman. 1984.A comparisonof 3 soil-testingpro-
ceduresfor estimatingthe plant availability of phosphorusin soils receiving either super-
phosphateor phosphaterock. N.Z.J. Agric. Res. 27:231-245.
Martin, 1.K. 1964. Soil organicphosphorus.1. Methodsfor the extractionand partial fractionationof
soil organicphosphorus.N.Z.J. Agric. Res. 7:723--735.
Martin, 1.K., and A.J. Wicken. 1966. Soil organicphosphorus.IV. Fractionationof organicphospho-
rus in alkaline soil extractsand the identification of inositol phosphates.N .Z.J. Agrjc. Res.
9:529--535.
Matar, A.E., S. Garabed,S. Riahi, and A. Mazid. 1988. A comparisonof four soil test proceduresfor
detenninationof available phosphorusin calcareoussoils of the Mediterraneanregion. Com-
Mattingly, G.E.G. 1970. Total phosphoruscontentsof soils by perchloric acid digestion and sodium
bicarbonatefusion. J. Agric. Sci. (Cambridge)74:79--82.
McKercher, R.B., and G. Anderson. 1968a.Contentof inositol penta-and hexaphosphates in some
Canadiansoils. J. Soil Sci. 19:47-55.
McKercher,RB., and G. Anderson.1968b.Characterizationof the inositol penta-and hexaphosphate
fractions of a numberof Canadianand Scottishsoils. 1. Soil Sci. 19:302-310.
McLean,E.O., T.O. OIoya, andS. Mostaghimi.1982.Improved correctivefertilizer recommendations
basedon a two-stepalternativeusageof soil tests:I. Recoveryof soil-equilibratedphospho-
rus. Soil Sci. Soc. Am. J. 46:1193--1197.
Mehlich, A 1978.Influenceof fluoride, sulfate,andacidity on extractablephosphorus,calcium, mag-
nesiumand potassium.Commun.Soil Sci. Plant Anal. 9:455-476.
Mehlich, A. 1984. Mehlich III soil test extractant:A modification of Mehlich II extractant.Comm.
Soil Sci. Plant Anal. 15:1409--1416.
Mehta, N.C., J.O. Legg, c.A.1. Goring, and C.A Black. 1954. Detenninationof organic phosphorus
in soils. 1. Extraction method.Soil Sci. Soc. Am. Proc. 18:443--449.
916 KUO
Mendoza,R.E., and N.J. Barrow. 1987. Ability of three extractantsto reflect the factors that deter-
mine the availability of soil phosphate.Soil Sci. 144:319-329.
Menon,RG., L.L. Hammond,and H.A. Sissingh.1988.Determinationof plant-availablephosphorus
by the iron hydroxide-impregnated filter paper(Pi) soil test. Soil Sci. Soc.Am. J. 52:110-115.
Moody, P.W., G.F. Haydon,andT. Dickson. 1983.Mineral nutrition of soybeansgrown in the South
Burnett region of south-eastern Queensland.2. Predictionof grain yield responseto phospho-
rus with soil tests.Aust. J. Exp. Agric. Anim. Husb. 23:38-42.
Moody, P.W., R.L. Aitken, B.L. Compton,and S. Hunt. 1988. Soil phosphorusparametersaffecting
phosphorusavailability to, andfertilizer requirementsof, maize(Zea mays). Aust. J. Soil Res.
26:611-622.
Motomizu, S., and M. Oshima.1987.Spectrophotometric determinationof phosphorusasorthophos-
phatebasedon solvent extractionof the ion associateof molybdophosphatewith malachite
greenusing flow injection. Analyst 112:295-300.
Motomizu, S., T. Wakimoto, and K Toei. 1983. Spectrophotometricdeterminationof phosphatein
river waterswith molybdateand malachitegreen.Analyst 108:361-367.
Murphy, J., and H.P. Riley. 1962.A modified single solution methodfor the determinationof phos-
phatein naturalwaters.Anal. Chim. Acta 27:31-36.
Murrmann,RP., and M. Peech.1969.Effect of pH on labile and solublephosphatein soils. Soil Sci.
Soc.Am. Proc. 33:205-210.
Nair, KP.P., and K Mengel. 1984. Importanceof phosphatebuffer power for phosphateuptake by
rye. Soil Sci. Soc.Am. J. 48:92-95.
Nelson,W.L., A Mehlich, and E. Winters. 1953. The development,evaluation,and use of soil tests
for phosphorusavailability. Agronomy 4:153-188.
Nesse,P., J. Grava, and P.R. Bloom. 1988. Correlation of severaltests for phosphoruswith resin-
extractablephosphorusfor 30 alkaline soils. Commun.Soil Sci. PlantAnal. 19:675-689.
Newman,RH., and K.R Tate. 1980.Soil phosphoruscharacterizationby 31p nuclearmagneticreso-
nance.Commun.Soil Sci. PlantAnal. 11:835-842.
Nieto, KF,. andW.T. Frankenberger, Jr. 1985.Singlecolumnion chromatography:I. Analysisof inor-
ganic anionsin soils. Soil Sci. Soc.Am. J. 49:587-592.
Nunozawa,F., and A Tanaka.1984. Comparisonof nutrient managementamongacid soils differing
in phosphorusfixation capacityand cation exchangecapacity.Soil Soil Plant Nutr. 30:51-61.
Olsen, S.R. 1952. Measurementof surfacephosphateon hydroxylapatiteand phosphaterock with
radiophosphorus. J. Phys.Chern.56:630-632.
Olsen,S.R, C.V. Cole, F.S. Watanabe,and L.A. Dean. 1954. Estimationof availablephosphorusin
soils by extractionwith sodiumbicarbonate.USDA Circ. 939. USDA, Washington,De.
Olsen,S.R, and L.A Dean. 1965. Phosphorus.p. 1035-1099.In C.A Black et al. (ed.) Methodsof
soil chemicalanalysis.Part 2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Olsen,S.R, and F.E. Khasawneh.1980. Use and limitations of physical-chemicalcriteria for assess-
ing the statusof phosphorusin soils. p. 361-410.In F.E. Khasawnehet al. (ed.) The role of
phosphorusin agriculture.ASA, CSSA,and SSSA,Madison,WI.
Olsen,S.R, and L.E. Sommers.1982. Phosphorus.p. 403-430.In AL. Pageet al. (ed.) Methodsof
Olsen,S.R, and F.S. Watanabe.1970.Diffusive supplyof phosphorusin relationto soil textural vari-
ations.Soil Sci. 110:318-327.
Olsen,S.R, F.S. Watanabe,and R.A Bowman. 1983. Evaluationof fertilizer phosphateresiduesby
plant uptakeand extractablephosphorus.Soil Sci. Soc.Am. 1. 47:952-958.
Omotoso,T.I., andA Wild. 1970a.Contentof inositol phosphates in someEnglishand Nigeriansoils.
J. Soil Sci. 21:216-223.
Omotoso,T.I., and A Wild. 1970b.Occurrenceof inositol phosphatesand other organicphosphate
componentsin an organiccomplex.1. Soil Sci. 21:224-232.
Ozanne,P.G., and T.e. Shaw. 1968. Advantagesof the recently developedphosphatesorption test
over the older extractant methods for soil phosphate.Trans. 9th Int. Congr. Soil Sci.
2:273-280.
Peterson,G.W., and R.B. Corey. 1966. A modified Changand Jacksonprocedurefor routine frac-
tionationof inorganicsoil phosphates.Soil Sci. Soc. Am. Proc. 30:563-565.
Pierzynski,G.M., T.J. Logan;S.J.Traina,andJ.M. Bigham. 1990. Phosphoruschemistryand miner-
alogy in excessivelyfertilized soils: Quantitativeanalysisof phosphorus-richparticles.Soil
Sci. Soc. Am. J. 54:1576-1583.
Pratt,P.F.,andM.J. Garber.1964.Correlations ofphosphorusavailability by chemicaltestswith inor-
ganicphosphorusfractions.Soil Sci. Soc.Am. Proc. 23:23-26.
PHOSPHORUS 917
Preston,C.M., 1.A Ripmeester,S.P. Mathur, and M. Levesque.1986. Application of solutoin and

solid-statemultinuclearNMR to a peat-basedcompostingsystemfor fish and crabscrap.Can.
J. Spectrose.31:6J-.{}9.
Randall, G.w., and 1. Grava. 1971. Effect of soil: Bray no. 1 ratios on the amount of phosphorus
extractedfrom calcareousMinnesotasoils. Soil Sci. Soc. Am. Proc. 35:112-114.
Reith, 1.W.S.,R.H.E. Inkson, N.M. Scott, K.S. Caldwell, JAM. Ross,andW.E. Simpson.1987. Esti-
matesof soil phosphorusfor different soil series.Fert. Res. 11:123-142.
Russell,R.S.,1.B. Rickson,and S.N. Adams. 1954. Isotopic equilibria between phosphates in soil and
their significancein the assessment of fertility by tracermethods.J. Soil Sci. 5:85-105.
Ryan,J., H.M. Hasan,M. Baasiri,and H.S. Tabbara.1985.Availability and transformationof applied
phosphorusin calcareousLebanesesoils. Soil Sci. Soc. Am. J. 49:1215-1220.
Ryden,J.c., and J.K. Syers.1977. Origin of the labile phosphatepool in soils. Soil Sci. 123:353-361.
Salcedo,LH., E Bertino, and E.Y.S.B. Sampaio. 1991. Reactivity of phosphorusin northeastern
Brazilian soils assessed by isotopic dilution. Soil Sci. Soc. Am. J. 55:140-145.
Saunders,W.M.H., and E.G. Williams. 1955.Observationson the determinationof total organicphos-
phorusin soils. J. Soili Sci. 6:254-267.
Schofield, R.K. 1955. Can a precise meaning be given to 'available' soil phosphorus?Soils Fert.
18:373-375.
Sharpley,AN., D. Curtin, and J.K. Syers. 1988. Changesin water-extractabilityof soil inorganic
phosphateinducedby sodium saturation.Soil Sci. Soc. Am. J. 52:637-640.
Sharpley,AN., and SJ. Smith. 1985. Fractionationof inorganicand organicphosphorusin virgin and
cultivated soils. Soil Sci. Soc. Am. 1. 49:127-130.
Sharpley,AN., U. Singh, G. Uehara, and J. Kimble. 1989. Modeling soil and plant phosphorus
dynamicsin calcareousand highly weatheredsoils. Soil Sci. Soc. Am. J. 53:153-158.
Sherrell,C.G., and W.M.H. Saunders.1966. An evaluationof methodsfor the determinationof total
phosphorusin soils. N.ZJ. Agric. Res. 9:972-979.
Shuman,L.M., P.L. Raymer,J.L. Day, and M.J. Cordonnier.1988. Comparisonof four phosphorus
extraction methods on three acid southeasternsoils. Commun. Soil Sci. Plant Anal.
19:579-595.
Sibbesen,E. 1978.An investigationof the anion-exchangeresin methodfor soil phosphateextraction.
Plant Soil 50:305-321.
Skogley,E.O., SJ. Georgitis,J.E. Yang, and B.E. Schaff. 1990. The phytoavailability soil tests-PST.
Smillie, G.w., D. Curtin, and J.K. Syers. 1987. Influence of exchangeablecalcium on phosphate
retentionby weakly acid soils. Soil Sci. Soc. Am. J. 51:1169-1172.
Smillie, G.W., and 1.K. Syers.1972. Calcium fluoride formation during extractionof calcareoussoil
with fluoride: II. Implications to the Bray-l test. Soil Sci. Soc. Am. Proc. 36:25-30.
Smith, D.H., and EE. Clark. 1951. Anion-exchangechromatographyof inositol phosphatesfrom soil.
Soil Sci. 72:353-360.
Soltanpour,P.N. 1991. Determinationof nutrient availability andelementaltoxicity by AB-DTPA soil
test and ICPS. Adv. Soil Sci. 16:165-190.
Soltanpour,P.N., E Adams,and AC. Bennett.1974. Soil phosphorusavailability as measuredby dis-
placed soil solutions,calcium-chlorideextracts,dilute-acid extracts,and labile phosphorus.
Soltanpour,P.N., R.L. Fox, and R.c. Jones.1987.A quick methodto extractorganicphosphorusfrom
Soltanpour,P.N., and A.P. Schwab.1977. A new soil test for simultaneousextractionof macro- and
micro-nutrientsin alkaline soils. Comm. Soil Sci. Plant Anal. 8:195-207.
Soltanpour,P.N., S.M. Workman, and AP. Schwab.1979. Use of inductively-coupledplasmaspec-
trometry for the simultaneousdeterminationof macro- and micro-nutrients in NH4HCOr
DTPA extractsof soils. Soil Sci. Soc. Am. J. 43:75-78.
Sommers,L.E., and D.W. Nelson. 1972.Determinationof total phosphorusin soils: A rapid perchlo-
ric acid digestionprocedure.Soil Sci. Soc. Am. 36:902-904.
Stevenson,FJ. 1982. Humuschemistry.John-Wiley & Sons,New York.
Steward,J.H., and M.E. Tate. 1971. Gel chromatographyof soil organicphosphorus.J. Chromatogr.
60:75-82.
Stewart, J.w.B., and H. Tiessen. 1987. Dynamics of soil organic phosphorus.Biogeochemistry
4:41--{)0.
Stoop,W.A 1983. Phosphateadsorptionmechanismsin oxidic soils: Implications for P-availability
to plants. Geoderma31:57--{)9.
918 KUO
Stott, D.E., and M.A. Tabatabai.1985. Identification of phopholipidsin soils and sewagesludgesby
high-performanceliquid chromatography.J. Environ. Qual. 14:107-110.
Susuki,A., K Lawton, and E.C. Doll. 1963. Phosphorusuptakeand soil testsas relatedto forms of
phosphorusin someMichigan soils. Soil Sci. Soc. Am. Proc. 27:401-403.
Syers,1.K, 1.D.H. Williams, A.S. Campbell, and T.W. Walker. 1967. The significance of apatite
inclusionsin soil phosphorusstudies.Soil Sci. Soc. Am. Proc. 31:752-756.
Syers,J.K., J.D.H. Williams, and T.W. Walker. 1968. The determinationof total phosphorusin soils
and parentmaterials.N.ZJ. Agric. Res. 11:757-762.
Tabatabai,M.A., and w.A. Dick. 1983. Simultaneousdeterminationof nitrate, chloride, sulfate,and
phosphatein natural watersby ion chromatography.J. Environ. Qual. 12:209-213.
Tate, KR. 1984. The biological transformationof P in soil. Plant Soil 76:245-256.
Tate, KR, and RH. Newman. 1982. Phosphorusfractions of a c1imosequenceof soils in New
Zealandtussockgrassland.Soil BioI. Biochem. 14:191-196.
Thomas,G.W., and D.W. Peaslee.1973. Testingsoils for phosphorus.p. 115-132.In L.M. Walsh and
J.D. Beaton(ed.) Soil testing and plant analysis.SSSABook Ser. 3. SSSA,Madison,WI.
Thomas,RL., and D.L. Lynch. 1960. Quantitativefractionationof organicphosphoruscompoundsin
someAlberta soils. Can. 1. Soil Sci. 40:113-120.
Thompson,EJ., A.L.E Oliviera, U.S. Moser, and c.A. Black. 1960. Evaluationof laboratoryindex-
es of absorptionof soil phosphorusby plants. II. Plant Soil 13:28-38.
Tiessen,H., 1.w.B. Stewart,and1.0. Moir. 1983. Changesin organicand inorganicphosphoruscom-
position of two grasslandsoils and their particle size fractions during 60-90 yearsof cultiva-
tion. J. Soil Sci. 34:815-823.
Tran, T. Sen,J.C. Fardeau,and M. Giroux. 1988. Effects of soil propertieson plant-availablephos-
phorus determinedby the isotopic dilution phosphorus-32method. Soil Sci. Soc. Am. J.
52:1383-1390.
Vaidyanathan,L.v., and O.Talibudeen.1968. Rate controlling processesin the releaseof soil phos-
phate.J. Soil Sci. 19:342-353.
Vaidyanathan,L. v., and O.Talibudeen.1970. Rateprocessesin the desorptionof phosphatefrom soils
by ion-exchangeresins.J. Soil Sci. 21:173-183.
van der Paauw,E 1971.An effective water extractionmethodfor the determinationof plant available
soil phosphorus.Plant Soil 34:467-481.
van der Zee,S.E.A.T.M., L.G.]. Fokkink, and W.H. van Riemsdijk. 1987.A new techniquefor assess-
ment of reversibly adsorbedphosphate.Soil Sci. Soc. Am. J. 51:599-604.
van Diest, A. 1963. Soil test correlationstudieson New Jerseysoils: Comparisonof sevenmethods
for measuringlabile inorganicsoil phosphorus.Soil Sci. 96:261-266.
van Lierop, W. 1988. Determinationof available phosphorusin acid and calcareoussoils with the
Kelowna multiple-elementextractant.146:284-291.
van Raij, B., J.A. Quaggio,and N.M. da Silva. 1986. Extraction of phosphorus,potassium,calcium,
and magnesiumfrom soils by an ion-exchangeresin procedure.Commun.Soil Sci. PlantAnal.
17:547-566.
Walbridge,M.R, and P.M. Vitousek. 1987.Phosphorusmineralizationpotentialsin acid organicsoils:
Processaffecting 32PO,t-isotopedilution measurements. Soil BioI. Biochem. 19:709-717.
Walker, T.W., and A.ER. Adams. 1958. Studieson soil organic matter: 1. Influence of phosphorus
contentof parent materialson accumulationsof carbon,nitrogen, sulfur, and organic phos-
phorusin grasslandsoils. Soil Sci. 85:307-318.
Watanabe,ES., and S.R. Olsen. 1965. Test of an ascorbicacid methodfor determiningphosphorusin
water and NaHC03 extractsfrom soil. Soil Sci. Soc. Am. Proc. 29:677~78.
Welch, L.E, L.E. Ensminger,and C.M. Wilson. 1957. The correlation of soil phosphoruswith the
yields of Ladino clover. Soil Sci. Soc. Am. Proc. 21:61~20.
White, RE. 1976. Conceptsand methodsin the measurement of isotopically exchangeable
phosphate
in soil. PhosphorusAgric. 67:9-16.
White, RE., and P.H.T. Beckett. 1964. Studieson the phosphatepotentialsof soils. I. The measure-
ment of phosphatepotential. Plant Soil 20:1-16.
White, RE., and A. W. Taylor. 1977. Effect of pH on phosphateadsorptionand isotopic exchangein
acid soils at low and high additionsof soluble phosphate.J. Soil Sci. 28:4~1.
Wild, A. 1964. Soluble phosphatein soil and uptakeby plants.Nature (London) 203:326-327.
Williams, J.D.H., T. Mayer, and J.O. Nriagu. 1980.Extractability of phosphorusfrom phosphatemin-
eralscommonin soils and sediments.Soil Sci. Soc. Am. J. 44:462-465.
Williams, J.D.H., J.K Syers,and T.W. Walker. 1967. Fractionationof soil inorganic phosphateby a
modification of Changand Jackson'sprocedure.Soil Sci. Soc. Am. Proc. 31:736-739.
PHOSPHORUS 919
Williams, J.D.H., J.K. Syers,T.w. Walker, and R.W. Rex. 1970. A comparisonof methodsfor the
determinationof soil organicphosphorus.Soil Sci. 110:13-18.
Williams, J.D.H., J.K. Syers, R.E Harris, and D.E. Armstrong. 1971a. Fractionationof inorganic
phosphatein calcareouslake sediments.Soil Sci. Soc. Am. Proc. 35:250-255.
Williams, J.D.H., J.K. Syers,D.E. Armstrong,and R.E Harris. 1971b.Characterizationof inorganic
phosphatein non-calcareouslake sediments.Soil Sci. Soc. Am. Proc. 35:556--561.
Williams, J.D.H., and T.W. Walker. 1967. Comparisonof 'ignition' and 'extraction'methodsfor the
determinationof organicphosphatein rocks and soils. Plant Soil 27:457-459.
Wolf, AM., D.E. Baker, and H.B. Pionke. 1986. The measurementof labile phosphorusby the iso-
topic dilution and anion resin methods.Soil Sci. 141:60-70.
Wolf, AM., D.E. Baker, H.B. Pionke,and H.M. Kunishi. 1985. Soil testsfor estimatinglabile, solu-
ble, and algae-availablephosphorusin agricultural soils. J. Environ. Qual. 14:341-348.
Wrenshall,c.L., andWJ. Dyer. 1941.Organicphosphorusin soils. II. The natureof the organicphos-
phoruscompounds.A Nucleic acid derivatives.B. Phytin Soil Sci. 51:235-248.
Yang, J.E., E.O. Skogley,and B.E. Schaff. 1990. Microwave radiationand incubationeffectson resin-
extractablenutrients:II. Potassium,calcium, magnesium,and phosphorus.Soil Sci. Soc. Am.
J. 54:1646--1650.
Yang, J.E.,E.O. Skogley,and B.E. Schaff. 1991. Nutrient flux to mixed-bedion-exchangeresin: Tem-
peratureeffects. Soil Sci. Soc. Am. J. 55:762-767.
Yost, R.S., G.c. Naderman,EJ. Kamprath,and E. Lobato. 1982. Availability of rock phosphateas
measuredby an acid tolerantpasturegrassand extractablephosphorus.Agron.l. 74:462-468.
Published 1996
Chapter33
Sulfur
Sulfur occursin soils in organicand inorganicforms, with the organicS account-
ing for >95% of the total S in mostsoils from humid and semihumidregions.The
proportion of organic and inorganic S in a soil sample,however,varies widely
accordingto soil type and depthof sampling.
The inorganic S fraction in soils may occur as sulfate (SO~-) and com-
poundsof lower oxidation statesuch as sulfide (S2-), polysulfide (S~-, where n
> 1), sulfite (SOh, thiosulfate (S20~-), and elementalS (SO). In well-drained,
well-aeratedsoils, most of the inorganic S normally occurs as sulfate, and the
amountsof reducedS compoundsare generally <1% (Freney, 1961). Under
anaerobicconditions,particularly in tidal swampsand poorly drainedor water-
loggedsoils, the main form of inorganicS in soils is sulfide and often elemental
S (Brummeret aI., 1971a,b;Harmsen,1954;Hart, 1959).Accuratedetermination
of the reducedforms of inorganicS in soils is difficult, partly becauseof the ease
with which they can be oxidized on exposureto air, but mainly becauseof the
limitations of current analytical methods.No procedurehas beenentirely satis-
factory for determinationof sulfide in soils. Smittenberget al. (1951) proposed
a method fordeterminationof sulfide, sulfite,thiosulfate,polysulfide, elemental
S, and organically bound S in soils. It involves digestionof a soil samplewith
HCI for determinationof S in the monosulfidic compoundsand digestion with
Sn and HCI for determinationof the total reducibleS. In both methodsthe H2S
releasedis determinedcolorimetrically as methyleneblue (MB). None of these
methods,however, gives accurateresults; digestion of soil with HCI does not
releasethe S from acid-insolublemetal sulfides,causingunderestimationof the
sulfide present;and digestionwith Sn and HCI releasesS from organicS, caus-
ing overestimationof the total oxidizableS (Melville et aI., 1971). The natureof
this S fraction, however,has not beenidentified. Much of the reducibleS frac-
tion in soils is extractablewith 0.5 M NaOH and is distributedbetweenboth the
fulvic acid andhumic acid fractions.Therefore,digestionof soil with Sn and HCI
may prove adequatewherethe amountsof recible S are relatively large, but this
methodis unsatisfactoryfor usewith soils containingsmall amountsof reduced
inorganic S compounds,especially with surface soils containing appreciable
amountsof organicS.
Book Seriesno. 5.
921
922 TABATABAI
Although well-drained,well-aeratedsoils donot containreducedinorgan-

ic S compounds,thiosulfateand tetrathionateare producedin soils when treated
with elementalS (Nor & Tabatabai,1976, 1977). Also, such reducedS com-
poundsare presentin tailing pond effluents from the mining industry, and it is
believedthat they may lead to acid pollution of rivers surroundingtheseindus-
tries. Concentrationof thiosulfatein tailing pond effluents have beenestimated
to be in the rangeof 500 mg L-l (Wolkoff & Larose,1975).
Severalmethodshave beenproposedfor determinationof thiosulfateand
tetrathionatein milligram quantities.Theseinclude iodimetric titration, reacting
thiosulfate with o-tolidine [3,3'-dimethyl-(l,l'-biphenyl)-4,4'-diamine] and
Cu(II) ions (Raff, 1970), reductionwith Zn and HCI and subsequentdetermina-
tion of the H2S producedas MB (Aspiraset aI., 1972),and cyanolysisunder var-
ious conditions(Sorbo, 1957; Kelley et aI., 1969). Similar methodshave been
proposedfor determinationof tetrathionate(Kelley et aI., 1969).Although some
of thesemethodsgive quantitativeresultswhen usedfor determinationof a spe-
cific reduced S compound, quantitative determination of thiosulfate and
tetrathionateindividually in the presenceof a relatively large amountof sulfate
is difficult. Someof thesemethodsare not specific, and othersrequire various
cooling andheatingconditionsthat maketheir applicationto soils or soil extracts
very difficult (Aspiras et aI., 1972; Kelley et aI., 1969). To overcomesomeof
theseproblems,Nor and Tabatabai(1976) developedcolorimetric methodsfor
determinationof thiosulfate and tetrathionatein soils. The methods involve
extractionof theseS forms from soils with 0.1 M LiCI or 0.15% CaCl2 solution
and alkaline cyanolysisof thiosulfate-Sin the presenceof Cu2+ and alkaline
cyanolysisof tetrathionate-Sin the absenceof Cu2+
Cu2+
S20j- + CN- --+ SOj- + SCN- [1]
Catalyst
and determinationof the thiocyanateproducedas a ferric thiocyanatecomplex
SCN- + Fe3+ --+ Fe-SCNcomplex. [3]
The absorbanceof the reddish brown color of the Fe-SCNcomplex formed is

measuredby using a spectrophotometer adjustedto a wavelengthof 460 nm.
This method is rapid, sensitive and accurate,and it permits determinationof
microgramquantitiesof thiosulfate,tetrathionate,or both in the presenceof large
amountsof sulfate.
Theoretically,for eachmole of tetrathionate,2 mol of thiocyanateare pro-
ducedby alkalinecyanolysisin the presenceof Cu2+ (Reactions[1] and [2]), and
1 mol of thiocyanate is produced in the absenceof Cu2+ (Reaction [2]).
Therefore,a factor of two hasbeenusedto calculatethe quantitiesof tetrathion-
ate presentin samplescontaining thiosulfate (Kelley et aI., 1969). Our work,
however, indicates that the theoretical factor two, is never achieved when
SULFUR 923
tetrathionateis analyzedby this methodand that the practicalfactor for calculat-

ing the quantitiesof tetrathionateis 1.75 (Nor & Tabatabai,1975).
Although it is well known that Sin soils is presentmainly in organiccom-
binations,very little is known about the identitiesof theseS compounds.Direct
determinationof the total organicS in soils is not possibleby presentanalytical
methods.The organic S fraction normally is estimatedfrom the difference be-
tween the total S and SO~--S values determinedindependently(Tabatabai&
Bremner,1972b; Neptuneet aI., 1975).
Although it hasbeenpossibleto identify, with somedegreeof certainty,the
chemicalnatureof the inorganicforms of S occurringin soils, the presentmeth-
ods are not suitable for identification of the chemical nature of the organic S
compoundsin soils with the samedegreeof certaintyas thoseof the inorganicS
fraction. Most of the reagentsusedfor determinationof the organicS compounds
are not specific, they attack a broad group of organic S compounds(Freney,
1958; Freneyet aI., 1970; Melville et aI., 1971). Among the componentsof the
organic S in soils, the amino acids cystine [3,3'-dithiobis(2-aminopropanoic
acid)] and methionine[2-amino-4-(methylthio)butyric acid] are the most readi-
ly identified. These amino acids have been detectedby ion exchangechro-
matographyof soil hydrolysate obtained by 6 M HCI. It has beenestimatedthat
about 10% of the total S in soils could be presentas cystine and methionine
(~tevenson, 1956; Kowalenko, 1978). The reliability of determinationof the S-
containing amino acids in soils is questionableunless special precautionsare
taken to prevent their decompositionduring extraction. Attempts to determine
cystineand methioninein soils by oxidation to stable compounds of cysteicacid
(3-sulfoalanine)and methioninesulfone beforecarrying out the hydrolysis with
6N HCI have beenreported(Beatonet aI., 1968). Treatmentwith performic acid
(methaneperoxoicacid) before acid hydrolysis is commonly usedfor this oxida-
tion. For two soils, an averageof 26% of the total S, equivalentto 46% of C-
bondedS, was estimatedto occur as amino acids (Freneyet aI., 1972).
Of the various organic S fractions in soils, two groups of S compounds
havebeenstudiedextensivelyThesegroupshavebeenclassifiedaccordingto the
natureof the reagentsusedor accordingto the groupsof S compoundsattacked
by the reagents.Thus, two distinct forms of S-containingcompoundshave been
identified: (i) organicS that is not bondeddirectly to C and is reducedto H 2S by
hydriodic acid (HI), and (ii) organic S that is directly bondedto C (C-S) and is
reducedto inorganic sulfides by Raney Ni in alkaline medium (Freney, 1961,
1967; DeLong & Lowe, 1961; Lowe & DeLong, 1963; Freneyet aI., 1970). The
first fraction is believedto consistlargely (if not entirely) of S in the form of ester
sulfates (organic sulfates containing C-O-S linkages, e.g., choline sulfate
[(CH3hWCH2CH20S03"],phenolicsulfates,sulfatedpolysaccharides).The sec-
ond fraction is believedto consistlargely of S in the form of S-containingamino
acids,suchas methionineand cysteine.
In determinationof total S in soils, the various S forms are convertedby
oxidation to oneform-mostoften to sulfate.The sulfateproducedis determined
by gravimetric,turbidimetric and nephelometric,titrimetric, or colorimetric pro-
cedures(for review of literature,seeBeatonet aI., 1968). Severalproceduresare
available for oxidation of the total soil S to sulfate. The most commonly used
924 TABATABAI
proceduresare digestionwith HN03 and HCI04 (Arkley, 1961),ashingwith sil-

ver oxide (Ag20) and sodium bicarbonate(NaHC03) (Steinbergset aI., 1962),
and oxidation with NaOBr (Tabatabai& Bremner,1970a).Among the methods
availablefor estimationof the sulfateproduced,the MB methodof Johnsonand
Nishita (1952) is the most sensitive and accurate.This method involves the
reductionof sulfate to H2S for subsequentMB color developmentby p-amino-
dimethylaniline and ferric ammoniumsulfate [FeNH4(S04)2 • 12H20] reagents.
Analytical instrumentssuch as ion chromatographs(IC) developedby
Dionex Corp. (Sunnyvale,CA) for determinationof anions, including sulfate,
and the LECO sulfur analyzerand the LECO SC-132automatedtotal S analyz-
er developedby LECO Corp. (St. Joseph,MI) for determinationof total S are
commerciallyavailable.The IC techniqueshasbeenevaluatedfor determination
of sulfatein soils, and it seemssuitablefor analysisof sulfatein a varietyof soil
extracts(Karmarkar& Tabatabai,1992; Krupa & Tabatabai,1986; Tabatabai&
Basta,1991;Tabatabai& Dick, 1979).For IC systems,seeTabatabaiand Frank:-
enberger(1996, Chapter8).
For separationof anion by the IC instrument,the separatorcolumn con-
tains a strongbaseanion exchangeresin in the HCOj" form, and the suppressor
column containsa strongacid cation exchangeresin in the H+ form. In the vari-
ant of the suppressorcolumn system, the resin in the suppressorcolumn is
replacedby an ion-exchangemembranein tubular form to condition the eluent
continuously (Stevenset aI., 1981). This membrane(sulfonatedpolyethylene
hollow fiber) acts exactly like the suppressorresin in that ions are exchanged
from the membranefor ions in the eluentstream.The innovationis that the mem-
brane is regeneratedcontinuously by a gravity-fed flow of low-concentration
H2S04 (in analysis of anions) that continuously replaces the ions that are
exchangedonto the fiber with ions from the regenerant.Thus, separate regener-
ation stepsare eliminated. The replacementof the conventionalion-exchange
resin bed suppressor column with the hollow fiber suppressorallows continuous
operationof an IC without varying interferencefrom baselinedips, ion-exclusion
effects,or chemicalreaction.
In the suppressed IC systems,the eluentis NaHCO:VNa2C03at a flow rate
of 3 mL/min and pump pressureof 31.6 kg/cm2 (450 psi). The elution time is
affectedby the concentrationsof NaHC03 and Na2C03 and by the pump pres-
sure used. When anions are separatedby the use of a solution containing
NaHC03 and Na2C03 as the eluent, the eluent Na ions and samplecationsex-
changefor H ions on the suppressorresin, leaving H2C03 as the effluent. The
suppressorcolumn also convertsthe sampleanionsto acidsbeforethey enterthe
detector.The suppressorreactionsare as follows
NaHC03 + Resin-H= Resin-Na+ H2C03
(H2C03 has very low conductivity)
Na+X- + Resin-H =Resin-Na+ H+X-
whereX- representsan anion.

SULFUR 925
From calibrationgraphsobtainedfor the linear relationshipbetweenpeak

height and concentration(partsper million) of SOi--s in standardsolutions,the
SOi- -S concentrationof samplescanbe calculated.Although the sampleloop of
the injection valve containsa volume of 100 ilL (the sampleloop can be adjust-
ed to give the desiredvolume), a 2-mL sampleof soil extract is convenientto
ensureproperflushing of the injection valve loop and lines.
Evaluation of a suppressed-type IC method showed that its results for
extractablesulfatein a variety of soils agreedclosely with thoseobtainedby the
MB method(Dick & Tabatabai,1979; Ajwa & Tabatabai,1993).The IC method
has a high sensitivity, precision, and accuracyfor analysisof SOi- -So It can
detectas little as 0.2 mg L-I of SOi- -S againsta backgroundof 10 mM chloride,
phosphate,or acetate.In the absenceof significant concentrationof other elec-
trolytes, it can detectas little as 5 Ilg L-I of SOi--S. Among the advantagesof
the IC methodis that sulfate,nitrateandotheranionsin soil extractscanbe deter-
mined simultaneouslyin about 10 min.
Other methodsare availablefor determinationof S in soils, especiallyfor
total Sand SOi- -S, and the readermay refer to the reviews by Beatonet a1.
(1968) and Blanchar(1986) for selectinga suitableprocedure.With any proce-
dure selected,caremustbe takento avoid contaminationfrom water, detergents,
rubber tubing and stoppers,filter paper,and chemicals-allof which may con-
tain S as impurities. Unexpectedresultsshould prompt a check of all possible
sourcesof contamination.With reasonableexperience,the analystnormally can
quickly identify the sourceof the contamination.
TOTAL SULFUR
Wet ChemicalMethods
Principles
Total S in mineralsoils may rangefrom <20 mg kg-I in sandysoils to >600
mg kg-I in heavy-texturesoils. Organic soils may contain as much as 0.5% S.
Most soils, however,containbetween100 and 500 mg of S kg-I.
The methodsavailable for accuratedeterminationof total S in soils in-
volvestwo steps:(i) conversionof the variousS compoundsin soils to one form,
either by oxidation to sulfate (dry or wet procedure)or by reductionto sulfide
(conversionto sulfateis morecommonthanconversionto sulfide); and(ii) deter-
mination of the sulfateor sulfide produced.
Various techniqueshavebeenemployedto oxidize soil S to sulfate.One of
the most widely acceptedproceduresis fusion with Na2C03 and an oxidizing
agent.The procedurerecommendedby the Associationof Official Agricultural
Chemists (1955) requires fusion of sample with Na2C03 and Na202. This
method is probably the best for all types of soils. A mixture of Na2C03 and
NaN03 hasbeenusedin otherprocedures(Bertrand& Silberstein,1927; Robin-
son, 1930).Sodiumcarbonatealonemay be adequateto oxidize and retainS dur-
ing fusion (Evans & Rost, 1945). Sodium peroxide also has been used alone
926 TABATABAI
(Hart & Peterson,1911; Steinbergs,1955). Any procedureused to accomplish

fusion before S analysisshouldbe adequate,but inclusion of an oxidizing agent
would add assuranceof completeoxidation during fusion.
Othertechniquesusedfor oxidationoftotal soil S to sulfatebeforeS analy-
sis include ignition with NaHC03 either alone or with AgzO (Steinbergset aI.,
1962). In this method, the soil sampleis mixed with NaHC03 and AgzO and
heatedat 550°Cfor 3 h beforesulfate analysis.A methodbasedon heatinga soil
samplein a streamof Oz and passingthe combustionproductsover a red-hot
NaZC03 also is available(Little, 1957).
Of the various acid treatmentsproposed,digestionwith HCI04 or a mix-
ture of HCI04 and HN03 hasbeenmost popular.The HCI04digestionprocedure
is lesstediousthen the NazCOrNazOzfusion method,but it may not decompose
all the mineral constituentsof some soils. In all HCI04 digestion procedures,
predigestionof soils high in organicmatterwith HN03 beforeadditionof HCI04
is necessaryto avoid the dangerof explosion and fire. Arkley (1961) recom-
mendedan acid oxidation procedure involvingHN03, HCI04, H3P04, and HCI.
He claims that inclusion of H3P04 with HN03 plus HCI04 in the digestionmix-
turesincreasedeaseof handlingand decreasedthe amountof HCI04 required.To
overcomethe unsafefeaturesof boiling HCI04, Tabatabaiand Bremner(1970a)
developeda methodthat involves oxidation of soil S with NaOBr. This method
is basedon wet oxidation of the soil S under alkaline conditions,becausewet
oxidation proceduresare more convenientthan ignition techniquesfor conver-
sion of soil S to sulfate(particularly for soil extracts),and useof an alkaline oxi-
dation should not involve the risk of gaseousloss of S (as H2S, S02, or S03)
associatedwith useof acid oxidantsundercertainconditions.
Methodsbasedon reductionof total S beforeits estimationalso are avail-
able. Direct reductionof the S in soils has beenproposedby Little (1957). This
methodemploysspecialheatingblocks and involves the useof/errum reductum
to reducethe soil S to sulfide. Sulfur is estimatediodimetrically after absorption
of the liberatedH2S in KOCI or colorimetrically as MB.
Thereareseveralmethodsavailablefor determinationof the oxidizedS. In
most of the oxidation proceduresdescribed,the product is sulfate,which can be
determinedgravimetrically as BaS04,(AOAC, 1955), turbidimetrically (Stein-
bergs, 1955; Bentley et aI., 1955), ion chromatographically(Tabatabaiet aI.,
1988) or colorimetrically after its reduction to HzS (Johnson& Nishita, 1952).
However, methods involvingconversionof soil S to SOz and determinationof
the S02evolvedalso are available.In a methoddevelopedby Bloomfield (1962),
the soil sampleis ignited with V Z05 in a streamof Nz gas,and the gasesevolved
are passedover hot CuO and hot metallic Cu to completeoxidation of volatile
productsand reduceS03 to SOz. The SOz in the gas streamis then absorbedin
a solution of sodium tetrachloromercurate(Na2HgCI4) and determinedcolori-
metrically with p-rosaniline(C2oHl~3) and formaldehyde(HCHO). Jenkinson
(1968) recommendedheatinga soil samplewith K2CrZ07 and H3P04. The sul-
fate formed is reducedby activatedcharcoalto SOz, which is absorbedin HzOz
and determinedby a titration procedure involvingaddition of BaCl2 to the result-
ing H2S04 to precipitateBaS04'The excessBa2+ is then determinedby titration
againsta standardK 2S04 solution, with sulphonazoIII as an indicator.
SULFUR 927
Of the various colorimetric methodsrecommendedfor determinationof

sulfate,the proceduredevelopedby Johnsonand Nishita (1952) is the most sen-
sitive and accuratemethodavailable.This methodinvolves reductionof sulfate
to H2S by a reducingmixture containingHI, HCOOH, and hypophosphorusacid
(H 3P02). The HzS thus liberatedis absorbedin a buffer containingZn(OAc)z and
NaOAc and subsequentlytreatedwith p-aminodimethylanilinesulfateand ferric
ammoniumsulfatereagentsfor MB color development.The intensity of the MB
color is determinedcolorimetrically at a wavelengthof 670 nm. The color devel-
opedis stablefor 24 h when the solutionis storedin a stopperedflask in subdued
light. The conversionof sulfate to H2S by the method of Johnsonand Nishita
(1952) after fusion, ignition, digestion,or oxidationof soil sampleobviatesmany
problems.This methodis capableof completelyconvertingall sulfate(solubleor
insoluble)to sulfide (Johnson& Nishita, 1952; Freney,1958). Methyleneblue is
formed by reactinga H2S solution with an acid solution of p-aminodimethylani-
line in the presenceof Fe3+. At first the solution is.red, but it changesto blue as
the dye is formed. The reactionmay be representedas
SO~- + reducingmixture ~ H2S
where A-representsan anion. Although the reaction is not quantitative, the

amountof MB producedis quite reproducibleif the conditionsof color develop-
ment are adheredto. St. Lorant (1929) obtained 68.7%, Skerrett and Dickes
(1961) obtained64.7%, and Gustafsson(1960a)obtained66.7% H 2S conversion
to MB.
Of the proceduresdescribed,oxidation of soil S to sulfate by NaOBr and
analysisof the entire residuefor sulfateby the Johnsonand Nishita (1952) reduc-
tion-distillation procedureare preferred.This is mainly becausethe MB proce-
dure has the sensitivity, accuracy,and precision neededfor total S analysisof
soils and soil extracts (Tabatabai& Bremner, 1970a). Steinbergset at. (1962)
have shown that the method is not subject to interferenceby other soil con-
stituentswhen usedfor determinationof sulfate in total S analysisof soils. Other
methods(e.g., turbidimetric) in which extractsof oxidation residuesare analyzed
for sulfatecan be subjectto seriouserrorsif the soil under analysiscontainssig-
nificant amountsof Ba or other metallic cationsthat form sparingly solublesalts
with sulfate (the Ba content of most soils is between100 and 3000 mg kg-I).
When the MB methodis usedfor determinationof the sulfateformed, the results
obtainedby NaOBr oxidation procedurewere similar to thoseobtainedby using
HCI04 digestionand NaHCOr Ag 20 ignition procedures(Tabatabai& Bremner,
1970b). Therefore,details of the digestion, ignition, and oxidation methodsare
describedhere for use with the MB method.
928 TABATABAI
~
28,115
CONDENSER - -
NITROGEN GAS
INLET TUBE-
em
DIGESTION - 0' , 1 '10
OISTILLATION
FLASK - - -
Fig. 33-1. Digestion-distillationapparatus.
Method
The methoddescribedis essentiallythat reportedby Johnsonand Nishita
(1952) and Johnsonand Urich (1959).
Special Apparatus
1. Modified Johnson-Nishitadigestion-distillationapparatus(Fig. 33-1):
This apparatusis designedso that a 50-mL boiling flask fitted with
standard-taper (19/22) ground-glassjoints can be usedas a reduction-
distillation chamber.The flasks are providedwith hooks so that they
can be fastenedto the reduction-distillationapparatusby spiral steel
springswith loop ends as shown in Fig. 33-1. The apparatuscan be
made by most local glass-blowingshops. It can be obtained from
Scientific GlassApparatusCo. Inc., Bloomfield, New Jersey.Before
use, the apparatusshouldbe conditionedto allow for any sulfide that
may be sorbed on the glass or dissolved in the pyrogallol
[C6H3(OH3)]-sodium phosphatewashcolumn. When conditioningthe
apparatus,be surethat wateris flowing throughthe condenserand that
N2 gasis bubbling through the system.For the procedureto condition
the apparatus,see"Comments"under"Total Sulfur."
2. Distillation flasks: Use 50-mL boiling flasks fitted with standard-taper
(19/22) ground joints and glass hooks, as shown in Fig. 33-1. The
flask dimensionsshouldbe suchthat when the flasks are connected to
the reduction-distillationapparatus,the distancebetweenthe tip of the
N2 inlet tube and bottom of the flask is approximately4 mm.
SULFUR 929
3. Cylinder of water-pumpedN2 with regulators:Place a gas-washing

bottle containing a solution of HgCl2 in 2% KMn04 between the
cylinder and the boiling flask to removetracesof S gases.
4. Microbumers,alcohol or gas.
5. Delivery tubes:Cut 4-mm (o.d.) glass tubing in 12-cm lengths, and
polish the endson a flame.
Reagents
The following reagentsare requiredfor the acid digestionprocedure:

1. Nitric acid, 69%.
4.. Hydrochloric acid, 37%.
5. Hydrochloric acid, 1 M: Add 83 mL of 37% HCI to about 800 mL of
water, and adjust the volume to 1 L.
The following reagentsare requiredfor the alkaline oxidation procedure:
1. Sodium hydroxide, approximately2M: Dissolve 8 g of NaOH in 100
mL of deionizedwater in a 125-mL Erlenmeyerflask, and cool under
running water.
2. Sodium Hypobromite solution: Prepare this reagent immediately
beforeuseby adding3 mL of Br2 slowly (approximately0.5 mL/min),
and with constantstirring, to 100 mL of 2 M NaOH in a 125-mL Er-
lenmeyerflask.
3. Formic acid (HCOOH), 90% (FisherScientific Co., Itasca,IL)
The following reagentsare requiredfor the dry-ashingprocedure:
1. Sodium bicarbonate.
2. Silver oxide
The following reagentsare requiredfor reductionand distillation of sulfate
to H2S and estimationas MB:
1. Reducingmixture: Mix 400 mL of HI (sp gr 1.7, methoxyl grade),100
mL of H3P02 (50%), and 200 mL of formic acid (90%) in a l-L, three-
necked round-bottomflask (Fig. 33-2). Place the flask in a heating
mantleconnectedto a rheostatin a well-ventilatedhood. Placea fitted
thermometergraduatedto 360°C in the center neck after lubricating
thejoint with a small amountof S-freeground-jointlubricant. Connect
the exhaustfumes outlet to one of the side necksand the N2 gas inlet
to the other neck. Placea 250-mL Erlenmeyerflask containingabout
150 mL of cold water under the fume exhaustoutlet. Allow a stream
of purified N2 gas to bubble through the acid mixture. Heat the mix-
ture slowly to 115°Cand hold at 115 to 117°Cfor 10 min. Caution: Do
not let the temperatureof the solution exceed117°C. Extremely poi-
sonousfumes of phosphine(PH3) may be liberatedfrom the reagentif
it is heatedabove 120°C or spilled on a hot surface. Coolthe mixture
930 TABATABAI
THERMOMETER - - em
L.L.L.LW
NITROGEN GAS
o [0
INLET TUBE_
Fig. 33-2. Apparatusfor preparationof the reducing

CORK RING./'
mixture.
by turning off the powerandallowing the mixture tostandin the hood.

To speedup the cooling time, removethe flask from the heatingman-
tle, and place it on a cork ring (N2 gas should continue bubbling
throughthe mixture during the cooling period). When the temperature
of the mixture is about room temperature,transfer it to a l-L bottle,
and storeit in the dark.
2. Nitrogen purification solution: Add an excessof mercuricchloride, 5
to 10 g1100 mL of 2% potassiumpermanganate solution.
3. Pyrogallol-sodiumphosphatesolution: Dissolve 10 g of sodium dihy-
drogen phosphate (NaH2P04 • H20) and 10 g of pyrogallol
[C6H3(OH3)] in 10 mL of S-free distilled water with the aid of a
streamof N2 bubbling through the solution. This reagentshould be
prepareddaily.
4. Sulfur-free distilled water: Distilled, deionizedwater is normally free
of S. If necessary,preparedby distilling watercontainingalkaline per-
manganatefrom an all-glassstill.
5. Zinc acetate-sodium acetate,H2S-absorbingsolution: Dissolve50 g of
zinc acetatedihydrate[Zn(CH3COO)2• 2H20] and 12.5 g of sodium
acetatetrihydrate (CH3COONa • 3H20) in about 800 mL of water,
adjust the volume to 1 L, and filter if turbid (Whatmanno. 42 filter
paper).
6. Aminodimethylanilinesolution: Dissolve 2 g of p-aminodimethylani-
line sulfate (no. 1333, EastmanKodak Co., Rochester,NY) in about
1500 mL of water in a bottle markedto 2 L. Placethe bottle in a pan
of cold water, and slowly add 400 mL of concentratedreagent-grade
sulfuric acid (H 2S04), This acid should be added in small portions,
mixing the solution, and allowing it to cool after each portion. Cool
and adjust the volume to 2 L.
7. Ferric ammoniumsulfatesolution: To 25 g of ferric ammoniumsulfate
[Fe2(S04)3(NH4hS04 • 24H20], add 5 mL of concentratedreagent-
gradesulfuric acid and 195 mL of distilled water. Allow it to standat
room temperatureuntil dissolved(normally it takesseveraldays be-
fore all the ferric ammoniumsulfate is dissolved.
SULFUR 931
8. Standardsulfatestocksolution: Preparethis stocksolutionby dissolv-

ing 5.434g of reagent-gradepotassiumsulfate in about800 mL of S-
free distilled water, and adjustthe volume to 1 L with water. One mil-
liliter of this solution contains1 mg of S.
9. Standardsulfate working solution: Dilute 1, 2, 3, 4, and 5 mL of the
standardsulfate stock solution to 100 mL of S-free distilled water.
Mix thoroughly. One milliliter of thesesolutionscontainsto, 20, 30,
40, or 50 /lg of SOi--S, respectively.
10. Sulfur-free ground-jointlubricant: In a 50-mL beaker, mix approxi-
mately 5 g of silicone stopcocklubricant (Dow Coming Corp., Mid-
land, MI) with 5 mL of hydriodic acid and 5 mL of hypophosphorus
acid. Gently heat to boiling on a hot plate in a well-ventilatedhood,
with frequentstirring, for about 45 min. A 1oo-mL beakerfitted with
a condensermadefrom a 50-mL round-bottomedboiling flask filled
with cold wateris convenientin the preparationofthe S-freelubricant.
At the end of the boiling period, pour off the acid mixture, and wash
the lubricantthoroughlywith S-freewater.
Procedures-Oxidation
of Total Sulfur to Sulfate
Acid Digestion. Place a 2.0-g sampleof finely ground «40 meshor 425
/lm) soil in a 125-mL Phillips beaker.Add 3 mL of 69% HN03, swirl the beaker
to mix the contents,coverthe beakerwith a watchglass,andheaton a steambath
(or steamplate) for 1 h. Removethe beakerfrom the steambath, uncover,and
add 3 mL of 60% HCI04 and 7 mL of H3P04 (theseacids may be premixed,if
desired,for easeof handling).Heat the beakeron a sandbath (or hot plate) situ-
atedin a hood and regulatedso that the temperatureof the sandis 190 to 210°C
until heavy white fumes of HCI04 are visible. Placethe watch glassand contin-
ue heatingfor 30 min. Removethe beakerfrom the sandbath,cool, uncover,and
add 2 mL of 37% HCI. Heat again until white fumes of HCI04 are againvisible.
Transferthe digestquantitativelyinto a loo-mL volumetricflask, and adjustthe
volume with 1 M HCI.
Alkaline Oxidation. Placea sampleof air-dried, finely ground(<100 mesh
or 150 )..lm) soil containing to to 50 )..lg of S (usually 0.1--0.2g of soil) in a 50-
mL dry digestion-distillationflask (for description of this flask, see "Special
Apparatus"under "Total Sulfur"), and add 3 mL of NaOBr solution. Swirl the
flask for a few secondsto mix the contents,and allow it to standfor 5 min. Then
swirl the flask againfor a few seconds,and placeit upright in a sandbath situat-
ed in a hood and regulatedso that the temperatureof the sandis 250 to 260°C.
Heat the flask until its contentsare evaporatedto dryness,and continueheating
for an additional 30 min. Removethe flask from the sandbath, allowit to cool
for about5 min, add 1 mL of water, and heatthe flask on the sandbathfor a few
secondsto bring the residue into suspension.Removethe flask from the sand
bath, swirl it for a few seconds,and allow it to cool to room temperature.
Dry Ashing. Place a sampleof air-dried, finely ground (<100 mesh) soil
containing to to 50 )..lg of S (usually 0.1--0.2g of soil) in a small porcelaincru-
cible (1.5 cm in diam. by 1.5 cm in height, crucible no. 00000, CoorsPorcelain
932 TABATABAI
Co., Golden, CO), and mix it thoroughly with 0.25 g of NaHC03 and 0.01 g of
Ag20. Placethe crucible in a cold electric muffle furnace,raise the temperature
to 550°C, and heatfor 3 h. Cool and transferthe mixture quantitativelyto a 50-
mL digestion-distillation flask (for description of this flask, see "Special
Apparatus"under"Total Sulfur").
Reduction of Sulfate to Sulfule and Estimation as Methylene Blue.
Lubricate allsphericaljoints of the reduction-distillationapparatuswith minimal
amountof the S-free lubricant.
Pipette10 mL of pyrogallol-sodiumphosphatesolution into the gas-wash-
ing column, and connect it to the apparatusholding it in place with a clamp.
Condition the apparatus(for conditioningthe apparatus,see"Comments"under
"Total Sulfur"). Add 10 mL of the Zn(OAc)z-NaOAc solution to a 100-mL,
glass-stoppered volumetric flask to be usedas a receiver,and add approximate-
ly 50 mL of distilled, deionizedwater. Connecta deliverytube to the side arm of
the gas-washingcolumn, using a piece of S-free rubber tubing. Insert the deliv-
ery tube into the liquid almostto the bottom of the receivingflask, and clamp the
receivingflask in place.
Dependingon the procedureusedto oxidize the sampleS to sulfate,pro-
ceedwith analysisof the total S as follows: If the acid digestionprocedurewas
used,transfera 1- to 5-mL aliquot of the digest into a 50-mL distillation flask.
With aliquots >2 mL, reducethe volume to about2 mL by heatingthe flask on a
sandbath. If the alkaline oxidation procedurewas used,add 1 mL of formic acid
to the flask containingthe oxidized sample,swirl the flask for a few secondsto
mix the contents,and allow it to standfor about 10 min. If the dry-ashingproce-
dure was used,add 2 mL of distilled, deionizedwater, and swirl the flask to mix
the contents.
To the flask containingthe sample,add by a rapid delivery pipette4 mL of
the reducingmixture, moisten thecondenser'sjoint with a drop of water, imme-
diately attachthe flask to the condenser,and adjust the N2 flow so that about 2
bubbles/sissuefrom the receivingflask. Make certainthat cold water is flowing
through the condenser.After about5 min of N2 flow, light the microburner,and
boil the contentsgently for 1 h. Then removethe receivingflask, leaving the con-
necting tube in the Zn(OAc)2-NaOAc solution (by disconnectingthe delivery
tube and leaving the small pieceof the rubbertubing attachedto the side arm of
the apparatus).Using a rapid-delivery pipette, add 10 mL of the p-aminodi-
methylaniline solution. Quickly stopper the flask, and mix the contentsthor-
oughly. Unstopperthe flask, and add 2 mL of ferric ammoniumsulfate solution.
Stopperthe flask, mix the contents,and adjustthe volume to 100 mL with water.
Mix the contentsthoroughlyby inverting the stopperedflask severaltimes. After
20 min, but within 24 h, measurethe absorbanceof MB color by using a spec-
trophotometeradjustedto a wavelengthof 670 nm. Calculatethe S contentof the
sample analyzed by referenceto a calibration graph plotted from the results
obtainedwith standardscontaining0, 10, 20, 30, 40, and 50 Ilg of S. To prepare
this graph, pipette a I-mL aliquot of the standardS working solutions into 50-
mL distillation flasks, add4 mL of the reducingmixture, andproceedwith analy-
sis of these standardsas describedfor the S content of soil sample. Plot the
absorbancereadingsagainstmicrogramsof S analyzed.
SULFUR 933
Controls should be performed with each soil analyzed to allow for S

derivedfrom the reagentsusedfor oxidation of S in the sample.
AutomatedInstrumentalMethods
Severalinstrumentalmethodsare availablefor determinationof total S in

soils. The LECO S Analyzer (LECO Corp., St. Joseph,MI) was designedfor
determinationof total S in steel,but becauseof its simplicity, speed,and conve-
nience,this analyzerhas beenusedfor S analysisof nonferrousas well as fer-
rous materials.In total S with this instrument,the sampleis treatedin a ceramic
cruciblewith combustionaccelerators(usually Fe plus Sn,Cu, or Sn-coatedCu),
and the crucible is coveredwith a porous cover and heat to high temperature
(approximately1600°C) in a steamof purified Ob the high temperaturebeing
generatedby using an induction furnaceto inducean electricalfield in the accel-
erator-treatedsample.The S02 thus liberatedis collectedin dilute HCI contain-
ing KI, starch,and a traceamountof potassiumiodate(KI0 3) and is determined
by titration with standardKI0 3 solution. The reactionsinvolved in this tittration
procedureare as follows
KI0 3 + SKI + 6HCI = 312 + 6KCI + 3H20
Before use, the buretteof the automaticS02 titrator is calibratedby ana-

lyzing S standards.The LECO Corp. suppliesstandardsfor this purpose.Eval-
uation of this analyzerfor rapid and precisedeterminationof total S in soils has
shownthat its resultsare usually higher, with poor precision,than thoseobtained
by wet or dry oxidation methods,followed by reduction and determinationof
MB (Tabatabai& Bremner,1970b).
The LECO Model SC-132automatedtotal S analyzerwas recently evalu-
atedfor determinationof total S in soils and sediments.In usingthis analyzer,the
exampleis mixed with combutionacceleratorsin a ceramicboat and combusted
in a resistancefurnace at 1371°C in an O2 atmosphere.The S02 thus produced
is passedthroughan infrared(IR) cell. The IR cell is usedasboth a referenceand
a measurechamber.It detectstotal S, asS02, continuously.It consistsof an IR
source,a choppermotor, a precisewavelengthfilter, a condensingcone, and IR
energy detector,and a cell body. The S02 collected absorbsthe IR energy at a
specific wavelengthof the IR spectrumwhile in the cell. David et al. (1989) used
this analyzerfor determinationof total S in soils and sedimentsand showedthat
the resultsobtainedby this instrumentagreedwith thoseby currentmethodsused
for determinationof total S in such materials.Sampleanalysistakes<3 min and
the instrumentprovidesa direct readoutof total S values.
Comments
The MB methoddescribedis very sensitive.Linear relationshipsbetween

the amountsof SOi--S taken for analysisand absorbanceof the resulting MB
934 TABATABAI
solutionsshould be expectedin the rangeof 2 to 50 Ilg of SO~- -So The absorp-

tion maximumis rathersharpat 670 nm(Fogo & Popowsky,1949); consequent-
ly, deviation from Beer's law may be expectedif instrumentsof low spectral
purity are used.If the amountof S analyzedis greaterthan the highestlimit of
the calibrationcurve, the solutionshouldbe diluted with a solutionof exactly the
sameacid concentration.This can be convenientlyaccomplishedby diluting the
concentratedsolution with distilled water containingthe sameconcentrationof
Zn(OAc)rNaOAc,p-aminodimethylaniline,and ferric ammonium sulfate rea-
gents, thus assuring identical conditions in solutions prepareddirectly from
smallerinitial amountsof S and thosepreparedby dilution. Dilution of the con-
centratedMB solution is satisfactoryfor amountsof S up to approximately300
Ilg. Amounts inexcessof 300 Ilg give low readings.The low readingsare due to
limiting amountsof p-aminodimethylanilinesolution addedwhen the color is
first developed(p-aminodimethylanilineaddedin the dilution stepdoesnot react
becauseexcesssulfide has previously beenoxidized by the addedFe3+).
If a stoichiometricreactionbetweensulfide andp-aminodimethylanilineis
assumed(i.e., 2 mol of p-aminodimethylanilinereact with 1 mol of sulfide), it
may be calculatedthat thereshouldbe sufficientp-aminodimethylanilineto react
with 433 Ilg of SO~- -So It is actually observedthat there is sufficientp-aminodi-
methylanilineto reactcompletelywith somewhatless sulfide, becausethe reac-
tion and productionof MB is less than quantitative(see"Principles" for "Total
Sulfur"). The calibration curvesshould be verified for eachtype of spectropho-
tometeror colorimetric used,becausethe spectralpurity obtainedwith different
typesof instrumentsmay vary considerably.Therefore,the extentand degreeof
linear responsemust be evaluatedfor eachset of operatingconditions.
Methyleneblue solutionsare stablefor at leastseveraldaysif storedin the
dark. Exposureto sunlight, however,causesrapid fading. The color is stablefor
at least 24 h in the laboratoryif the solution is kept away from direct sunlight.
The rate of heatingof the digestion mixture is not extremelycritical, pro-
vided free flame is not allowed to contactthe surfaceof the flask abovethe level
of the reducingmixture. Best resultsare obtainedby the use of an asbestosmat
on the top of the flame shield.
All tubing in the N2 lines to the boiling flasks must be tygon or of similar
inert material. Rubbertubing should not be usedfor this purpose.
The rate of N2 flow through the apparatusshould be sufficiently rapid to
sweepthe systemcompletely free of H2S in a reasonabletime. Ratesof 100 to
200 mLimin are adequate.Slower ratesrequire excessivelylong time to ensure
completetransferof H2S.
Regenerationof the used reagentsshould be avoided.The used reducing
mixture should be collectedin a wastebottle and disposedof properly. The use
of other reducingmixtures hasbeendiscussedby Gustafsson(1960b).
Resultsobtainedby the three methodsdescribedfor oxidation of total soil
S to sulfate and subsequentreduction-distillationare very similar (Tabatabai&
Bremner,1970b).The NaOBr oxidation method,however,is less time-consum-
ing, and sinceit doesnot include transferof samples,samplecontaminationdur-
ing handling is minimized. When the NaOBr method is used for oxidation of
organic S in soils, it is important that the NaOBr solution be preparedimmedi-
SULFUR 935
ately before use. Tests with several soils have shown that the total S values
obtainedby the proceduredescribedwith NaOBr solution that had beenstored
for 24 h were Sto 12% lower than thoseobtainedwith freshly preparedsolution
(Tabatabai& Bremner, 1970a). The stability of hypobromitesolutions is gov-
ernedby such factors as the concentrationof hypobromiteand hydroxide,light,
temperature,and the presenceof foreign substances (Polaket aI., 1966).To avoid
breathingthe fumes evolved,the NaOBr solution, oxidationof sampleswith this
reagent,digestionof samplewith HCI04, and preparationof the reducing mix-
ture usedfor conversionof sulfate to H2S should be made in a well-ventilated
hood.
Resultsobtainedby the proceduresare not affectedby samplemeshsize;
resultswith 40- (42S-l1m), 100- (ISO-11m), and 300-mesh(SO-11m) are identical
(Tabatabai& Bremner,1970a).It is recommended,however,that soil samplesbe
ground to passthrough at least a lOO-mesh (ISO-11m) screenbefore analysisby
the alkaline oxidation or ignition methoddescribedand that samplesground to
passthrough a ISO-mesh(lOO-l1m) screenbe usedif they are suspectedto con-
tain significant amountsof gypsumor other S-rich minerals.
When the NaOBr oxidation method is used, formic acid is added after
completion of the NaOBr treatmentto destroy residual NaOBr and acidify the
oxidation residuebeforeaddition of the reducingmixture. The formic acid-treat-
ed residueshould not be stored more than a few hours before distillation; the
residuesometimessolidifies and preventsmixing with the reducing mixture if
storedovernight. Reductionof sulfateto H2S is affectedif the oxidation residue
is not acidified and the residualNaOBr is not destroyedbeforeaddition of the re-
ducing mixture. Similar to the soil digest obtainedwith HCI04, if the NaOBr-
oxidizedsamplecontainsan excessof oxidizing agent,a brown color due to oxi-
dation of 1- to 12 is producedin the reactionflask after addition of the reducing
mixture to convertsulfateto H2S. Under such conditions,an excessof the reduc-
ing mixture shouldbe added.
The NaOBr oxidationstepin the methoddescribedis simple andrapid, and
many oxidations can be performedat one time. The sandbath requiredcan be
constructedreadily by placing a layer of silica sand(approximately4 cm deep)
on a thermostaticallycontrolled electric hot plate. A wire frame assemblywith
approximately6-cm squareopeningsis convenientfor holding the flasks used
for the NaOBr treatmentin an upright position when theyare heatedon the sand
bath.
Johnsonand Nishita (1952) recommendedthat the ground-glassjoints of
the digestion-distillation flask be lubricated with purified silicone grease,but
work by TabatabaiandBremner(1970a)showedthat no lossof H2S occursif this
joint is lubricatedwith water (or H3P04) insteadof silicone greaseand the flask
is attachedto the condenserby spiral steelspringsas shown in Fig. 33-1. Use of
water insteadof greasefacilitatescleaningof the condenserand distillation flask.
The N2 gas inlet tube can be readily cleanedafter severaldistillations by a jet of
water from a washbottle.
Becauseof absorptionand solubility of H2S in the apparatusand wash
solution, it is necessaryfor work of highestaccuracyto condition the apparatus.
To accomplishthis, a preliminary run shouldbe madewith a sampleor standard
936 TABATABAI
containinga small amountof reducibleS. The sampleneednot be takenthrough

the developmentand measurement of color step.The digestion-distillationappa-
ratus and wash solution are equilibratedafter the first run with respectto H2S,
and determinationof the total S in the samplemay be carried on in the usual
manner.
The delivery tube leading to the receiving vesselis attachedto the con-
necting arm of the wash column by a short piece of rubber tubing. The rubber
tubing used should be freed of S compoundsby boiling the rubber piecesin a
mixture containing 10 mL of HI and 10 mL of H3P02 in a 100-mL beakerfor
about45 min (a 50-mL boiling flask containingcold water placed onthe beaker
may serve as a condenser).BecauseZnS adheresto the portion of the delivery
tube dipping in the Zn(OAch solution, the tube must be disconnectedand
allowed to remain in the volumetric flask during the developmentand measure-
ment of the absorbanceof MB color. The tubesmay be removedwith washing
after developingthe MB color and before making the volume. If the delivery
tubesare not removed,the volumetric flask shouldbe selectedso that the marks
are approximatelyin the sameposition on the neck. When the delivery tubesare
of the samediametersandsufficiently long to extendabovethe graduationmarks
of the volumetric flasks, negligible relative error is encounteredwhen the tubes
are left in the flasks throughoutthe whole procedureof color developmentand
color absorbancemeasurement.
Distilled water used for preparationof the wash solution and Zn(OAc)2
absorbingsolution sometimescontainsas much as 0.1 mgIL of Cu. Under these
conditions, sulfide is quantitatively precipitatedin the wash solution, and the
Zn(OAc)2 solution is not availableto form MB. Copperin the sampleor in the
digestion mixture is without effect on the determination(Johnson& Arkley,
1954).Therefore,it is recommendedthat distilled, deionizedwater,or waterpre-
paredby distillation from an all-glassstilI be usedin preparationof the reagents
indicated.
When samplescontain large amountsof nitrate (e.g., nitrate remaining
from digestionof soil with HN03 and HCl04), the resultsshow low apparentS
values.Evidenceof nitrate interferenceis a rapid and pronounceddarkeningof
the sodium phosphate-pyrogallolwash solution. In the absenceof nitrate in the
sample,a freshly preparedsodium phosphate-pyrogallolwash solution remains
almostcolorlessfor as many as six or eight runs.
The volumetric flask should be stopperedafter disconnectingit from the
apparatus,and the contentsshouldbe mixed by shakingafter addition of eachof
the reagentsusedfor MB color development.The final solution shouldbe mixed
thoroughly by inverting the stopperedflask severaltimes before measuringthe
absorbanceof the color developed.
An alternative to the MB color finish describedwas proposedby Dean
(1966). In this method the H2S evolved is adsorbedin a solution containing
NaOH and treated with Bi reagent to develop the colloidal bismuth sulfide
(Bi 2S3). The color is then measuredat 400 nm. Standardcurvesobtainedby this
method obey Beer's law up to 200 Ilg of S. Studiesby Kowalenko and Lowe
(1972) showedthat the bismuthsulfide and MB colorimetric finishes for sulfate
determinationby the Johnson-Nishitamethoddescribedgive comparableresults
SULFUR 937
when applied to soils and soil extracts.Theseauthorsclaim that the Bi finish is

more rapid and more reliable. It has been my experience,however, that the Bi
finish is difficult to reproduceand haslesssensitivity. Similar to the MB method,
the bismuth sulfide method is affectedby excessnitrate. A reductionof 75% of
sulfatedeterminedwith nitrate in excessof 6 mg hasbeenobserved(Kowalenko
& Lowe, 1972).
A methodbasedon determinationof total S in soils by IC also is available
(Tabatabaiet aI., 1988). It involves ignition of a mixture of soil and NaHC03
containingAg20 at 550°C for 3 h. The residueis dissolvedin 1 M HOAc, dilut-
ed with deionizedwater, filtered and analyzedfor SO~- by a suppressed-type Ie.
Resultsby the IC method for sevenIowa soils and three Chilean soils agreed
closely with those obtained by the MB method after alkaline oxidation with
NaOBr.
The LECO S analyzerinvolves the assumptionthat the combustiontech-
nique convertsS quantitativelyto S02 and the titration proceduredescribeddoes
not recoverS evolved as S03' Studiesby Searle(1968) to overcometheseas-
sumptions indicated that the precision of the LECO method could not be
improved even when a soil samplewas heatedwith Fe powder, molybdic acid
(Mo03), and chromiumtrioxide (Cr03) in a LECO high-frequencyinduction fur-
naceand the S-containinggasesevolvedwere absorbedin 1 M NaOH. This solu-
tion was subsequentlyanalyzedfor S by the MB methodof Johnsonand Nishita
(1952). Lowe (1969) also reportedpoor precision in analyzingsome Canadian
soils for total S by the LECO S analyzer.Similarly, Kaplan et aI. (1963) found
that the LECO S analyzerdid not give reproducibleresultswhen usedfor total S
analysisof marine sedimentsand that the resultsobtainedby this analyzerwith
surfacesamplesof thesesedimentswere much lower than thoseobtainedby wet
combustionmethod.The LECO S analyzeris attractive when it comesto sim-
plicity and speed(40-50 analyses/d),but it is unsatisfactoryfor researchrequir-
ing accurateand precisedeterminationof total S in soils. It is useful, however,
for total S analysisin sometypesof soil investigationswhen only an estimateof
the total S is required(Bremner& Tabatabai,1971; Tabatabai& Bremner,1991).
Calibrating the LECO Model SC-132 instrumentwith soil or vegetation
standardsby using two combustionaccelerators(iron powder, LECO Part 501-
078, or LECOCEL, LECO Part 763-266)improvedthe accuracyand recoveryof
S by this instrumentalmethod.The upper99% confidenceinterval relative stan-
dard deviation for duplicate samplesusing this LECO analyzerwere <15% for
mineral soils, <8% for forest floor, and <3% for sedimentsamples(David et aI.,
1989).
Methodsfor analysisof total S in soils basedon x-ray fluorescencespec-
troscopy also are available (Roberts & Koehler, 1968; Tabatabai& Bremner,
1970b;Darmodyet aI., 1977). Evaluationof the x-ray fluorescenceprocedurefor
determinationof total S in mineral soils has shown that this methodgives satis-
factory results. In one such study, the results by the x-ray fluorescencemethod
was greatly different from those obtained by the NaOBr oxidation or HCI04
digestionmethodsdescribed,indicating that the methodcan be usedsatisfactori-
ly for routine determinationof soil S in laboratorieshaving facilities for x-ray
fluorescencespectroscopy(Tabatabai& Bremner, 1970b). This method would
938 TABATABAI
not be particularlyconvenientif only S analyseswere required,becausethe pro-

cedureis rathertediousandtime-consuming.It hasconsiderableattraction,how-
ever, if analysesfor K and P also were needed,becausethe samesoil pellet can
be usedfor S, P, and K analysis(Stanceet aI., 1968). When the x-ray fluores-
cencemethodis usedfor determinationof total S in organicsoils (soils having a
weight loss on ignition >25%), however, it is necessaryto apply an empirical
correctionfactor derivedfrom the percentage weightlosson ignition at 450°Cto
correctfor matrix effects.
INORGANIC SULFATE
Principles
Inorganic sulfate may occur in soils as water-solublesalts, may be ad-

sorbedby soil colloids, or may occuras insolubleforms. The fractions of inor-
ganicsulfatepresentin eachof thesestatesare dependenton severalsoil prop-
erties.Surfacehorizonsof mostwell-drainedtemperatesoils containonly small
amountsof water-solublesulfate. Larger quantities of water-solublesulfate,
however,accumulatein soils of arid regions.Becauseof the effect of seasonal
conditionson mineralizationof organicS, leachingof solublesulfate,andsulfate
uptakeby plants,it is very commonto observea considerableseasonalfluctua-
tion in the amountsof water-soluble sulfatein surfacesoils. Other factors that
may affect the concentrationof water-solublesulfate in soils are applicationof
fertilizer materialsandthe sulfatecontentof rain andirrigation waters(Tabatabai
& Laflen, 1976a,b).
Severalfactors affect inorganic sulfateadsorptionby soils. Among these
clay content,natureof the clay mineral, pH, and exchangeablecationsare the
most important. Adsorption of sulfate by soils is negligible abovepH 6.5 and
increaseswith decreasingpH below this value (Kamprathet aI., 1956; WIliams
& Steinbergs,1962). Sulfate adsorption is greater in soils containing large
amountsof aluminum and iron oxides (Berg & Thomas, 1959; Chao et aI.,
1962a,b,1964; Ensminger,1954); the former generallybeing more important.
Adsorptionof sulfateby soil constituentsoccursat a numberof energeticallydif-
ferent reactionsites.This was demonstratedby the work reportedby Alymore et
al. (1967) showingthat sulfateadsorptionon kaolinite clay is weakly held rela-
tive to that adsorbedon iron and aluminumoxides.
In addition to the water-solubleand absorbedsulfatesfractions, several
forms of insolublesulfatesare likely to occurin soils. The insolubleforms may
includebariumandstrontiumsulfates,sulfatecoprecipitatewith CaC03, andba-
sic iron and aluminum sulfates under certain conditions. Basic iron sulfate,
jarosite[KFe3(OHMS04)2],andcoquinbite[Fe2(S04)J• 5H20], havebeeniden-
tified in separateteststakenfrom tidal marshsoils (Clark et aI., 1961; Fleming
& Alexander,1961).Although no report is availableon the detectionof basalu-
nite [A4(0H)lOS04 • 5H20] in soils, this compoundhas been preparedfrom
Al(0H)3 and sulfateions in the presenceof clay (Singh, 1967; Singh & Brydon,
1967,1969).
SULFUR 939
The solublesulfatefraction in soils is extractablewith water and with salt

solutionscontainingNaCl, LiCI, or CaCI2. A salt solution, however,is frequent-
ly preferredbecauseof dispersiondifficulty associatedwith aqueousextractions.
Arkely (1961) introducedthe use of 0.1 M LiCI, and Williams and Steinbergs
(1959) suggestedthe use of 0.15% CaCl2 solution.The concentrationsof elec-
trolytes in thesesolutionsare sufficient to keep the soil flocculated, but neither
the hydratedLi nor the CI ions would influencegreatly the chemistryof the soil
surfacedominatedwith much more strongly adsorbedions (Arkely, 1961). The
use of LiCI salt solution has the additional advantagethat microbial activity is
slowedor stoppedduring extractionof sulfate,or storageof the extract,because
it hasbeenshown that Li ion is a metabolicinhibitor (King, 1953; Laties, 1959).
Two approacheshave beenusedfor estimationof the amountof adsorbed
sulfate in soils. The first approachmakesthe use of the findings by Chao et al.
(1962a)that adsorbedsulfate is in kinetic equilibrium with solution sulfate and
hencecan be measuredby isotopeexchange.This approachwas usedby Hasan
et al. (1970) to study sulfate desorptionfrom soils. This method,however,over-
estimatesthe amountof adsorbedsulfatepresentin soils, becausethe radioactive
sulfatealso may exchangewith S in soil organicmatter(Freneyet aI., 1971).The
secondapproachinvolves displacementof the adsorbedsulfate in soils by shak-
ing with an appropriatesolution. Severalreagentshavebeenproposedfor extrac-
tion of soluble sulfate plus a fraction of the adsorbedsulfates.These include
NaHC03 (Kilmer & Nearpass,1960), CaC03 suspensions (Williams& Stein-
bergs, 1962), neutral, 1 M NH40Ac (McClung et aI., 1959), acid NH40Ac
(Bardsley & Lancaster,1960; Stanford& Lancaster,1962), and Morgan'ssolu-
tion (Chesnin & Yien, 1950). The adsorbedsulfate also may be displacedby
solutionscontaininganionsof greatercoordinatingability accordingto the fol-
lowing series(Binghamet aI., 1965; Chang& Thomas,1963; Chaoet aI., 1964).
Hydroxyl> Phosphate>Sulfate == Acetate>Nitrate == Chloride.
Alkaline solutions,such as the NaHC03 solution of Kilmer and Nearpass
(1960) and the CaC03 suspensionof Williams and Steinbergs(1962), are effec-
tive, but results obtainedby theseextractantsare difficult to interpret because
thesereagentsalso extractpart of the organicS. Calcium carbonatesuspensions,
however,extractsless organic S than NaHC03• A solution of KH 2P04 contain-
ing 500 mg kg-I of P was recommendedby Ensminger(1954) for extractionof
soluble and adsorbedsulfate in soils. This solution extracts adsorbedsulfate
effectively from most soils, but it has the disadvantagethat the extractsare dif-
ficult to filter and often resultsin turbid extracts;it also may extractsomeS from
organicS fraction. Thesedisadvantages are overcomeby the 0.01 M Ca(HZP04h
introducedby Fox et ai. (1964). This solution containssufficient phosphateto
displacemost of the adsorbedsulfateof many soils. It is necessaryto use a wide
solution/soil ratio or repreatedextractionsto displace the adsorbedsulfate in
soils having strong adsorptioncapacities.The amountsof adsorbedsulfate ex-
tractedby this reagentcan vary from <1 mg kg-I to severalthousandparts per
million. Somestrongly sulfate-adsorbingsurfacesoils of Hawaii contain>1000
mg kg-I and the subsurfacesoils >7000 mg kg-I, of adsorbedSO~--S (Hasanet
aI., 1970).
940 TABATABAI
The sulfatethus extractedcan be determinedby oneof a numberof meth-

ods. Gravimetry,turbidimetry,titrimetry, and colorimetryarethe mostcommon-
ly usedmethodsfor estimationof sulfatein soil extracts.Becausesulfateis pre-
sent in soils as salts of variousmetals,and becausedifferent metalsassociated
with sulfateaffect its determination,recentlyAjwa and Tabatabai(1993) evalu-
ated four analytical methodsbasedon different principles for determinationof
sulfatein different metalsaltsand in soil extractsobtainedwith threeextractants
[0.1 M LiCl, 0.15%CaCI2, and500mg P/L asCa(H2P04)2]. The analyticalmeth-
odswere: (i) a MB colorimetricmethodafter the reductionof sulfateto H2S, (ii)
an IC method,(iii) a turbidimetric (TO) method,and (iv) an indirect Ba atomic
absorptionspectrophotometric(SP) method.They showedthat the recoveryof
sulfateassociatedwith variousmono-, di-, and trivalent metalswas quantitative
by the MB method.But, trivalent metals,such as AI, In, La, and Sc, decreased
the recoveryof sulfateby the otherthreemethods.The MB andIC methodsgave
similar valuesfor sulfatein soils by using the threeextractants.The TO and SP
methodsgavevariableresultsand,in general,underestimated the amountsof sul-
fate in soils. Among the four methods,the MB and IC methodswere the most
accurateand precise.
In anotherstudy, Maynard et al. (1987) evaluatedeight extractantsfor
extractionand determinationof S01--S and total extractableS in surfaceorgan-
ic horizonsof five forest soils. The analysesfor S01--S and total extractableS
were performedby a single-columnIC and inductively coupledplasmaatomic
emissionspectrometry(ICP-AES), respectively.They reportedthat the precision
rangedfrom 3.7 to 8.9% for the IC ana13sesexcludingone aspen(Populus tre-
mulcides Micbx) sample and from 1.9 to 4.8% for the ICP-AES analyses.
Quantitativerecoveriesfrom 102 to 108%andfrom 97 to 108%were reportedin
0.01 M NH4CI extractsby the IC and ICP-AES method,respectively.
Selectionof a procedurefor sulfateanalysisof aqueoussolutionsis depen-
dent on the accuracyand precisiondesired.The advantagesand limitations of
theseandothermethodsusedfor determinationof sulfatehavebeenreviewedby
Beatonet a1. (1968)andis not coveredhere.Among thesemethods,however,the
MB colorimetric methoddevelopedby Johnsonand Nishita (1952) and the IC
suppressed systemarethe mostsensitiveand accurate.Therefore,thesemethods
are describedherefor usein determinationof sulfateextractedfrom soils.
Methods
MethyleneBlue Method
Special Apparatus
1. Suction funnels (polyethylene) and filtering funnel stand (Soil
Moisture EquipmentCo., SantaBarbara,CA).
2. Digestion-distillation apparatusdescribed in "Special Apparatus"
under"Total Sulfur" (Fig. 33-1).
3. Distillation flasks: The flasks employedare 50-mL boiling flasks fit-
ted with standard-taper (19/22)ground-glassjoints andglasshooks,so
that they can be connectedto the reduction-distillationapparatusby
SULFUR 941
spiral steelspringswith endsasshownin Fig. 33-1. Their dimensions

shouldbe suchthat whenthe flasks areconnectedto the reduction-dis-
tillation apparatus,the distancebetweenthe tip of the N2 inlet tube and
the bottom of the flask is approximately4 mm.
4. Cylinder of water-pumpedN2, microburners,delivery tubes,andspec-
trophotometerdescribedin "SpecialApparatus"under"Total Sulfur."
Reagents
1. Calcium phosphatemonohydratesolution [Ca(H2P04h• H20], 500
mg L-l of P: Dissolve 2.03 g of Ca(H2P04h• H20 in about 700 mL
of water, and make to a volumeof 1 L with water.
2. Lithium chloride, 0.1 M: Dissolve 4.2 g of LiCl in about 700 mL of
water, and make to a volumeof 1 L with water.
3. Calcium chloride dihydrate(CaCI2 • 2H20), 0.15%: Dissolve 1.5 g of
CaCl2 • 2H20 in about700 mL of water,and maketo a volumeof 1 L
with water.
4. Reagents1 to 10 describedin "Reagents"under"Total Sulfur" for re-
duction and distillation of sulfateto H2S and estimationas MB.
Procedure.Extract the inorganicsulfate from soils by shaking5 g of soil
«2 mm) with 50 mL of 500 mg L-l P, 0.1 M LiCI, or 0.15% CaCl2 for 30 min
and by filtering (Whatman no. 42 filter paper) the resulting suspension.For
extractionof the adsorbedand soluble sulfate, use 50 mL of the solution con-
taining 500 mg L-l of P; for extractionof soluble sulfate, use 50 mL of 0.1 M
LiCI or 0.15% CaCI2.
Placean aliquot of soil extract(usually 2-20 mL) containing10 to 50 Ilg
of soi"-s in a 50-mL reduction-distillationflask (if >2 mL of aliquot is taken,
place the flask in an oven at 100°C, heat the sampleto dryness,cool the flask,
and add2 mL of water), add 4 mL reducingmixture, and proceedin reduction
and distillation of the SOi"-S to HzS and estimationas MB. For commentscon-
cerning theMB determination,see"Comments"under"Total Sulfur."
Ion ChromatographicMethod
For instruments,reagents,and procedureusedin suppressed-type
IC sys-
tems,seeTabatabaiand Frankenberger(1996, Chapter8).
Comments
The chief advantageof the MB method describedis the rapidity with

which the colorimetric determinationmay be completedand the small sample
size (1-2 mL) requiredfor determinationof the extractedsulfate.The procedure
can detectas little as 2 Ilg of S. If an aliquot of 20 mL is takenfor analysisand
reducedin sizebeforeanalysis,the 2 mg of S is equivalentto O.lllg of S per mil-
liliter. Work by Pirela and Tabatabai(1987) showed that sulfate also can be
reducedto H2S by reagentcontaining Sn and H3P04. Similar to the reducing
mixture used in the 10hnson-Nishita(1952) method, the Sn-H3P04 reagent
942 TABATABAI
reducesa variety of organic S and reducedS compounds(Pirela & Tabatabai,

1987, 1988).
Freney(1958) suggestedthat to obtain a measureof the true water-soluble
sulfate contentof a soil by using the reduction methodof Johnsonand Nishita
(1952), it is necessaryto preparean extract free from colloidal material. Other
inorganicS compoundsthat would be estimatedas sulfatemustbe removedfrom
the solution. This can be achievedby precipitating the sulfate as BaS04 and
removing all soluble compoundsby use of the Pregl filter stick. The reagents
described,however,seemnot to extract significant amountsof organic S redu-
cible by the reducing mixture, becausestudiesby Dick and Tabatabai(1979)
showedthat the resultsobtainedby the MB methodfor inorganicsulfate in soil
extractsobtainedwith severalreagents,including the reagentsdescribedin "Ion
ChromatographicMethod", are similar to thoseobtainedby the IC method.
The amountsof inorganicsulfate extractedfrom neutral and slightly alka-
line soils by the reagentdescribedare almost identical (Tabatabai& Bremner,
1972a; Ajwa & Tabatabai,1993). When the soil pH is <5, however, the phos-
phatereagentdescribedextractsmore inorganicsulfatethan that extractedby the
other two reagents(0.1 M LiCI or 0.15% CaCI2). Consequently,the phosphate
reagentshould be usedfor extractionof total inorganicsulfate. In soils contain-
ing high amountsof adsorbedsulfate,severalextractionsshouldbe made.
The other two reagents,0.1 M LiCI and 0.15% CaClz, extract the soluble
inorganicsulfate in soils. The distinction betweensolubleand adsorbedinorgan-
ic sulfateis especiallyimportantwhen fractionationof the total S in subsoilsam-
ples is desired,becausesubsoilscommonlycontaingreateramountsof adsorbed
sulfatethan do surfacesoils. This differencemay be due to a lower pH in the sub-
soils or to the saturationof adsorption sitesin the surfacesoils with phosphateor
other anions.The presenceof adsorbedsulfate in Brazilian soils comparedwith
Iowa soils has beendemonstratedby Neptuneet al. (1975).
Air drying of soils normally increasesthe amount of sulfate extractedby
the reagentsdescribed(Freney, 1958; Williams & Steinbergs,1959, 1964; Bar-
row, 1961;Tabatabai& Bremner,1972a).An averageincreaseof 20% of extract-
able sulfate has beenreportedfor 12 Iowa soils (Tabatabai& Bremner, 1972a).
Therefore, it is important to use field-moist soil· sampleswhen appropriate.
Resultsreportedby Freney(1958) show that the extractshouldbe madeon sam-
ples taken without delay directly from the field, becausethe degreeof desicca-
tion of the sampleswill influence the value obtainedfor water-solublesulfate.
In determinationof sulfate in soil extract by the MB methoddescribed,it
is importantthat the volume takenfor analysisdoesnot exceed2 mL. With larg-
er volumes, the soil extract should be reducedto 2 mL by heatingor by taking
the aliquot to drynessin the reduction-distillationflask and then adding 2 mL of
water beforeadding thereducingmixture. The conditionsdescribedfor determi-
nation of S by the MB methodshould be observed.
Several IC instrumentspermitting the simultaneousdeterminationof a
variety of anions, alkali metals, alkaline earth metals, and many other metals
have beenevaluated forthe analysisof soils, plant materials,and natural waters
(Tabatabai& Basta,1991; Basta& Tabatabai,1990, 1991; Karmarkar& Tabata-
bai, 1992). The Ie is well suited tothe analysisof complex mixturesof ions, but
SULFUR 943
care must be taken to obtain the samplesin a suitablesolution. This is because

many of the proceduresemployedin soil analysisresult in matricesthat are not
compatiblewith the chemistry of the IC systems.Therefore,such procedures
have to be modified beforeusing them in conjunctionwith the IC system.
Other methods are available for determinationof the extractedsulfate.
These include gravimetric, TB and nephelometric,titrimetric, x-ray fluores-
cence,and atomic absorptionSP methods(for review of literature,seeBeaton et
aI., 1968). None of thesemethods,however, is as sensitiveand accurateas the
MB and IC methods.
If the soil extract under analysis contained reduced inorganic S com-
pounds,the quantitiesof suchcompoundsshouldbe determinedand allowed for
in calculation of results, becausethe MB method describedrecoversreduced
inorganicS compoundsas well as sulfate. In analysisof well-drained,well-aer-
atedsoils, however,reducedinorganicS compoundsare not expected.
The soil extracts obtained by the proceduredescribedare stablefor sever-
al days if storedin a refrigeratorat 4°C.
ORGANIC SULFUR
Although the chemicalnatureof organicS in soils hasbeenunder investi-

gation for the pastthreedecades,little is known aboutthe organicS compounds
presentin soils. Early work showedthe presencein soils of organicS compound
suchas trithiobenzaldehyde(C7H60S3) (Shorey,1913) and free cystine(Putnam
& Schmidt,1959). Other reports(Stevenson,1956; Bremner,1967; Kowalenko,
1978) showedthat cystine and methionineare presentin small amountsin soil
hydrolyzates,and the reports indicate that they may occur as constituentsof
polypeptidesin soils.
Although the chemical nature of organic S in soils cannot be defined as
well as that of inorganic S, three broad groupsof S compoundshave beenrec-
ognized:
1. Organic S that is reducedto H2S by HI. This S is not bondeddirectly
to C and is believed to be largely in the form of estersulfates(e.g.,
phenolicsulfates).
2. Organic S that is reducedto inorganic sulfide by Raney Ni and that
seemsto consistalmost entirely of S in the form of amino acid (e.g.,
cystine and methionine).
3. Organic S that is not reducedby either HI or Raney Ni. This fraction
is assumedto consistof S bondeddirectly to C but not recoverableby
currentmethodsusedfor estimationof C-bondedS (Fraction2).
Unfortunately,no satisfactorydirect methodis availablefor determination
of the total organicS fraction or the organic S fraction reducible by HI. These
fractions, however,can be determinedindirectly. The total organicS can be de-
terminedin a soil residueafter extractionof inorganicS or by differenceinvolv-
ing subtractionof the inorganic S fraction from total S. The organic S reducible
by HI can be estimatedby subtractingthe extractableinorganic S fraction from
944 TABATABAI
the fraction of Sreducibleby HI (Tabatabai& Bremner,1972b; Neptuneet aI.,

1975).
Work by Pirela and Tabatabai(1988) showedthat Sn-H3P04 reagentcan
reducea numberof S model compoundsand organicS in soils. Expressedas a
percentageof total organicS in 13 Iowa surfacesoils, the amountsof Sn-H3P04-
reducible S rangedfrom 15 to 64% (avg. = 35%) and from 42 to 91% (avg. =
60%) for 1 and 10 h of distillation, respectively.The correspondingpercentages
of sevenChileansurfacesoils were from 8 to 75% (avg. =25%) and from 29 to
100%(avg. =55%). The amountsof organicS potentiallyreducible(Sr) with this
reagentrangedfrom 35 to 86% (avg. =61%) of organicS in Iowa surfacesoils.
The correspondingvaluesfor sevenChileansoils rangedfrom 33 to 100% (avg.
=66%). The timesrequiredto distill 50% of Sr rangedfrom 0.3 to 1.7 h (avg. =
0.8 h) for the Iowa soils, and from 0.4 to 9.2 h (avg. =3 h) for the Chileansoils.
Studieswith five Iowa soil profiles showedthat the percentagesof organicS
reducedwith Sn andH3P04in 1 and 10 h of distillation variedwith depthof sam-
pling and that thesevalueswere similar, lower, or greaterthan those obtained
with the HI-reducingmixture.
Hydriodic Acid-Reducible Sulfur
Principles
The basic principles of the Johnsonand Nishita (1952) method is the
reductionof all inorganicSandH2S by a strongacid reducingmixture contain-
ing ill, formic acid, and H3P02. The S evolvedas H2S is determinedcolorimet-
rically as MB. This method does not recover the S in organic compoundsin
which the S is directly bondedto C, such as cysteine,cystine, methionine,and
taurine (H002SCH2CH2NH2) (Johnson& Nishita, 1952; Freney,1958; Arkley,
1961). The acid mixture, however,reducesa variety of organicS compoundsin
which S is presentas estersulfates(C-O-S linkage), suchas diethyl sulfate(sul-
furic acid diethyl ester),p-nitrophoenylsulfate (CJI4N06SK),and chondroitin
sulfate. Application of the Johnsonand Nishita method directly to analysisof
soils hasshown that, on the average,about50% of the total S in soils of humid
and temperateregions canbe convertedto H2S, althoughanalysescoveringwide
rangesof soils have indicated that the percentagemay range from 30 to 80%
(Freney, 1961; Tabatabai& Bremner, 1972b; Neptune et aI., 1975; Williams,
1967; Biederbeck,1978). Becauseof the reductionof S in severalmodel com-
poundsand becauseof presenceof someof these compounds in soils, it is be-
lieved that the major portion of ill-reducible S in soils is presentassulfatedpoly-
saccharides,phenolicsulfates,choline sulfate [(CH3)3WCH2CH20S0j"],or sul-
fated lipids (Freney, 1967). The fraction of S in soils reducedby HI includes
inorganicS (mainly sulfate).Therefore,it is essentialto correctthe HI-reducible
fraction 'for inorganic S when estimationof the estersulfate fraction is desired.
This can be accomplishedby subtractingthe value of inorganicS obtainedby a
separateanalysis(asdescribedin "Methods"under"InorganicSulfate")from the
HI-reducibleS value.The fraction of estersulfatein soils is sometimesdescribed
as organicSOi--So This may be a more accuratedescriptionbecausethe proce-
SULFUR 945
dure usedto determineHI-reducible S recoversS in organicsulfatescontaining

C-N-S linkages(Freney,1961),and thereis no evidencethat suchsulfatesdo not
occur in soils.
Becausethe S in this fraction can be readily hydrolyzedto inorganic sul-
fate by acid or alkali, HI-reducible S is consideredto be the most labile fraction
of soil organic S (Spencer& Freney, 1960; Lowe, 1965; Freney et aI., 1971;
Cooper,1972).This fraction of S is thought to be largely associatedwith certain
side chain componentsof fulvic and humic materials (Bettany et aI., 1973).
Extracts obtainedfrom soil with NaHC03 at pH 10 and with chelating resin
(Freneyet aI., 1969)indicatedthat much of the HI-reducibleS occursin the high
molecularweight (Humic acid) fraction of the soil organicmatter.
Methods
Special Apparatus
1. Digestion-distillation apparatus(Fig. 33-1), and 50-mL distillation
flasks, cylinder of water-pumpedN2, microbumers,delivery tubes,
and spectrophotometer describedin "SpecialApparatus"under"Total
Sulfur."
Reagents
1. Reagents1 to 10 describedin "Reagents"under "Total Sulfur" for
reductionand distillation of sulfate to H2S and estimationas MB.
Procedure.Placea sampleof finely ground «100 mesh) soil containing
20 to 100 ~g of total S (usually 0.1-0.5 g) in a dry 50-mL distillation flask, add
2 mL of water and 4 mL of the reducingmixture, and proceedwith reductionof
the organic and inorganic S04-S to H2S and estimationas MB as describedin
"Reduction of Sulfate to Sulfide and Estimation as Methylene Blue" under
"Total Sulfur."
Comments
The resultsobtainedfor the HI-reducible S fraction are very reproducible.
For estimationof the organicsulfate (ester sulfate)fraction, subtractthe amount
of inorganicsulfate and anyreducedinorganicS compoundspresentin the sam-
ple from the value obtainedfor HI-reducible S. A significant portion of the or-
ganicS in soils can be reduced toH2S by digestionand distillation with a reagent
containingSn and H3P04 (Pirela & Tabatabai,1988). The amountof S reduced
is a function of distillation time. The curvesobtained forcumulativeamountsof
S reducedin surfaceand subsurfacesoils as a function of distillation time sug-
gest that the chemical natureof S is dissimilar amongsomesoils. The time re-
quired to reducethe potentially reducibleorganicS by this reagentdependson
the soil type and depthof sampling.The possibility that S-containingamidesare
presentin soils merits attentionbecauseit is known that soils containamidesand
that this form of N is a major sourceof NHt in soils. It could be a sourceof SO~
as well. The Sn-H3P04 reagentcan be usedin conjunctionwith HI and RaneyNi
in studiesof the reducibleorganic S fraction in soils. As is the casewith HI and
RaneyNi, however,the resultsindicatethat the Sn-H3P04 reagentreducessome
946 TABATABAI
of the organicS compoundsin soils, but the chemical natureof theseis uncer-
tain. For commentsabout the MB color measurement,see"Comments"under
"Total Sulfur."
Carbon-Bonded Sulfur
Principles
In 1925 Murray Raney was granteda patent covering a new method of
preparationof a Ni catalyst.A pulverizedNi-Si alloy was reactedwith aqueous
NaOH to produce a pyrophoric, brownish Ni residue with superior catalytic
properties.On investigationof otheralloys ofNi and alkali-solublemetals,it was
found that the aluminum alloy could be madewith easeand was easily pulver-
ized. The catalyst,which is preparedby the action of aqueousNaOH on this Ni-
AI alloy, is known as Raney Ni. Raney Ni has beenshown to remove S from
organicand inorganiccompounds(Lieber & Morritz, 1953). The reductionwith
Ni-AI alloy and aqueousalkali is probably due to the activation by the freshly
formed Ni catalystof the H2 liberatedby the action of the alkali on the AI com-
ponentof the alloy.
Application of RaneyNi to analysisof S directly bondedto C in soils was
first proposedby DeLong and Lowe (1961), and since then it has beenusedby
many workersfor estimationof C-bondedS in soils. The methodinvolves desul-
furization by RaneyNi in the presenceof alkali in the Johnsonand Nishita appa-
ratus, liberation of H2S by acidification, and estimationof the sulfide released
colorimetrically as MB. Application of the methodto a variety of organicS com-
poundsshowedthat the methoddevelopedby DeLong and Lowe (1961) recov-
ers all forms of organic S exceptS in organic sulfate and alkyl sulfones(Lowe
& DeLong, 1963; Freneyet aI., 1970). It also recoverselementalS and reduced
inorganic S (e.g., S20J-,S201-, S40g-, and SOJ-),but well-drainedsoils usual-
ly do not contain measurableamountsof theseinorganic forms of S (Tabatabai
& Bremner, 1972b; Nor & Tabatabai,1976, 1977). Lowe and DeLong (1963)
found that >95% of the total S in four of five Quebecsoils could be accounted
for as C-bondedS and HI-reducibleS, but only 57% ofthe total S in one ofthese
soils could be accountedfor in thesetwo forms, and they postulatedthe existence
in this soil of an "inert" form of S that is not recoveredby the proceduresused
to determine HI-reducible and C-bonded S. Freney et ai. (1970), however,
showedthat Fe and Mn can interferewith determinationof C-bondedS in soils
by the methodof Lowe and DeLong (1963) and that a substantialfraction (avg.
=23%) of the organicS in 15 Australiansurfacesoils could not be accountedfor
as (HI-reducible + C-bonded)-Seven when the method used to determineC-
bondedS was modified to reduceinterferenceby Fe and Mn. Other factors that
may affect the results obtained for C-bondedS by the method of Lowe and
DeLong (1963) are the amount of Raney Ni and concentrationand amount of
NaOH used(Freneyet aI., 1970).
It hasbeensuggestedthat C-bondedS shouldbe estimatedas beingthe dif-
ferencebetweentotal and HI-reducibleS (Freney,1967),andsomeworkershave
adoptedthis simple procedure.The assumptionthat all organicS in soils that is
SULFUR 947
S
18/15
CONDENSER .............
FUNNEL~
NITROGEN GAS
INLET TUBE
f2
em
I...J...J...J..
o 10
Fig. 33-3. Digestion-distillation apparatusfor determina-

tion of C-bondedS.
not HI-reducible consistsof C-bondedS, however,is probably an oversimplifi-

cation and is not justified in view of the existencein soils of sizeableamountsof
unidentified S with characteristicsdiffering from those of HI-reducible and C-
bondedS (Lowe, 1965; Freneyet aI., 1970; Tabatabai& Bremner,1972b; Nep-
tune et aI., 1975). The presenceof this unidentified or residualfraction of soil
organicS was first suggestedby Lowe (1964) and later supportedby resultsfrom
other studiesshowingthat this fraction accountedfor between3 and 59% of the
total organic S in mineral soils (Biederbeck,1978).
Basedon analysesof soil hydrolyzates,it has been estimatedthat about
half of the C-bondedS occursas amino acids(Freneyet aI., 1972).The natureof
the remainder(unidentified)of this fraction is unknown.A review by Biederbeck
(1978) showsthat C-bondedS accountsfor between5 and 54% of the total S in
some mineral soils of Canada,Australia, USA, and Brazil. Lowe and DeLong
(1963) reportedconcentrationsof 47 to 58% C-bondedS in severalorganicsoils.
Methods
Special Apparatus
1. Digestion-distillationapparatus(Fig. 33-3). This apparatusis similar
to that describedin Fig. 33-1, but a funnel connectedto a two-way
Teflon stopcockis attachedto the N zgasdelivery tube outsidethe con-
denser.The two-way stopcockis used to regulatethe Nz flow and to
deliver HCI into the flask.
2. Distillation flask: The flasks employedare 150-mL boiling flasks fit-
ted with standard-taper (19/22) ground-glassjoints andglasshooks,so
that they can be connectedto the reduction-distillationapparatusby
spiral steelspringsas shownin Fig. 33-3. Their dimensionsshouldbe
such that when the flasks are connectedto the reduction-distillation
apparatus,the distancebetweenthe tip of the Nz inlet tube andthe bot-
tom of the flask is approximately4 mm.
948 TABATABAI
3. Cylinder of water-pumpedN2, microbumers,delivery tubes,andspec-

trophotometerdescribedin "SpecialApparatus"under"Total Sulfur."
Reagents
1. RaneyNi catalyst:Preparein the reductionflask from RaneyNi alloy
(The British Drug HousesLimited, B.D.H., Poole,England).
2. Sodiumhydroxidesolution, 5%: Dissolve50 g of NaOH in about800
mL of water, and adjust the volume to 1 L with water.
3. Hydrochloric acid, 1:1: Mix 100 mLof HCI with 100 mLofwater.
4. Reagents2 to 7 describedin "Reagents"under "Total Sulfur" for
reductionand distillation of sulfate to H2S and estimationas MB.
Procedure.Place a sampleof finely ground «80 mesh or 180 !lm) soil
containing 10 to 50 !lg of C-bondedS (usually 0.1-0.5 g) in a digestion flask,
add about 0.1 g of Ni-Al alloy, 5 mL of 5% NaOH, and 25 mL of deionized
water. Moisten the joint of the condenserwith a drop of water, and connectthe
flask to the condenser.Digest over a low flame for 30 min. Tum off the flame,
and allow the flask to cool somewhat.Attach a delivery tube to the side arm of
the washingcolumn, and immersethe other end of the tube in a 100-mL volu-
metric flask (receiving flask) containing Zn(OAc)2 and NaOAc "Reduction of
Sulfate to Sulfide and Estimationas MethyleneBlue." Add excessHCI (e.g., 5
mL of 1:1 HCI) to the funnel, and tum the two-way stopcockto deliver the HCI
into the boiling flask. Then tum the stopcockback to its original position (to
allow N2 bubbling in the mixture). Heatgently on a low flame for 30 min to dis-
till the H2S evolved,and proceedwith estimationof the H2S evolvedas MB after
addition of p-aminodimethylanalineand ferric ammonium sulfate reagentsas
describedin "Reduction of Sulfate to Sulfide and Estimation as Methylene
Blue." The methoddescribedis essentiallythat reportedby Lowe and DeLong
(1963).
Comments
The resultsobtainedby the proceduredescribedfor estimationof C-bond-
ed S are affectedby samplesize, amountof Raney Ni added,and amount and
concentrationeachof NaOH and HCI used.Boiling soil and RaneyNi in a solu-
tion containing NaOH as describedmay lead to volatilization of some organic
compoundsthat lead to adsorption,inside the condenser and V-tube, of the H2S
evolved after addition of HCl. Therefore,when the resultsare less than expect-
ed, the apparatusshouldbe disassembledand washedwith dilute HCI to remove
the adsorbedsulfides. The apparatusmust be washed thoroughly with water
before it is reassembled.
If the sample under analysis is suspectedto contain sulfides or other
reducedinorganicS compounds,a separateanalysismust be performedby boil-
ing a samplein 20 mL of watercontaining5 mL of 5 M HCI and determiningthe
H2S evolvedas describedin "Reductionof Sulfate to Sulfide and Estimationas
MethyleneBlue." The valueobtainedfor C-bondedS shouldbe correctedfor the
amountof sulfide and other reducedinorganicS compoundsthat may evolve as
H2S upon boiling in HCl.
SULFUR 949
The conditionsreportedfor MB color developmentand absorbancemea-

surementdescribedin "Comments"under"Total Sulfur" shouldbe adheredto.
For commentsaboutusing a Sn-H3P04 reagent,see"Comments"under"Hydro-
idic Acid-ReducibleSulfur."
It is generallyacceptedthat S is takenup by plantsin the sulfateform. The

information availablefrom nutrient culture experimentsindicatesthat organicS
compoundsmay be utilized by plants (Miller, 1947; Bardsley,1960); however,
availability of soil organicS dependsprimarily on its mineralization.This in tum
dependson climatic factors suchas temperatureand moisture(Williams, 1967),
which affect the rate of mineralizationas well as chemical nature of the soil
organicS itself.
Determinationof the plant-availableS in soils is complicatedby the fact
that severalsourcesbesidesthe soil itself contribute to the S needsof crops.
Thesesourcesinclude S in rainfall and irrigation water, atmosphericS, and S in
fertilizers and pesticides.Of thesesourcesof S, the contributionsof S in rainfall
and direct absorptionby soils and plants from the atmosphereare perhapsthe
most difficult to evaluate.Annual additionsby rainfall rangingfrom 2.2 to 234
kg of S perhectarehavebeenreportedby Eriksson(1960).In addition,both plant
and soils absorbS02' and probablyother S gases,directly from the atmosphere
(Fried, 1947;Ulrich et aI., 1967;Allway et aI., 1937).A study in Wisconsinwith
35S showedthat underoptimum yield, 14% of the S in alfalfa (Medicago sativa
L.) was derivedfrom atmosphericsources(Hoeft et aI., 1973).
Numerousprocedureshave been proposedfor evaluation of the plant-
availableS in soils. The proceduresusedinclude extractionwith water, extrac-
tion with various salts and acids, S mineralizationduring incubation,microbial
growth, and plant growth and composition,including "A" value determinations.
The S removedby the variousextractantstendsto fall into the following groups:
(i) readily soluble sulfate, (ii) readily soluble and portions of adsorbedsulfate,
and (iii) readily solubleand portionsof adsorbedsulfateand portionsof organic
S (Reisenaueret aI., 1973).
Reviews of the proceduresavailable indicate that no one procedurehas
provedconsistentlysuperiorin predictingresponsesto S fertilization (Reisenau-
er et al., 1973).The S extractedby many of the procedurescorrelateswell, how-
ever,with S uptakeby plantsin greenhouse experiments.Extractantsthat remove
the readily soluble plus portionsof adsorbedsulfatetend to be bettercorrelated
with S uptakeby plants than those removing only the readily soluble sulfate.
Many extractantshavebeeninvestigatedto evaluateplant-availableS, including
acetate(Harward et aI., 1962), NaHC03 (Kilmer & Nearpass,1960), cold and
hot water(Spencer& Freney,1960),dilute alkali and salt solutions(Williams &
Steinbergs,1959), and NH4HC03-DTPA (diethylenetriaminepentaacetic acid)
(Vaughan,1991).HeatingreleasesS from soils, andthis hasbeenmadethe basis
of the heat-solubletest developedby Williams and Steinbergs(1959). Although
there are reportson the correlationbetweenplant-availableS and organic S or
950 TABATABAI
total S, such relationshipsare not expectedwith the various fractions of the

organicS pools in soils (e.g., HI-reducibleS RaneyNi-reducibleS). Greenhouse
experimentsinvolving tracer techniquesby Freneyet aI. (1975) showedthat S
uptake by Sorghum vulgare resulted in changesin both HI-reducibleand C-
bondedS fractions, 60% of the S taken up by plantswas derivedfrom the latter.
The changesin C-bondedS involved changesin both Raney Ni-reducible and
nonreducibleS fractions. Freneyet aI. (1975) concludedthat all the S fractions
they investigatedcontributedavailableS for plant uptake,and none of them is
likely to be of any value for predictingthe S requirementsof plants.
Little information is available about the nature of the mineralizableor
readily availablefraction of soil S. Most of the proceduresusedfor estimationof
availableS are basedon arbitrary analytical methods.Therefore,as mentioned
previously,numerousreagentshavebeenproposedfor extractionand estimation
of the availablesoil S fraction (Beatonet aI., 1968). However,there is no gener-
al agreementon the best methodor methodsfor use as an index of plant avail-
ability. Most of the soil testsdevelopedup to 1966were reviewedby Ensminger
and Freney(1966), and little progresshasbeenmadesincethat time. The proce-
dure usedgenerallyfalls into two groups:(i) plant analysesand (ii) soil analyses.
The use of plant analysesfor assessingS deficienciesin soils is basedon the
notion that an essentialelementshouldbe presentin the plant at a concentration
just sufficientfor unrestrictedplant growth. The discussionof thesemethodsis
beyondthe scopeof this chapterand is not presentedhere.The readeris referred
to the summaryof the procedureson diagnosisof S deficiency by plant analysis
preparedby Ensmingerand Freney(1966).
Severalmethodsbasedon analysisof soils havebeenproposedfor estima-
tion of plant-availableS. These include extraction of soi--s, extraction of
organicS, incubationtechniques,and microbial assays.
Extractionof Sulfate-Sulfur
Numerousreagentshave beenproposedfor measuringextractableSOi--S

in soils. Theseinclude water (Spencer& Freney,1960; Fox et aI., 1964; Walker
& Doomenbal,1972), salt solutions such as 0.15% CaCl2 (Williams & Stein-
bergs,1959; Barrow, 1961),0.1M LiCI (Tabatabai& Bremner,1972a),as mM
MgCl2 (Roberts& Koehler, 1968),500mg L-l of PasCa(H2P04)2 or KH 2P04
(Fox et aI., 1964;Joneset aI., 1972); and acidic solutionssuchas 0.5 M NH40Ac
+ 0.25 M aceticacid (CH3COOH) and Bray P-1 (Rehm& Caldwell, 1968; Hoeft
et aI., 1973). Severalof theseand otherextractantshavebeenevaluatedby May-
nard et ai. (1987)for extractionand determinationof S in organichorizonsof for-
est soils. They reportedquantitative recoveriesof total and SOi- -S in 0.01 M
NH4CI extracts.Generally,phosphatesolutionsextract more SOi--S from soils
than water or CaCl2 or LiCI solution, becausephosphateions displace the
adsorbedsulfate, which is known to be readily available to plants (Spencer&
Freney,1960; Williams & Steinbergs,1964). Seealso "Principles" under"Inor-
ganic Sulfate."
Recenttracer experimentsshowedthat the sulfate extractedwith 0.01 M
Ca(H2P04)2 is derivedfrom the samepool of soil S as that usedby plants(a trop-
SULFUR 951
icallegume,Stylosanthes guyanensis); this led Probert(1976) to suggestthat the

reagentscontainingCa(H2P04)2 are the bestextractantsfor assessingthe S-sup-
plying power of soils. Also, since near full recovery of added 35S0~- was
obtainedinitially by this reagent,Probert (1976) found that this S fraction is
comparablewith the "L" value. On the other hand, 0.5 M NaHC03 removed
some soil S that was not available to plants. The finding reportedby Probert
(1976) on Ca(H2P04h-extractable sulfateis in agreementwith findings reported
by other workers. Significant correlation coefficients for the relationship
betweenphosphate-extractable S and plant uptakeof S havebeenreportedfor a
wide rangeof soils (Fox et aI., 1964; Barrow, 1969; Joneset aI., 1972; Hoeft et
aI.,1973).
Severalfactors are involved in evaluatingchemicalmethodsfor assessing
the plant-availableS. Theseincludeanalyticalfeasibility, reactionsof the reagent
with different soils, and the probablesignificanceof the forms of S extractedto
plant nutrition. To be useful for routine analysis,the reagentusedfor extraction
of sulfatefrom soils shouldgive a clearfiltrate. In this regard,Ca(H2P04hseems
to be one of the bestextractants,becausephosphateions displacethe adsorbed
sulfate,and Ca ions flocculatethe soil colloids. Use of KH 2P04normally results
in a moreturbid solutionthandoesCa(H2P04h, andwateror Cl- saltsdo not dis-
place the adsorbedsulfate from soils (Fox et aI., 1964; Harward & Reisenauer,
1966).
Severalmethodsare availablefor determinationof the extractedsulfate.
Theseinclude gravimetric,TB, and nephelometric,titrimetric, colorimetric, x-
ray fluorescence,and atomic absorptionSP, IC methods,and ICP-AES (Beaton
et aI., 1968; Krupa & Tabatabai,1986; Maynardet aI., 1987;Tabatabai& Basta,
1991). Of thesemethods,the TB determinationof sulfateas BaS04is the most
widely used.This is becauseof the simplicity of the procedure,but the results
obtainedby thesemethodsinvolve a high risk of errors(Reisenaueret aI., 1973).
To overcomethe error associatedwith TB methods,BettanyandHalstead(1972)
havedevelopedan automatedprocedureinvolving TB-nephelometricdetermina-
tion of sulfatepresentin 0.01 M CaCl2 extractsin soils. In this methodthe sul-
fate in the extract is precipitatedas BaS04in an acidic aqueousmedium using
polyvinyl alcohol as a suspension-stabilizing agent. Interferencefrom organic
matteris preventedby clarifying the soil extractwith Na202' The determination
of sulfate in the clarified extract utilizes Technicon autoanalyzerequipment
(TechniconIndustrial Systems,Tarrytown, NY) in conjunction with a Turner
Model III Fluorometer(Turner, Mountain View, CA).
The Johnsonand Nishita (1952) reduction-distillationmethod,which mea-
suresorganicas well as inorganicsulfate,is bestsuitedfor determinationof sul-
fate in soil extracts.The resultsby this methodare not affectedby the turbidity
of the soil extracts,but they include any organicsulfatereleasedfrom soil dur-
ing extraction(Freney,1958). Recentevaluationsof severalreagentscommonly
usedfor extractionof sulfatefrom soil [e.g., 0.15% CaCI2, 0.1 M LiCI, 500 mg
L-1 of P as Ca(H2P04)2]in conjunctionwith the MB methodof Johnsonand
Nishita (1952) showed,however,that the resultsby this methodagreedclosely
with thoseobtainedby IC, indicating that very traceamounts,if any, of organic
sulfateare releasedfrom soils by theseextractants(Dick & Tabatabai,1979).
952 TABATABAI
Severalfactors affect the extraction of sulfate from soils. Theseinclude

samplepreparation,soil extractantratio, and shakingtime. Oven drying marked-
ly increasesthe amount of extractableS (Barrow, 1961). Similarly air drying
increasesthe amountof sulfateextractedfrom soils (Freney,1958; Barrow, 1961;
Williams & Steinbergs,1964; Williams, 1967; Tabatabai& Bremner, 1972a,b).
In one such study, for example,air drying of 12 field-moist Iowa surfacesoils
increasedthe amountof SO~--S extractedby 0.1 M LiCI, and the increaseranged
from 0.2 to 1.6 mg kg-1 and averaged0.9 mg kg-1 (Tabatabai& Bremner,
1972a).Theseincreasesare similar to thoseobservedin studiesof the effect of
air drying on the amountof SO~--S extractedfrom Australian soils by 0.15%
CaCl2 solution (Barrow, 1961; Williams & Steinbergs,1964; Williams, 1967).
Otherwork hasshownthat storageof air-dried organichorizonsof forest soils at
room temperature(20 ± 4°C) for 1.5 yr resultedin increasedtotal extractableS
and SO~--S concentrations(Maynard et aI., 1987).
Extractionof OrganicSulfur
Since the extractable SO~--S is an index of readily plant-availableSand

doesnot include the portion of the labile fraction of soil organicS that is miner-
alized during the growing season,some workers have developedmethodsto
extract the sulfate S plus a fraction of the labile organic S. 1)te significanceof
the organicS fraction extractedwith the sulfate as an index of plant-availableS
is mainly related to level of inorganic SO~--S initially presentin soils. When
soils containvery low amountsof sulfate,the amountsof S mineralizedmay be
significant in preventingS deficiency in plants. In a recent study, Hamm et al.
(1973) showedthat, if the contributionsof mineralizedS are neglected,53% of
the soils testedin the Grey soil zoneand 18% of the Black soils in Saskatchewan
can be consideredas potentially S deficient for alfalfa and rapeseed(Brassica
Napus L.). The importanceof the mineralizableS in predictingS requirementof
crops also was emphasizedby Bettany et ai. (1974), who found that most of the
S takenup by alfalfa from severalSaskatchewan soils was obtainedfrom S min-
eralizedduring plant growth.
The following methodshavebeenusedfor extractionand estimationof the
inorganicsulfateand a fraction of the labile soil organicS that contributesto the
plant-availableS.
Heat-SolubleSulfur
This methodmeasuresthe amountof water-solubleS releasedduring gen-
tle hydrolysis as a result of sequentialwet and dry heatingof soils (Williams &
Steinbergs,1959). It involves evaporationto drynessof a mixture of 5 g of soil
and 20 mL of water in a silica basin placedon a gently boiling water bath and
heatingthe dried soil in a hot-air oven at 102°C for 60 min. After cooling, the
soil is transferredinto a 50-mL centrifugetube andextractedwith 33 mL of 0.1%
NaCl. The total S extractedthen is determinedTB after ignition. The total S in
this fraction also can be estimated by the reduction-distillation method of
Johnson and Nishita (1952). Spencer and Freney (1960) reported that this
method extracts amountsof S from soils similar to those obtained by direct
SULFUR 953
extractionwith hot water. Heat-solubleand autoclavemethods,however,extract

more than twice and four-times as much soil S, respectively,as that extractedby
a solution containing 500 mg L-I of P (Fox et aI., 1964). Greenhouseexperi-
ments in Australia (Williams & Steinbergs,1959; Spencer& Freney, 1960) and
field studiesin Nebraska(Fox et aI., 1964) have demonstratedthat heat-soluble
S is highly correlatedwith S uptake by plants. This very labile fraction of soil
organic S seemsto provide a satisfactoryindex of plant-availableS in a variety
of soils.
Acetate-ExtractableSulfur
This method was proposedby McClung et aI. (1959) for estimation of
plant-availableS in Brazilian soils. It involves extraction of S from soils with
neutral,1 M NH40Ac, evaporationof the extractto dryness,ashingof the residue
by ignition in the presenceof Mg(N03)z, and determinationof the sol- -S in the
ignited sample by the method of Johnsonand Nishita (1952). Generally, this
methodextractsamountsof S similar to thoseextractedby a solution containing
500 mg L-I of P as Ca(HZP04)z, but it does not displacethe adsorbedsulfate.
Although the fraction of S extractedby NH40Ac containsrelatively little organ-
ic S, it was found to provide a reasonableindex of the S statusof someAustralian
soils (Spencer& Freney,1960).
Bicarbonate-Extractable
Sulfur
This methodwasproposedby Kilmer and Nearpassin 1960,and sincethen
it has becomeone of the most widely acceptedtests for plant-availableS. It
involves determinationby the MB methodof organicand inorganic sulfatein an
aliquot of extractobtainedby shaking10 g of soil with 40 mL of 0.5 M NaHC03
(adjustedto pH 8.5 with NaOH) and filtering the resultingsuspension(Whatman
no. 42 filter paper).The pH of the NaHC03 reagentis important becauseat pH
10 bicarbonatesolution extracts considerablymore soil S than at the recom-
mendedpH 8.5 (Kilmer & Nearpass,1960). This methodmeasures adsorbed sul-
fate togetherwith free sulfateand somelabile organicS (Williams & Steinbergs,
1964; Freney et aI., 1971). Evaluations of this method have shown that the
amountsof S extractedare generallytwice thoseextractedwith phosphatesolu-
tions (Rehm & Caldwell, 1968; Probert, 1976). Other studieswith this method
have shown that it provides a reliable index of plant-availableS. For example,
Kilmer and Nearpass(1960) reportedthat the results by this method were sig-
nificantly correlated(r = 0.89) with S "A" valueswhenusingyoungcotton plants
(Gossypium hirsutum L.) grown on 30 soils from the southeasternUSA. Studies
by Rehm and Caldwell (1968) showedthat plant uptake of S from Minnesota
soils was more closely correlatedwith bicarbonate-extractable S (r = 0.95) than
the acetate-or phosphate-extractable S. The amountsof S determinedby the
bicarbonateextractionmethodare generallygreaterthan those extractedby the
acetatemethodsof McClung et al. (1959) and Bardsley and Lancaster(1960).
Analysis of soils beforeand after cropping indicatedthat decreasesin bicarbon-
ate-extractableS during plant growth accountedfor most of the S taken up by
plants (Williams & Steinbergs,1964). Contrary to thesefindings, in a study of
954 TABATABAI
available and isotopically exchangeableS in severalAustralian soils, Probert

(1976) reportedthat the bicarbonatemethodmeasureda much larger pool of S
than that which is availableto plants. He concludedthat this fraction doesnot
representthe potentially mineralizableS sinceit was not significantly correlated
with the "L" valuesof the soils studied.Since the bicarbonatemethodextracts
somecolored soil materials,it is not recommendedto be usedwith procedures
involving TB determinationof sulfate. A review by Reisenaueret al. (1973)
shows that with the exception of few state soil testing laboratories,the TB
methodof analysisis the most widely usedprocedurefor providing availableS
soil tests.The extractantsusedin conjunctionwith the TB methodsof analysis
are water, acid NaOAc, Ca(H2P04)2,KCI, HCI, CaCI2, and acid Ca(H2P04)2
solution, all of which gaverelatively clear extractswith certainsoils.
OtherMethods
Incubation proceduresfor assessingplant-availableS paralleling those
used in estimationof available N have met with little success(Barrow, 1961;
Hesse,1957;Tabatabai& Bremner,1972a),becauseprecisedeterminationof the
small amountof sulfateproducedduring incubationis difficult. In addition, the
amountof S mineralizedis affectedby presenceor absenceof plant (Nicolson,
1970).
Most studies to evaluate the S mineralization potentials of soils have
involved incubatingsoils under aerobicconditionsand determiningthe amount
of sulfatereleasedafter certaintime. Suchstudiesnormally show that very little
SO~--S is mineralized under laboratory conditions. For example, studies by
Tabatabaiand Bremner(1972a)showedthat the averageamountof S mineral-
ized in 12 Iowa surfacesoils during lO wk of aerobicincubationwas2.8 mg kg-1
and represented 1.3% of the total S in 12 surfacesoils. They also showedthat
mineralizable S (lO-wk value) was not significantly correlatedwith total S,
SO~- -S, organicC, total N, or mineralizableN (lO-wk value). Otherwork, how-
ever,showedthat incubationof soils in leachingcolumnswith intermittentleach-
ing with 0.01M KCI resultedin mineralizationof significantamountsof SO~- -So
Results reported by Tabatabaiand Al-Khafaji (1980) show that from 3.7 to
12.6%(avg. =7.7%) andfrom 8.7 to 33.2%(avg. =22.1%)of the soil organicS
was mineralizedin 12 Iowa surfacesoils incubatedfor 26 wk at 20 and 30°C,
respectively. The mineralization of S during incubation of soils in leaching
columnswith intermittentleachingdeservesfurther investigation,becauseTaba-
tabai & Al-Khafaji (1980) showedthat the cumulativeamountsof SandN min-
eralizedduring 26 wk were significantly correlatedat 20°C (r =0.95,P < 0.001)
and at 35°C (r =0.79,P < 0.01). For use as a diagnosticaid, the time involved
in obtainingthe quantitativedatawould be a seriousshortcoming.
Biological evaluationusing growth of Aspergilus niger on soils has been
shown to correlateas well as the better chemical extractants(e.g., phosphate
solutions)with plant response and S uptakein greenhouse experiments.This fun-
gus can remove amountsof S from soils similar to thosethat can be extracted
with phosphatesolutions. Spencerand Freney(1960) found that the growth of
Aspergilus on severalAustraliansoils was significantly correlatedwith S uptake
SULFUR 955
by plants(r =0.83). becauseof the absenceof any markedadvantage,however,

the procedurallimitations of such methodswould precludetheir use as routine
laboratory procedures.The current information of the methodsused as plant-
availability indicesindicatesthat thereis a real needfor a simple and quick lab-
oratory procedurethat adequatelyestimatesthe S supplyingpower of soils.
Recently,Qian et al. (1992) developedand assessed a methodfor simulta-
neousextractionof plant-availableN, P, S, and K by using anion- and cation-
exchangemembranes(ACEM). They reportedthat this techniqueis highly suit-
able for routine soil testing due to its simplicity, rapidness,and accuracy.The
study comparedthe amountof nutrientsextractedby ACEM with conventional
chemical-based extractantsfor P and K (0.5 M NaHC03) and Nand S (0.001 M
CaCI2) for 135 soils samplesrepresentinga wide rangeof soil types in Western
Canada.The nutrient availability predictedby ACEM was significantly correlat-
ed with the conventionalmethods.The availability of all four nutrientscan be
assessed in a single ACEM extraction.
REFERENCES
Ajwa, H.A., and M.A. Tabatabai.1993. Comparisonof somemethodsfor determinationof sulfate in
soils. Commun.Soil Sci. Plant Anal. 24:1817-1832.
Allway, F.J., A.w. Marsh, and W.J. Methley. 1937. Sufficiency of atmosphericsulfur for maximum
crop yields. Soil Sci. Soc. Am. Proc. 2:229-238.
A1ymore, L.A.G., M. Karim, and J.P. Quirk. 1967. Adsorption and desorptionof sulfate ions by soil
constituents.Soil Sci. 103:10-15.
Arkley, T.H. 1961. Sulfur compoundsof soil systems.Ph.D. diss. Univ. California, Berkeley.
Aspiras,RD., D.R Keeney,and G. Chesters.1972. Determinationof reducedinorganicsulfur forms
as sulfide by zinc-hydrochloricacid distillation. Anal. Lett. 5:425-432.
Associationof Official Agricultural Chemists.1955. Official methodsof analysis.8th ed. AOAC,
Washington,DC.
Bardsley,C.E. 1960.Absorption of sulfur from organicand inorganicsourcesby bushbeans.Agron.
J.52:485-486.
Bardsley,C.E., andJ.D. Lancaster.1960. Determinationof reservesulfur and solublesulfatesin soils.
Barrow, N.J. 1961. Studieson mineralizationof sulphurfrom soil organicmatter.Aust. J.Agric. Res.
12:306-319.
Barrow, N.J. 1969. Effects of adsorptionof sulfate by soils on the amountof sulfate presentand its
availability to plants. Soil Sci. 108:193-201.
Basta, N.T., and M.A. Tabatabai.1990. Ion chromatographicdeterminationof total metals in soils.
Soil Sci. Soc. Am. J. 54:1289-1297.
Basta,N.T., and M.A. Tabatabai.1991. Determinationof total metalsin sewagesludgesby ion chro-
matography.J. Environ. Qual. 20:79-88.
Beaton,J.D., G.R Burns, and J. Platou. 1968. Determinationof sulphur in soils and plant material.
Tech. Bull. 14, SulphurInst., Washington,DC.
Bentley, C.F., DJ. Hoff, and D.B. Scott. 1955. Fertilizer studieswith radioactivesulphur. II. Can. J.
Agric. Sci. 35:264-281.
Berg, w.A., and G.w. Thomas.1959. Anion elution patternsfrom soils and soil clays. Soil Sci. Soc.
Am. Proc. 23:348-350.
Bertrand,G., and L. Silberstein.1927. Sur la dosagedu soufrecontenudansla terre arable.Ann. Sci.
Agron. Fr. Estrang.43:71-77.
Bettany,J.R,and E.H. Halstead.1972. An automatedprocedurefor the nephelometricdetermination
of sulfate in soil extracts.Can. J. Soil Sci. 52:127-129.
Bettany, J.R., J.W.B. Stewart,and E.H. Halstead.1973. Sulfur fractions and carbon, nitrogen, and
sulfur relationshipsin grassland,forest, and associatedtransitionalsoils. Soil Sci. Soc. Am.
Proc. 37:915-918.
956 TABATABAI
BettanY3I.R., 1.w.G. Stewart,and E.H. Halstead.1974. Assessmentof available soil sulphur in an

5S growth chamberexperiment.Can. 1. Soil Sci. 54:309-315.
Biederbeck,V.O. 1978. Soil organicsulfur and fertility. p. 273-310.In M. Schnitzerand S.M. Khan
(ed.) Soil organicmatter. ElsevierNorth-Holland,Inc., New York.
Bingham, ET., I.R Sims, and AL. Page.1965. Retentionof acetateby montmorillonite. Soil Sci.
Soc. Am. Proc. 29:670-672.
Blanchar,R.w. 1986.Measurementof sulfur in soils and plants.p. 455-490.In M.A. Tabatabai(ed.)
Sulfur in agriculture.Agron. Monogr. 27. ASA, CSSA, and SSSA,Madison,WI.
Bloomfield, C. 1962. A colorimetric methodfor determiningtotal sulfur in soils. Analyst (London)
87:586-589.
Bremner,I.M. 1967. Nitrogenouscompounds.p. 19-66. In AD. Mclaren and G.H. Peterson(ed.)
Soil biochemistry.Vol. 1. Marcel Dekker, Inc., New York.
Bremner,I.M., and M.A Tabatabai.1971. Use of automatedcombustiontechniquesfor total carbon,
total nitrogen, and total sulfur analysisof soils. p. 1-15. In L.M. Walsh (ed.) Instrumental
methodsfor analysisof soils and plant tissue.SSSA,Madison,WI.
Briimmer, G., H.S. Grunwaldt,and D. Schroeder.1971a.Contributionsto the genesisand classifica-
tion of Marsh soils. II. On the sulfur metabolism of muds and salt marshes. Z.
Pflanzenernaehr. Dueng.Bodenkd.128:208-220.
Briimmer, G., H.S. Grunwaldt,and D. Schroeder.1971b.Contributionsto the genesisand classifica-
tion of Marsh soils. III. Contents,oxidation status,and mechanismsof boundingof sulfur in
poldersoils. Z. Pflanzenernaehr. Dueng. Bodenkd.129:92-108.
Chang,M.L., and G.W. Thomas.1963. A suggestedmechanismfor sulfate adsorptionby soils. Soil
Sci. Soc. Am. Proc. 27:281-283.
Chao, T.T., M.E. Harward, and S.c. Fang. 1962a. Movement of 35S tagged sulfate through soil
columns.Soil Sci. Soc.Am. Proc. 26:27-32.
Chao,T.T., M.E. Harward,and S.c. Fang. 1962b. Soil constituentsand propertiesin the adsorption
of sulfate ions. Soil Sci. 94:276-283.
Chao, T.T., M.E. Harward, and S.C. Fang. 1964. Iron and aluminum coatingsin relation to sulfate
adsorption.Soil Sci. Soc. Am. Proc. 28:632-635.
Chesnin,L., and C.H. Yien. 1950. Turbidimetric determinationof availablesulfates.Soil Sci. Soc.
Am. Proc. 15:149-151.
Clark, J.S.,C.A Gobin, andD.N. Sprout. 1961. Yellow mottles in somepoorly drainedsoils of the
Lower FraserValley, British Columbia. Can. J. Soil Sci. 41:218--227.
Cooper, PJ.M. 1972. Aryl sulphataseactivity in northern Nigerian soils. Soil BioI. Biochem.
4:333-337.
Darmody,RG., D.S. Fanning,WJ. Drummond,Jr., and J.E. Foss. 1977. Determinationof total sul-
fur in tidal marshsoils by x-ray spectroscopy.Soil Sci. Soc. Am. J. 41:761-765.
David, M.B., MJ. Mitchell, D. Aldcorn, andR.B. Harrison. 1989. Analysis of sulfur in soil, plant
and sediment materials:Sample handling and use of an automatedanalyzer. Soil BioI.
Biochem. 21:119-123.
Dean,G.A 1966.A simple colorimetric finish for the Johnson-Nishitamicro-distillation of sulphur.
Analyst (London) 91:530-532.
DeLong, W.A, and L.E. Lowe. 1961. Note on carbon-bondedsulphur in soil. Can. I. Soil Sci.
42:223.
Dick, W.A, and M.A Tabatabai.1979. Ion chromatographicdeterminationof sulfate and nitrate in
soils. Soil Sci. Soc. Am. 1. 43:899-904.
Ensminger,L.E. 1954. Some factors affecting the adsorptionof sulfate by Alabamasoils. Soil Sci.
Soc. Am. Proc. 18:259-264.
Ensminger,L.E., and l.R Freney.1966. Diagnostictechniquesfor determiningdeficienciesin crops
and soils. Soil Sci. 101:283-290.
Eriksson,E. 1960.The yearly circulation of chloride and sulfur in nature:Meterological,geochemi-
cal, and pedologicalimplications.Part II. Tellus 12:63-109.
Evans, C.A., and C.O. Rost. 1945. Total organic sulfur and humus in Minnesota soils. Soil Sci.
59:125-127.
Fleming, J.E, and L.T. Alexander.1961. Sulfur acidity in South Carolinatidal marshsoils. Soil Sci.
Soc. Am. Proc. 25:94-98.
Fogo, 1.K., and M. Popowsky.1949. Spectrophotometricdeterminationof hydrogensulfide. Anal.
Chern. 21:732-734.
Fox, RL., RA Olson, and H.E Rhoades.1964. Evaluatingthe sulfur statusof soils by plants and
soil tests.Soil Sci. Soc. Am. Proc. 28:243-246.
Freney,l.R. 1958. Determinationof water-solublesulphatein soils. Soil Sci. 86:241-244.
SULFUR 957
Freney,J.R. 1961. Someobservationson the natureof organic sulphurcompoundsin soils. Aust. J.

Agric. Res. 21:424-432.
Freney,J.R. 1967. Sulfur-containingorganics.p. 220--259.In AD. McLaren and G.H. Peterson(ed.)
Soil biochemistry.Vol. 1. Marcel Dekker, Inc., New York.
Freney,J.R., G.E. Melville, and C.H. Williams. 1969. Extraction,chemicalnature,and propertiesof
soil organicsulphur.J. Sci. Food Agric. 20:440--445.
Freney,J.R., G.E. Melville, and C.H. Williams. 1970. The determinationof carbonbondedsulfur in
soil. Soil Sci. 109:310--318.
Freney,J.R., G.E. Melville, and C.H. Williams. 1971. Organicsulphurfractions labeledby addition
of 35S-sulphate to soils. Soil BioI. Biochem.3:133-141.
Freney, J.R., G.E. Melville, and C.H. Williams. 1975. Soil organic matter fractions as sourcesof
plant-availablesulphur.Soil BioI. Biochem.7:217-221.
Freney, J.R., F.J. Stevenson,and AH. Beavers. 1972. Sulfur-containing amino acids in soil
hydrolysates.Soil Sci. 114:468-476.
Fried, M.l. 1947. The absorptionof sulfur dioxide by plants as shownby the use of radioactivesul-
fur. Soil Sci. Soc. Am. Proc. 13:135-138.
Gustafsson,L. 1960a.Determinationof ultramicro amountsof sulphateas methyleneblue. I: The
colour reaction.Talanta4:227-235.
Gustafsson,L. 1960b. Determinationof ultramicro amountsof sulphateas methyleneblue. II: The
reduction.Talanta4:236-243.
Haff, L.v. 1970. Oxygen-containinginorganic compounds.p. 183-283.In J.H. Karchmer(ed.) The
analytical chemistryof sulfur and its compounds.Wiley-Intersci., New York.
Hamm, J.w., J.R. Bettany,and E.H. Halstead.1973. A soil test for sulphurand interpretativecriteria
for Saskatchewan. Commun.Soil Sci. Plant Anal. 4:219-231.
Harmsen,G.w. 1954. Observationson the formation and oxidation of pyrite in the soil. Plant Soil
5:324-348.
Hart, E.B., and W.A Peterson.1911. Sulfur requirementsof farm cropsin relation to the soil and air
supply. WisconsinAgric. Exp. Stn. Res. Bull. 14.
Hart, M.G.R. 1959. Sulphuroxidation in tidal mangrovesoils of SierraLeone.PlantSoil 11:215-236.
Harward, M.E., T.T. Chao, and S.c. Fang. 1962. The sulfur statusand sulfur supplying power of
Oregonsoils. Agron. J. 54:101-106.
Harward, M.E., and H.M. Reisenauer.1966. Movement and reactionsof inorganic soil sulfur. Soil
Sci. 101:326-335.
Hasan, S.M., R.L. Fox, and c.c. Boyd. 1970. Solubility and availability of sorbed sulfate in
Hawaiian soils. Soil Sci. Soc. Am. Proc. 34:897-901.
Hesse,P.R. 1957. Sulphurand nitrogen changesin forest soils of EastAfrica. Plant Soil 9:86-96.
Hoeft, R.G., L.M. Walsh, and D.R. Keeney.1973. Evaluationof variousextractantsfor availablesoil
sulfur. Soil Sci. Soc. Am. Proc. 37:401-404.
Jenkinson,D.S. 1968. A titrimetric methodfor determiningtotal sulphur in mineral soils. Analyst
(London) 93:535-539.
Johnson,C.M., and T.H. Arkley. 1954. Microestimationof sulfur in plant materials,soils, and irri-
gation waters.Anal. Chern.26:1525.
Johnson,C.M., and H. Nishita. 1952. Microestimationof sulfur in plant materials,soils, and irriga-
tion waters.Anal. Chern. 24:736-742.
Johnson,C.M., and A.U1rich. 1959. Analytical methodsfor use in plant analysis.Calif{)rnia Agric.
Exp. Stn. Bull. 766.
Jones,L.H.P., D.W. Cowling, and D.R. Lockyer. 1972. Plant-availableand extractablesulfur in some
soils of Englandand Wales. Soil Sci. 114:104-114.
Kamprath,E.L., w.L. Nelson,and J.W. Fitts. 1956. The effect of pH, sulfate and phosphateconcen-
trationson the adsorptionof sulfateby soils. Soil Sci. Soc. Am. Proc. 20:463-466.
Kaplan, I.R., K.O. Emery, and S.c. Rittenberg.1963. The distribution of isotopic abundanceof sul-
phur in recent marine sediments off southern California. Geochim. Cosmochim.Acta
27:297-331.
Karmarkar,S.V., and M.A Tabatabai.1992.Eluentcompositioneffect on ion chromatographicdeter-
mination of oxyanionsin solutionsequilibratedwith soils. Chromatographia34:643-648.
Kelley, D.P., L.A Chambers,and P.A Trudinger. 1969. Cyanolysisand spectrophotometricestima-
tion of trithionate in mixture with thiosulfateand tetrathionate.Anal. Chern.4:898-901.
Kilmer, V.J., and D.C. Nearpass.1960. The determinationof availablesulfur in soils. Soil Sci. Soc.
Am. Proc. 24:337-340.
958 TABATABAI
King, R.C. 1953. Effects of alkali metal ions on developmentof drosophilla,with specialreference
to lithium inducedabnormalities.Proc. Natl. Acad. Sci. U.S.A 39:403-406.
Kowalenko,C.G. 1978. Organicnitrogen,phosphorusan dsulfur in soils. p. 95-126.In M. Schnitzer
and S.M. Khan (ed.) Soil organicmatter.ElsevierNorth-Holland,Inc., New York.
Kowalenko,e.G.,and L.W. Lowe. 1972.Observationson the bismuthsulfide colorimetric procedure
for sulfateanalysisin soil. Commun.Soil Sci. Plant Anal. 3:79-86.
Krupa, S.Y., and M.A. Tabatabai.1986. Measurementof sulfur in the atmosphereand in natural
waters.p. 491-548.In M.A. Tabatabai(ed.) Sulfur in agriculture.Agron. Monogr. 27. ASA,
CSSA, and SSSA,Madison,WI.
Laties, G.G. 1959. Thedevelopmentand control of coexistingrespiratorysystemsin slicesof chico-
ry root. Arch. Biochem. Biophys. 79:378-391.
Lieber, E., and F.L Morritz. 1953. Theuse of Raneynickel. Adv. Catal. 5:417-454.
Little, R.C. 1957. Sulphur in soils II. Determinationof the total sulphurcontentof soil. 1. Sci. Food
Agric. 8:271-279.
Lowe, L.E. 1964.An approachto the study of the sulphurstatusof soils and its applicationto select-
ed Quebecsoils. Can.1. Soil Sci. 44:176-179.
Lowe, L.E. 1965. Sulphurfractionsof selectedAlberta soil profiles of the chemozemicand podzolic
orders.Can. 1. Soil Sci. 45:297-303.
Lowe, L.E. 1969. Sulfur fractionsof selectedAlberta profiles of the gleysolic order. Can.1. Soil Sci.
49:375-381.
Lowe, L.E., and W.A DeLong. 1963. Carbonbondedsulfur in selectedQuebecsoils. Can. 1. Soil
Sci. 43:151-155.
McClung, Ae., L.M. DeFreitas,and w.L. Lott. 1959.Analysesof severalBrazilian soils in relation
to plant responsesto sulfur. Soil Sci. Soc. Am. Proc. 23:221-224.
Maynard,D.G., Y.P. Kalra, and EG. Radford. 1987.Extractionand determinationof sulfur in organ-
ic horizonsof forest soils. Soil Sci. Soc. Am. 1. 51:801-806.
Melville, G.E., I.R. Freney,and e.H. Williams. 1971. Reactionof organic sulfur compoundsin soil
with tin and hydrochloricacid. Soil Sci. 112:245-248.
Miller, L.P. 1947. Utilization of DL-methionineas a sourceof sulphur by growing plants. Contrib.
Boyce ThompsonInst. 14:443-456.
Neptune, AM.L., M.A. Tabatabai,and U. Hanway. 1975. Sulfur fractions and carbon-nitrogen-
phospnorus-sulfurrelationshipsin some Brazilian and Iowa soils. Soil Sci. Soc. Am. Proc.
39:51-55.
Nicolson, A.1. 1970. Soil sulfur balancestudiesin the presenceand absenceof growing plants. Soil
Sci. 109:345-350.
Nor, Y.M., and M.A Tabatabai.1975. Colorimetric determinationof microgramquantitiesof thio-
sulfate and tetrathionate.Anal. Lett. 8:537-547.
Nor, Y.M., and M.A Tabatabai.1976. Extraction and colorimetric determinationof thiosulfateand
tetrathionatein soils. Soil Sci. 122:171-178.
Nor, Y.M., and M.A. Tabatabai.1977. Oxidation of elementalsulfur in soils. Soil Sci. Soc. Am. 1.
41:736-741.
Pirela, H.I., and M.A Tabatabai.1987. Determinationof sulfate in soils by reduction with tin and
phosphoricacid. Commun.Soil Sci. Plant Anal. 18:1131-1141.
Pirela, H.1., and M.A Tabatabai.1988. Reductionof organic sulfur in soils with tin and phosphoric
acid. Soil Sci. Soc. Am. 1. 52:959-964.
Polak, H.L., G. Feenstra, and 1. Siagman. 1966. Stability of hypobromite solutions. Talanta
13:714-724.
Probert, M.E. 1976. Studiesof "available" and isotopically exchangeablesulphur in some North
Queenslandsoils. Plant Soil 45:461-475.
Putnam,H.D., and E.L. Schmidt. 1959. Studiesof the free amino acid fraction of soils. Soil Sci.
87:22-27.
Qian, P., 1.1. Schoenau,and W.Z. Huang. 1992. Use of ion exchangemembranesin routine soil test-
ing. Commun.Soil Sci. Plant Anal. 23:1791-1804.
Rehm, G.W., and Ae. Caldwell. 1968. Sulphur supplying capacityof soils and the relationshipto
soil type. Soil Sci. 105:355-361.
Reisenauer,H.M., L.M. Walsh, and R.G. Hoeft. 1973. p. 173-200.In L.M Walsh and I.D. Beaton
(ed.) Soil testingand plant analysis.SSSA,Madison,WI.
Roberts,S., and EE. Koehler. 1968. An x-ray spectrographicmethodof determiningtotal sulfur in
soil extracts.Soil Sci. 106:164-171.
Robinson,W.O. 1930. Method and procedureof soil analysisused in the divison of soil chemistry
and physics.USDA Circ. 139. U.S. Gov. Print. Office, Washington,DC.
SULFUR 959
St. Lorant, I. 1929. Uber eine neuecolorimetrischemikromethodezur bestimmungdes schwefelsin

sulfiden, sulfatenusw. Z. Physiol. Chern. 185:245-266.
Searle,P.L. 1968. Determinationof total sulfur in soils by using high-frequencyinduction furnace
equipment.Analyst (London) 93:540-545.
Shorey, E.C. 1913. Some organic soil constituents.U.S. Div. Soils Bull. no. 88. U.S. Gov. Print.
Singh,S.S. 1967.Sulfateions and ion activity product(AJ)(OH)3 in Wyoming bentonitesuspensions.
Soil Sci. 104:433-438.
Singh, S.S.,and1.E. Brydon. 1967. Precipitationof aluminum by calcium hydroxide in the presence
of Wyoming bentoniteand sulfate ions. Soil Sci. 103:162-167.
Singh, S.S., and J.E. Brydon. 1969. Solubility of basic aluminum sulfatesat equilibrium in solution
and in the presenceof montmorillonite. Soil Sci. 107:12-16.
Skerrett,E.1., and G.1. Dickes. 1961. The determinationof microgramamountsof sulphur-contain-
ing compounds.Analyst (London) 86:69-71.
Smittenberg,J., G.w. Harmsen,A Quispel,and D. Otzen. 1951. Rapid methodsfor determiningdif-
ferent typesof sulphurcompoundsin soil. Plant Soil 3:353-360.
Sorbo, B. 1957. A colorimetric methodfor the determinationof thiosulfate. Biochim. Biphys. Acta
23:412-416.
Spencer,K., and J.R. Freney.1960. A copmarisonof severalproceduresfor estimatingthe sulphur
statusof soils. Aust. J. Agric. Res. 11:948-959.
Stance,H.C.T., et al. 1968. A handbookof Australian soils. Rellim Tech. Publ., Glenside,South
Australia.
Stanford,J.O., and1.0. Lancaster.1962. Biological and chemicalevaluationof the readily available
sulfur statusof Mississippisoils. Soil Sci. Soc. Am. Proc. 26:63-65.
Steinbergs,A 1955. A method for the determinationof total sulphur in soils. Analyst (London)
80:457-461.
Steinbergs,A, O. Iismaa,1.R. Freney,and N.J. Barrow. 1962. Determinationof total sulphurin soil
and plant material.Anal. Chim. Acta 27:158-164.
Stevens,T.S., I.C. Davis, and H. Small. 1981. Hollow fiber ion-exchangesuppressorfor ion chro-
matography.Anal. Chern. 53:1488-1492.
Stevenson,F.J. 1956. Isolation and identification of someamino compoundsin soils. Soil Sci. Soc.
Am. Proc. 20:201-204.
Tabatabai,M.A, and AA A1-Khafaji. 1980. Comparisonof nitrogen and sulfur mineralization in
Tabatabai,M.A., and N.T. Basta. 1991. Ion chromatography.p. 229-259.In K.A. Smith (ed.) Soil
analysis.2nd ed. Dekker, New York.
Tabatabai,M.A, N.T. Basta,and H.J. Pirela. 1988. Determinationof sulfur in soils and plant mate-
rials by ion chromatography.Commun.Soil Sci. Plant Anal. 19:1701-1714.
Tabatabai,M.A, and J.M. Bremner.1970a.An alkaline oxidation methodfor determinationof total
sulfur in soils. Soil Sci. Soc. Am. Proc. 34:62-65.
Tabatabai,M.A, and I.M. Bremner.1970b.Comparisonof somemethodsfor determinationof total
sulfur in soils. Soil Sci. Soc. Am. Proc. 34:417-420.
Tabatabai,M.A, and I.M. Bremner. 1972a.Distribution of total and availablesulfur in selectedsoil
and soil profiles. Agron. J. 65:40-44.
Tabatabai,M.A, and 1.M. Bremner. 1972b. Forms of sulfur, and carbon, nitrogen, and sulfur rela-
tionships,in Iowa soils. Soil Sci. 114:380-386.
Tabatabai,M.A, and J.M. Bremner.1991. Automatedinstrumentsfor determinationof total carbon,
nitrogen, and sulfur in soils by combustiontechniques.p. 261-285.In K.A Smith (ed.) Soil
analysis.2nd ed. Dekker, New York.
Tabatabai,M.A, and W.A Dick. 1979. Ion chromatographicanalysisof sulfate and nitrate in soils.
p. 361-370. In 1.0. Mulik and E. Sawicki (ed.) Ion chromatographicanalysis of environ-
mental pollutants.Vol. 2. Ann Arbor Sci. Publ., Inc., Ann Arbor, MI.
Tabatabai,M.A, and W.T. Frankenberger,lr. 1996. Liquid chtomatography.p. 225-245. In D.L.
Sparkset al. (ed.) Methodsof soil analysis. Part 3. Chemical methods.SSSA Book Ser. 5.
Tabatabai,M.A, and 1.M. Laflen. 1976a. Nitrogen and sulfur content and pH of precipitation in
Iowa. 1. Environ. Qual. 5:108-112.
Tabatabai,M.A, and I.M. Laflen. 1976b. Nutrient contentof precipitationover Iowa. Water Air Soil
Pollut. 6:361-373.
Ulrich, A, M.A Tabatabai,K. Ohki, and C.M. Johnson.1967. Sulfur contentof alfalfa in relation to
growth in filtered and unfiltered air. Plant Soil 26:235-252.
960 TABATABAI
Vaughan,B. 1991. Evaluationof AB-DTPA as a soil extractantfor plant availablesulfur. p. 304. In

Agronomy abstracts.ASA, Madison,WI.
Walker, D.R.,and G. Domenbal.1972.Soil sulfate:II. As an index of the sulfur availableto legumes.
Can. 1. Soil Sci. 52:261-266.
Williams, C.H. 1967.Somefactors affecting the mineralizationof organicsulphurin soils. Plant Soil
26:205-223.
Williams, C.H., and H. Steinbergs.1959. Soil sulphurfractions as chemicalindicesof availablesul-
phur in someAustralian soils. Aust. J. Agric. Res. 10:340-352.
Williams, C.H., and A. Steinbergs.1962. The valuation of plant-availablesulphur in soils: I. The
chemicalnatureof sulphatein someAustralian soils. Plant Soil 17:279-294.
Williams, C.H., and A. Steinbergs.1964. The evaluationof plant-availablesulphur in soils: II. The
availability of adsorbedand insoluble sulphates.Plant Soil 21:50-62.
Wolkoff, A.w., and R.H. Larose.1975.Separationand detectionof low concentrationsof polythion-
atesby high speedanion exchangeliquid chromatography.Anal. Chern.47:1003-1008.
Published 1996
Chapter34
Total Carbon, Organic Carbon,

and Organic Matter
D. W. NELSON, University of Nebraska, Lincoln, Nebraska
L. E. SOMMERS,Colorado State University, Fort Collins, Colorado
GENERAL INFORMATION
This chapteris an updated,revisedversion of the material containedin Chapter

29, in Volume 2, of Methods of Soil Analysis, 2nd edition (Nelson & Sommers,
1982). Much of the material presentedin the original chapterhasbeenmodified
or replacedby more modem proceduresand recent literature pertaining to the
methodshasbeenincluded. In addition, the total C sectionhasbeenmodified to
include the latestinformation on automatedinstrumentsfor analysisof C.
Total C in soils is the sum of both organic and inorganic C. Organic C is
presentin the soil organicmatter fraction, whereasinorganic C is largely found
in carbonateminerals. Not all soils contain inorganic C becauseof dissolution
during soil formation of carbonatemineralsoriginally presentin parentmaterial.
However,organicC is presentin all agriculturalsoils. In soils formed from cal-
careousparentmaterial under arid conditions,it is not unusualfor the inorganic
C concentrationto exceedthe amountof organicC present.
Organic C is containedin the soil organic fraction, which consistsof the
cells of microorganisms,plant and animal residuesat various stagesof decom-
position, stable"humus" synthesizedfrom residues,and highly carbonizedcom-
poundssuchas charcoal,graphiteand coal (elementalforms of C). OrganicC in
soil may be estimatedas the differencebetweentotal C and inorganicC. Organ-
ic C can be determineddirectly by total C proceduresafter removalof inorganic
C or by rapid dichromate,oxidation-titrationtechniques.In the absenceof inor-
ganic C, a total C analysiscan be used to determineorganic C and recover all
forms of organic C in soils. However, organic C methodsbasedon dichromate
oxidation recover variable proportionsof elementalC (e.g., charcoal) and, in
someprocedures,variableamountsof organicC containedin "humus."
Book Seriesno. 5.
961
962 NELSON & SOMMERS
Calcite and dolomite are the principal carbonatemineralspresentin soil,

and most inorganic C is associatedwith thesecompounds.However, in some
alkali soils, significant amountsof inorganic C may be presentin soluble car-
bonateand bicarbonatesalts.The amountsof soluble carbonatespresentin soil
may be determinedby proceduresoutlined in Chapter15 (Loeppert& Suarez,
1996),anda variety ofmethodsfor the estimationof total inorganicC in soils are
presentedin Chapter15 (Loeppert& Suarez,1996).
Total C analysisof soil involvesconversionof all forms of C in soils to car-
bon dioxide (C02) by wet or dry combustionand subsequentquantitationof
evolved CO2 by gravimetric, titrimetric, volumetric, spectrophotometric,or gas
chromatographictechniques.Dry combustionis conductedby heating(-I000°C)
a soil-catalystmixture in a resistancefurnaceor induction furnacein a streamof
O2 or COrfreeair, followed by quantitationof evolvedCO2. Wet combustionis
normally carriedout by boiling a soil samplewith a mixture of potassiumdichro-
mate(K2Cr207)' sulfuric acid (H2S04), and phosphoricacid (H3P04) in a closed
systemflushed with a streamof COrfree air and absorbingevolved CO2 in a
tared weighting bulb filled with Ascarite (Arthur H. ThomasCo., Philadelphia,
PA) (Allison, 1960).Alternatively, wet combustionmay be carriedout in a Van
Slyke-Neil apparatusand evolved CO2 estimatedby manometricprocedures
(Bremner,1949). 1\vo dry combustionand one wet combustionproceduresfor
total C analysisare describedin this chapter.
Soil organic matter hasbeendefmedasthe organicfraction of soil, includ-
ing plant, animal, and microbial residues,fresh and at all stagesof decomposi-
tion, and the relatively resistantsoil humus(SSSA,1979). Soil organicmatteris
normally restrictedto only thoseorganicmaterialsthat accompanysoil particles
through a 2-mm sieve. It is difficult to quantitatively estimatethe amount of
organicmatterpresentin a soil. Proceduresusedin the past involve determina-
tion of the changein weight of a soil sampleresultingfrom destructionof organ-
ic compoundsby H20 2 treatmentor by ignition at high temperature.Both tech-
niques are subject to error. The H20 2 method does not quantitatively remove
organicmatterand the ignition methodgives an overestimatebecauseboth inor-
ganic and organic constituentsin soils during ignition can be minimized by
removingaluminosilicateswith hydrofluoric acid (HF)/hydrochloricacid (HCI)
prior to heatingor ignition at temperaturesthat decomposeorganicmatterwith-
out appreciabledehydroxylationof inorganicmaterials.Alternatively, the organ-
ic mattercontentof a soil may be estimatedby multiplying the organic C con-
centrationby a constantfactor basedon the percentageof C in organic matter.
PublishedorganicC-organicmatterconversionfactorsfor surfacesoils havevar-
ied from 1.724to 2.0. The appropriatefactor mustbe determinedexperimentally
for eachsoil by independentanalysisof organicmatterand organicC. Although
neitherthe direct determinationof organicmatter nor the calculationof organic
mattercontentis completelyaccurate,the mostusefulprocedurescurrentlyavail-
able are describedin this chapter.Becauseof the problemassociatedwith deter-
mining the organicmattercontentof a soil, it is strongly suggestedthat investi-
CARBON AND ORGANIC MATIER 963
gatorsdetermine and report the organicC contentasan index of the organicmat-

ter in a soil.
TOTAL CARBON
Introduction
Analytical proceduresusedfor determiningtotal C in soils must quantify
both inorganicand organicforms. In humid regionswhereextensiveleachingof
the soil profile hasoccurred,organicC will be the predominantform present.In
arid or semiaridregions,carbonateminerals(e.g., calcite, dolomite) along with
solublecarbonatesalts may constitutea significant percentageof the total C.
Dry combustionand wet combustionare the two basicapproachesusedto
quantify total C in soils. In both instances,the CO2 liberatedfrom organic and
inorganic C is determinedthrough spectrophotometric,volumetric, titrimetric,
gravimetric, or conductimetrictechniques.An apparatusfor performing total C
analysis by dry combustion can be fabricated from conventional laboratory
glasswareand a medium-temperature(-1000°C) resistancefurnace. Dry com-
bustion proceduresusing either high-temperature(> 1500°C) or induction fur-
nacesare found in commercially available, automatedtotal C analyzers.The
major~ty of dry combustionmethodsemploy gravimetric determination.of CO2
althoughtitrimetric techniquesalso are described.Wet combustionmethodsfor
total C employ a strong oxidant, such as K2Cr207, in an acid digestionmixture
for quantitativeoxidation of organicC and dissolutionof carbonateminerals.A
comparisonof principles, advantages,and disadvantagesof commonly used
methodsfor total C determinationis given in Table 34-1.
The developmentsin instrumental methods in recent years should be
assessed beforechoosinga procedurefor determiningtotal C in soils. The major-
ity of instrumentsare automatedversionsof primarily dry combustionproce-
dures. The relative advantagesand disadvantagesof manual and instrumental
methodsshould be consideredbefore initiating total C analysis. From a cost
standpoint,manual procedurescan be set up, in many cases,with apparatus
already present in most laboratories; however, they are time-consumingand
tediousand require use of careful analytical technique. Incontrast,instruments
are costly, typically greaterthan $20 000, but they are capableof analyzing a
large numberof sampleswith minimal variability due to operatorerror. Nearly
all commercialunits are availablewith autosamplersand computerinterfacesto
aid in dataacquisitionand handling.In addition, severalcommercialunits enable
the simultaneousdeterminationof elements(C, H, N, or S).
The methodspresentedfor total C are essentiallyidentical to those pro-
posedby Allison et ai. (1965) in the first edition of Methods of Soil Analysis
(Black et aI., 1965) and subsequentlyupdatedin the secondedition (Nelson &
Sommers,1982). Much of the text presentedis usedwith only minor alterations
to updatethe equipmentavailableand the literaturecited. A brief descriptionhas
t
Table 34-1. Comparisonof methodologiesusedfor determinationof total C in soils.

Method Principle CO2 determination Advantages Disadvantages
Dry combustion Sampleis mixed with CuO and heated Gravimetric, Referencemethodwidely usedin other Time-consuming,leakfree,O2 sweep
(resistance to -lOOO°C in a streamof O2 to titrimetric disciplines,variable samplingsizes, train is required,slow releaseof
furnace) convert all C in sampleto CO2 variable samplesize CO2 from alkaline earth carbonates.
Dry combustion Sampleis mixed with Fe or accelera- Gravimetric, Rapid combustion,high temperature LeakfreeO2 sweeptrain is required,
(induction tors and rapidly heatedto > 1650°C titrimetric ensuresconversionof all C to CO2 induction furnaceis expensive.
furnace) in streamof O2 to convert all C in
to CO2•
Dry combustion Sampleis mixed with catalystsor ac- Thermal conductivity, Rapid and simple, good accuracyand Expensiveequipment,slow releaseof
(automated celeratorsand heatedwith resistance conductimetric,in- precision CO2 from alkaline earth carbonates
methods) or induction furnace in a streamof frared detector, with resistancefurnace
O2 to convertall C in sampleto CO2 gravimetric
Wet combustion Sampleis heatedwith K2Cr207-H2S04- Titrimetric, gravi- Equipmentreadily available,good ac- Time-consuming, gravimetric deter-
(combustion H3P04 mixture in a COrfreeair metric curacy,easily adaptedto analysisof mination of CO2 requirescareful
train) streamto convertall C in sample solutions,titrimetric analysisof CO2 analytical techniques,titrimetric
to CO2 less subjectto operatorerror determinationof CO2 lessprecise.
~
Z
~
C"I.l
I
CARBON AND ORGANIC MATTER 965
Fig. 34-1. Block diagramof dry combustiontrains. Option A is basedon Allison (1965). Option B is
adaptedfrom Rabenhorst(1988).
to updatethe equipmentavailableand the literaturecited. A brief descriptionhas

beenaddedon the principles employedin selectedcommerciallyavailabletotal
C analyzers.
Total Carbonby Dry Combustion
Introduction
The dry combustionmethodis basedon oxidation of organic C and ther-
mal decompositionof carbonateminerals in a medium-temperatureresistance
furnace.The CO2 liberatedis commonly trappedin a suitablereagentand deter-
mined titrimetrically or gravimetrically. Spectrophotometric,volumetric or con-
ductimetric proceduresare used to determineCO2 in some commercialinstru-
ments. Alternatively, the CO2 releasedcan be reducedto CH4 and quantitated
with a gas chromatographfitted with a flame ionization detector (Geiger &
Hardy, 1971). The following descriptionof a medium-temperature dry combus-
tion is that presentedby Allison et al. (1965).
Principles
In the dry combustionproceduresdescribedhere, thesampleis burnedin
a stream of purified O2 and CO2 in the effluent gas stream is absorbedby
Ascarite or someother suitableabsorbentand weighed.Other absorbablegases
formed during combustionare removedfrom the O2 streambeforethey reachthe
CO2 absorptionbulb. A typical combustiontrain is comprisedof 10 basic ele-
mentsas diagrammedin Fig. 34-1. The make-upof the individual elementsare
modifications (AOAC, 1975, p. 924-926; ChemistsU.S. Steel Corp., 1938, p.
40-54; Salter,1916; Winters & Smith, 1929) of thoserecommendedby Fleming
(1914) for the rapid determinationof C in Fe and steel.
NELSON & SOMMERS
The O2 supply (commercialcompressedO2) is first scrubbedby passage

througha train consistingof concentratedH2S04 to removeammonia(NH3) and
hydrocarbons,an absorbentsuch as soda lime to remove CO2, and anhydrous
magnesiumperchlorateMg(CI04)z to removewater vapor. The rate of O2 flow
is controlledby a needlevalve and is measuredby a flow meter.
A furnaceprovidesthe heat necessaryfor combustionof the organicC to
CO2 and for decompositionof carbonates.In a resistancefurnace,the sampleis
heatedby radiation,conduction,and convection ina tube surroundedby heating
elementsmadeof high-resistancematerialssuch as Nichrome (in medium-tem-
peraturemodels)or silicon carbide(in high-temperaturemodels).In an induction
furnace,the sourceof energyis high-frequencyelectromagneticradiation. Fer-
rous metals and certain other materialscan be heatedto high temperaturesby
electromagneticinductionif enoughenergyis present.Materialssuchassoil that
do not heat by induction can be heatedindirectly by radiation, conduction,and
convectionfrom susceptors(materialsthat do heat) in the induction field. The
susceptormay take the form of iron or tin chips that are mixed with the sample
to be burned, or a radiator [e.g., the Pt cage (Simons et aI., 1955), quartz-
enclosedcarboncrucible(Allison et aI., 1965)] that will surrounda cruciblecon-
taining the sampleto be burned.
The type of furnacedeterminesthe packingof the combustiontube. With
medium-temperaturefurnaces, cupric oxide (CuO) or another acceleratoris
mixed with the soil to aid in combustionof the organicmatterand elementalC.
With high-temperaturefurnaces,the organic and elementalC is generally oxi-
dized to CO2 by gaseousO2 without specialassistance.When medium-tempera-
ture furnacesare used,catalystsmust be included in the combustiontube at the
rear of the heated zone to ensure essentially complete oxidation of carbon
monoxide(CO) or othervolatile C compounds.Platinizedasbestosor CuO wire
may be usedasa catalyst.However,with any type of furnace,someCO may pass
through.A low-temperature(-250°C)catalystfurnace,with catalystsuppliedby
the manufacturer,may follow the main combustiontube to convert any CO to
CO2•
Medium-temperaturecombustionis not entirely satisfactoryfor soils con-
taining alkaline-earthcarbonatesbecausethesemineralsreleaseCO2 slowly at
950°C(C02 may not be releasedcompletelyin 30 min). High-temperaturecom-
bustion, on the other hand, causesrapid and quantitativereleaseof CO2 from
both Na2C03and alkaline-earthcarbonates.
The gasstreamleaving the furnaceis freed of particulatematterby a dust
trap in the exit endof the combustiontube. The removalof nitrogenoxides,sul-
fur oxides,and halogengasescan be effectedin severalways. Activated Mn02
appearssatisfactoryas a dry absorberfor the oxidesof Nand S and the halogens
(Robertsonet aI., 1958). To protect the catalystsin the catalyst furnace from
beingpoisonedby thesesubstances, a trapof activatedMn02 mustbe insertedat
the combustiontube outlet. Liquid absorbersfor theseinterfering gasesinclude
solutions of H2S04-Cr03,Ag2S04, and KI but they are not recommendedfor
insertion aheadof the catalystfurnace. Most of the water vapor formed during
combustionis removedby a concentratedH2S04 tower immediatelyfollowing
the catalystfurnace.The little watervaporpassingthroughis trappedby a tower
of anhydrousMg(CI04h next in line. The CO2 is finally absorbedin a suitable

bulb containingAscariteor other absorbentbackedby anhydrousMg(CI04h to
ensurethat water vapor pressureis the samein exit gas as in enteringgas.
Rabenhorst(1988) evaluatedmany of the parametersinvolved in deter-
mining organic and inorganicC in soils by dry combustionusing gravimetric
determinationof C collectedin an absorptionbulb. He concludedthat sequential
combustionof the samesampleat 575°C for 15 min followed by combustionat
1000°Cfor 10 min would quantitativelyrecoverorganicand inorganicC, respec-
tively, from soils. He also describeda simplified analytical apparatusthat con-
sistedof the following components:(i) compressedO2; (ii) concentratedH 2S04;
(iii) Ascarite;(iv) Mg(CI04h; (v) quartzor ceramiccombustiontube to hold the
sampleboat andcontainingCuO wire and glasswool at distal end; (vi) Drierite;
(vii) Mg(CI04)2 and; (viii) Nesbitt bulb containinglayers of glasswool, ascarite
and Mg(CI04)2. This simplified gashandlingtrain is listed as Alternative C (see
"Alternative Arrangements"below; Fig. 34-1) in the following. method and
should be consideredif dry combustionwill be usedfor total C analysis.
Medium-TemperatureResistanceFurnaceMethod
Special Apparatusl
1. Oxygencylinder and pressureregulator(A).
2. Oxygenpurifying train consistingof concentratedH 2S04 for removal
of NH3 and hydrocarbons,Ascaritefor removalof CO2 and acid gases,
and anhydrousMg(CI04)2 for removal of water vapor (B).
3. Flow indicator and needlevalve for O2 control (C).
4. Furnaceunit (D): (a) Resistancefurnace equippedwith temperature
controller and indicator (Lindberg multiple-unit combustion-tubefur-
naceor equivalent)for operationat 900 to 1000°C;(b) Sampleinsert-
er (LEC02 no. 501-062,Alpha3 AR-061 or equivalent);(c) Combus-
tion tube, 2.5-cm diam. by 75 cm (zircon ceramicor equivalent).
5. Dust trap (LECO no. 501-010or equivalent)insertedin the exit end of
the combustiontube (E).
6. Sulfur trap filled with activatedMn02 (LECO no. 503-033or equiva-
lent) (F).
7. Catalystfurnace and tube (LECO no. 507-010or equivalent)(G).
8. Gas scrubber(H): (a) Sulfuric acid tower to absorbmost of the water
vapor and to prolong the life of the anhydrousMg(CI04h trap that fol-
lows (especially desirablewhen combustingorganic soils and other
organic materials; (b) Water vapor trap filled with anhydrous
Mg(CI04h (LECO no. 598-157or equivalent).
9. Carbon dioxide absorptiontube, a Nesbitt, Fleming, or Turner bulb
packedwith an indicating CO2 absorbentand anhydrousMg(CI04)(I).
The bulb containsfrom bottom to top: (i) glasswool, (ii) 3-cm layer
1 Capital lettersin parenthesesrefer to units in Fig. 34-1.

2 LECO Corporation,3000 Lakeview Ave., SI. Joseph,MI 49085-2396.
3 Alpha Resources, Inc., 3090JohnsonRd., Stevensville,MI 49127-0199.
of 8- to 14-mesh(1.4-2.36mm) absorbent(e.g., Ascarite), (iii) 2-em

layer of 14- to 20-mesh(0.85-1.4mm) absorbent,(iv) l-cm layer of
anhydrousMg(CI04)z, and (v) glasswool (also describedin "Special
Apparatus"under"Wet Combustion Method").
10. Bubblertrap to seal thetrain from the atmosphereand indicate flow of
exit gas(J).
11. Alternative arrangements;(i) The O2 purifying train (B) and the flow
indicator(C) are availableas a combinedunit (LECO no. 516-000);(b)
The scrubber-absorption train describedin "SpecialApparatus"under
"Wet CombustionMethod" (Units F-K) can substitutefor units H, I,
andJ (Items 8-10); (c) Items 4-8 can be replacedas follows. Item 4,
pack combustionwith 7 to 10 cm of cupric oxide wire followed by
plug of glasswool; Items 5 to 7 are deleted;Item 8, replacewith by
trap of Drierite followed by trap of anhydrousMg(CI04)z.
12. Accessory items: (i) Combustion boats, ceramic (Alundum, zircon,
etc.), (LECO no. 528-053 or equivalent); (b) Boat puller with eye
shield (LECO no. 501-062or equivalent);(c) Combustiontube clean-
ing brush(LECO no. 501-082or equivalent);(d) Plastictubing (Tygon
or equivalent) for connectingcomponents.(Rubber is permeableto
CO2.) Any depositsdue to malfunctionscan readily be observedin
transparenttubing); (e) Analytical balance(Mettler H31AR, Mettler
Instrument,Hightstown,NJ; or equivalent)on groundedmetal plate.
Reagents
1. Oxygengas.
3. Manganesedioxide (Mn02), activated(LECO no. 501-060or equiva-
lent).
4. Platinized asbestos,5% Pt (J. T. Baker Chemical Co.; no. 0922,
Philipsburg,NJ; or equivalent);or CuO, wire or granular,low in C for
combustiontube catalyst.
5. Cupric oxide powder,low in C, to serveas an acceleratorwhen mixed
with soil in the boat.
6. Alundum or Sinderiteor equivalent,refractory grade,60- or 90-mesh
(165-250~m) size, C free.
7. Anhydrousmagnesiumperchlorate(Anhydrone,Dehydrite,etc.).
8. Carbondioxide absorbent,indicating, 14- to 20-mesh(0.85-1.4mm)
size [Ascarite, Caroxide (Fisher Scientific, Pittsburgh,PA), Indicarb
(Fisher Scientific, Pittsburgh,PA), or Mikhobite (G. FrederickSmith
ChemicalCo., Columbus,OH).
9. StandardC source [dextrose(CJI1206) or benzoic acid (C2H60 2) of
reagent-gradeor primary-standardquality].
Procedure
Looselypackthe combustiontubewith a 7.5-emcoreof platinizedasbestos
so that it will comewithin the exit end of the heatedzoneof the furnace.Alter-
natively, use coarselygranular CuO, held in position by two plugs of asbestos

fiber packedno more tightly than is necessaryto hold the CuO in place.
Bring the furnace to 950°C to lO00°C. Connectthe train, and sweepthe
apparatuswith O2 at the rate of 100 mL/min for 10 min. Removeand weigh the
CO2 absorptionbulb. Repeatthis stepuntil the CO2 absorptionbulb hasattained
a weight constantto ±0.2 mg. Replacethe CO2 absorptionbulb in the train, and
introducewell within the heatedzoneof the combustiontube a ceramicboat con-
taining 1.0 g of finely divided CuO. Admit O2 and continue the flow at 100
mL/min for 10 min. Close the stopcockson the CO2 absorptionbulb, disconnect
the bulb, and weigh it. The increasein weight of the bulb representsthe blank.
Remove the boat from the combustiontube. Repeatthe determinationuntil a
reproducibleblank is obtained.With a properly preparedtrain and high-quality
reagents,the blank shouldbe negligible, i.e., within limits of weighing error.
Grind soil to be analyzedfor C to passthrough a 100- or 140-meshsieve
(0.149-or 0.105-mmopenings).Mix 1.000g of mineral soil of known watercon-
tent with 1.0 g of finely divided CuO in a combustionboat, cover the mixture
lightly (-2 mm) with Alumdum or Sinderite,and follow the procedureusedfor
the blanks. For soils high in organic matter, use a 0.500-gsample.The increase
in weight, correctedfor blank, should representthe CO2 from the sample.The
calculationis as follows
[g CO2, sample] - [g CO2, blank]

Total C, % = x 0.2727 x 100 [1 ]
g water-freesoil
Comments
Cleanlinessis proverbial in the C laboratory,especiallywhen one is deal-
ing with low C samplesor doing work of the highestorder of accuracy.In such
instances,freedomfrom dust, dirt, or fumes is essential.It is advisablehabitual-
ly to ignite all boatsat -900°C before use and to store boatsout of contactwith
the atmosphere.Handling boats and covers only with tongs is consideredgood
practice.
Someexperienceis usually necessarybefore reproducibleweights of CO2
absorptionbulbs can be obtained.Thermal equilibrium with the atmosphereis
important. The surfacemust be kept clean and free of static charge.The follow-
ing points may be helpful:
1. Before each weighing, the absorptionbulb should be wiped slowly
with a lint-free papertissue,such as Kimwipes or napkin stock. Paper
is superiorto cloth. Rapid wiping builds up the staticcharge.Handling
the absorptionbulb with cleancotton glovesmay be helpful.
2. After the CO2 absorptionbulb has beenwiped, it shouldbe touchedto
a groundedplate to remove the static chargebefore weighing. If the
bulb is in temperatureequilibrium with the atmosphere,repeated
weighing shouldagreewithin ±0.2 mg. The position of the absorption
bulb on the balancepan may be critical. With somebalances,accura-
cy is dependenton careful centeringof the objectsto be weighed.
3. The CO2 absorptionbulb is brought to constantweight by inserting it

in the combustiontrain and operatingthe train as a blank. Cold bulbs
characteristicallygain severalmilligrams on blank runs before attain-
ing a weight constantto ±0.2 mg. If a steadygain in weight is obtained
in repeatedtrials with the bulb properly handledand placedon the bal-
ancepan, one or more of the absorbersin the train may not be func-
tioning properly.
Fine grinding «150!lm or <100 mesh)of soil samplesis especiallyimpor-
tant in obtaining reproducibleresults in determinationsof C. Only from finely
ground, homogeneoussamples,can uniform small subsamplesbe drawn. Poor
replication often can be tracedto poor sampling.Fine grinding is a task at best.
Wheremany samplesare to be processed,a mechanicalmortar and pestleunit, a
high-speedimpact shaker,or a ball mill will savemuch time and labor.
When C in soil extractsor other liquids is determined,the samplemay be
evaporatedand dried, preferably under vacuum at 60°C in porcelain or nickel
boatsof 5- or 10-mL capacity.Liquids will slowly seepthrough the usual grade
of ceramicboats.Porcelainboatsare short lived, evenat 950°C. Unglazedboats
may be renderedleakproofby treatingwith a glazing mixture and firing in a lab-
oratory furnace(Lindbeck & Young, 1964). An alternativemethodfor analyzing
soil extractsinvolves filling ceramiccombustionboatswith siliceousearth, i.e.,
Kieselgur(Tiessenet aI., 1981). SinceKieselguhrwill absorbaboutfour times its
weight in liquid, multiple 3-mL samplesof a soil extractcan be addedto the boat
if water is evaporatedon a hot plate for 10 min at 80°C betweenadditions.
Combustiontubeseventuallydevelopfine cracksin the hottestregion and
needto be replaced.Erratic results are one indication of a crackedcombustion
tube. After every 50 or 100 analyses,the tube should be testedfor leaks under
operatingpressureby stopperingthe exit and observingif O2 passesthe H2S04
tower in the purifying train. Combustiontube life is prolongedif, during con-
templateddaily use, the furnaceis kept on continuously.
A standardC source,such as analytical reagentor primary standard-quali-
ty glucoseor benzoicacid, should be run from time to time to checkthe appara-
tus. The organic standardshould be diluted and coveredwith Alundum or Sin-
derite to preventexplosion.
Explosivecombustionwill blow stoppers,and eventhe boat,from the com-
bustiontube. After an explosion,it is essentialto bum off the C depositsfrom the
cooler areasinside the tube before additionalanalysesare made.
A two-tube furnaceis advantageous eventhoughonly one tube is usedrou-
tinely. The secondtube servesas a reservein the eventthe other cracksduring a
seriesof analysesor when C depositsresultingfrom an explosionmust be burned
out.
The insert dust trap shouldbe cleanedand refilled with glasswool after 40
or 50 determinationsor more frequently if depositsappearin the exit tubing.
The Mn02 usedto removeS02 from the combustionproductsbefore they
enter the catalyst furnace should be changedafter about 50 determinationsor
beforeall the granulesappeargray or agglomerated.Peterson(1962) pointedout
that the accumulationof combustionreactionproductson the Mn02 changesits
CO2 absorption-desorption patternso that longerflushing times must be used,as
CARBON AND ORGANIC MAITER 971
in the gasometricdeterminationof C. Petersonrecommendeda specially pre-

pared Pb02 as a substitutefor Mn02 to increasethe efficiency of S02 removal
but no evidence waspresentedconcerningits capacityto absorbnitrogen oxides
or the halogens.
Air-dry samplesare preferred to oven-dry samples,becauseoven drying
may result in lower valuesfor total C in somesoil samples.
A slight pressurein the combustiontube will be noticed when the stopper
is removedafter a determination.If this pressurebecomespronounced,it indi-
cates increasedresistanceto gas flow in the S trap or in the water vapor trap.
Thesetrapsshouldbe examinedand repackedor replacedas necessary.
Temperature> 1OOO°Cmustbe avoided.Heatingelementswill be subjectto
burnout, andfusion of CuO is likely to occur and causeslagging andtube rupture
on cooling. Attention to this is especiallyimportantif a temperaturecontroller is
not used.
A supply of boatscan be renderedC free by preliminary ignition in a muf-
fle furnace at 850 to 900°C. Theseignited boatsshouldbe kept in dust-freestor-
age until used.
High-TemperatureInduction FurnaceMethod
Special ApparatusS
1. Oxygencylinder and regulator(A).
2. Oxygen purifying train (B).
3. Flow meterand needlevalve (C).
4. Furnaceunit: (i) Induction (high-frequency)furnace (Alpha no. AR-
521 or equivalent) for operation at 1400 to 1600C; (b) Combustion
tube (LECO no. 550-122or equivalent);and (c) Crucible (LECO no.
528-018)with cover.
S. Dust trap for induction furnace,external(LECO no. 501-0lD or equiv-
alent).
6. Sulfur trap filled with activatedMn02 (LECO no. 503-033or equiva-
lent).
7. Catalystfurnaceand tube (G).
8. Gasscrubber(H).
9. Carbondioxide absorptiontube (I).
10. Bubble trap (1).
11. Alternative arrangements:(a) The O2 purifying train (B) and the flow
indicator(C) are availableas a combinedunit (LECO no. 516-000);(b)
The inductionfurnaceactually is a complex unitthat combinesthe fur-
nace(D), dust trap (E), Strap(F), and catalystfurnace(G) (Items 4-5
above),in a single unit; and (c) Various combinationsof the 10 basic
elementsthat comprisethe train (Fig. 34-1) are available under vari-
ous trade namessuch as Leco. The output of the furnaces(resistance
or induction) can be put througha watervapor trap (H, a single V-tube
filled with anhydrone)into a CO2 absorptiontube (I). No exact com-
5 Capital letters in parentheses

refer to units in Fig. 34-1.
bination of units is prescribedhere since many suitablecombinations

are possible.
12. Analytical balance(Mettler H31AR or equivalent).
Reagents
1. Reagents1, 2, 3, 7, 8, and 9 describedin "Reagents"under"Medium-
TemperatureResistanceFurnaceMethod."
2. Tin metal accelerator(LECO no. 501-076or equivalent).
3. Iron chip accelerator,C free (LECO no. 501-077or equivalent).
4. Tin-coatedcopperaccelerator(LECO no. 501-263or equivalent).
5. Scoop for adding 1 g of accelerators(LECO no. 503-032or equiva-
lent).
Procedure Using Iron, Tin, and Tin-coated Copper Accelerators

Weigh the CO2 absorptionbulb on the analyticalbalance,insert the bulb in
the train, and openthe stopcocks.Set the O2 flow at the rate of 1.5 Umin. Place
an empty crucible in the induction furnace.Fire the furnace,following the man-
ufacturer'sinstructions,for 5 min. At the end of the combustionperiod, remove
the crucible, tum off the O2 flow, and then closeoff and removethe CO2 absorp-
tion bulb. Weigh the CO2 absorptionbulb (see "Comments"under "Medium-
TemperatureresistanceFurnace Method"). Repeat this processuntil a blank
reproducible±0.2 mg is obtained. Alternatively,ignite two or more blanks (cru-
cible containing1 scoopeachof iron chip, tin and tin-coatedcopperaccelerators)
until a blank reproducibleto ±0.2 mg is obtained.
Transfera 0.5000-gsampleof soil that passesthrougha 100- or 140-mesh
(106-150 /-lm) sieve to a crucible. Add one scoopof tin metal accelerator,one
scoopof iron chip accelerator,and one scoopof tin--coatedcopperaccelerator,
and cover the crucible. Insert the coveredcrucible in the induction furnace. Set
the O2 flow at a rate of 1.5 Umin. Fire the furnaceaccordingto a manufacturer's
instructions.At the end of the combustionperiod, removethe crucible, tum off
the flow of O2, and close off, removeand weight the CO2 absorptionbulb (see
"Comments" under "Medium-TemperatureResistanceFurnaceMethod"). The
increasein weight of the CO2 absorptionbulb, after correction for the blank,
shouldbe due to CO2 releasedfrom the soil sample.Flush the train (without the
CO2 absorptionbulb) with O2 for about 1 min betweensuccessiveruns. Deter-
mine the blank for the crucible and acceleratorsusing the sameprocedure.
nt [g COz, sample]- [g COz, blank]

Total C, 70 = x 0.2727x 100 [2]
g water-freesoil
Comments
Commentsin "Comments"under "Medium-TemperatureResistanceFur-
naceMethod" on weighing of absorptionbulbs and on samplegrinding are fully
applicableto this procedure.Other commentsin "Comments"under "Medium-
TemperatureResistanceFurnaceMethod" also are applicable.
Under optimum conditions, the two proceduresyield comparableresults.
Adequatelyhigh temperature(> 1650°C)can be developedwith the properaddi-
CARBON AND ORGANIC MAITER 973
tions of Sn and Fe, but the temperaturemaximum is held only briefly. The tem-
peraturerises steadily until the susceptorsmelt and fuse with the sample, and
thereafter, it falls rapidly. Occasionally this temperaturerise and fall occurs
before thermal decompositionof C is complete,perhapsbecauseof inadequate
contactbetweenthe sampleand the susceptormaterial. For most soils, this does
not appearto be a major problemsinceFe, Sn, and tin-coatedcopperaccelerators
(-1 g of each/sample)havebeenfound to yield accuratetotal C valuesin a range
of calcareousandnoncalcareous soils and standardcarbonateminerals(Tabatabai
& Bremner,1970).
If the organic matter contentof the soil is high, the sampleweight should
be reducedappropriately.Organic materialscan be analyzedby this technique,
but sampleweightsmustbe reducedto 20 or 30 mg if explosionsare to be avoid-
ed. Alternatively the organic material in amountsup to 60 mg can be mixed and
coveredwith Alundum or Sinderiteas describedin "Procedure"under"Medium-
TemperatureResistanceFurnaceMethod."
The gravimetricdeterminationof CO2 following combustionwith the Leco
induction furnace was found by Carr (1973) to yield total C levels comparable
with manualwet and dry combustionmethods.In addition to gravimetry,an auto-
matedCO2 analyzerbasedon thermal conductivity measurements of the effluent
gaseswas applicableto soil analysis(Tabatabai& Bremner,1970).
Alternatively, a titrimetric method was developedto allow estimationof
both total C and 14C in soil samplesamendedwith 14C compounds(Cheng& Far-
row, 1976). A bypassvalve and a 12S-mL gas washing bottle (e.g., Corning
31760) are used in place of the CO2 absorption bulb of Fig. 34-1. All CO2
releasedby combustionis trappedin SO mL ofO.S M NaOH followed by removal
of one aliquot for liquid scintillation counting to quantify 14C2 and a second
aliquot for titration with standardHCI to determinetotal C. The total C data
obtained were comparableto those obtained by a wet combustionprocedure.
Recentdata also indicatesthat titrimetric and thermal condl'ctivity methodsare
comparablefor the determinationof CO2 (Winter et aI., 1990).
InstrumentalMethods
The following sectiondescribesrepresentativecommercialinstrumentsfor
determining total C in soils. They were chosen to illustrate the principles
involved in instrumentingtotal C analysis.The inclusion of the following three
instrumentsdoes not imply that they are superioror inferior to otherscurrently
beingmarketed.As with all instruments,variousevaluationproceduresshouldbe
usedto determineand to confirm the validity of data obtainedin comparisonto
accepted,standardmethods.Tabatabaiand Bremner(1991) describedautomated
instrumentsavailablefor determiningtotal C in soils aswell aswhich instruments
are capableof simultaneousdeterminationof N or S.
Carbo-ErbaNA 1500.Carlo-ErbaInstruments(Milan, Italy) developedan
automatedinstrumentcapableof simultaneousdeterminationof C, H, and N in
geologic materials, soils, and other environmental samples. The principles
involved, developmentof the instrumentand a descriptionof modesof operation
are presentedby Pella(1990a,b).A sampleis placedin a tin samplecup, crimped
Helium (continuous flow)
Sample in Sn
container
Reference Thermal
Conduct-
ivity
Sample Detector
Cu
NiO
Chro atographic
colum
J
Co,Oj Ag
Fig. 34--2. Schematicdiagramfor Carlo-ErbaModel NA 1500analyzer.
to confineit, andintroducedinto a quartzreactor.For mineralsoils, a typical sam-

ple size is 5- to 10 mg, necessitatingthe use of samplesfinely ground in a ball
mill or similar apparatus.The quartz reactoris maintainedat 1050°Cwith a con-
stant flow of He. Flashcombustionwill occurif a pulseof O2 is injectedinto the
quartz reactorshortly after introduction of the sample.Under thesetemperature
and O2 conditions, the tin is oxidized to SnOz resulting in the temperature
increasingto 1700 to 1800°Cand the completecombustionof soil organicmat-
ter. The combustionproducts(C02, N oxides,and HzO) are swept by the helium
carrier gas through chromium dioxide (Cr02) to catalyzeoxidation of organic
fragmentsand C030 4 coatedwith Ag to removehalogensand sulfur oxides.The
gasesthen flow through a heatedCu (650°C) column to removeexcessoxygen,
Mg(CI04)z to removeH20 and into a chromatographiccolumn for separationof
N2 and COz. The different gasesare detectedwith a thermal conductivity detec-
tor. A generalizedflow diagramfor this instrumentis shownin Fig. 34-2.
Scheperset al. (1989) evaluatedthe NA 1500 coupledwith a massspec-
trometerfor simultaneousdeterminationof C, N, and 15N in soil and plant mate-
rials. The NA 1500 yielded Nand 15N datacomparableto that obtainedwith con-
ventional manual methods(Kjeldahl digestion followed by mass spectroscopy
analysis).A detailedcomparisonof C data was not conductedalthoughrealistic
values for total C in soils and plant materialswere obtained.An evaluationof
samplepreparationmethods(soil grindinglWiley mill vs. ball milling) indicated
that homogeneous soil and plant sampleswere essentialto reduceanalyticalvari-
ability, especially in view of the typical sample size of 10 mg. Verardo et al.
(1990) have describedproceduresfor using the NA 1500 to determineC and N
in marine sediments.Due to the analysisof 5 to 10 mg, careful sampleprepara-
tion and grinding are neededon insure that a representativesampleis analyzed.
A" with any analyticalmethod,the inclusion of appropriatestandardsand blanks
is essentialtinsure valid total C data. The capability of coupling this instrument
with a massspectrometeris a potentialbenefit as well.
CARBON AND ORGANIC MATfER 975
Leco Instruments.The LECO Corporation(St. Joseph,MI) has marketed

instrumentsfor automatedanalysisof total C in soils and othersolid materialsfor
the pastseveraldecades.
A descriptionof Leco instrumentsis presentedby Tabatabaiand Bremner
(1991). Earlier results with the Leco automatic 70-s C analyzer (Tabatabai&
Bremner, 1970; Carr, 1973) indicate that reliable soil total C data are obtained
using Fe, Sn, and tin-coatedcopperacceleratorsin an induction furnacefollowed
by thermal conductivity to quantitateCO2, The newer LECO IR-12 instrument
involves combustionof a soil samplein an induction furnaceusing an O2 atmos-
pherefollowed by passingthe gas mixture over a catalystto convert CO to CO2
and CO2 quantitation with an infrared detector. A related instrument, model
DC-12 Duo-Carb,involves combustionof samplesmixed with V 205 (vanadium
pentoxide) in an induction furnace heatedto 1000°C under an O2 atmosphere.
The COz producedis measuredwith a thermal conductivity detector.
In 1993, LECO marketedtwo instrumentsfor determiningtotal C in soils.
Both instrumentsutilize resistancefurnacesto combustsamplesat >950°C. In the
Model CR-412, a sample containedin a ceramic boat is placed in a specially
designedhorizontalresistancefurnacemaintainedat a constanttemperaturein the
rangeof 950 to 1400°CunderOz flow. After a delay, Oz is directedonto the sam-
ple and carriesthe COz releasedthrough dust and water vapor traps and into an
infrared detection system. Merry and Spouncer (1988)evaluated the earlier
model CR-12 and found that it gave reasonablesoil organicC valueswhen oper-
ated at 1200°C. In an evaluationof combustiontemperatureon C recoveryfrom
noncalcareousand calcareoussoils, it was found that both inorganic and organic
C were recoveredbetween600°C and 1000°C.Total C determinedby the CR-12
and the Allison method(Allison, 1960; see"Wet CombustionMethod") were in
close agreement for20 Iowa soils (Yeomans& Bremner,1991).
Chichesterand Chaison(1992) evaluatedthe CR-12for determiningorgan-
ic C and inorganicC by combustingsamplesat 575°C and 1000°C,respectively.
They recommendedcombustionat 575°C for 250 to 360 s to determineorganic
C followed by combustionat 1000°Cfor 250 s to determineinorganicC. In addi-
tion, the time requiredfor analysisof inorganicC could be reducedfrom 250 to
60 s by increasingthe combustiontemperaturefrom 1000°Cto 1371°C.In sum-
mary, total C could bedeterminedby a single combustionat 1371°C as found by
other investigators.
A secondLECO instrument is the model CHN 600 which is capableof
simultaneousanalysisof C, H, and N. A flow diagramfor the CHN 600 is shown
in Fig. 34-3. The successorto the CHN 600 is the model CHN 1000. A soil sam-
ple «200mg) is placedin a tin capsuleand combustedin a resistancefurnace at
950°C using Oz as a carriergas.The gasesformed are scrubbedto removeS gases
and equilibratedin a ballast chamber.Mter equilibration, the gas mixture flows
through two infrared detectorsset to detectCOz and HzO. An aliquot of the gas
mixture is analyzedfor N z by thermal conductivity after reduction of N oxides
and removal of COz and HzO.
The CHN 600 has beenused in severalsoils studies.Total C resultsfrom
20 Iowa soils were essentiallythe sameusing the CHN 600 and a standardwet-
combustionmethod(Yeomans& Bremner,1991). A comparisonof dataobtained
950C
Dual Heat Zone

Furnace
Exhaust
Fig. 34-3. Schematicdiagramfor LECO Model CHN 600 analyzer.
by the CHN 600 with a LECO induction furnaceinstrumentand a wet-oxidation

method indicated that the CHN 600 was the most precise total C technique
(0.014>.12%C) and enableda technicianto perform 90 to 100 analysesin an 8-
h d (Sheldrick, 1986). The CHN 600 hasbeenshown to recover 100% of the C
in a range of pure organic C compounds[acetanilide(N-acetylaniline),sucrose
(C12H 220 1), sulfanilic acid (C6H7N03S),and EDTA (ClOH16N20S)] and, as ex-
pected,to yield soil organic C values 16 to 59% greaterthan those obtainedby
the Walkley-Black method (McGeehan& Naylor, 1988). In general, the CHN
600 has shown to be a reliable and accurateinstrumentfor the determinationof
total C in soils.
Perkin-Elmer CHN2400. The Perkin-Elmer (Perkin-ElmerCorp., Instru-
ment Division, Norwalk, CT) simultaneouslymeasuresC, H, and N using the
principles employed in the traditional Pregl and Dumas procedures.A sample
containedin a platinum boat is oxidized with O2 at -lOOO°C for 2 min in a com-
bustion tube in the absenceof carrier gas (He) flow. Mter combustion,He flow
is initiated and the CO2, H20, and N2 basesproducedby combustionare passed
over CuO to convertCO to CO2 and silver mesh(silver vanadateon silver wool)
to remove S and halogengases.The gasesthen flow into a tube maintainedat
650°C and packedwith coppergranules between end plugs of silver wool, where
quantitative reductionof N oxidesto N2 occurs.The gasesare broughtto constant
pressureand volume in a gas mixing chamberand then allowed to expandinto
the analyzerportion of the instrument.The analyzerconsistsof threethermalcon-
ductivity (TC) detectorsconnectedin series and separatedby two traps. The
sequenceof TC detectorsand traps enablingquantificationof H, C, and N is as
follows:
1. TC detector1 (output equalstotal gas composition).
2. Magnesiumperchloratetrap to removeH 20.
3. TC detector2 (decreasein output from detector1 is proportionalto H
content).
4. Sodaasbestosplus Mg(Cl04)2 trap to removeCO2.
5. TC detector3 (decreasein output from Detector2 is proportionalto C

content).
6. The remaininggasesin the sampleare N2•
All operations withinthe instrumentare automatic.Additional work is neededto
evaluatethis instrumentfor soil analysis.
The above discussionis an overview of instrumentalmethodsfor total C
analysisof soils. At present,the LECO instrumentshave beenthe most widely
usedfor soil analysis.Severalresearchlaboratorieshavebegunusing Carlo-Erba
instrumentsfor total C analysisof soils. Due to rapid changesin technologyand
instrumentation,it is essentialthat manufacturersbe contactedfor currently avail-
able modelsfollowed by an evaluationof the instrument.
Total Carbonby Wet Combustion
Introduction
The wet combustionanalysisof soils by chromic acid digestion has long
beena standardmethodfor determiningtotal C, giving resultsin good agreement
with dry combustion.The main advantagesfor wet combustionare that the cost
of apparatusis but a small fraction of the cost for dry combustionequipmentand
that the parts neededto assemblethe apparatusare standardequipmentin most
laboratories.The chief disadvantageof the earlier wet combustionprocedures
(e.g., Heck, 1929) is that they use macro equipment,which is tediousto assem-
ble and disassemble,and which occupiesconsiderablebenchspacemore or less
permanently.Wet combustion also is used when the special manometricVan
Slyke-Neil apparatus(Van Slyke & Folch, 1940; Bremner,1949) is employedto
estimatetotal C in soils.
The wet combustionmethodof Allison (1960), describedhere, embodies
important refinementsfrom publishedprocedures,such as simple and effective
digestionacid mixture (Clark & Ogg, 1942),a simple purification and absorption
train assembledon a small panel (McCready& Hassid,1942), and a more rapid
procedurethan formerly used(Heck, 1929; Jackson,1958, p. 211). The signifi-
cant featuresof this apparatus(Fig. 34--4) are as follows: (i) it can be assembled
from simple parts and requiresno ground-glassconnections,(ii) the small inter-
nal volume precludesthe necessityfor preaerationunder most laboratorycondi-
tions, (iii) it requiresonly a short period of aerationfollowing digestion,and (iv)
the entire assembly(F-K) occupiesonly a small area.This methodis satisfacto-
ry for salt-affectedsoils high in Cl- and also for the dry residuesof soil extracts
rich in organicmatter.A rapid treatmentto removecarbonatesdescribedin "Pre-
treatmentPrior to Wet Combustion"permits determinationof organic C on the
residueof a pretreatedcalcareoussoil. The following descriptionof wet combus-
tion methodologywas presentedby Allison et al. (1965).
Principles
The soil sampleis digestedin a 60:40 mixture of H2S04 and H3P04 con-
taining K2Cr207' The boiling temperatureof this mixture, 210°C, is high enough
to ensurecompleteoxidation of carbonaceous matter, yet low enoughto prevent
To Trap
I or"
Ga,-
.2
~
Trop I
MglCIO ) JFiber Qloss

Ascarite-
Fibtr Qloss
K
Go.-·
2t
T'OP ~I
.
.
"
•
250 ml Side. arm
mon
Mods
+
NoOH
Nesbitt Erlenmeyer
bulb
Fig. 34-4. Diagram of apparatususedto determineC by the wet combustionmethod.Trap I or II is

usedfor determinationof CO2 evolved by gravimetricor titrimetric techniques,respectively(dia-
gram is not drawn to scale).
excessivefuming in the condenser.The CO2 evolved is absorbedby a suitable

absorbentandweighed,althoughit may be absorbedin a standardbaseand titrat-
ed.
A combination of fuming H2S04, phosphoric acid (H3P04), iodic acid
(HI03) [added to potassiumiodate (KI0 3)], and cr03 has been used for deter-
mining C in organic compounds(Van Slyke & Folch, 1940) and in soil (Mc-
Cready & Hassid, 1942).The reportedadvantagesof this oxidation mixture are
that it vigorously attacksand dehydratesresistantforms of C, thereby reducing
boiling time for completeoxidation, and that it facilitates conversionof CO to
CO2, Carbonmonoxideis often producedwhen readily oxidizablecarbohydrates
are presentin the sample.Extensivecomparisonsof the Van Slyke-Folchand the
60:40 H2S04-H 2P04 oxidizing mixtureson many soils indicatethat the two mix-
turesare equallyeffective in convertingtotal soil C to CO2, The more rapid diges-
tion with the Van Slyke-Folch mixture, resulting in a saving of 3 or 4 min per
determination,is not sufficient advantageto offset the difficulties of preparing
and maintaininga digestion acid that containsfuming H2S04, Moreover it was
found that the needfor HI0 3 in the digestionmixture doesnot exist, which indi-
catesthat soil organicmattercontainslittle or no active carbohydratecapableof
producingCO during digestion(Allison, 1960).
Salt-affectedsoils frequently contain sufficient Cl- to give errors by wet
combustionanalysiswhetherthe CO2 is determinedtitrimetrically (Clark & Ogg,
1942) or gravimetrically (Allison, 1960). When soil high in Cl- is heatedin a
digestion mixture containingCr20-r-, chromyl chloride (CrOzCI2) is formed by
the following reactionbeforeboiling begins
The reddishCr02Cl2 decomposesat about 190°C, releasingfree C12, with

a color changeto pale green.Any Cl2 and tracesof undecomposedCr02CI2 that
passthe purification train are retainedin the CO2 absorptionbulb to give a posi-
tive error.
In the methodsdescribed,Cl2 interferenceis preventedby including two
traps in the purifying train, one containingKI and one containingsilver sulfate
(Ag2S04) (TrapsF and G in Fig. 34-2). The use of Ag2S04 alone gives protec-
tion up to about0.2% Cl- (Allison, 1960), but its protectivevalue becomesques-
tionableat higher Cl- concentrations,SinceKI hasa very high capacityto absorb
free Cl 2 by the reaction
2KI + Cl 2 =2KCl + 212, [4]
the use of a KI trap is recommendedfor soils high in Cl-. With both traps in the
system,Cl- up to 5% of the sampleweight does not interfere, provided proper
precautionsare observedduring the early stagesof sampledigestion.Use of the
Ag2S04 trap in conjunctionwith the KI trap servesto indicatewhen the latter is
exhausted.For soils containing trace or low amountsof Cl-, the carrier stream
may flow directly into the Ag2S04 trap.
Wet CombustionMethod
The wet combustionmethodwas describedby Allison (1960).
Special Apparatus
The apparatusis shown in Fig. 34-4. Assemblethe apparatusfrom the fol-
lowing parts: (A) Hoke needlevalve: (B) 25-cm high soda-limetower; (C) 100-
mL Kjeldahl flasks to fit a no. 2 stopper;(D) Allihn four-bulb condenser,fitted
with a no. 2 stopperat the delivery end; (E) 60-mL open-topseparatorfunnel;
(F-H) 25- by 90-mm shell vials with no. 4 stoppers;(I and1) I5-cm long CaCl2
U-tube; and (K) Nesbitt absorptionbulb. Use neoprenestoppersand gum rubber
tubing for all connections.Coat all rubbertube connectionslightly with silicone
lubricant.
Items C through E can be ground-glassjoint glasswareif desired (Fig.
34-4). All joints are standard-taper24/40. The following parts are needed:(C)
100-mL round-bottomflask (Coming 4320); (C-I) distilling adaptertube (Com-
ing 9421), which containsinlet tube for bubbling C0z-freeair into digestionacid
mixture; (D) Allihn condenser,-300-mmjacketlength(Coming2480); (E-1) dis-
tilling tube with suction sidearm (Coming 9420) (side arm is connectedto puri-
fying traps); (E) graduatedseparatorfunnel (Coming 6382A). A heatingmantle
and rheostatare usedto heat the 100-mL digestionflask.
Providea C0z-freecarrier streamby releasingair from an air pressureline
through Valve A and passingit through soda-limeTower B. ConnectB in a glass
tube 4-mm o.d. that extendsdownwardthroughCondenserD and dips about1 cm
below the surfaceof the oxidizing acid in DigestionFlask C. Shortenthe stemof
FunnelE to a length of about 9 em, and reducethe tip openingof the stem to a
diameterof about 2 mm. Adjust the position of the FunnelE to extendinto D at
least 5 cm below the stopperto avoid contactbetweenoxidizing acid and stop-

per. LubricateStopcockE with the digestionacid mixture or with syrupy H3P04•
Regularstopcocklubricant should not be usedon stopcocks.
Assemblethe purifying traps,F to J, on a panelto provide stability. Fit the
vials of trapsF, G, and H with no. 4 stoppersthey have approximately6 cm of
the bottom cut off to provide a tight sealwith the vials. Reducethe tip openings
of the inflow tubesin F and G, but do not makethem smallerthan 1 mm in diam-
eter, or sealingmay occur. Fill trapsF and G approximatelytwo-thirds full with
50% KI solution and saturatedAg2S04, respectively.Adjust the inflow tubesso
that they extend into the solutionsnot more than 3.8 cm for Trap F and 1.3 cm
for Trap G; otherwiseback pressuremay developand causeleaks in the system.
Fill Trap H not more than one-thirdfull with concentratedH2S04, Prepare
the inflow tube for H from the barrel of a 5-mL pipettewith the tip extendingnot
more than 1.3 cm into the acid (note that Trap H connectsdirectly to Trap I).
Placea fiberglassdisc in the bottom of the V-tube; and fill the right side, Trap [,
with 30-mesh(600 !lm) granularZn for absorbingany acid fumes that escape
pastH. Fill the left trap, TrapJ, with anhydrousMg(CI04)2, which absorbswater
from the carrier streamcontainingevolvedCO2 beforeit entersK.
Fill the Nesbitt absorptionbulb K with any good, self-indicatingabsorbent
having a high capacityfor absorbingCO2, Indicarb and Mikhobite are excellent
for this purpose.When filled as shown in Fig. 34--4, the bulb containssucces-
sively a 3-cm layer of 8- to 14-mesh(1.4-2.36mm) absorbent,a 2-cm layer of
14- to 20-mesh(0.85-1.4 mm) absorbent,and a l-cm overlayer of anhydrous
Mg(CI04)2, with a wad of glasswool aboveand below the column.
Reagents
1. Digestion acid mixture: Pour 600 mL of concentratedH2S04 into 400
mL of 85% H3P04, cool the mixture, and store it in a glass-stoppered
bottle. Keep the bottle well stopperedto prevent absorptionof water
vapor.
2. Potassiumdichromate,reagentgrade.
3. Potassiumiodide solution, 50%: Dissolve 100 g of KI in 100 mL of
water.
4. Silver sulfatesolution, saturated.
5. Carbondioxide absorbent,self-indicating,7- to 14- (1.4-2.8 mm) and
14- to 20-mesh(0.85-1.4mm) size; Suitablematerialsare Mikhobite
(G. FrederickSmith ChemicalCo., Columbus,OH), Caroxiteor Indi-
earb(FisherScientific, Pittsburgh,PA), or Ascarite(Arthur H. Thomas
Co., Philadelphia).
6. Sodalime, 8- to 14-meshsize (0.85-1.4mm).
7. GranularZn, <30-mesh«600!lm)size.
8. Anhydrousmagnesiumperchlorate(Anhydrone,Dehydrite,or equiva-
lent).
Procedure
Place a finely ground soil samplecontaining 20 to 40 mg of C (usually
0.5-3 g of oven-dry soil) into digestionflask C, and addabout 1 g of K2Cr207'
Wash down the neck of the flask with 3 mL of distilled water, and connectthe
flask to CondenserD. Weigh the Nesbitt bulb ("Comments"under "Medium-
TemperatureResistanceFurnaceMethods" and the following "Comments"sec-
tion), attachit to the system,and immediatelyopen the valve at the top of the
bulb. Pour 25 mL of the digestion acid mixture into Funnel E above the con-
denser,andcoverthe funnel with a small beaker.OpenStopcockE, allow the acid
to flow throughD into Flask C, and close the stopcockimmediatelyto prevent
loss of CO2, Adjust the air delivery tube that passesthroughD into C so that its
tip extendsnot more than 1 cm into the acid during digestion.
At this point, tum on the cooling water. Adjust the carrierstreamto a flow
rate of about2 bubbles/s,and maintainthis rate during digestion.Placethe heat-
ing mantlearoundthe flask or apply a flame 5 to 6 cm high, and bring the sam-
ple to boiling in 3 or 4 min. If Cl- is high, heatthe mixture slowly at first, and
bring it to boiling in about 5 min. Continuegentle boiling, avoiding excessive
frothing, for a total heatingperiod of 10 min. Reducethe rate of healingif visi-
ble white fumes of S03 occurabovethe secondbulb of D during digestion.
Removethe heatingmantleor flame at the endof the digestionperiod,and
aeratethe systemfor 10 min at the rateof 6 to 8 bubbles/soWhenaerationis com-
plete, shut off the air stream,and disconnectthe digestionflask from the con-
denser.Closethe stopcockon the Nesbittbulb, anddisconnectit from the system.
Brushthe bulb with camel'shair to removeany lint anddust,andweight it imme-
diately. Make a blank determination,using the identical procedure,but without
sample.Add four to five glassbeadsto the blank to preventbumping.The calcu-
lation is as follows
[g CO2, sample]- [g CO2, blank]

Total C, % = x 0.2727x 100 [5]
g water-freesoil
Comments
Soil samplesshould be ground to passthrough a sievewith openings0.5
mm or smallerin diameter.This is necessaryto reduceerrorsdue to the presence
of occasionalfragmentsof carbonatemineralsin a predominantlynoncalcareous
matrix.
A singleanalysis,involving all operationsfrom weighingthe sampleto cal-
culationof results,requires25 min. By usingtwo setsof apparatus,onemay ana-
lyze two samplesconcurrently,therebyreducingthe overall time requiredto 15
min perdetermination,providedthe digestionphaseof onesamplecoincideswith
the aerationphaseof the other.
BecauseCO2 absorptionbulbs changeweight on standingovernightor for
longerperiods,it is necessaryto bring the bulb to constantweight by the follow-
ing procedure before beginning C or blank determinations.Without being
weighed,the bulb shouldbe connectedto the system,all reagents(but no soil)
shouldbe addedto the digestionflask, and the apparatusshouldbe operatedas
directedfor sampledeterminations.After aeration,the bulb shouldbe detached
andweighed,andthis weight shouldbe usedasthe initial (constant)weight of the
bulb. See"Comments"under"Medium-TemperatureResistanceFurnaceMeth-
ods" for additionalcommentson careand useof CO2 absorptionbulbs.
Blank determinationshaverangedfrom 0.8 to 1.2 mg of CO2, for which an

averagevalue of 1.0 mg has beenused.If blanks are found to be high, preaera-
tion may be necessary.The systemmay be preaeratedby placing the digestion
flask (containing all materialsexcept the digestion acid) in position for diges-
tion, disconnectingthe rubber tube betweenD and F, opening Valve A, and
directing a streamof C0z-freeair (about 10 bubbles/s)into C and throughD for
2 min (spatteringof the contentsin C mustbe avoided).The air flow is then read-
justedto about2 bubbles/s,D is connectedto F, and the analysisis performedas
directed.
The H2S04 in the Trap H shouldbe renewedat the beginningof eachday's
operationor more often if frothing occurs.The KI solution in Trap F has a high
capacity for absorbingCl-, and the needfor its renewal is indicatedby the first
trace of an AgCI precipitatein Trap G.
The Nesbitt absorptionbulb, when filled as described,weighs about 125 g
and will absorbabout 10 g of CO2, equivalentto about 100 determinationsaver-
aging 100 mg of CO2 each.
When the apparatusis idle overnightor for longer periodsand the Nesbitt
bulb is detached,the tube connectingJ and K should be clampedoff to prevent
entranceof water vapor into the desiccantin TrapJ.
A titrimetric procedurefor CO2 determinationis readily adaptableto the
above procedure(Fig. 34-4). Replacethe Nesbitt bulb with a 250-mL sidearm
Erlenmeyer(filtering flask) fitted with a no. 61/2 stoppercontaininga 22-cm by
14-mmdiam. glasstube. This bubble towershouldextendto within 0.5 cm of the
flask bottomand shouldbe filled with glassbeads.Throughthe glasstube, 25 mL
of 1 M KOH should be added, and the soil sample should be oxidized as de-
scribedpreviously. The acid-baseindicator Tropaeolin0 (e.g., Sigma Chemical
Co., St. Louis, MO; Aldrich Chemical Co., Milwaukee, WI; FisherScientific,
Pittsburgh, PA) can be added to the KOH to ensure that sufficient alkalinity
remains after trapping the CO2 evolved. After oxidation, the KOH is washed
from the bubble tower with distilled water, treatedwith 5 mL of saturatedBaCl2
and severaldropsof phenolphthalein,and titratedwith standardHC!. The dataare
calculatedfrom
'".0tIC
a ,70
Of -
-
m4,lank - mi-sample
. X
NHCl X
0• 6 [6]
g soIl
The basic principle of the above wet-combustionprocedurehas been used in

developingtubelflask digestion methodsfor total C analysis. In essence,these
methods involve mixing a soil sample, solid K2Cr07, and 3:2 concentrated
H2S04:concentrated85% H3P04 in a sealedvesselcontainingNaOH to trap CO2
evolved from oxidized organic C and solubilizedcarbonates.Snyderand Trofy-
mow (1984) and Coughtreyet a!. (1986) describemethodsusingmodified culture
tubesand Erlenmeyerflasks, respectively.The samplecontainersare heatedin a
digestionblock or on a hot plate at about 120°C for 2 h followed by diffusion of
CO2 into an alkali trap for 12 h. The amountof OH- remainingis determinedby
titration. These methods also have the advantageof being readily adaptedto
determinationof both 12C and 14C in the samesample.In addition, they are rapid,
provide comparable data

to establishedmethodsand use relatively inexpensive
equipment.
ORGANIC CARBON
Introduction
Carbonis the chief elementpresentin soil organicmatter,comprisingfrom

48 to 58% of the total weight. Therefore,organicC determinationsareoften used
as the basisfor organicmatterestimatesthroughmultiplying the organicC value
by a factor. For many yearsthe Van Bemmelenfactor of 1.724wasusedbasedon
the assumptionthat organicmattercontains58% organicC. However,a number
of studieshave shown that the proportion of C in soil organicmatter is highly
variablefor a rangeof soils andthereis no factor appropriatefor all soils. If a fac-
tor must be selectedfor convertingorganic C concentrationsof organic matter
contents,valuesof 1.9 and 2.5 for surfaceand subsoils,respectively,are most
appropriate(Broadbent,1953). The factor varies not only from soil to soil but
also betweenhorizons in the samesoil. This finding suggeststhat it is most
appropriateto determineand reportthe organicC in a soil ratherthanconvertthe
analytically determinedorganicC value to organicmattercontentthroughuseof
an approximatecorrectionfactor.
o.rganicC may be determinedby: (i) analysisof a soil for total C and inor-
ganic C and subtractionof the inorganicC concentrationfor the total C content,
(ii) a total C determinationon the sampleafter destructionof inorganic C, and
(iii) oxidationof organicC compoundsby Cr20r- and subsequentdetermination
of unreducedCr2o.1-by oxidation-reductiontitration with Fe2+ or by colorimet-
ric methods.Table 34--2 summarizesthe principles, advantages,and disadvan-
tagesof severalmethodsfor determinationof organicC in soils.All currentmeth-
odshaveinherentproblemsassociatedwith them,andthe investigatorshoulduse
the methodmost applicablefor the soils to be analyzedand the requiredaccura-
cy of the results.
In this section,proceduresare describedfor the determinationof organicC
in both calcareousand noncalcareous soils basedon the differencebetweentotal
C andinorganicC concentrations.Two methodsalsoaregiven for organicC esti-
mationsbasedon destructionof inorganicC compoundsprior to total C determi-
nations. In addition, two rapid dichromateoxidation proceduresare described.
The Walkley and Black (1934) methodthat oxidizesorganicC through heat-of-
dilution of H2SO.4 is given becauseit is simple, rapid, widely used,and requires
minimal equipmenteventhoughthe resultsobtainedcannotbe consideredquan-
titative. Many soil testingand soil survey personnelhaveneedfor a methodthat
gives an approximateorganicC concentration.A tube digestiontechniques(Nel-
son & Sommers,1975) that involves extensiveheating of the chromic acid-soil
mixture is given becauseit is quantitative,rapid, and representsthe bestcombi-
nationof digestionreagents,heatingprocedure,andtitration reagentsof the mod-
em dichromatemethods.Dichromateproceduresare widely usedin soil investi-
gationsbecauseof their simplicity and rapidity comparedwith wet or dry com-
!
Table 34-2. Comparisonof methodologiesusedfor determinationof organicC in soils.

Method Principle Advantages Disadvantages
Difference between Total C and inorganicC are determinedon separate Useful if total C and Two separateanalysesare required,total C determination
total C and inor- samples:organicC =total C - inorganicC inorganicC are rou- requiresspecialequipment,organicC calculatedby
ganic C tinely determined differencehas someinherenterror
Determinedas total C Total C is determinedin soil sampleafter removal Accurateif dolomite Not all dolomite in soil may be removedby acid treat-
after removal of of inorganicC with an acid pretreatment: is absentfrom soil ment, specializedequipmentneeded.
inorganicC organicC = total C
Dichromateoxidation DichromateoxidizesorganicC to CO2 in acid Very rapid and simple, Incompleteoxidation of organicC necessitates use of a
without externalheat medium; amountsof Cr20:r- reducedis quanti- no specialequip- correctionfactors, which often resultsin erroneous
tatively relatedto organicC present;not all ment needed values;chloride, Fe2+, and Mn02 interfere with meth-
organicC in samplesis oxidized when external od; it assumessoil organicC has an averagevalence
heat is omitted, and a correctionfactor is required of 0; variable recoveryof C from carbonizedmaterials
Dichromateoxidation This is the sameas the dichromatemethodabove Rapid and simple, com- Chloride, Fe2+, and Mn02 interfere with method;some
with externalheat except thatall organicC in the sampleis oxidized, plete oxidation of or- specializedequipmentis needed;it assumessoil organ-
and no correctionfactor is required ganic C occurs ic C has an averagevalenceof 0; variable recoveryof
C in carbonizedmaterials
e
o
z
Roo
~
i
bustion. However,the rapid K2Cr207 methodsare subjectto interferenceby oxi-

dizable or reduciblesoil constituentssuch as Cl-, Fe2+, and Mn02'
OrganicCarbonas Calculatedfrom Total CarbonDeterminations
Methodspreviously describedfor total C are basicfor many of the proce-

dures used to determineorganic C in soils. However, soils may contain both
organicand inorganicC and, thus, total C analysisproceduresrecoverboth forms
of C. In noncalcareous soils and soils not recently limed, the total C can be con-
sideredto be organic C. With calcareousor recently limed soils, organic C may
be estimatedas the differencebetweentotal C and inorganicC concentrations.
OrganicCarbonin NoncalcareousSoils
Preparesoil samples,and conduct a total C determinationby dry or wet
combustionusing titrimetric, gravimetric, volumetric, infrared, or thermal con-
ductivity techniquesto quantitateevolved CO2 as describedin "Total Carbon."
Reportthe total C determinedas percentageorganicC in the sample(i.e., total C
=organicC).
OrganicCarbonin CalcareousSoils
Preparesoil samples,and conducta total C determinationon the sampleby
dry or wet combustiontechniquesas describedin "Total Carbon."Determine
inorganicC on a separatesampleby one of the quantitativemethodsdescribedin
Chapter15 (Loeppert& Suarez,1996). Calculatethe percentageorganicC in the
samplefrom the relationship
organicC, % =% total C - % inorganicC [7]
Wet and Dry CombustionTechniquesfor Direct Measurement

of OrganicCarbonin CalcareousSoils
In contrastto noncalcareoussoils, inorganicC must be removedfrom cal-

careousor recently limed soils before the analysisif wet or dry combustiontech-
niquesare usedto directly measurethe organicC present.
InorganicC is convenientlyremovedbeforewet combustionby pretreating
the samplecontainedin a digestionflask with a mixture of dilute H2S04 and fer-
rous sulfate (FeS04)'The FeS04is addedto the mixture to minimize oxidation
and decarboxylationof organic matter by added H2S04 or by Mn02 presentin
soil (Allison, 1960). After pretreatment,the digestion flask containing soil is
transferredto the combustiontrain, and a total C determinationis carried out as
describedin "Total Carbonby Wet Combustion."
Inorganic C removal is generally more difficult before determinationof
organicC by dry combustiontechniques.Treatmentof soil at room temperature
with sulfurousacid (H 2S03) followed by heatingto removeexcessH 2S03 is nor-
mally used to decomposeinorganic C compounds(Piper, 1942, p. 221-222;
Bremner, 1949); however, severaldifficulties are apparentwith the procedure.
Little destructionof organicmatteroccursduring room temperaturetreatmentof

sampleswith HZS03, but somedecarboxylationis possibleas the sampleis heat-
ed (Bremner, 1949). It is difficult to decide when all inorganic C has been
removedand when H ZS03 treatmentshould be discontinued.It is doubtful that
dolomite is completely decomposedby the relatively mild HzS03 treatment
employed(Allison, 1965). Nommik (1971) suggestedthat inorganic C may be
effectively removedfrom soil samplesby treatmentwith a metaphosphoricacid
solution for 30 min at room temperatureand 30 min at 130°C. However, Nom-
mik's procedurehasnot beenevaluatedwith a variety of soils.
Test for Presenceof InorganicCarbon

Placefinely ground soil on a spotplate, and moistenwith a few drops of
water. Add 4 M HCI dropwise to the wetted sample,and observeany efferves-
cence.Allow sufficient time for dolomite to react (-5 min). If inorganic C is
absentfrom the soil, proceedwith organicC (total C) analysisas per the section
on "Total Carbon."If inorganic C is presentor the test is not definitive, proceed
as describedbelow.
PretreatmentPrior to Wet Combustion

Special Apparatus6
Reagents
1. Digestion reagentfor carbonates(HzS04-FeS04):Dissolve 57 mL of
concentratedHzS04 and 92 g of ferrous sulfateheptahydrate(FeS04•
7HzO) in 600 mL of deionizedwater, cool, and dilute to 1 L.
2. Potassiumdichromate,reagentgrade,pulverized.
3. Other reagentsas describedin "Reagents"under "Wet Combustion
Method."
Procedure
Preparesoil samplesas describedin "Procedures"under"Wet Combustion
Method." Transfera sampleof known water contentand containing20 to 40 mg
of C (but not more than 2 g of soil) to the flask usedfor the wet combustionappa-
ratus (e.g., a 100-mL Kjeldahl digestion flask or standard taperround bottom).
Using 3 mL of the HZS04-FeS04digestionacid, washdown any soil that adheres
to the neck of the flask. Placethe flask in a rack or beaker,and allow the sample
to digest at room temperaturewith occasionalturning of the flask for at least20
min or until effervescenceappearsto cease.Then hold the flask upright over a
flame 2 cm high, and boil the contentsslowly for 1.5 min to destroyany remain-
ing carbonate.Rotate the flask continuouslyduring boiling to avoid excessive
frothing. Allow the sampleto cool.
Insert a long-stemmedfunnel into the flask, and add 2 g of pulverized
KZCrZ07. Immediatelyconnectthe flask to the reflux condenser(Fig. 34-4), and
6 Seethe specialapparatuslisted in "SpecialApparatus"under"Wet CombustionMethods."

proceedwith the determinationof organic C as directed in "Procedure"under

"Wet CombustionMethod" beginningwith the third sentence.
Reportthe C presentin the pretreatedsampleas percentageorganicC.
Comments
The 3 mL of 1 M (2N) HZS04-5% FeS04used in this procedurereplaces
the 3 mL of distilled waterusedin the total C proceduredescribedin "Procedure"
under"Wet CombustionMethod." ThreemL of this reagentadds3 millimoles (6
meq) W, which will neutralize0.3 g of CaC03 (Le., 15% CaC03 in a 2-g soil
sample).An appreciableexcessof acidity must be presentto ensurecomplete
decompositionof carbonates.Rather than using >3 mL of the 1 M reagentfor
soils containingmore than -10% CaC03 equivalent,it is preferableto use 3 mL
of a 1.5 M or evena 2 M HZS04-5% FeS04reagent.
PretreatmentPrior to Dry Combustion

Special Apparatus'
Reagents
1. Sulfurousacid, approximately5%: Bubble SOz through distilled water
until a saturatedsolution is obtained.Keep the bottle well stopperedto
preventrapid loss of SOz.
2. Sodiumhydroxide(NaOH), pellets.
Procedure
Transfer a soil samplethat passesthrough a 100- or 140-mesh(106-150
/lm) sieve (see "Comments" under "Medium-TemperatureResistanceFurnace
Method") and of known water contentto a nonporouscombustionboat that has
beenpreviously ignited and cooled.Basedon an estimateof inorganicC present,
treat the sample with an excessof a 5% HZS03 solution. After several hours,
removethe water and excessHZS03 by leaving the boat overnightin an evacuat-
ed desiccatorcontainingNaOH pellets.Repeatthe treatmentuntil evolutionceas-
es on addition of HZS03•
Proceedwith the determinationof organicC by one of the dry combustion
methods (see "Medium-TemperatureResistanceFurnace Method" or "High-
TemperatureInduction FurnaceMethod"). Report the C presentin the pretreated
samplesas percentageorganicC.
OrganicCarbonin Soil Extracts
SpecialApparatus
Seethe special apparatuslisted in "Special Apparatus"under "Wet Com-
bustionMethod."
7 Seethe specialapparatuslisted in "Special Apparatus"

under"Medium-TemperatureResistance
FurnaceMethod" and "SpecialApparatus"under"High-TemperatureInduction FurnaceMethod."
Reagents
Seethe reagentslisted in "Reagents"under"Wet CombustionMethod."
Procedure
Placean aliquot of the extract (10 to 50 mL, dependingon the organicC
content)in a l00-mL Klejdahl digestionflask, and add1 mL of the H2S04-FeS04
reagent.Immersethe bulb of the flask in boiling water,anddirect a streamof dry,
dust-freeair onto the surfaceof the liquid in the flask. Reducethe volumeof solu-
tion in the flask to 3 mL or less.Add five or six glassbeadsand 1 g of K2Cr207
to the flask, andproceedwith the determinationof organicC asdirectedin "Pro-
cedure"under"Wet CombustionMethod."
Comments
Drying of extractsis bestaccomplishedin l00-mL flasks of the Kjeldahl
type. A 2-L beakerconvenientlyholds four flasks.
Rapid DichromateOxidationTechniques
Introductionand Principles
Schollenberger(1927)first proposedthat the organicmatterin soil may be
oxidizedby treatmentwith a hot mixture of K2Cr207andH2S04accordingto Eq.
[8].
2 Cr20~- + 3 CO + 16 H+ =4 cr3+ + 3 CO2 + 8 H20 [8]
After the reaction,the excessCr20~- is titratedwith Fe(~h(S04)2 • 6H20, and

the Cr201-reducedduring the reactionwith soil is assumedto be equivalentto
the organicC presentin the sample.It mustbe emphasizedthat all methodsbased
on determinationof Cr201- remaining or cr3+ formed assumethat C in soil
organicmatterhasan averagevalenceof zero.Although mostdichromateoxida-
tion proceduresdescribed since the original Schollenbergermethod have
involved chromicacid solutionsor mixturesof concentratedH2S04and aqueous
K2Cr207 solutions(Table 34-3), the use of other oxidants has beenproposed.
Degtjareff(1930) suggestedthat a mixture of H20 2and chromicacid be usedto
oxidize organicmatter.However,Walkley and Black (1934) conclusivelyestab-
lishedthat the additionof H20 2to chromicacid proceduresgavefictitiously high
valuesfor organicC becauseH20 2 reducedCr201-in acid solution. Edsonand
Mills (1955)suggestedthat organicC be oxidizedby Cl2(1% solution)andresid-
ual Cl2 determinedcolorimetrically by reactionwith o-tolidine (C14Hl~2)' The
intensity of yellow color was proportionalto organic C oxidized. Othershave
suggestedthat organicC in aqueousextractsof soil canbe determinedby oxida-
tion with a Mn(III)-pyrophosphatecomplex(Bartlett & Ross,1988).The loss of
color from Mn(III) is proporti~nal to the amountof organicC oxidized. Tmsley
(1950)andKalembasaandJenkinson(1973)proposedthat the chromicacid mix-
ture usedto oxidize organicC compoundsbe 3 and 1.5 M (9 and 4.5 N), respec-
~
o
z
~
I::'
o
Table 34-3. Characteristicsof dichromatemethodsfor determiningorganic C in soils. ~

(=)
Digestion reagentconcentrations
Method K 2Cr20 7 H2SO4 H3P04 Ratio of H2O/acid Digestion conditions Reportedprecision
N v:v CV,%
~
tol
::c
Schollenberger(1927) 0.058 18 Tube heatedby flame at 175°Cfor 90 s 1.4-1.9
Tyurin (1931) 0.066 9 1.00 Flask with funnel boiled at 140°C for 5 min 8.5
Walkley-Black (1934) 0.055 12 0.50 Flask with no externalheat, max. temp is 120°C 1.6-4.2
Anne (1945) 0.027 11 0.46 Flask with condensorrefluxed at 178°C for 5 min 1.3
Tinsley (1950)t 0.027 7.2 3 0.67 Flask with condenserrefluxed for 2 h at 150°C 0.8-3.1
Mebius(1960) 0.045 10 0.42 Flask with condensorrefluxed for 30 min at 159°C 1.2-1.8
Kalembasa& Jenkinson(1973) 0.033 9 1.67 0.067 Flask with condenserrefluxed for 20 min at 165°C 0.8
Nelson& Sommers(1975) 0.066 10.8 0.67 Tube heatedin block at 150°C for 30 min 3.5
Modified Mebius:j: 0.033 10.8 0.67 Flask with condenserrefluxed at 150°Cfor 30 min 1.0-3.6
Heanes(1984) 0.055 12 0.50 Tube heatedin block at 135°C for 30 min 4.1
Yeomans& Bremner(1988) 0.066 10.8 0.67 Tube heatedin block at 170°C for 30 min 1.0-4.4
Ciavattaet al. (19898) 0.145 10.2 0.77 Specialflask heatedover flame at 1160°Cfor 10 min 5.4
Soon& Abboud (1991) 0.066 10.8 0.67 Tube heatedin block at 155°C for 30 min 2.7
t Reagentsusedby Bremnerand Jenkinson(1960a).
:j: As describedby Nelson and Sommers(1982).
!
tively, with respectto H3P04 (Table34-3). Thereis no evidence,however,to sug-

gest that oxidation mixtures containing H3P04 are more efficient in oxidizing
organic matter than K2Cr20TH2S04mixtures. The oxidizing mixtures used in
most publishedmethodsare between0.0267and 0.0583M (0.16 and 0.35 N) in
K2Cr207 and 7.5 and 12.5 M (15 and 25 N) in H2S04 (Table 34-3). However,
Tyurin (1931) Tinsley (1950), Nelsonand Sommers(1975) and Yeomansand
Bremner(1988) usedan aqueousH2S04-water mixture that was 0.145 M (0.87
N) in K2Cr207. Schollenberger(1927) usedconcentratedH2S04 (-18 M or -36
N) as the solventfor K2Cr207.
Rapid dichromateoxidation techniqueshave employedheatingtimes and
temperaturesthat vary from no externalheatto extensiveboiling of chromic acid
mixtures. Schollenberger(1927) suggestedthat the soil-H2S04-K2Cr207mixture
be heatedin a Pyrex test tube over a flame until the solution temperaturereached
175°C at which time heatingwas discontinued.Later investigatorsrealizedthat
the time and temperatureof heating were critical and must be standardizedto
insure that a constantproportion of soil organic matter was oxidized and that a
consistentamountof dichromatewasthermally decomposedduring the digestion.
Degtjareff(1930),Tyurin (1931), Schollenberger(1945) andJackson(1958) sug-
gestedthat the soil-chromic acid mixtures be heatedfor defined periods (5-10
min) in test tubessubmergedin H2S04 or oil bathsmaintainedat prescribedtem-
peratures(140-170°C).
Walkley and Black (1934), however,proposedthat the heat of dilution of
H2S04 (120°C) was satisfactoryfor oxidizing 75% of the organicC in soils and
that a correctionfactor could be usedto accountfor incompletedigestion.Sever-
al investigatorshave found that an extendedperiod of heating is required to
obtain quantitative oxidation of soil organic C by chromic acid (Anne, 1945;
Tinsley, 1950; Mebius, 1960; Kalembasa& Jenkinson,1973; Heanes,1984).
High digestiontemperatures(>145°C) lead to thermal decompositionof dichro-
mateand resultanthigh blank values(Tinsley, 1950; Metsonet aI., 1979;Heanes,
1984). Clay mineralshave beenreportedto catalyzethe thermal decomposition
of Cr20,y-(Walkley, 1947) buta recentstudy has shown little thermaldecompo-
sition when high clay soils free of organic matter were heatedwith K2Cr20T
H2S04 for 60 min at 125 or 145°C(Heanes,1984). Digestiontemperatureis nor-
mally regulatedby the ratio of water/H2S04in the mixture (Table 34-3) and the
temperaturerisesaswatervapor is lost during heating.Tinsley proposedthat cold
finger condensersfitted on Erlenmeyerflasks be used to prevent loss of water
during digestion,whereasother investigatorshave usedErlenmeyerflasks fitted
with Liebig condensers.Heating times employedin reflux methodshave varied
from 20 min to 2 h. Ciavattaet ai. (1989) have recently proposedthat a chromic
acid mixture successfullyoxidizes soil organic C when samplesare heatedin a
200 mL narrow-neckeddigestionflask by direct flame at 160°Cfor 10 min. The
neck of the flask servesto reflux the digestionreagents.
Nelson and Sommers(1975) proposedthat organicC could be determined
by heating soil-chromicacid mixtures under reflux in 50 mL Folin-Wu nonpro-
tein nitrogentubesplacedin an aluminumblock on ahot plate. Heatingtime and
digestiontemperaturerecommendedwere 30 min and 150°C, respectively.Sub-
sequently,a numberof other similar tube digestionmethodshave beenproposed
for estimationof organic C all employing 100-mLtubes. Heanes(1984), Yeo-

mansand Bremner(1988), and Soon and Abboud (1991) recommendedheating
for 30 min at 135°C, 170°C, and 155°C, respectively.Yeomansand Bremner
(1988) and Soonand Abboud (1991) specify the samedigestionreagentsas Nel-
son and Sommers(1975), i.e., 5 mL of 0.167M K2Cr207 and 7.5 mL of concen-
trated H2S04, whereasHeanes(1984) recommends10 mL of 0.167 M K2Cr207
and 20 mL of H2S04, Tube digestion procedureshave been reported to yield
organic C values that approximatethose from dry and wet combustiontech-
niques.
Diphenylaminewas the first oxidation-reductionindicator used for the
titration of excessCr20j- with Fe2+ (Schollenberger,1927, 1931, 1945; Allison,
1935). Later studies suggestedthat the diphenylamine end point could be
improved by addition of H3P04, NaF, or HF before titration (Schollenberger,
1931, 1945; Walkley & Black, 1934), and thesesubstanceswere widely usedin
dichromatetitrations. Peechet al. (1947) establishedthat barium diphenylamine
sulfonate(diphenyl-4-sulfonicacid) in combinationwith H3P04 was as effective
and more stablecomparedwith diphenylamine(C12H ll N) and has beenusedas
an indicator in other procedures(Tinsley, 1950). Jackson(1958) recommended
that o-phenanthroline(C12HgN2) be used as an indicator in Cr20j- titrations
becausethe color change(formation of the complexwith Fe2+) occursat higher
oxidation-reductionpotential comparedwith diphenylamine.A mixture of 0-
phenanthrolineand H3P04 is normally usedto give a good end point; however,
the indicator hasbeensuccessfullyusedwithout H3P04 addition. A problemwith
o-phenanthrolineis that the indicator tends to be absorbedby some suspended
soil materials,therebyobscuringthe color changeat the end point. therefore,the
diluted chromic acid-soil mixture is often passedthroughan acid fast filter paper
on Buchnerfunnel before titration. Simakov (1957) proposedthat N-phenylan-
thranilic acid (C13HnHO)be usedas an indicator in Cr20j- titrations with Fe2+.
Mebius (1960) confirmed that N-phenylanthranilicacid gives a very sharp and
cleanend point and this compoundis currently the indicatorof choicefor Cr20j-
titrations.
Other methods of titration not involving oxidation-reductionindicators
havebeen usedto estimatereactedCr20j-. Oneapproachis to add a slight excess
of Fe2+ to the Cr20j--H2S04-Soilmixture and then back-titrate the Fe2+ with
KMn04 (Smith & Weldon, 1941).In this titration procedure,the only reagentthat
requiresstandardizationis KMn04 if the sameamountsof Fe2+ and Cr20j- are
addedto both samplesand blanks. The end point in the titration of Cr20j- with
Fe2+ also may be estimatedvery accuratelyby monitoring the oxidation-reduc-
tion potential with platinum and calomel electrodesattachedto a potentiometer
(Raveh& Avnimelech, 1973). The end point of the titration involves a potential
changeof -400 m V with 0.02 mL of titrant.
The amount of Cr20j- remaining after reaction with soil organic matter
also may be estimatedby colorimetry after removal of soil by filtration or cen-
trifugation (Carolan,1948). Perrierand Kellogg (1960) proposedthat any possi-
ble interferenceof cr3+ in Cr20j- determinationbe eliminated by dilution and
subsequentreactionof excessdichromatewith s-diphenylcarbazide (C13H14N40)
to yield a violet colored complex with an absorptionmaxima at 540 nm. Con-
NELSON & SOMMERS
versely, colorimetry has been widely used to determinethe amountsof cr3+

formed from the reactionof Cr20.y-with soil (Wilde, 1942; Graham,1948; Car-
olan, 1948; Datta et aI., 19862; Sinha & Prasad,1970; Sims & Haby, 1971;
DeBolt, 1974; Guptaet aI., 1975; Baker, 1976; Heanes,1984). The greencolor
due to cr3+ is normally quantitatedat wavelengthsof 590 to 625 nm and the
absorbanceis usually relatedto organic matterconcentrationsin soil by a stan-
dard curve preparedfrom sucrose(Graham,1948; Sims & Haby, 1971; DeBolt,
1974; Heanes,1984). Baker (1976) used a probe colorimeterto measurecr3+
absorbancedirectly in the reactionvesselafter centrifugation,therebyavoiding a
transferinto a spectrophotometer cuvette.From a comparisonof the methodsthat
quantitateCr20.y- and thosethat determinecr3+, Metson (1965) concludedthat
measurement of cr3+ is the preferredprocedure.
Dichromatemethodsthat use heat of dilution or minimal heatingdo not
give completeoxidation of organiccompoundsin soil althoughthe most active
forms of organicC are convertedto CO2, Walkley and Black (1934) found that
on the averageabout76% of the organicC in 20 soils was recoveredby the heat
of dilution procedure,and they proposedthat a correctionfactor of 1.32 be used
to accountfor unrecoveredorganicC. However,the actualrecoveriesof organic
C from the soils testedvaried from 60 to 86%. Schollenberger(1945) reported
that the Walkley and Black procedureoxidized an averageof 79% (range
70-86%) of organic C in soils he studied. Allison (1960) reviewed available
informationon the recoveryof organicC in a wide variety of soils by the Walk-
ley and Black procedureand showedthat the averagerecovery with different
groupsof soils varied from 63 to 86% and that the correctionfactor varied from
1.16 to 1.59. Table 34-4 gives dataon the correctionfactor found to be required
for the Walkley and Black procedurein investigationscarriedout during the past
30 yr. Recoveriesof organicC by the Walkley and Black techniquewere highly
variable,andthe correctionfactor appropriatefor individual soils variedfrom 1.0
to 2.86.The averagecorrectionfactor appropriatefor a groupof soils variedfrom
1.03 to 1.41. This data clearly show that Cr20~--H2S04 methodsthat involve
minimal heatinggive variablerecoveryof organicC from soils. An averagecor-
rection factor found for a group of soils may be applicableto the "average"soil
in the group but will give erroneousvaluesfor many soils in the group. There-
fore, proceduressuch as the Walkley and Black should be consideredto give
approximateor semi-quantitativeestimatesof organic C in soil becauseof the
lack of an appropriatecorrectionfactor for eachsoil analyzed.If an experimen-
tally determinedcorrectionfactor is not availablefor a particulargroupsof soils,
the use of 1.3 as the factor appearsmost reasonableover a rangeof soils. Meth-
ods that involve extensiveheating, such as those of Tinsley (1950), Mebius
(1960), Nelson and Sommers(1975), Heanes(1964) or Yeomansand Bremner
(1988) do not requirea correctionfactor becauseall of the organicC in the soil
is oxidizedto CO2. However,methodsthat involve minimal heating(e.g.,Schol-
lenberger,1927; Tyurin, 1931) require a small correctionfactor (e.g., 1.15) to
accountfor unoxidizedorganicC.
The rapid dichromatemethodsare subjectto interferencesby certainsoil
constituentsthat lead to spuriousresultswith somesoils (Walkley, 1947). Chlo-
ride, ferrous iron and higher oxidesof Mn have beenshownto undergooxida-
CARBON AND ORGANIC MAllER 993
Table 34-4. Correction factors for organic C in surface soils not recoveredby the Walkley-Black
method.
Number Organic C recovery,% Average
Origin of soils correction
Reference of samples studies Range Average factor
Tinsley (1950) England 10 77-92 83.6 1.20
Bremner& Jenkinson(1960a) England 15 27-92 84 1.19
Kalembesa& Kenkinson(1973) England& Wales 22 46--80 77 1.30
Orphanos(1973) Cyprus 12 69-79 75' 1.30
Richter et at. (1973) Argentina 12 79-87 83 1.20
Nelson & Sommers(1975) Indiana 10 44-88 79 1.27
Bornemiszaet at. (1979) CostaRica 50 69-81 75 1.33
Rhodeset at. (1981) Sierra Leone 10 93-100 97 1.03
Richardson& Bigler (1982) North Dakota 21 35-91 88 1.33t
1.13*
Heanes(1984) Australia 12 85-98 92 1.09
Amacheret at. (1986) Louisiana 179 46--87 71 1.41
Gillman et at. (1986) Queensland 450 65-95 76§ 1.32
8111 1.24
Willet & Beech(1987) Australia 30 60-144 85 1.18
Lowther et at. (1990) Australia 38 74-102 88 1.14
Soon& Abboud (1991) Alberta 39 62-87 71.4 1.40
t Low C samples.
* Other samples.
§ Soils derivedfrom basalt,alluvium, or beachsand.
'II Soils derivedfrom graniteor metamorphicrocks.
tion-reductionreactionsin chromic acid mixtures leadingto incorrectvaluesfor

organicC. The presenceof significant amountsof Fe2+ or Cl- in soil will lead to
a positive error, whereasreactiveMn02 in soil sampleswill result in a negative
error and low valuesfor organicC.
Chloride interferes with dichromate methods through the formation of
chromyl chloride, as indicated in Eq. [3], which results in consumption of
Cr20?-. Chloride interferencemay be eliminatedby washingthe soil free of Cl-
before analysisor by precipitatingthe Cl- as AgCl by addition of Ag2S04 to the
digestion acid (Walkley, 1947; Quinn & Salomon, 1964; Gupta et aI., 1975).
Alternatively, Walkley (1947) found that Eq. [9] may be used to correct organic
C valuesfor soils having CUC ratio of ~5.1
organicC in soil (%) = (apparent% C in soil) - (% CU12) [9]
It has recently been reportedthat Ag2S04 addition did not eliminate Cl-
interferencein a low temperaturetube digestionmethodand that an assayfor Cl-
coupledwith stoichiometriccorrectionfor chromyl chloride loss is necessaryfor
accurateestimatesof organicC (Heanes,1984).
When presentin soil, Fe2+ will be oxidized to Fe3+ by Cr20?-,as indicated
in Eq. [10], resulting ina positive error in the analysis,i.e., giving high valuesfor
organicC content
[10]
AppreciableFe2+ may be presentin highly reducedsoils, and errors may

resultwhendichromatemethodsare appliedto undriedsamplesof anaerobicsoils
beforedrying (Lee, 1939).However,Walkley (1947) found that thoroughair-dry-
ing of reducedsoils beforeanalysisresultedin oxidationof Fe2+ to Fe3+ and accu-
rate determinationof the organicC present.The amountsof Fe2+ presentin well-
aeratedsoils are so small relative to the amountsof organic C presentthat no
detectableinterferenceis likely. Pyrite also is oxidized during treatmentof soils
with dichromate and samplescontaining pyrite sulfur concentrations>0.29%
result in significant over estimation of organic C by the Walkley and Black
method(Willett & Beech,1987). Metallic iron (FeO) presentin soil samplesalso
may lead to positive interferencesin dichromate methods, (Allison, 1935;
Heanes,1984).Therefore,careshouldbe takento ensurethat soils are not ground
with iron or steel equipmentbefore analysis.The higher oxides of Mn (largely
Mn02) competewith Cr20j- for oxidizable substanceswhen heatedin an acid
medium accordingto Eq. [11].
[11]
Therefore,any reactive Mn02 presentwill give a negativeerror when soils are

analyzedby dichromatetechniques.Although soils contain substantialamounts
of Mn02 and otherhigher oxidesof Mn, Walkley (1947) and Heanes(1984) con-
cludedthat in most soils the quantity of reactive(reducible)oxidesofMn is small
becauseonly the freshly precipitatedMn02 will take part in redox reactions.
Even in highly manganiferoussoils, only a small fraction of the Mn02 presentis
able to competewith Cr20j- for oxidation of organic C compounds.Therefore,
interferencefrom Mn02 is not thought to be a seriouserror in the vast majority
of soils. In soils with large amountsof reactiveMn02' Walkley (1947) suggested
pretreatmentof sampleswith the exactamountof FeS04necessaryto reducethe
amountof reactiveMn2 presentprior to treatmentwith K2Cr207 and H2S04,
Other problemsassociatedwith dichromatemethodsinvolve assumptions
about the averageoxidation stateof organicC in soils (i.e., equivalentweight of
C) and recoveryof highly reducedforms of organicC from soils. All dichromate
methodsassumethat the organic C in soil has an averageoxidation stateof zero
and an equivalentweight of 3 g per equivalentwhen reactedwith dichromate
accordingto Eq. [8] eventhoughno studieshavebeenconductedto evaluatethis
assumption.However, the fact that dichromatemethodsusing extensiveheating
give organicC valuessimilar to thoseobtainedwith wet or dry combustionwhere
CO2 is determineddirectly suggeststhat this assumptionis reasonablycorrect.
Dichromate methodsthat involve little or no external heating give very
poor recovery of organic C present in carbonized materials (e.g., charcoal,
graphite,coal, coke,andsoot). For example,Walkley (1947) found that the Walk-
ley and Black methodrecoveredonly 2 to 11% of the organic C presentin such
materials. In a detailed study, Bremner and Jenkinson(1960b) found that the
Walkley and Black methodgave low recovery (0-57%) of organic C from car-
bonized materials, whereasmethodsinvolving external heat such as those of
Tinsley gavesubstantial(64-104%)andvariablerecoveryor organicC from such
materials.Conversely,Heanes(1984) found that very little organicC in charcoal
or coke was oxidized by a tube digestionprocedureat 135°C.Otherinvestigators

have found that the Walkley and Black procedurecompletely recovers C in
weatheredcoal seams,i.e., coal "blooms" (Kalisz & Sainju, 1991).Thesecon-
flicting results suggestthat recovery of organic C from carbonizedmaterialsis
highly dependentupon the characteristicsof the materialsand digestioncondi-
tions (i.e., temperature,reagentconcentrations).It is appropriateto concludethat
dichromatemethodscannotbe usedto quantitativelyrecovercarbonizedmateri-
als from soils or to discriminatebetweenC in carbonizedmaterialsand C in soil
organicmatterbecauseorganic C recoveryvarieswith type of carbonizedmate-
rial and time and temperatureof heatingof the chromic acid mixture. Therefore,
unreliable results for organic C will be obtained if dichromate methods are
appliedto soils containingsignificant amountsof carbonizedmaterials.Dry com-
bustion methodsare most appropriatefor soils containinglarge amountsof ele-
mentalC.
Walkley-Black Method
The Walkley-Black Method was describedby Walkley (1946), Peechet al.
(1947) and Grewelingand Peech(1960).
Reagents
1. Potassiumdichromate,0.167 M (1 N): Dissolve 49.04 g of reagent-
gradeK2Cr207 (dried at 105°C) in water, and dilute the resclutionto a
volume of 1000 mL.
2. Sulfuric acid, concentrated(not lessthan96%): If Cl- is presentin soil,
add AgzS04 to the acid at the rate of 15 g per liter.
3. Phosphoricacid, concentrated.
4. o-Phenanthroline-ferrous complex, 0.025 M: Dissolve 14.85 g of 0-
phenanthrolinemonohydrateand 6.95 g of ferroussulfateheptahydrate
(FeS04·7H20) in water. Dilute the solution to a volume of 1000 mL.
The o-phenanthroline-ferrous complex is availableunder the nameof
Ferroin from the G. FrederickSmith ChemicalCo. (Columbus,OH).
5. Barium diphenylaminesulfonate:Preparea 0.16% aqueoussolution.
This reagentis an optional substitutefor no. 4.
6. Ferroussulfate heptahydrate(FeS04• 7HzO) solution, 0.5 M (0.5 N):
Dissolve 140 g of reagent-gradeFeS04• 7HzO in water, add 15 mL of
concentratedsulfuric acid, cool the solution, and dilute it to a volume
of 1000 mL> Standardizethis reagentdaily by titrating it against10
mL of 0.167M (1 N) potassiumdichromate,as describedbelow.
Procedure
Grind the soil to passthrough a 0.5-mm sieve, avoiding iron or steel mor-
tars. Transfera weighedsample,containing10 to 25 mg of organicC, but not in
excessof 10 g of soil, into a 500-mL wide-mouthErlenmeyerflask. Add 10 mL
of 0.167 M (1 N) KZCrZ07, and swirl the flask gently to dispersethe soil in the
solution. Then rapidly add 20 mL to concentratedHZS04, directing the stream
into the suspension.Immediatelyswirl the flask gently until soil and reagentsare
mixed, then more vigorously for a total of 1 min. Allow the flask to standon an
NELSON & SOMMERS
insulatedsheetfor about30 min. Thenadd200 mL of waterto the flask, and fIl-

ter the suspensionusing an acid resistantfIlter paper(e.g., Whatman540), if
experienceshowsthat the end point of the titration cannototherwisebe clearly
discerned.Add three to four drops of o-phenanthrolineindicator and titrate the
solution with 0.5 M (0.5 N) FeS04.As the end point is approached,the solution
takeson a greenishcastand then changesto a dark green.At this point, add the
ferrous sulfate heptahydratedrop by drop until the color changessharply from
blue to red (marooncolor in reflectedlight againsta white background).Make a
blank determinationin the same manner, but without soil, to standardizethe
K2Cr207. Repeatthe determinationwith less soil if >75% of the dichromateis
reduced.
Calculatethe resultsaccordingto the following formula, usinga correction
factor ''/' =1.30 or a more suitablevalue found experimentally
2
OrganicC, % = .-.:. (m_4.. . ; .; : 18=nk,--_m_Ls.. ; ; 8=m=ple: .:. .). .;.(M.. . . ;+.. ;:F. e_. )..:....(0_.0_0-'3)'-('-lO_0....:.,.) x f [12]
wt. • water-freesoil, g
Comments
The coefficient of variation for the Walkley-Black procedurehas been
reportedto vary between1.6 and 4.2% (Table 34-3). Ferrousammoniumsulfate
also is a suitabletitrant for excessCr20?-in conjunctionwith the Walkley-Black
method.The Smith and Weldon (1941) modification involving completereduc-
tion of Cr20:r- with Fe2+, and subsequentback-titration of excessFe2+ with
Mn04" solutionalso may be usedto estimateunreactedCr20:r-. Otheroxidation-
reduction indicators that have provided satisfactory results include barium
diphenylaminesulfonateand N-phenylanthranilicacid. The amountsof Cr20?-
reducedto Cr3+ by reactionwith soil organicmatteralso may be estimatedcol-
orimetrically or by potentiometrictitration with a ferrousammoniumsulfatesolu-
tion. Grinding samplesto <0.2 mm hasbeenshownto reducesamplingerrorsand
the coefficientof variationevenwhen relatively largesamplesizes(1 g) are used
(Metsonet aI., 1979).Heanes(1984) reportedthat reductionin particlesizefrom
0.5 to 0.15 mm significantly increasedrecoveryof organicC in 12 soils.
Thbe DigestionMethod
Special Apparatus
1. Pyrex digestiontubes(lOO mL) sizedfor block digestor.
2. Block digestor:40-tubeKjeldahl block digestorsuppliedby Technicon
InstrumentsCorp., Tarrytown, NY, or TecatorInc., Herndon,VA, or
equivalent.
Reagents
1. Potassiumdichromatesolution, 0.167M (1.0 N)-dissolve49.025 of
K2Cr207 (dried at 140°C) iIi 800 mL of distilled water and dilute the
solutionwith waterto a volumeof lOoo mL in a volumetricflask. This
is the primary standardfor the procedure.
2. Concentratedsulfuric acid-specificgravity 1.84.

3. Ferrousammoniumsulfate solution 0.2 M (0.2 N)-Dissolve 156.8 g
of ferrous ammoniumsulfate [Fe(NH4)z(S04)2• 6H20] in 100 mL of
concentratedsulfuric acid and dilute the solution with water to a vol-
ume of 2 L in a volumetric flask. This solution must be standardized
daily becauseit undergoesslow oxidation.
4. Indicatorsolution-Dissolve0.1 g of N-phenylanthranilicacid and 0.1
g of Na2C03in 100 mL of distilled water.
Procedure
Weigh an amountof soil air dried and ground to <0.15 mm containingnot
greaterthan 8 mg of organicC (usually 100--500mg) into a clean, dry digestion
tube and add 5 mL of 0.167M (1.0 N) K2Cr207 solution and 7.5 mL of concen-
trated H2S04. Placethe tube in the digestionblock preheatedto 150°C for exact-
ly 30 min. Removethe digestion tube from the block and allow the samplesto
cool for 30 min at room temperature.Quantitatively transferthe contentsof the
tube to a 125-mL Erlenmeyerflask and titrate the samplewith 0.2 M (0.2 N) fer-
rous ammoniumsulfate solution using 0.2 mL of the N-phenylanthranillicacid
solution as the indicator.The color change atthe end point is from violet to bright
greenand is very rapid. An illuminated backgroundis recommendedfor easein
observing the end point and the titration should be performed using a 25-mL
burettecalibratedat O.l-mL intervals and a variable speedmagneticstirrer and
teflon coatedstiffing bar.
Each set of soil samplesshould be analyzedwith two unheatedreagent
blanks and two reagentblanks that are heatedat the sametime as the samples.
The unheatedblanks are usedto standardizethe ferrous ammoniumsulfate solu-
tion. The difference in titration values betweenheatedand unheatedblanks is
usedto correctall sampletitration valuesfor the amountof dichromateconsumed
by thermal decompositionduring the heatingprocess.
Computationof the organic C content of soil is performedas follows: (i)
subtractsample titration values (mLsoi/) from the averagetitration value of the
heated(boiled) blank (mL bb ), (ii) correct the resulting [mLbb - mL,.oil] value for
thermal decompositionof dichromateby dividing the differencein averagetitra-
tion value for unheatedand heatedblanks by the averagetitration value for the
unheatedblank, multiplying the correction factor (normally 0.04-0.08) by the
[mLt,b - mL,.oil] value, and adding the product to the [mLt,b - mLsoi1] value (Eq.
[13]). The resulting value,labeled "A" is proportionalto the amountof organicC
presentin the soil, (iii) completethe calculationof organic C content using Eq.
[14]
where ub is unboiledblank and bb is boiled blank.
(,A....:.-)-,-(M---,F=e2~+)-'..(0_.0_O--,3)-O.(1_0---"-.0)
OrganicC, % = - [14]
wt. water-freesoil, g
Comments
The coefficientof variation for the method hasbeenreportedas 3.5% (Nel-
son& Sommers,1975).Coefficientsof variation reportedfor othertube digestion
methods have ranged from 1.1 to 4.4% (Heanes,1984; Yeomans & Bremner,
1988; Soon & Abboud, 1991).The precisionof the methodcan be improved by
using a computer-aidedautomatictitration system(Yeomans& Bremner,1988).
Colorimetric analysisof cr3+ also can be usedto estimatethe amountof dichro-
mate that has reactedwith organicC during tube digestion(Heanes,1984; Soon
& Abboud, 1991).The potassiumdichromatesolution is the primary standardfor
the methodand careshouldbe taken in its preparation.This solution is quite sta-
ble and may be storedat room temperatureindefinitely. The ferrous ammonium
sulfate solution oxidizes slowly and thus must be standardizedeach time it is
used. Small particle size reducesthe sampling error and increasesrecovery of
organic C. Heanes(1984) found that organicC valuesincreasedby about 2% as
particle size was reducedfrom 0.5 to 0.15 mm.
Thermal decompositionof dichromateoccurs at temperaturesexceeding
136°C (Heanes,1984) and the degreeof decompositionis quite dependentupon
the heatingconditions.Therefore,it is recommendedthat the digestion tubesby
dry before use to eliminate differencesin acid/waterratio and that the heating
temperatureand time be accuratelycontrolled.A variety of temperaturesvarying
from 135 to 170°C have been recommendedfor tube digestion methods(Table
34-3). When thermal decompositionof dichromate is accurately taken into
accountwith a heatedblank, the four tube digestionmethodshave quantitatively
determinedorganicC in a variety of soils.
Interferencespresentin the Walkley-Black procedurealso are a problem
with tube digestionmethods.As a result of extensiveheating,the tube digestion
methodsgive completerecoveryof organicC from soils and, thus, do not require
a factor to accountfor incompleteoxidation of organic matter. Heanes(1984)
reported that little organic C in charcoal and coke was recoveredby the tube
digestionprocedurethat he described.
The tube digestion techniquecan be used to estimateorganic C in soil
extractsby carrying out the digestionwith 1 or 2 mL of extractand 4 or 3 mL of
dichromatesolution, respectively.It is essentialthat the acid/waterratio be main-
tained at 1.5 in the digestso the volume of dichromatesolution must be reduced
as the volume of extractis increased.Both heatedand unheatedblanksshouldbe
preparedusing the samevolume of blank extractingsolution and the dichromate
solution as that employedfor the extracts.
The modified Mebius methoddescribedby Nelsonand Sommers(1982) is
recommendedas an accurateand precise dichromateoxidation procedurefor
thoseinvestigators nothaving accessto a block digestor.The major advantageof
the tube digestionprocedureis the decreasedanalysistime per samplebecauseof
the relatively large numberof samples(40) that can be heatedat one time.
Comparisonof Methodsfor DeterminingOrganicCarbon
Most studieshave shown that very good agreementis obtainedwhen wet

combustion,dry combustion,and Van Slyke-Folch (1940) methodsare used to
determineorganic C in soils (Bremner& Jenkinson,1960a; Kalembasa& Jenk-

inson, 1973; Nelson& Sommers,1975). For this reason,wet and dry combustion
methodsare normally consideredto yield absolutevaluesfor organic C in soils
and other methodsare calibratedagainstthe combustionprocedures.
A numberof studieshavebeenconductedto comparerapid dichromateoxi-
dation methodswith dry or wet combustionmethods(Table 34-5). Many studies
haveshownthat the Walkley-Black methodyields variable recoveryof organicC
from soil, i.e., in somesoils >95% of organicC may be oxidized but in othersoils
<60% of the organic C is convertedto CO2• For example,Bremnerand Jenkin-
son (1960a)showedthat for 15 soils the recoveryof organic C by the Walkley-
Black method(using a correctionfactor of 1.3) varied from 73 to 119% of wet
combustionvalues.Kalembasaand Jenkinson(1973) observedthat with 22 soils
the recoveryof organic C by the Walkley-Black procedureswith correctionvar-
ied from 60 to 122% of wet combustionvaluesalthoughthe averagerecoveryby
the Walkley-Black method was 102%. Nelson and Sommers(1975) found that
the Walkley-Blackmethodwith correctiongave organicC valuesfor 10 soils that
varied from 57 to 114% (averageof 102%) of thoseobtainedwith wet combus-
tion. More recent studieshave found that organic C in soils from SierraLeone
and Australia is much more susceptibleto oxidation by dichromatein the Walk-
ley-Black procedurethan was expected,e.g., averageuncorrectedrecoveriesof
organicC varied from 88 to 97% of wet or dry combustion(Rhodeset ai., 1981;
Heanes,1984; Lowther et ai., 1990). The Walkley-Black method employing a
correctionfactor for unoxidizedorganic C is not highly accuratefor an individ-
ual soil, but for a group of soils the averagerecoveryis goodwhen comparedwith
organicC valuesdeterminedby dry or wet combustion.This is particularly true
when the appropriatecorrectionfactor hasbeendeterminedfor the group of soils
understudy. Furthermore,the simplicity and rapidity of the Walkley-Black meth-
od, in part, compensatefor the lack of accuracyinherentin the procedure.
Dichromatemethodsthat involve limited periodsof heating(Schollenberg-
er, 1927;Tyurin, 1931) havebeenshownto yield good recoveriesof organicC if
appropriatecorrectionfactors are applied(Allison, 1935; Crowther, 1935; Smith
& Weldon, 1941; Kalembasa& Jenkinson,1973). The usual correction factors
are 1.15 and 1.08 for the Schollenbergerand Tyurin methods,respectively.Cia-
vatta et al. (1989) found that no correction factor was neededwhen a modified
Schollenbergerprocedurewas usedwith heatingat 160°C for 10 min.
Modem dichromateoxidation methodsthat involve an extendedperiod of
heating,often underreflux, (e.g., Anne, 1945; Tinsley, 1950; Mebius, 1960; Nel-
son & Sommers,1975; Heanes,1984; Yeomans& Bremner,1988) give organic
C values equivalentto those obtainedby dry or wet combustion(Table 34-5).
Bremnerand Jenkinson(1960a) found that the Tinsley method gave organic C
valuesof 15 soils that varied from 88 to 106% (avg. 101%) of wet combustion
values. Kalembasaand Jenkinson(1973) showed that the Tinsley and Mebius
methodsyielded organic C recoveriesfrom 22 soils that averaged95 and 94%,
respectively,of wet and dry combustionvalues. Nelson and Sommers(1975)
reportedthat the Mebius procedurerecoveredfrom 92 to 110% (avg. 103%) of
the organic C found in 10 soils by the wet combustionmethod.Tube digestion
methodshavegiven excellentaveragerecoveriesof organicC in soils [Nelson &
~
Table 34-5. Comparisonof methodsfor determiningorganicC in soils.
No. of Dry Wet Walkley & Modified Tube
Reference samples combustion combustion Schollenbergert Black:j: Tinsley II§ Mebius digestion Tyurin.
- Avg. mg C/kg soil - Avg. % of wet or dry combustion

Allison (1935) 71 12.5 100
Crowther(1935) 8 37.6 100 100 100
Smith & Weldon (1941) 69 16.5 96 101
Bremner& Jenkinson(1960a) 15 114.9 109 94
Nelson & Sommers(1975) 10 35.0 102 103 99
Kalembasa& Jenkinson(1973) 22 25.6 25.6 104 95 94 105
Metsonet aJ. (1979) 7 238.7 235.9 109
Rhodeset aJ. (1981) 10 26.0 125
Heanes(1984) 12 19.6 120 100
Yeomans& Bremner(1988) 15 40.4 98 99
Ciavattaet aJ. (1989) 2 21.8 99# 92 84
Lowther et aJ. (1990) 38 12.4 115 99
Soon& Abboud (1991) 12 21.1 95tt 96:j::j: 97
t Valuescorrectedfor incompleteoxidation of organicC using a factor of 1.15.
:j: Valuescorrectedfor incompleteoxidation of organicC using a factor of 1.30.
§ Reagentconcentrationsas describedby Bremnerand Jenkinson(1960a).
'V Values correctedto incompleteoxidation of organicC with a factor of 1.08.
z
# Method was similar to Schollenbergerexceptdigestionreagentconcentrationsdiffered and sampleswere heatedfor 10 min.
tt Digestion carried out in 100 mL Pyrex tube. ~
Z
:j::j: Digestion carried out in 100 mL Pyrex tubesin heatingblock.
Ro
CIl
o
::
~
Sommers(1975),99%;Heanes(1984),100%,Yeomans& Bremner(1988),99%;
Soon & Abboud (1991), 97%].
ORGANIC MATTER
Introduction
The organic matter content influencesmany soil properties,including (i)

the capacityof a soil to supply N, P, and S and trace metalsto plans; (ii) infiltra-
tion and retentionof water, (iii) degreeof aggregationand overall structurethat
affect air and water relationships;(iv) cation exchangecapacity; (v) soil color,
which in turn affects temperaturerelationships;and (vi) adsorptionor deactiva-
tion (or both) of agricultural chemicals.Determinationof organicmattercontent
is a routine procedurecarriedout in soil analysisand testinglaboratoriesthrough-
out the world becauseof the importanceof organic matter in supplying plant
available N and deactivating pesticides.However, no completely satisfactory
method exists for determinationof the matter content of soils. The ignition
method describedbelow is a modification of that presentedby Ben-Dor and
Banin (1959).This methodprovidesa reasonableestimateof organicmattercon-
centrationsin soils but cannotbe consideredquantitative.
Calculationof OrganicMatter Content
The organicmattercontentof soil may be indirectly estimatedthroughmul-

tiplication of the organicC concentration(as determinedby proceduresoutlined
in "OrganicCarbon")by the ratio of organicmatterto organicC commonlyfound
in soils. The organic matter content is normally arrived at by multiplying the
organicC concentrationby 1.724. However, a numberof studieshavesuggested
that this factor is too low for many soils, and consequentlythe organic matter
contentis underestimated.For example,Robinsonet al. (1929) and Lunt (1931)
showedthat the correctfactor for peatsis 1.86 to 1.89. In summarizingmuch of
the early work, Broadbent(1953) concludedthat conversionfactors of 1.9 and
2.5 would be appropriatefor surfacesoils and subsoils,respectively.Otherwork-
ers have found that factors of 1.9 to 2.0 were satisfactoryfor surfacelayers of
mineral soils (De Leenheeret aI., 1957; Howard, 1965; Ponomareva& Platniko-
va, 1967; Christensen& Malmros, 1982). Loftus (1966) reportedthat the appro-
priate factors for mineral and organic soils in Pennsylvaniawere 2.2 and 1.8,
respectively,whereas,Ranney (1969) found that the organic matter content of
Pennsylvaniasurfacesoils may be accuratelyestimatedby the equation
organicmatter, % = .35 + 1.80 x % organicC [15]
It is evidentthat estimationof organicmattercontentfrom organicC con-

centrationsis not highly accurate,becausethe organicC contentof organicmat-
ter is variable from soil to soil and with depth in the profile. Accurate organic
mattercontentestimatesrequire a knowledgeof the factor for the particularsoil
studied.However,if an estimateof organicmattercontentof surfacesoils must

be basedon organicC dataandno informationon the exactfactor is available,a
factor of two appearsto be more universallyacceptable.
Direct Estimationof OrganicMatter

Principles
To achievea direct determinationof soil organicmatter,one must separate
it from the inorganicmaterial,which in mostsoils makesup 90% or more of the
weight of the soil. Extractionproceduresthat bring part of the organicmatterinto
solution while leaving the inorganicmaterialundissolvedhave only qualitative
value,sirice no solventhasbeenfound that will dissolveall or evena major por-
tion of the organicfraction. The alternativeis to destroythe organicmatter,after
which the loss in weight of the soil is takenas a measureof the organiccontent.
The requirementsof a suitablemethodare that the treatmentusedto destroythe
organicmattershouldnot destroyor alterthe othersoil constituentsin sucha way
that their weight is changedand that the organicmattershouldbe quantitatively
removed.
The two mostcommonlyusedmethodsfor achievingdestructionof organ-
ic matterare: (i) oxidationof the organicmatterwith H20 2 and(ii) ignition of the
soil at high temperature.The H20 2 method(Robinson,1927) hasseriouslimita-
tions in that the oxidationof organicmatterby this reagentis incomplete,andthe
extent of oxidation varies from one soil to another.This method is therefore
unsatisfactoryasa meansof determiningtotal organicmatterof soil, but it canbe
useful as a meansof comparingthe readily oxidizablematerialin different soils.
The loss-on-ignition(LOI) methodcarriedout at high temperaturegives quanti-
tative oxidation of organicmatter,but inorganicconstituentsof the soil, chiefly
the hydratedaluminosilicates,lose structuralwater and carbonatemineralsand
somehydratedsaltsare decomposedupon heating.Dehydroxylationanddecom-
positionof inorganicconstituentsby heatingresultsin weight lossesconsiderably
in excessof the actualorganicmattercontent.This problem is particularly pro-
nouncedwith high clay soils containinglow amountsof organicmattersuchas
subsoils(Christensen& Malmros, 1982;Howard & Howard, 1990).
Studieshaveshown,however,thattemperatures exceeding750°Careneed-
ed to decomposecarbonatesand that little dehydroxylationof phyllosilicates
occursat temperatures below 450°C(Ball, 1964;Ben-Dor& Banin, 1989).Gibb-
site is an exceptionbecausethis clay mineral hasbeenreportedto lose structural
waterwhen heatedat 300 to 350°C (Ranney,1969; Gallardoet aI., 1987). Some
investigatorshave ignited soils and attemptedto correct LOI valuesfor dehy-
droxylation by multiplying a weight loss factor by the clay contentof samples
(Howard, 1966; Ranney,1969; Spain et al., 1982). Ranney(1969) also useda
low-temperature(100--200°C) ashing procedure(organic matter was oxidized
underreducedpressureby activatedO2 excitedby a radio frequencyelectromag-
netic field) for LOI estimationof organic matter content. He found that some
dehydroxylation(1.5% of soil weight) occurredwhen subsoilswere subjectedto
low-temperatureashingandthat the procedurerequired5 d for completeremoval
of organicmatter.
The need in soil characterizationand testing laboratoriesfor a rapid and

semiquantitativetechniquefor routine estimationof organic matter content has
resultedin developmentof a numberof LOI method(Table 34-6). Environmen-
tal hazardsassociatedwith useand disposalof Cr has accelerated interestin alter-
natives to dichromate oxidation methods for estimation of organic matter
(Schulte et aI., 1991). Most investigatorshave attemptedto obtain complete
removal of organic matter without dehydroxylatingaluminosilicatesor decom-
posingcarbonatesby heatingsamplesat or below 450°C for extendedperiodsof
time. Excellentcorrelationshave beenobtainedbetweenLOI valuesand organic
matter contentcalculatedfrom organic C data (Table 34-6). In Table 34-6, val-
ues of regressioncoefficient "b" below one suggestthat someconstituentother
than organicmatterwas lost duringheating,whereascoefficientsgreaterthan one
indicatethat incompleteremovalof organicmatterhasoccurred.In general,low
temperature(36(}-375°C) proceduresparticularly thosewith short heatingtimes
had coefficients greaterthan one whereashigh temperature(>5OO°C) procedures
much less than one. Thesefindings suggestthat ignition of soils at 400 to 450°C
will removeall organic matter and causeminimal dehydroxylationof clay min-
erals.A heatingtime of 8 to 16 h at 400°C resultsin nearmaximum weight loss
(Ben-Dor & Banin, 1989).
Reflectanceand absorptionspectroscopyalso have beenused to estimate
the organic matter contentof soils. AI-Abbas et al. (1972) showedthat relative
reflectanceat 0.72 to O.80llm was relatedto the organicmattercontentof Indiana
soils. A curvilinearrelationshipwasobtainedbetweenreflectanceas measured by
Table 3~. Relationshipbetweensoil organic mattercontentand weight loss-on-ignition(LOI).

Ignition conditions Regressioncoefficientst
Number
Reference of soils Temperature Time b a ,.z
°C h
Ball (1964) 65 375 16 0.916 -0.8
Howard (1965) 36 550 To constant 0.94 -0.96 0.99
weight
Ramney(1969) 48 375 28 1.11 +0.35 0.99
Davies(1974) 17 430 24 0.983 -0.64 0.99
Christensen& 85 550 4 0.985 -0.23 0.99
Malmros (1982)
Spainet al. (1982) 766 950 0.5 0.796 -0.60
Storer(1984) 215 500 4 0.937 -1.70 0.96
Goldin (1987) 60 600 6 0.810 -1.42 0.86
David (1988) 174 450 12 1.04 -0.03 0.92
Ben-Dor & 91 400 8 0.972 -0.37 0.97
Banin (1989)
Howard & 564 550 3 0.840 -1.68 0.98
Howard (1990)
Lowther et al. 38 450 16 0.914 0 0.99
(1990)
Donkin (1991) 45 450 6 0.568 0 0.98
Schulteet al. (1991) 316 360 2 1.126 -0.38 0.90
t Regressionmodel usedwas: soil organicmatter= (b • LOI) + a, where units are g/100 of soil. Soil
organic mattercontentwas estimatedas two times organicC concentration.Organic C was deter-
mined by Walkley & Black, Tinsley, or dry combustionmethods.calculationsassumethat organic
matterwas 50% C.
a color differencemeterand organicmattercontent(Page,1974). Krishnanet al.

(1980) found thatthe organicmattercontentof Illinois soils was highly correlat-
ed with reflectanceat wavelengthsof 0.624 and 0.564 um. Near infrared diffuse
reflectanceat wavelengthsof 1744,1870and 2052 nm hasbeenrelatedto organ-
ic mattercontentof Australiansoils (Dalal & Henry, 1986). Several investigators
have proposedthat the organic matter content of soils be estimatedby partial
extraction with alkaline reagentsand determinationof humic materials in the
extractby absorptionspectrometryat 550 to 650 nm (Mehlich, 1984; Streket aI.,
1990; Bowman et aI., 1991). Moore (1985) used absorbanceat 330 nm to esti-
matedissolvedorganicmatter in peatwater samples.Although eachof the tech-
niquesdescribedabovehas promise as methodfor determiningorganic matter
content,nonecan be recommendedat this time becauseof a lack of evaluationof
their usefulnessand applicability to a wide rangeof soils.
Loss-On-IgnitionMethod
The Loss-On-IgnitionMethod is a modification of a methoddescribedby
Ben-Dor and Banin (1989).
Special Apparatus
1. Pyrex beakersor porcelaincrucibles(20 mL).
2. Muffle furnacecapableof :!:5°C temperaturecontrol.
3. Drying oven (105C) with :!:5°C temperaturecontrol.
4. Analytical balancecapableof weighing :!:0.1 mg.
Procedure
Heat beakersor crucibles in muffle furnace at 400°C for 2 h, cool, and
determinetare weight to 0.1 mg. Add 1 to 3 g of air-driedsoil groundto <0.4 mrn
to a taredbeakerand heatat 105°Cfor 24 h. Cool the beakerin a dessicatorover
CaCl2 and determineweight of beakerplus sampleto 0.1 mg. Obtain weight of
oven-driedsampleby subtraction.Ignite samplesin a muffle furnaceat 400°Cfor
16 h. Cool beakersin a desiccatorover CaCl2 and determineweight of beaker
plus ignited sampleto 0.1 mg. Calculateweight of ignited sampleby subtraction.
The LOI contentof the sampleis calculatedas
Weight105 - Weight400
LOI, % = x 100 [16]
Weight105
where"weight 105" is weight of soil sampleafter heatingat 105°Cand"weight4oo"

is weight of soil sampleafter ignition at 400°C. The organic matter content is
assumedto equal the LOI in most surfacesoils. The LOI can be correctedfor
dehydroxylationof inorganicconstituentsthrough regression analysis.Determine
the LOI and the organicC contentof representativesampleshaving organicmat-
ter levels coveringthe rangeexpectedin soils understudy. Regressorganicmat-
ter content [organic C x 2 (or other suitable correctionfactor)] on LOI and use
the resulting relationshipto convert LOI of test samplesto organic matter. In
many casesthe interceptof the regressionline (LOI on Yaxis and organicmatter
onX axis) will be greaterthan zero indicatingweight loss due to dehydroxylation

of clay mineralsduring heating.
Comments
Control of the heatingof samplesat 105°Cis as importantas at 400°C.The
useof an automaticbalanceconnectedto a laboratorycomputeras an automated
data acquisitionsystemwill greatly speedup analysesand tend to improve the
precisionof the method(Storer,1984).The coefficientof variationfor LOI meth-
odsappliedto soils containing>1% organicmatterhasbeenestimatedto be about
3.3% (Storer, 1984),5.0%(David, 1988),2.0%(Lowther et aI., 1990), and 3.5%
(Donkin, 1991). The coefficient of variation tendsto decreaseas the samplesize
increasesbecauserelative weighing errors becomeless(Lowther et aI., 1990).
A more accurateestimation of organic matter content of soils may be
obtainedby using the Rather(1917) methoddescribedby Nelson and Sommers
(1982). This methodinvolves pretreatingthe samplewith a mixture of HCI and
HF to removehydratedmineral matterand carbonatesprior to ignition. This pre-
treatmentdissolvespart of the organicmatterso that a correctionfor the soluble
material is necessary.Both weight loss and CO2 evolution are measuredduring
ignition so that the C contentof organicmattercan be calculated.The methodhas
a coefficient of variation of about 2%. The Rather method is undoubtedlythe
most accurateavailable for determinationof organic matter since it does not
involve the use of an arbitrary conversion factor. The tedious nature of the
methodcoupledwith the requirementfor pretreatmentwith HF limits its useful-
nessfor routine organicmatterdeterminations.
Expressionof Soil OrganicMatter Content
Due to the difficulty in directly estimatingor calculating the amount of

organicmatterpresentin soil, it appearsthat a more appropriateprocedurewould
be to determineand expressthe soil organic C contentas a measureof organic
matter. Organic C concentrationsin soil may be accuratelyand precisely mea-
sured by a variety of procedures,whereasorganic matter content may be only
estimated.Therewould be little confusionin the amountsof organicmatter. Fur-
thermore,the analystwould be in a position to convertorganicC data to organic
mattercontentsusingthe correctionfactor deemedmost appropriate.On the other
hand,when organicmatterconcentrationsare given, they are extremelydifficult
to convert to organicC valuesunlessthe correctionfactor originally usedalso is
supplied.At the currentpoint in the developmentof soil science,a uniform sys-
tem for expressionof the amountsof soil constituentsis necessary.Therefore,
organic C concentrationis preferableto the term soil organic matter content,
becausethe latter is not an appropriateor an accuratelymeasurableentity.
ACKNOWLEDGMENTS
A joint contribution of the University of NebraskaAgricultural Research

Division, JournalSeriesno. 10742,Lincoln, NE 68583-0704;and The Colorado
Agricultural Experiment Station, Colorado State University,Fort Collins, CO

80523.Mention of a trademarkor productdoesnot constitutea guaranteeor war-
ranty of the productby the University of Nebraskaor ColoradoStateUniversity,
nor doesit imply its approvalto the exclusionof other suitableproducts.
REFERENCES
AI-Abbas, AH., P.I. Swain, and M.F. Baumgardner.1972. Relatingorganic matter and clay content
to the multispectralradianceof soils. Soil Sci. 114:477-485.
Allison, L.E. 1935. Organicsoil carbonby reductionof Cr03' Soil Sci. 40:311-320.
Allison, L.E. 1960.Wet-combustionapparatusand procedurefor organicand inorganiccarbonin soil.
Allison, L.E. 1965. Organiccarbon.p. 1367-1378.In C.A. Black et al. (ed.) Methodsof soil analy-
Allison, L.E., W.E. Bollen, and C.D. Moodie. 1965. Total carbon.p. 1346-1366.In e.A Black et al.
(ed.) Methodsof soil analysis.Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Amacher,M.e., R.E. Henderson,R.H. Brupbacher,and J.E. Sedberry,Jr. 1986. Dichromate-oxidiz-
able and total organic carbon contents of representativesoils of the major soil areas of
Louisiana.Commun.Soil Sci. Plant Anal. 17:1019-1032.
Anne, P. 1945. Sur Ie dosagerapide du carboneorganiquedessols. Ann. Agron. 15:161-172.
Associationof Official Analytical Chemists. 1975. Official methodsof analysis. 12th ed. AOAC,
Washington,De.
Baker, K.F. 1976. The determinationof organic carbonin soil using a probe-colorimeter.Lab. Pract.
25:82-83.
Ball, D.F. 1964. Loss-on-ignitionas an estimateof organic matterand organiccarbonin noncalcare-
oussoils. J. Soil Sci. 15:84-92.
Bartlett, R.I., and D.S. Ross.1988. Colorimetricdeterminationof oxidizablecarbonin acid soil solu-
tions. Soil Sci. Soc. Am. J. 52:1191-1192.
Ben-Dor, E., and A. Banin. 1989. Determinationof organic mattercontentin arid-zonesoils using a
simple "loss-on-ignition"method.Commun.Soil Sci. Plant Anal. 20:1675-1695.
Black, e.A, D.D. Evans,J.L. White, L.E. Ensminger,and F.E. Clark. 1965. Methodsof soil analysis.
Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Bornemisza,E., M. Constenla,A Alvarado, E.J. Ortega, and AJ. Vasquez.1979. Organic carbon
determinationby the Walkley-Black anddry combustionmethodsin surfacesoils and Andept
profiles from CostaRica. Soil Sci. Soc. Am. J. 43:78-83.
Bowman, R.A, W.D. Guenzi, and D.J. Savory. 1991. Spectroscopicmethod for estimationof soil
organiccarbon.Soil Sci. Soc. Am. J. 55:563-566.
Bremner,J.M., and D.S. Jenkinson.1960a.Determinationof organiccarbonin soil. I. Oxidation by
dichromateof organicmatterin soil and plant materials.J. Soil Sci. 11:394-402.
Bremner,J.M., and D.S. Jenkinson.1960b.Determinationof organiccarbonin soil. II. Effect of car-
bonizedmaterials.J. Soil Sci. 11:403-408.
Bremner,J.M. 1949. Use of the Van Slyke-Neil manometricapparatusfor the determinationof organ-
ic and inorganic carbon in soil and of organic carbon in soil extracts. Analyst (London)
74:492-498.
Broadbent,F.E. 1953. The soil organicfraction. Adv. Agron. 5:153-183.
Broadbent,F.E. 1965. Organic matter.p. 1397-1400.In e.A Black et al. (ed.) Methodsof soil analy-
Carolan,R. 1948. Modification of Graham'smethod for determiningsoil organic matter by colori-
metric analysis.Soil Sci. 66:241-247.
Carr, e.E. 1973. Gravimetric determinationof soil carbon using the Leco induction furnace.J. Sci.
Food Agric. 24:1091-1095.
Chemistsof the United StatesSteel Corporation. 1938. Samplingand analysisof carbon and alloy
steels.Van NostrandReinhold Co., New York.
Cheng,H.H., and F.O. Farrow. 1976. Determinationof 14C-labeledpesticidesin soils by a dry com-
bustion technique.Soil Sci. Soc. Am. J. 40:148-150.
Chichester,F.W., and R.F. Chaison,Jr. 1992.Analysis of carbonin calcareoussoils using a two tem-
peraturedry combustioninfrared instrumentalprocedure.Soil Sci. 153:237-241.
Christensen,B.T., and P.A. Malmros. 1982. Loss-on-ignitionand carboncontentin a beechforest soil

profile. Holartic Ecol. 5:376-380.
Ciavatta,c., L.V. Antisari, and P. Sequi. 1989. Determinationof organiccarbonin soils and fertiliz-
ers. Commun.Soil Sci. Plant Anal. 20:759-773.
Clark, N.A., and c.L. Ogg. 1942.A wet-combustionmethodfor determiningtotal carbonin soils. Soil
Sci. 53:27-35.
Coughtrey,DJ., DJ. Nancarrow,and D. Jackson.1986.Extractionof carbon-14from biological sam-
ples by wet oxidation. Commun.Soil Sci. Plant Anal. 17:393-399.
Crowther,E.M. 1935. First report of the organiccarboncommittee.p. 114--127.In Trans. Third Int.
Congr. Soil Sci., Vol. 1. Oxford, England.ThomasMubry & Co., London.
Dalal, R.C., and R.I. Henry. 1986. Simultaneousdeterminationof moisture,organiccarbonand total
nitrogenby near infrared reflectancespectrophotometry. Soil Sci. Soc. Am. 1. 50:120--123.
Datta, N.P., M.S. Khera, and T.R. Saini. 1962. A rapid colorimetric procedurefor determinationof
organiccarbonin soils. J. Indian Soc. Soil Sci. 10:67-74.
David, M.B. 1988. Use of loss-on-ignitionto assesssoil organiccarbonin forest soils. Commun.Soil
Davies, B.E. 1974. Loss-on-ignitionas an estimateof soil organic matter. Soil Sci. Soc. Am. Proc.
38:150--151.
De Bolt, D.C. 1974.A high samplevolumeprocedurefor the colorimetricdeterminationof soil organ-
ic matter. Commun.Soil Sci. Plant Anal. 5:131-137.
De Leenher,L., 1. Van Hove, and M. Van Rurjmbeke.1957. Determinationquantitativede la matiere
organiquedu sol. Pedologie7:324--347.
Degtjareff, W.T. 1930. Determining soilorganicmatterby meansof hydrogenperoxideand chromic
acid. Soil Sci. 29:239-245.
Donkin, MJ. 1991.Loss-on-ignitionasan estimatorof soil organiccarbonin A-horizon forestry soils.
Edson,S.N., and R.H. Mills. 1955. Colorimetric field test for organic matter in mineral soils. Agric.
Food Chern. 852-853.
Fleming, W.R. 1914. Rapid determinationof carbonin steel. Iron Age 93:64--66.
Gallardo,1.F.,1. Saavedra,T. Martin-Patino,andA. Milan. 1987. Soil organicmatterdeterminations.
Geiger, PJ., and J.P. Hardy. 1971. Measurementof organic carbon in arid soils using a hydrogen-
flame ionization detector.Soil Sci. 111:175-181.
Gillman, G.P., D.F. Sinclair, and T.A. Beech.1986. Recoveryof organic carbonby the Walkley and
Black procedurein highly weatheredsoils. Commun.Soil Sci. Plant Anal. 17:885-892.
Goldin, A. 1987. Reassessing the useof loss-on-ignitionfor estimatingorganicmattercontentin non-
calcareoussoils. Commun.Soil Sci. Plant Anal. 13:1111-1116.
Graham,E.R. 1948. Determinationof soil organic matter by meansof a photoelectriccolorimeter.
Soil Sci. 65:181-183.
Greweling,T., and M. Peech.1960. Chemicalsoil tests. Cornell Univ. Agric. Exp. Stn. Bull. 960.
Gupta, U.S., S.M. Gorantiwar,and G.P. Verma. 1975. A new colorimetric procedurefor the determi-
nation of soil organiccarbon.1. Indian Sol. Soil Sci. 23:328-331.
Heanes,D.L. 1984. Determinationof total organic-Cin soils by an improvedchromic acid digestion
and spectrophotometricprocedure.Commun.Soil Sci. Plant Anal. 15:1191-1213.
Heck, A.F. 1929. A methodfor the determiningof total carbonand also for the estimationof carbon
dioxide evolvedfrom soils. Soil Sci. 28:225-231.
Howard, PJ.A., and D.M. Howard. 1990. Use of organiccarbonand loss-on-ignitionto estimatesoil
organicmatterin different soil types and horizons.BioI. Fert. Soils. 9:306-310.
Howard, PJ.A. 1966. The carbon-organicmatterfactor in varioussoil types. Oikos 15:229-236.
Jackson,M.L. 1958. Soil chemicalanalysis.Prentice-Hall,Inc., EnglewoodCliffs, N1.
Kalembasa,SJ.,and D.S. Jenkinson.1973.A comparativestudy of titrimetric and gravimetricmeth-
ods for the determinationof organiccarbonin soil. 1. Sci. Food Agric. 24:1085-1090.
Kalisz, PJ., and U.M. Sainju. 1991. Determinationof carbon in coal "blooms." Commun. Soil Sci.
Plant Anal. 22:393-398.
Krishnan,P., J.D. Alexander,B.J. Butler, and J.W. Hummel. 1980. Reflectancetechniquefor predict-
ing soil organic matter. Soil Sci. Soc. Am. J. 44:1282-1285.
Lee, C.K. 1939. The determinationof organic matterin paddy soils. The reliability of rapid titration
methods.Ind. Eng. Chern. Anal. Ed. 11:428.
Lindbeck, M.R., and J.L. Young. 1964. Glazing technic for leak-proofingcombustionboats.Chern.
Anal. 53:18.
Loeppert,R.H., andD.L. Suarez.1996.Carbonateand gypsum.p. 437-475.In D.L. Sparkset al. (ed.)

Methodsof soil analysis.Part 3. Chemicalmethods.SSSA Book Ser. no. 5. SSSAand ASA,
Madison,WI.
Loftus, N.S., Jr. 1966.The contributionof extractablehumic colloids to the exchangecapacityof sur-
face soils. Ph.D. diss. PennsylvaniaState Univ., Univ. Microfilms, Ann Arbor, MI (Diss.
Abstr. 27:2752B).
Lowther, 1.R., PJ. Smethurst,J.e. Carlyle, and E.K.S. Nambiar. 1990. Methods for determining
organiccarbonin podzolic sands.Commun.Soil Sci. Plant Anal. 21:457-470.
Lunt, H.A 1931. The carbon-organicmatterfactor in forest soil humus.Soil Sci. 32:27-33.
McCready,R.M., and W.T. Hassid. 1942. Semi-microdeterminationof carbonusing the Van Slyke-
Folch oxidation mixture. Ind. Eng. Chern.Anal. Ed. 14:525-526.
McGeehan,S.L., and D.V. Naylor. 1988.Automatedinstrumentalanalysisof carbonand nitrogen in
plant and soil samples.Commun.Soil Sci. Plant Anal. 19:493-505.
Mebius, LJ. 1960.A rapid methodfor the dtterminationof organiccarbonin soil. Anal. Chim. Acta
22:120-121.
Mehlich, A 1984. Photometricdeterminationof humic matterin soil, a proposedmethod.Commun.
Merry, R.H., and L.R. Spouncer.1988. The measurementof carbonin soils using a microprocessor-
controlled resistance furnace.Commun.Soil Sci. Plant Anal. 19:707-720.
Metson, AJ. 1956. Methods of chemical analysis for soil survey samples.N.Z. Dep. Sci. Indust.
Resour.,Soil BureauBull. 12.
Melson, AJ., L.e. Blakemore,and D.A Rhodes.1979. Methodsfor the determinationof soil organ-
ic carbon:A review, and applicationto New Zealandsoils. N.Z. J. Sci. 22:205-228.
Mitchell, J. 1932. The origin, nature, and importance of soil organic constituentshaving base
exchangeproperties.J. Am. Soc. Agron. 24:256-275.
Moore, T.R. 1985.The spectrophotometric determinationof dissolvedorganiccarbonin peatwaters.
Soil Sc. Soc. Am. 1. 49:1590-1592.
Nelson,D.W., andL.E. Sommers.1975.A rapid and accurateprocedurefor estimationof organiccar-
bon in soil. Proc. IndianaAcad. Sci. 84:456-462.
Nelson,D.W., andL.E. Sommers.1982.Total carbon,organiccarbon,andorganicmatter.p. 539-579.
In AL. Pageet aI. (ed.) Methodsof soil analysis.Part 2. Agron. Monogr. 9. 2nd ed. ASA and
SSSA,Madison,WI.
Nommik, H. 1971. A modified procedurefor determinationof organic carbonin soils by wet com-
bustion.Soil Sci. 11:330-336.
Orphanos,P.1. 1973. On the determinationof soil carbon.Plant Soil 39:706-708.
Page,N.R. 1974. Estimationof organic matterin Atlantic CoastalPlain soils with a color-difference
meter.Agron. J. 66:652-653.
Peech,M., L.A Dean, and J. Reed. 1947. Methods of soil analysis for soil fertility investigation.
USDA Circ. 757. U.S. Gov. Print. Office, Washington,DC.
Pella, E. 1990a.Elementalorganic analysis.Part 1. Historical developments.Am. Lab. 22:116-125.
Pella, E. 1990b.Elementalorganicanalysis.Part 2. Stateof the art. Am. Lab. 22:28-32.
Perrier, E.R., and M. Kellogg. 1960. Colorimetric determinationof soil organic matter. Soil Sci.
90:104-106.
Peterson,W.M. 1962. Removalof sulfur fumes by lead dioxide in the combustionmethodfor carbon
in iron and steel.Anal. Chern.34:575-579.
Piper, C.S. 1942. Soil and plant analysis.Intersci., New York.
Ponomareva,v.v., and T.A Platnikova. 1967. Data on the degreeof intra-molecularoxidation of
humusin varioussoil groups(problem of the carbonto humusconversionfactors). Sov. Soil
Sci. 1967(7):924-933.
Quinn, J.G., and M. Salomon. 1964. Chloride interference in the dichromate oxidation of soil
hydrolyzates.Soil Sci. Soc. Am. Proc. 28:456.
Rabenhorst,M.e. 1988. Determinationof organicand carbonatecarbonin calcareous soils using dry
combustion.Soil Sci. Soc. Am. J. 52:965-969.
Ranney,R.W. 1969. An organiccarbon-organicmatterconversionequationfor Pennsylvaniasurface
soils. Soil Sci. Soc. Am. Proc. 33:809-811.
Rather,J.B. 1917. An accurateloss on ignition methodfor determinationof organic matter in soils.
ArkansasAgric. Exp. Stn. Bull. 140.
Raveh,A., and Y. Avnimelech. 1973. Potentiometricdeterminationof soil organic matter. Soil Sci.
Soc. Am. Proc. 36:967.
Rhodes,E.R., P.Y. Kamara,and P.M. Sutton. 1981. Walkley-Black digestionefficiency and relation-
ship to loss on ignition for selectedSierraLeone soils. Soil Sci. Soc. Am. J. 45:1132-1135.
Richardson,J.L., and R.I. Bigler. 1982. Comparisonof Walkley-Black and dry combustionorganic
carbon determinationsin calcareouswater-loggedNorth Dakota soils. Commun. Soil Sci.
Plant Anal. 13:175-183.
Richter, M., G. Massen,and I. Mizuno. 1973. Total carbonand "oxidizable" organiccarbonby the
Walkley-Black procedurein somesoils of the Argentine Pampa.Agrochimica 17:462-472.
Robertson,G.I., L.M. Jett, and L. Dorfman. 1958. Microdeterminationof carbonand hydrogenby a
rapid combustionprocedure.Anal. Chern.30:132-135.
Robinson,G.w., W. McClean,and R. Williams. 1929. The determinationof organiccarbonin soils.
J. Agric. Sci. 19:315-324.
Robinson,W.O. 1927. The determinationof organicmatterin soils by meansof hydrogenperoxide.
J. Agric. Res. 34:339-356.
Salter, R.M. 1916. A rapid methodfor the accuratedeterminationof total carbon in soils. Ind. Eng.
Chern.8:637-639.
Schepers,J.S.,D.O. Francis,and M.T. Thompson.1989. Simultaneousdeterminationof total C, total
N, and tSN in soil and plant material.Commun.Soil Sci. Plant Anal. 20:949-959.
Schollenberger,CJ. 1927.A rapid approximatemethodfor determiningsoil organicmatter.Soil Sci.
24:65-68.
Schollenberger,CJ. 1931.The determinationof soil organicmatter.Soil Sci. 31:483-486.
Schollenberger,CJ. 1945. Determinationof soil organicmatter. Soil Sci. 59:53-56.
Schulte,E.E., C. Kaufman, and J.B. Peter. 1991. The influence of samplesize and heatingtime on
soil weight loss-on-ignition.Commun.Soil. Sci. Plant Anal. 22:159-168.
Sheldrick,B.H. 1986. Test of the Leco CHN-600 determinatorfor soil carbonand nitrogen analysis.
Can. J. Soil Sci. 66:543-545.
Simakov, V.N. 1957. The use of phenylanthranilicacid in the determinationof humus by Tyurin's
method.Pochvovedenie8:72-73.
Simons,E.L., J.E. Fagel, and E.W. Balis. 1955. Combustionof tungstencarbideby high frequency
inducedradiantheating.Anal. Chern.27:1123-1125.
Sims, J.R., and V.A. Haby. 1971. Simplified colorimetric determinationof soil organic matter. Soil
Sci. 112:137-141.
Sinha, H., and R.N. Prasad.1970. A new colorimetric methodfor the determinationof organic car-
bon in soils. J. Indian Soc. Soil Sci. 18:83-87.
Smith, H.W., and M.D. Weldon. 1941. A comparisonof somemethodsfor the determinationof soil
organicmatter.Soil Sci. Soc. Am. Proc. 5:177-182.
Snyder,J.D., andJ.A. Trofymow. 1984.A rapid accuratewet oxidation diffusion procedurefor deter-
mining organicand inorganiccarbonin plant and soil samples.Commun.Soil Sci. Plant Anal.
15:587-597.
Soil ScienceSociety of America. 1979. Glossaryof soil scienceterms.Rev. ed. SSSA,Madison,WI.
Soon,Y.K., and S. Abboud. 1991.A comparisonof somemethodsfor soil organiccarbondetermina-
tion. Commun.Soil Sci. Plant Anal. 22:943-954.
Spain,A.v., M.E. Probert,R.E Isbell, and R.D. John. 1982. Loss-on-ignitionand the carboncontent
of Australian soils. Aus!. 1. Soil Res.20:147-152.
Springer,U., and1. KIee. 1954. Profungder Leistungfahigkeitvon einigenWichtigerenzur Bestiim-
mung der Kohlenstoffs mittels Chromschwefelsaure sowie Vorschlag einer neuen Schnell-
methode.Z. Pflanzenernahr.Dung. Bodenk.64:1-26.
Storer,D.A. 1984. A simple high samplevolume ashingprocedurefor determinationof soil organic
matter.Commun.Soil Sci. Plant Anal. 15:759-772.
Strek, HJ., J.J. Dulka, and AJ. Parsells.1990. Humic mattercontentvs. organic matter contentfor
making herbiciderecommendations. Commun.Soil Sci. Plant Anal. 21:1985-1995.
Tabatabai,M.A., and J.M. Bremner.1970. Use of the Leco automatic70-secondcarbonanalyzerfor
total carbonanalysisin soils. Soil Sci. Soc. Am. Proc. 34:608-610.
Tabatabai,M.A., and J.M. Bremner.1991. Automatedinstrumentsfor determinationof total carbon,
nitrogen,and sulfur in soils by combustiontechniques.p. 261-286.In K.A. Smith (ed.) Soil
analysis.Marcel Dekker, New York.
Takahashi,Y., R.T. Moore, and R.J. Joyce.1972. Direct determinationof organiccarbonin water by
reductivepyrolysis. Am. Lab. 4:31-38.
Tiessen,H., J.R. Bettany,and J.w.s. Stewart. 1981. An improved methodfor the determinationof
carbon in soils and soil extracts by dry combustion. Commun. Soil Sci. Plant Analysis
12:211-218.
TInsley, J. 1950. Determinationof organic carbon in soils by dichromatemixtures. p. 161-169.In
Trans.4th Int. Congr. Soil Sci., Vol. 1. HoitsemoBrothers,Gronignen,the Netherlands.
Tyurin, LV. 1931. A new modification of the volumetric methodof determiningsoil organic matter
by meansof chromic acid. Pochvovedenie26:36-47.
Van Slyke, D.D., andI. Folch. 1940. Manometriccarbondetermination.1. BioI. Chern. 136:509-541.
Verardo,D.I., P.N. Froelich,and A. Mcintyre. 1990. Determinationof organiccarbonand nitrogenin
marine sedimentsusing the Carlo Erba NA-1500 analyzer.DeepSeaRes.37:157-165.
Walkley, A. 1947. A critical examinationof a rapid methodfor determiningorganic carbonin soils:
Effect of variations in digestion conditions and of inorganic soil constituents.Soil Sci.
63:251-263.
Walkley, A., and LA. Black. 1934. An examinationof the Degtjareff method for determiningsoil
organic matter and a proposedmodification of the chromic acid titration method. Soil Sci.
37:29-38.
Wilde, S.A. 1942. Rapid colorimetric determinationof soil organic matter. Soil Sci. Soc. Am. Proc.
7:393-394.
Willett, LR, and T.A. Beech.1987. Determinationof organiccarbonin pyritic and acid sulfatesoils.
Winter, I.P., E.G. Gregorich,RP. Voroney, and RG. Kachanoski.1990. Comparisonof two sample
oxidation methodsfor quantitativemeasurementof 12C and 14C in plant and soil. Can. I. Soil
Sck. 70:525-529.
Winters, E., and RS. Smith. 1929. Determinationof total carbonin soils. Ind. Eng. Chern.Anal. Ed.
1:202-203.
Yeomans,I.C., and I.M. Bremner. 1988. A rapid and precisemethod for routine determinationof
organiccarbonin soils. Commun.Soil Sci. Plant Anal. 19:1467-1476.
Yeomans,I.C., and I.M. Bremner. 1991. Carbon and nitrogen analysisof soils by automatedcom-
bustion techniques.Commun.Soil Sci. Plant Anal. 22:843-850.
Published 1996
Chapter35
Organic Matter Characterization
ROGERS. SWIFT, CSIRO Division of Soils, Adelaide, Australia
INTRODUCTION
It is generallyacceptedthat the term soil organic matter refers only to the non-
living organic materialin the soil, which makesup by far the major portion of the
total organiccomponents.The living organic components,which are part of the
soil biota, comprisea minor portion of the total organic material, and they will
not be consideredin this chapter.The soil organicmatter canbe of plant, animal
or microbial origin and may be relatively fresh or highly decomposedand trans-
formed. It is to the characterizationof this material to which this chapterwill be
devoted.For in-depthreviews onthis topic, the readershouldconsultHayesand
Swift (1978), Stevenson(1994), Aiken et al. (1985), and Hayeset al. (1989).
In chemicalterms, it is possibleto identify in soil organic matter, compo-
nents belonging to the main classesof naturally occurring organic compounds
found in plants and animals. Each of thesecompoundscan be found in a wide
variety of physical environmentsand physicochemicalassociations.In addition
to theseidentifiable compounds,there are much larger amountsof organic mat-
ter which are not amenableto current methodsof chemicalcharacterization.To
bring somesemblanceof order to this diverse and complex system,it is neces-
sary to establishand superimposea set of classificationsand definitions in order
to establisha commonframework for discussionand investigation.
Given the complexity of the soil system,any attemptto rigorously catego-
rize soil organiccomponentsis likely to be, at best, imperfect. Quite clearly the
most likely basisfor classificationlies in readily observablephysical, chemical
and/orbiochemicaldifferencesbetweenthe variouscomponents.A useful delin-
eation based on physical characteristicsis that drawn between recognizable
remainsof plants (or animal) debris and the highly degradedand transformed
materialswhich contain no recognizableplant, animal or microbial structures.
Although this classificationis essentiallybasedon visual observationof physical
differences,in essence,it purposesto differentiate betweenthe results of bio-
chemical transformations.As such it is unlikely to be wholly successful.For
example,the sameclasses of organic compounds(e.g., carbohydrates,peptides
and amino acids) can be found in both fractions.
Book Seriesno. 5.
1011
1012 SWIFf
In an attemptto overcomethis problememphasiscan be placeddirectly on

the chemicalcharacteristicsthemselvesas the basisof classification.Using this
approach,the nonliving organicmattercan be divided into humic and nonhumic
substances(Hayes& Swift, 1978). In this scheme,all of the recognizableplant
debris plus all of the identifiable classesof organic compounds(such as carbo-
hydratesand peptides)in their natural or transformed state are categorizedas
nonhumic substances.The remaining amorphous,highly transformed, darkly
colouredmaterialwhich cannotbe identified asbelongingto an establishedgroup
of organiccompoundare classifiedas humic substances.
As well as the classificationsreferred to above, it would be possible to
deviseothers based, for instance,on the distinction betweenfree, discreteorgan-
ic particlesin the soil as opposedto bound organic matterwhich is adsorbedby
clays, oxides and other mineral surfacesin the soil. Once again, the actual ar-
rangementsin the soil are not quite so simple, as it is difficult to distinguish
absolutelybetweenfree and bound organic matter. For example,organic matter
which is apparently bound may be sorbedto the surfaceof a mineral particle, or
may be involved in other processes,such as occlusion, clustering or aggregate
formation of mineral particles.Onceorganiccomponentsare involved in sorption
or occlusion,the physical and chemicalconditionsin which they exist may pro-
tect them from other soil processes,such as degradationand/or complexation,
possibly for long periodsof time (Thenget aI., 1992).
As can be deducedfrom the above discussion,the order which we may
appearto superimposeon the systemby the use of definitions and classification
systems,although helpful, is, nonetheless,illusory. Soil organic matter is, and
will, remain a complex mixture of organic compoundsin a wide variety of
physicochemicalenvironmentsand subjectto a wide raIlge of associationswith
minerals,metal cations,and anthropogenicorganiccompounds.It is this system
which we must study either in situ in all its complexity, or we may simplify the
material to be studied by undertakingextraction, purification and fractionation
procedures.
Soil organicmatteris an importantpool of C in the global C budgetand is
a major componentof the more active fractions involved in the earth'sC cycle in
terms of both total amountand annualflux. The levels of soil organicmatter are
sensitiveto changesin temperature,rainfall and atmosphericCO2 concentrations
and will be both an important indicator of, and possible contributor to, global
climate change.
It has beencommonly estimatedthat soil organic matter containsapproxi-
mately 1500 x 1015 g C (Schlesinger,1984). Although this figure is basedon a
large number of measurements,it is a very difficult quantity to estimateaccu-
rately comparedwith, for instance,the amountof C as CO2 in the atmosphereor
dissolvedin the surfacewatersof the oceans.This is due to the spatial variabili-
ty of soils and the paucity of datafrom someregionsof the world. Most estimates
for soil organicC rangefrom 1100x 1015 to 3000 X 1015 g C (Schlesinger,1984),
but all estimatesindicatethat thereis more C in the soil than thereis in the atmos-
phereor in living biomass.
Just as important as the total amountof C is the flux of CO2 from the soil
organicmatterwhich is estimatedat 75 x 1015 g C yr-1 (Schlesinger,1984).This
ORGANIC MATIER CHARACTERIZATION 1013
is roughly equivalentto twice the amountof plant materialenteringthe soil each

year. There are, of course,much larger reservoirsof C on the earth than soil or-
ganic matter (e.g., carbonateand fossil fuel depositsor CO2 dissolved in the
oceans)but mostof theseare involved to a lesserextentin the annualfluxes pass-
ing through the C cycle.
As indicatedabove,the physically and chemically heterogeneous mixture
of materialswhich make up soil organic matter varies substantiallyin terms of
both their amount and resistanceto biological decomposition.In most mineral
soils under equilibrium conditions the highly decomposedand biochemically
transformedmaterials are the dominant component,whereasthe recognizable
plants and other biological fragmentsrepresentonly a few percentagepoints of
the total organicmatter.Of the transformedmaterials,the humic substances gen-
erally constitutetwo-thirds to three-quarters.Most of the remainderis made up
of much lesser amounts of carbohydrateand proteinaceousmaterials. Lesser
amountsof a wide range of other compoundscan also be identified, including
lipophilic compoundssuch as fats and waxes.
On enteringthe soil, some of the more labile plant materialsare decom-
posedvery rapidly, sometimeswithin days or weeks,whereasthe more resistant
compoundsmay survive for severalmonthsor even years. Similarly, the trans-
formed materialswill consistof substances with widely varying ratesof decom-
position. One of the characteristicsof the decompositionprocessis that many of
the compoundswhich are producedare more resistantto decompositionthan
their precursors.This is one of the essentialfeaturesof the humification process.
Part of this stability is due to the chemical changeswhich result from the bio-
logical transformationswhich occur during decompositionand part is due to the
associationof organic macromoleculeswith mineral surfaces and with one
another.
The overall or meanturnovertime of organicmatterin mineral soils is fre-
quently several tens to a few hundred years but, due to the factors outlined
above,this is madeup of turnovertimes of particularcomponentsfrom as low as
hours/daysthrough to severalthousandsof yearsor longer for the more intract-
able, charcoal-likematerials.Although peatsand a numberof other soils contain
large amountsof organic matter, most mineral soils containjust a few percent-
age points of organic matter (usually 1-5% C) in the surfacelayers. Neverthe-
less, this relatively small amount of organic matter has a profound effect on a
wide range of soil properties,such as: cation exchangecapacity,soil structure,
the retention and cycling of nutrients,the binding of heavy metals and organic
pesticides,etc. Given the importanceof theseeffects, it is not surprisingthat the
natureand propertiesof soil organicmatterhaveattractedthe interestof soil sci-
entists for many years. Although considerableprogresshas been made in the
understandingof many of the propertiesof thesematerials,it is, perhaps,disap-
pointing that more progresshas not beenmadein other aspects,such as the elu-
cidationof structure.As we shall seebelow, therearegood reasonsfor this which
are inherentin the propertiesof the moleculeswhich make up soil organic mat-
ter in general,and humic substancesin particular.
The purposeof this chapteris to provide an outline of the experimental
methodology currently being applied to the study of organic matter in soils.
1014 SWIFT
Thosemethodswhich are commonly used,or able to be implementedwith min-

imal cost, are reportedin detail. Othermethodswhich require expensiveor spe-
cializedequipmentare mentioned,but the readeris referredto more detailedref-
erencetexts for further information.
Someof the chemicalmethodsare somewhatdated,but have not changed
in recentyears.However,the instrumentalmethodsgenerallyhavechangedsig-
nificantly since the last edition. The methodsoutlined and/or referredto in this
chapterare meantto be usedas a guide to the methodswhich havebeenreport-
ed in the literature. Scientific researchis a dynamic study and the researcher
shouldbe flexible with ideasand methods,and be able to adjustthesewhen nec-
essary.
EXTRACTION OF SOIL ORGANIC MATTER

Although a numberof techniquesare now availablewhich allow organic
matterto be studiedin situ in its unaltered,naturalstate,mostof thesetechniques
still require the organicmatterto be removedor extractedfrom the soil. Among
other things, extraction results in separatingthe organic from the mineral soil
components,removingother inorganicinterferences,increasingthe organicmat-
ter concentrationand renderingthe organic matter soluble. The application of
many chemicalproceduresrequiresthat the material being studiedshould be in
solution.
One criticism of extraction proceduresis that, as a consequenceof the
extraction process,the organic matter constituentsare modified to a greateror
lesserextent. It is, of course,inevitable that associationswith mineral surfaces
and otherorganicconstituentswill be disrupted,as this is a necessarypart of the
extractionprocess.Of more concernis the possibility of chemical alterationof
the organic matter itself, resulting in artifact formation causedby the extraction
process.It is importantthat suchchemicalmodification shouldbe minimized or
avoided,if at all possible.This is more likely to be achievedthrough the use of
mild extractionreagents[e.g., neutral pyrophosphate(Na4PZ07)],as opposedto
strong reagents[e.g., sodium hydroxide (NaOH)]. However, this usually results
in a substantialdecline in the overall yield of extractedorganicmatter.
Having decidedto use an extractionprocedure,it is as well to optimize its
usefulnessand efficiency by incorporatingother steps,such as purification and
fractionation into the overall procedure.Although the intention of an extraction
proceduremay be to isolatea particularcomponent,the reality is that a complex
mixture is usually obtainedand, by the useof appropriateprocedures,this can be
turned to advantage.Thus, it is possibleto obtain samplesof humic substances
and soil polysaccharides from a single extractionand to further fractionatethem
both. Indeed,it is highly desirablethat suchproceduresare employed,in order to
properly purify the particularcomponentbeing studied.
Extractionand Purification of Humic Substances

The proceduresused to extract, purify and fractionate humic substances
exploit the basic chemical and physical propertiesof these materials (Hayes,
1985). If we are to understandhow the various proceduresoperateand whether
ORGANIC MATTER CHARACTERIZATION 1015
they can be improvedor refmed,then it is necessaryto havea reasonableunder-

standingof the composition,propertiesand structureof humic substances.
Humic substancesare a closely related family of naturally occurring
macromolecules,with a chemicallyill-defined structure.For a detailedreview of
the compositionand structureof humic substancesthe readeris referredto the
book Humic Substances II: In Search of Structure (Hayeset aI., 1989),only sali-
ent pointswill be noted here.
In terms of the chemicalcompositionof humic substances,the following
statementscan now be madewith somedegreeof certainty.The presenceof sub-
stituted aromatic rings in humic substanceshas now been independentlycon-
firmed by a rangeof chemicaland instrumentaltechniques.The substituentson
the aromaticrings are predominantlycarboxyl groups,hydroxyl groups,carbo-
nyl groups, and aliphatic units. Multiple substitutionon the aromatic rings is
common.Significant amountsof aliphatic C are also presentwith chain lengths
of 1 to 20 C atomsbut with smallerchain lengthspredominating.The aliphatic
units may be presentas side chainsor as units within the main polymer chain
attachedto aromatic unitsat both ends.The functional groups,which aredistrib-
uted throughoutthe length of the molecule, are certainly attachedto aromatic
groupsand they may also be attachedto the aliphatic moieties.
The various substitutedaromatic and aliphatic units describedabove are
assembledtogetherin a somewhatrandomor, at least,a largely disorderedarray,
andjoined mainly by strongC-C bondsandperhapsetherlinkagesas well. Such
assembliesmake up the molecularbackbone of soil humic substances.Carbohy-
drates and peptidescan be attachedto this backbone in lesserbut significant
amounts.
In terms of physical properties, humic substancesare variable-charge
materialsin which the carboxyl and, to a lesserextent,phenolichydroxyl func-
tional groupsdissociateprogressivelywith increasingpH. The resultantnegative
chargegives rise to cation-exchangesitesand to numerousinter- and intramole-
cular chargeinteractions.Soil humic substancesare extremelypolydisperseand
havemolecularweights rangingfrom a few thousandto well over a million dal-
tons (Swift, 1989).Togetherwith the wide rangeof molecularweightstherewill,
of course,be concomitantvariation in molecularsize but this will also be depen-
dent on the molecularshapeor conformation.Of the variouspossiblemodelsfor
macromolecularconformationit has beensuggested(Swift, 1989) that the ran-
dom coil structureis most appropriatefor humic substances.Unlike other possi-
ble conformationsthis model accommodates the disorderedchemicalstructure,
the solubility and solvation propertiesand the chargecharacteristicsof these
materials.
In summary,the moleculesof humic substances consistof hydrophilic and
hydrophobicgroupings,chargedsites and counterions, the identity and propor-
tion of which vary from moleculeto molecule.In addition, thereis a substantial
molecularweight range to superimposeon the variation in chemical composi-
tion. The limitations imposedby thesepropertieson the applicationand success
of extractionand fractionationproceduresneedto be recognizedat the outset.
Soil humic substancesare largely insoluble and may remain in the soil
system for long periods of time before they are degradedby relatively slow
1016 SWIFT
chemical and biological oxidative decompositionprocesses.Humic substances

in soil are renderedinsoluble by: their own inherent chemical properties,the
compositionof balancingcations, interactions with otherorganicmolecules,and
interactionwith mineral surfaces.In order to extract the humic substances,it is
necessaryto displacethe insolubilizing cationsand to disrupt the various intra-
and intermolecularassociationsand surfaceinteractions.
Generally,the cationsresponsiblefor the insolubility of humic substances
are a combinationof di- and trivalent ions (e.g., Ca2+, Mg2+, Fe3+, and Al3+) as
well as H+. All of theseions are strongly held on the organicexchangesites so
that there is only a small amountof dissociationand, as a consequence, very lit-
tle intramolecularchargerepulsion.In addition, the multivalent cationssimulta-
neouslysatisfy chargedsites at different parts of the molecule,or betweenmol-
ecules,therebyforming cationicbridges.Both of theseeffectscausethe organic
moleculeto adopta condensedconformationfrom which solvent(usually water)
is largely excluded.The net effect of theseprocessesis to causethe moleculesto
be insoluble.
Humic substancescan form associationswith other humic moleculesor
with different organic macromoleculessuch as lignin or partially decomposed
plant materials.Theseassociationsmay be via cationic bridges,polar interac-
tions, hydrogenbonding or van der Waals forces. Whatevertheir nature these
interactions willtend to confer insolubility on the humic macromolecule.
Similar bonding mechanismsare also possiblewith mineral surfaces.In
this case,however,someof the associationsmay be stronger.For instance,humic
substancesmay bond directly with Fe or Al atomsat a clay or sesquioxidesur-
face. The net result of the many interactionsis strongattachmentto a surfaceand
resultantinsolubility. In addition to the effects listed above,the inherentchemi-
cal propertiesof humic substances suchas the relative amountsof polar and non-
polar groupsand the molecularweight will greatly affect their solubility proper-
ties.
The selectionof both the extractantand the methodusedfor the extraction
processshould be made with an understandingof the chemical and physical
characteristicsof the fraction(s) of humic substancesrequired to be separated
from the soil matrix. Whiteheadand Tinsley (1964) have proposedfour criteria
for solventsfor humic substances.In their view, an effective extractant should
have:
1. A high polarity and a high dielectric constantto assistthe dispersionof

the chargedmolecules.
2. A small molecularsize to penetrateinto the humic structures.
3. The ability to disrupt the existing hydrogenbondsand to provide alter-
native groupsto form humic-solventhydrogenbonds.
4. The ability to removeand immobilize metallic cations.
Stevenson(1994) haslisted four criteria for the effective extraction method:
1. The methodleadsto the isolation of unalteredmaterials.

2. The extractedhumic substances are free of inorganiccontaminantssuch
as clay and polyvalentcations.
ORGANIC MAlTER CHARACTERIZATION 1017
3. Extraction is complete, thereby ensuring representationof fractions

from the entire molecularweight range.
4. The methodis universallyapplicableto all soils.
Although the four requirementsfor an extractant are achievable,the four
requirementsfor an extraction method representmore an ideal than a set of cri-
teria that are achievable.
Extractionwith AqueousSolutions
In terms of the desirablecriteria for an extractantlisted above,water is a
very good solvent. It is polar, has a high dielectric constant,is able to form
hydrogenbondsand therebydisrupt other hydrogenbonds,and is small and pen-
etrative. However, an accompanyingsolute or additional step is requiredto dis-
place and immobilize the insolubilizing, multivalent metal ions by use of a
reagentwhich is capableof forming soluble complexeswith thesecationsor of
removing them as precipitates.In this regard, solutions of pyrophosphateand
EDTA {N,W-1,2-ethanediylbis[N-(carboxymethyl)glycine]}have becomepop-
ular extractantsbecauseof their ability to form stable,soluble complexeswith
metal ions at nearneutralpH values.Alternatively, the multivalentcationscanbe
removedby prior washingwith dilute acid which gives a H+ -saturatedsoil. How-
ever, the acidic conditionsare not conduciveto extractionof humic substances
and the pH needsto be increasedbefore proceedingfurther with the extraction
process.
Having removedthe polyvalentcationsfrom the negativelychargedorgan-
ic exchangesites, it is importantto replacethem with a counter-ionthat dissoci-
atesvery readily, and Na+ and K+ are the ions most commonly usedfor extrac-
tion. The resultant high degree of dissociation causesintramolecular charge
repulsionand leadsto molecularexpansiontogetherwith ingressof waterwhich
effectively solvates polar groups and disrupts intermolecularbonds. The net
result is that the humic substancesbecomesoluble and are extracted.
One of the oldest and most frequently used methodsof extractinghumic
substances from soils usessodiumhydroxideas the extractant.In this procedure
many of the polyvalentcations(if not previouslydisplacedby acid pretreatment)
are removedthrough the formation of insoluble hydroxides at high pH values
and are replacedby sodium. The high pH of sodium hydroxide solutions also
causesmany organicfunctional groupsto ionize resultingin a higherchargeden-
sity on the molecules.The importanceof this effect canbe judgedby the fact that
sodium hydroxideextractsfar larger amountsof humic material at pH valuesof
12 and abovethan the sodiumsaltsof complexingagentsusedat nearneutralpH
values (Table 35-1). The problems of artifact formation by oxidation of the
humic substanceswhich can occur at high pH values can be substantially
reducedby performingthe extractionsundera Nz atmosphere.
Extractsobtainedby using sodium hydroxide have higher averagemolec-
ular weight and lower functional group content than those extractedfrom the
samesoil by sodium pyrophosphate(Na4PZ07)at neutral pH values. Observa-
tions suchas theseclearly indicatethat, at any given pH value, the solubility lim-
itations of humic substancesare predominantlydeterminedby charge density
and molecularweight considerations.
1018 SWIFf
Table 35-1. Yields and compositionsof humic acid (HA) and fulvic acid (FA)t fractions extracted
with different solventsfrom H+ soil (adaptedfrom Hayeset aI., 1975).
Elementalcomposition
Extractant Fraction Yield C H N S Ash

%
OMF* HA 16 53.4 4.5 2.6 1.7 1.8
OMF* FA 2.0 49.9 4.0 3.1 1.5 4.7
Sulpholane HA 10.0 54.0 4.8 3.2 2.4 0.8
Sulpholane FA 12.0 51.8 4.3 3.2 1.7 3.3
OMSO* HA 17.0 53.5 4.1 3.2 1.9 2.7
OMSO* FA 6.0 51.5 4.1 2.1 1.2 6.5
Pyridine HA 34.0 54.7 5.0 4.3 NO§ 2.2
Pyridine FA 2.0 45.3 5.1 5.8 NO 3.8
EO~ HA 49.0 54.7 5.7 6.2 NO 3.7
EO~ FA 14.0 48.2 5.4 10.5 NO 5.8
O.5MNaOH HA 58.0 52.0 5.9 2.8 NO 2.0
O.5MNaOH FA 2.0 43.9 5.9 4.2 NO 2.5
1MEOTA HA 12.5 50.8 4.0 NO NO 2.6
1MEOTA FA 3.8 45.7 4.0 NO NO 5.6
t Excluding material lost during dialysis.
* OMSO - dimethylsulfoxide,OMF =dimethylformamide,EOA =1,2-diaminoethane
§ NO =not determined.
InternationalHumic SubstanceSociety Method. A numberof methods

for the extractionof humic substances from soil using sodiumhydroxidesolution
havebeenpublished.Thesemethodsare generally successful and yield compara-
ble results.The following is a methodwhich hasbeendevelopedby the Interna-
tional Humic SubstanceSociety (IHSS) as an acceptablemethodfor the extrac-
tion of humic substances from soils.
It has been clearly statedby IHSS that this is not meant to be a recom-
mended or approved method,but a methodthat hasbeenfound to be satisfactory
for most soil typesand one which can be performedin most laboratories.It pro-
ducesrelatively high yields and can be usedas a standardmethod for compar-
isonsbetweenandwithin laboratories.An importantcomponentof this methodis
the use of an adsorbentresin in the purification process.This can be replacedby
dialysis if the resin is unavailable.
Materials
1. Hydrochloric acid (HCI), 1 M, 6 M
2. Sodiumhydroxide, 1 M, 0.1 M
3. Potassiumhydroxide(KOH), 0.1 M
4. Potassiumchloride (KCI)
5. Hydrofluoric acid (HF) concentrated,0.3 M
6. XAD-8 resin (Rohm & HaasCo., Philadelphia,PA)
7. Visking dialysis tubing(Visking Co., Chicago,IL) [MWCO (molecular
weight cut-oft)] 10 000 dalton
Method
Removeroots and sievethe dried soil sampleto passa 2.0-mmsieve.Equi-
librate the sampleto a pH value between1 to 2 with 1 M HCI at room tempera-
ture. Adjust the solution volume with 0.1 M HCI to provide a final concentration
that has a ratio of 10 mL liquid/1 g dry sample.Shakethe suspensionfor 1 hand
then separatethe supernatantfrom the residueby decantationafter allowing the
solution to settleor by low speedcentrifugation.Savethe supernatant (FA Extract
1) for the isolation of fulvic acid using XAD-S.
Neutralize the soil residuewith 1 M NaOH to pH = 7.0 then add 0.1 M
NaOH under an atmosphereof N2 to give a final extractantto soil ratio of 10:1.
Extract the suspensionunderN2 with intermittentshakingfor a minimum of 4 h.
Allow the alkaline suspensionto settle overnight and collect the supernatantby
meansof decantationor centrifugation.Acidify the supernatantwith 6 M HCI
with constantstirring to pH = 1.0 and then allow the suspensionto standfor 12
to 16 h. Centrifugeto separatethe humic acid (precipitate)and fulvic acid (super-
natant- FA Extract 2) fractions.
Redissolvethe humic acid fraction by addinga minimum volume of 0.1 M
KOH under N2. Add solid KCI to attain a concentrationof 0.3 M [K+] and then
centrifugeat high speedto removethe suspendedsolids. Reprecipitatethe humic
acid by adding6 M HCI with constantstirring to pH = 1.0 and allow the suspen-
sion to standagain for 12 to 16 h. Centrifuge and discard the supernatant.Sus-
pendthe humic acid precipitatein 0.1 M HClIO.3 M HF solution in a plastic con-
tainer and shake overnight at room temperature.Centrifuge and repeat the
HCI/HF treatment,if necessary,until the ash content is below 1%. Transferthe
precipitateto a Visking dialysis tube by slurrying with water and dialyze against
distilled water until the dialysis water gives a negativeCl- test with silver nitrate
AgN03• Freezedry the humic acid.
Passthe supernatantdesignated"FA Extract I" through a column of XAD-
S (0.15 mL of resin per gram of initial sampledry weight at a flow rate of 15 bed
volumesper h). Discardthe effluent, rinse the XAD-S column containingsorbed
fulvic acid with 0.65 column volumes of distilled H20. Back elute the XAD-S
column with 1 column volume of 0.1 M NaOH, followed by 2 to 3 column vol-
umesof distilled H20. Immediately acidify the solution with 6 M Hel to pH =
1.0. Add concentratedHF to a final concentrationof 0.3 M HE The solution vol-
ume should be sufficient to maintain the fulvic acid in solution.
Passthe supernatantdesignated"FA Extract 2" through a column of XAD-
8 (1.0 mL of resin per gram of initial sampledry weight). Repeatthe backelution
and acidification as for "FA Extract I" above. Combine the final eluatesfrom
eachof the fulvic acid extractsand passthis solution through XAD-8 resin in a
glasscolumn (column volume shouldbe one-fifth of samplevolume). Rinsewith
0.65 column volumesof distilled H20. Back elute with 1 column volume of 0.1
M NaOH followed by two column volumes of distilled H20. Pass the eluate
through W-saturatedcation exchangeresin [Bio-Rad AG-MP-5 (Bio-Rad, Rich-
mond, CA) using threetimes the mole of Na ions in solution]. Freezedry the elu-
ate to recoverthe H+ -saturatedfulvic acid.
Comments
XAD-S is a nonionic, macroporous(pore size 25 Ilm), methyl methacrylate
esterresin (see"Fractionationof Humic SubstancesAdsorption"). Becauseit is
sometimesdifficult to obtain it may be necessaryto use an alternativeresin such
1020 SWIFT
as Polyclar, which is a cross-linkedpoly(vinylpyrrolidone)(PVP) (Watanabe&

Kuwatsuka1991; De Nobili et aI., 1990a)or other equivalentresin.
Extensive purification proceduresof the resins are required before use.
Thesemethodsand methodsusedto store the resin are detailedby Thurman&
Malcolm (1981).
If it is not possibleto purify the fulvic acid using resin treatments,exhaus-
tive dialysis againstdistilled H20 is an alternativebut lesssatisfactorymethodof
purification. If there is a significant concentrationof polyvalent cationssuch as
Al3+ present,thesemay form insoluble metal-humatecomplexesas the solution
is neutralized.Therefore,the dialysisshouldbe carriedout againstdilute HCI ini-
tially until the concentrationof any polyvalent cations has been significantly
reduced,before finally dialyzing againstdistilled H20. Technically, a fraction
obtainedin this way should be referredto as a fulvic fraction, rather than fulvic
acid, as it is likely to containsignificantamountsof unboundsoil polysaccharide.
SequentialExtraction. By extractingthe humic substancesfrom the soil
at only one pH value as in the above method,variations in the physiochemical
propertiesof the humic substanceswhich reflect the environmentin which they
exist in the soil, may not be apparent.Sequentialextractionof humic substances
at a numberof different pH may be a more sensitivemethod for differentially
extractingthe humic substancesand for determiningrelative distribution in the
soil and their degreeof interactionwith soil colloidal particles.This can be done
by changingthe pH aloneor changingthe natureof the extractantanion (Posner,
1966; Skjemstad,1992). The sequenceoutlined by Posner(1966), 0.1 M pyro-
phosphate(PH = 7), cold 0.5 M NaOH followed by hot 0.5 M NaOH, was found
to producefractionswhich were distinctly different with respectto their molecu-
lar size, functional group contentand infrared spectralcharacteristics(Posneret
aI., 1968; Swift & Posner,1971; Cameronet aI., 1972a). The method outlined
below is basedon that of Posner(1966) and involves three different extractants
at two pH values. This methodologycould be refined and developedto vary
and/or increasethe numberof extractantsor pH valuesusing similar procedures
as outlined by Posner(1966).
Materials
1. Sodiumdihydrogenpyrophosphate/tetrapotassium
pyrophosphate
(Na2H2P207~P207)' 1:1, 0.1 M, pH = 7.0.
2. Hydrochloric acid, 0.1 M, 5 M.
3. Sodiumhydroxide,0.5 M, 5 M.
4. Visking dialysis tubing.
Method
Preparethe soil by crushingand air drying beforepassingit througha 2.0-
rom sieve. Pretreatthe soil with 0.1 M HCI (with stirring) for two consecutive
periodsof 24 h beforecommencingthe extractionprocess.
Combinethis soil samplewith the first extractant(pyrophosphate),in the
soil to solution ratio of 1 g/5 mL. Allow the suspensionto standfor 48 h at the
desiredtemperature(20 or 60°C) underN2. Separatethe soil andextractant(Frac-
ORGANIC MATI'ER CHARACTERIZATION 1021
tion 1) by centrifugationat 1200g for 30 min. Washthe soilresidue withdistilled

water severaltimes. Repeatthe aboveextractionfor eachextractant(e.g., 0.5 M
NaOH) changingthe temperatureas required.
The humic acid componentsof thesefractions can be obtainedas follows.
Acidify the fraction to pH =1 with HCI and allow it to standfor 24 h beforecen-
trifugation. Adjust the solution volume to 600 mL with distilled water and raise
the solution to pH = 7 using a suitable alkali (e.g., 5 M NaOH). Centrifuge the
solution at 6000 g at 2 to 5°C for 30 min. Repeatthis precipitationand dissolu-
tion procedurea further four times, increasingthe centrifugationtimes by 30 min
eachtime, to removethe clay and huminsfrom the solution. Retainall the super-
natantsolutionsfor fulvic acid fractionation.
Finally, precipitatethe humic acid at pH =1, allow the solution to standfor
24 h, centrifugeand then exhaustivelydialyze the precipitatedhumic acid against
distilled water in Visking dialysis tubing until chloride free. Freezedry the slur-
ry to recoverthe humic acid.
Separatethe fulvic acid fraction from the combined supernatantsusing
XAD-8, PVP or dialysis tubing, as outlined in the previousmethod. Freezedry
the fulvic acid.
Extractionwith Non-AqueousSolvents
Non-aqueoussolventshave been used by a numberof workers to extract
humic substances from soils. The most successfulsolventstried so far beingpyri-
dine and dipolar aprotic solventssuch as dimethylsulfoxide(DMSO, C2H60S)
and dimethylformamide(DMF, C3H7NO) (Hayes,1985; Hayeset aI., 1975; Pic-
colo, 1988; Piccolo & Mirabella, 1987; Ma'shumet aI., 1988).
Thesedipolar aprotic solventshavebeenfound to work mosteffectively for
humic substanceswhere the ionization of the humic moleculeshas been sup-
pressedso that they will behaveessentiallyas if they are unchargedmolecules.
This is achievedby first washingthe samplein dilute acid to remove the metal
cationsand then maintainingthe solution pH at moderatelyacidic levels during
the extractionwith the aprotic solvent.Under theseconditionsthe DMSO is able
to efficiently solvatethe humic molecules,extractinghumic substances with sim-
ilar yields and molecular weightand cation-exchangecapacityas thoseobtained
when extractingin aqueoussolutionsat neutralpH. By using an amphiphilic sol-
vent consistingof a mixture of ammonium(NUt)/isopropanolMa'shum et al.
(1988) were able to isolate the hydrophobicorganic matterfrom a soil.
A methodof supercriticalgas extractionof organicmatter using an organ-
ic solvent has beenapplied by Schnitzeret al. (1986) to extract specific organic
componentsfrom a soil matrix. The methodhas beenfurther extendedto sequen-
tially extractorganicmatterfrom soils using supercriticalfluid extraction,togeth-
er with a rangeof organicsolvents(Schnitzer,1990).
Extractionand Purification of Soil Polysaccharides
As the interestin understandingthe role of soil organicmatterhasincreased

the focus has frequently beenon the humic substancesbecausethey accountfor
a large percentageof the soil organicC. The soil polysaccharides,which arecom-
1022 SWIFf
posedof a wide rangeof monosaccharides in both simple and complex molecu-

lar structures,form a smaller but nonethelessimportant part of the soil organic
matter.
Polysaccharides in the soil may occur as an original fragmentof once liv-
ing organicmatteror may have beenformed as a result of microbial activity as
part of the decompositionprocess.Polysaccharides exist in associationwith inor-
ganic or organic colloidal components,either through sorption (van der Waals
forces, H-bonding)or throughchemisorption(e.g., phenolicglycosidelinkages).
Polysaccharideshave a numberof effects on soil propertiessuch as the cation
exchangecapacity(uronic acid group), C metabolism,biological activity and the
complexingof metals.But more importantly, it has beenfound that polysaccha-
rides are involved with stability of aggregatesin soils.
The percentageof the total soil polysaccharidecontentinvolved in aggre-
gation of soil particlesis probably quite low as it requiresthe interactionof the
polysaccharidewith multiple soil components.This interactionwill dependon
the type of functional groupson the polysaccharideand the conformationalstruc-
ture of the sugar groups, as well as the nature of the other soil components
(Williams et aI., 1967). The correlation between total polysaccharideand degree
of aggregationhas beenfound to be poor (Oades,1967). However, many of the
polysaccharidesare not active (e.g., partly decomposedcellulose)and so do not
contribute to aggregationof the soil particles. More recent work (Haynes &
Swift, 1990) hasshownthat thereare particularly activefractions of polysaccha-
ride involved in the aggregationprocess.It has been noted (Greenlandet aI.,
1962) that the aggregatingeffectsof soil polysaccharidesare more noticeablein
soils with low organicmattercontent.
As indicatedabove,one of the main reasonsfor studying polysaccharides
in soils is associatedwith their ability to bind and stabilize soil particles.Ironi-
cally, the polysaccharidesinvolved in this processare probably thosemost firm-
ly held in the soil systemand, therefore,the most difficult to remove without
degradingthem. Any extractionmethodshould, ideally, be able to extract all of
the polysaccharidesfrom the soil matrix and, if this is not possible,then be able
to extract a representativefraction of the polysaccharides.It is possibleto esti-
matethe efficiencyof the extractionprocedureby comparingthe amountof prod-
uct from the extractionwith that which is obtainedusing hydrolysis procedures
to determinethe total carbohydratecontentof the soil (Cheshire,1979). Howev-
er, sucha comparisondoesnot clearly indicatewhetherpreferentialextractionof
somepolysaccharides is occurringin the extractionprocedure,and hencewhether
the extractedfraction is representativeof that in the original soil sample.
Extraction of polysaccharidesfrom soil utilizes similar methodsto those
applied to extractionof humic substances,and often is part of the procedureof
their isolation. None of the methodsusedare ideal and to someextent the most
suitablemethodwill dependon the natureand origin of the soil. The most effi-
cient methods for general application are those using 0.1 or 0.5 M sodium
hydroxide.Large amountsof other fractions of organicmatterare simultaneous-
ly extractedfrom the soil using this methodand the efficiency of separatingthe
polysaccharides from the bulk of the extract(especiallythe humic acid fraction)
is doubtful. It has beenfound that yields are significantly improvedby pretreat-
ment of the soil with dilute HCI or HF (Swincer et aI., 1968), or sulfuric acid
(H2S04) (Barkeret aI., 1967) to acidify the soil organiccomponents.Methylation
(Cheshireet al. 1983)of the soil as a pretreatmentis anotheralternativealthough
this methodmay be unsatisfactoryas a preparativemethodfor someanalyses.
Other methodsof extraction include using extractantssuch as hot water,
which tendsto havelower yields and is thoughtto be degradative,and dilute min-
eral acids, which give lower yields but significantly reducethe amountof humic
substancebeing extracted,so simplifying the purification of the polysaccharide.
Organicreagentssuchas DMSO give a high percentageof polysaccharides in the
product. The polysaccharidecan be separatedfrom the humic substances,by
sorbing the latter on XAD-8 resin, but separatingthe polysaccharidesfrom the
DMSO is difficult. Complexingagentssuchas EDTA give low yields and hence
a greaterprobability of unrepresentative fractions.
Resins such as XAD-8 or Polyclar AT [cross-linked poly(vinylpyrroli-
done)] (Sanderson& Perera,1966; Swinceret aI., 1968; Drijber & Lowe, 1990)
may be used as a meansof removing humic substancesfrom the soil extract to
obtain the polysaccharidefraction. With an appropriateeluant, the humic sub-
stancesare adsorbedon the resin and the polysaccharideis elutedthroughthe col-
umn. Purification of this polysaccharidefraction may then be achievedby dialy-
sis to removesalts,solventsand otherlow molecularweight material.Adsorption
of the coloured compoundsonto charcoal is another method of purification,
althoughit may be difficult to recoverthe polysaccharidesfrom the charcoal.
Gel chromatographycan be usedto fractionatepolysaccharides on the basis
of molecularsize. Ion exchangechromatographyusing diethylaminoethyl-cellu-
lose {(C6H70)..{OHhx.a[OCH2CH2N(CH3CH2)]a}(DEAE-cellulose)can then be
usedto fractionateon the basisof chargedensity differences.In gel chromatog-
raphy care must be taken to select a gel that isolates the polysaccharidesas a
group without losing a specific part of the fraction; suchas the smallermolecules
that bypasswith the largest molecules in the case of using SephadexG-lOO
(Pharacia,Uppsala,Sweden).The gel chromatographymethod given below is
basedon that of Swinceret al. (1968).
Materials
2. Sodiumhydroxide,0.5 M.
3. Dowex 50 (W) (Dow Co., Midland, MI).
4. Polyclar AT resin.
5. SephadexG-25.
Method
Pretreatthe soil for 16 h with 1 M HCI at 20°C, then extractthe soil twice
for 16 h with 0.5 M NaOH at 20°C. Passthe extractsupwardsthrough a column
containingDowex 50 (H+) to remove the humic acids and then concentratethe
eluatesin vacuo at 45°C. Passthe eluatesthrough Polyclar AT to remove the
residualcolouredmaterials.Removethe salts and low molecularweight materi-
al by separationin a column of SephadexG-25.
1024 SWIFT
Comments
The cation exchangeresin in the H+ form was usedinsteadof precipitating
the humic acid in acid solution, as it overcomesthe problemof coprecipitationof
the polysaccharides.New resins, such as XAD-4 (polystyrenedivinylbenzene)
can be usedin place of Polyclar AT.
FRACTIONATION OF SOIL ORGANIC MATTER
Fractionationof Humic Substances
Becauseof the complexity of structureand interactionsof soil humic sub-

stances,the physical and chemical propertiesof thesenatural organic mixtures
are difficult to define precisely.In order to simplify the study of humic sub-
stances,a variety of techniqueshave beendevelopedto fractionatesamplesinto
distinctive and hopefully less complex parts. Fractionatinga sample of humic
substancesdoesnot result in pure homogeneouscompoundsbut ratherfractions
in which one or more of the physicalor chemicalpropertieshasa narrowerrange
of valuesthan the original sample.
The particularmethodchosenfor the fractionation processwill dependon
the chemicaland/orphysicalcharacteristicsbeing studied.Commonlyusedfrac-
tionation proceduresare basedon characteristicssuch as differencesin solubili-
ty, molecular size and electrostaticcharge of the moleculeswithin the system
(Swift, 1985). Someof the techniquescan be classedas preparativeas they result
in fraction samplesof sufficient size to be usedin further study (e.g., thosebased
on solubility and separationaccording to molecular size). Other fractionation
techniques,such as some of those basedon chargecharacteristics(e.g., electro-
focusing) are more appropriatefor fingerprinting. Such techniquesare able to
characterizea samplefor purposesof comparisonwith other sampleschemicals,
but do not easily producesufficient samplefor further study.
FractionationBasedon Solubility
The solubility of humic substancesis not only dependenton the pH of the
solution, but is also dependenton the type and concentrationof the cationsand
other solutespresent,and on the natureof the solvent system.Theoretically,all
of thesepropertiescould be usedto fractionatea humic substancesample.
Use of pH. Precipitationusing changesin pH is the basis of the humic
acidlfulvic acid fractionation(Fig. 35-1). The procedurecan be refined by using
smaller changesin pH to obtain more narrowly defined fractions (Flaig et aI.,
1975). Unfortunately,it is difficult to obtainfractionswith limited overlapdue to
significant coprecipitationwhich occursin the experimentalprocedure.Howev-
er, becauseof the variety of structuraland functional groupswithin a sampleof
humic substance,and of the variety of interactionsbetweenthesegroups,frac-
tionation of humic substanceson this basisis unlikely to yield subfractionswith
substantiallydifferent chemicalcharacteristics.
FRACTIONATION OF SOIL HUMIC SUBSTANCES

e.g. recognizable plant debris;
plus polysaccharides, proteins.
lignins, etc. in their natural or
transformed states.
fractionation on the
basis of jOIUbility
soluble in acid insoluble in acid insoluble in acid

soluble in alkali soluble in alkali insoluble in alkali
~
....I [ f - - - - - Decreasing molecular weight - - - - -
.....f - - - - - - Decreasing carbon content - - - - -
......f - - - - - - Increasing oxygen content - - - - -
I [ f - - - - - Increasing acidity and CEC - - - - -
....
I( Decreasing nitrogen content
.....! - - - - - Decreasing resemblance to lignin - - - - -
Fig. 35-1. Diagramshowingthe categorisationof soil organicmatterinto humic and nonhumicsub-

stances,the fractionationof the humic substances
and the propertyvariationswithin thesefractions.
A variation in usingthe rangein solubility as a meansof obtainingfractions

is to sequentiallyextracta soil sampleusinga rangeof solutionsof increasingpH,
and ability to solubilize thehumic substancespresentin the sample.This type of
extractionis discussedin "SequentialExtraction."
Salting Out. Increasingthe salt concentrationof a humic substancesolu-
tion decreases the intramolecular chargerepulsionandcausesthe polyelectrolyte
macromoleculesto shrink and to excludethe solvent.Simultaneously,increasing
the salt concentrationcausesa decreasein the extensionof the diffuse double
layer of charge, thereby decreasingintermolecular charge repulsion allowing
moleculesto approacheachother more closely, and henceto coagulate.It is pos-
sible to fractionatehumic substances basedon this behaviour(Thenget aI., 1968)
as outlined below.
Materials
1. Sodiumhydroxide, 1.0 M.
2. Ammonium sulfate [(NH4)2S04], solid.
3. Sulfuric acid (H2S04), 1.0 M.
Method
Prepare 150 mL of 0.33% K+ -humate solution. Add 1.0 g of solid
(NH4)2S04,readjustthe pH to sevenusing 1 M NaOH andshakefor 5 to 6 h. Sep-
arate the precipitateby centrifugationand shakethe solution overnight without
1026 SWIFT
adding more salt. Centrifuge the suspensionand combine the precipitatesand

retain.
Add successiveincrementsof 1.0 g of (NH4)zS04and repeatthe procedure
until the amount of precipitateobtainedafter centrifugation is negligible. For
eachfraction obtainedfrom the salting out procedure,dissolvethe precipitatein
distilled water or dilute alkali, adjust the pH to one using 1 M H2S04 and then
removethe excesssalt by exhaustivedialysis. Freezedry the productandweigh.
Commen~ther Precipitation Methods

The interactionbetweenheavymetal ions and humic substances in solution
can affect the solubility of the humic substances.Using the insolubility of the
metal-humicsubstancescomplex is one way of isolating that fraction of humic
substancewhich interactswith the metal concerned.MacCarthyand O'Cinneide
(1974a)usedthis methodto study complexingof humic substances with both Cu
and Co underboth acidic and alkaline conditions.Metal-humateinteractionsare
of interestagriculturally as they may influencethe availability of trace metalsin
soiVplant systems.The ability of humic substances to interactwith heavy metals
is also of great interest environmentallywith respectto the transportof heavy
metalsthroughsoiVwatersystemsand, hence,the fate of toxic metalsin soils and
groundwaters.
Fractionationon the basis of metal-humatesolubility, and that basedon
salting out, are not usedregularly as they are rathertediousand the fractions are
ill-dermed due to problemsof coprecipitation.
Use of OrganicSolvents.Humic substances haverelatively low solubility
in many conventionalorganic solventswhich can be usedas a methodof frac-
tionation (Hayes,1985). Historically, ethanolwas usedto extractthe hymatome-
lanic acid fraction from precipitatedhumic acid (Stevenson,1994),but it canalso
be usedto fractionally precipitatealkalinesolutionsof humic acid (Kyuma, 1964;
Kumada& Kawamura,1968).The methodbelow is basedon that of Kumadaand
Kawamura (1968). Other water-miscible organic solvents, such as acetone
[(CH3)zCO)andmethanol(CH30H), may be usedin a similar way to obtainfrac-
tions of humic substances.
Materials
1. Sodiumhydroxide,0.1 M, 0.2M.
2. Ethanol(C2HsOH), absolute.
3. Hydrochloric acid, 1:3.
Method
Dissolve 150 mg of humic acid in 20 mL of 0.2 M NaOH. Add absolute
ethanol(~HsOH) to give a concentrationof 10% (v/v), allow to standovernight
and then centrifugethe suspensionat 7000 rpm for 20 min. Dissolvethe precipi-
tate in 0.2 M NaOH, add ethanolto the same concentration as aboveand recover
the precipitateby centrifugation.
ORGANIC MAITER CHARACTERIZATION 1027
Add ethanolto the combinedsupernatantto give an ethanolconcentration

of 20% and repeatthe aboveprocedure.Continuethe processby increasingthe
alcoholconcentrationin stepsof 10% until the fmal concentrationis 80% ethanol
and eight fractionatedprecipitateshavebeenobtained.
Dissolveeachprecipitatein 0.1 M NaOH, precipitateby addingHCI (1:3)
andthenwashthe precipitatewith ether(C4HlOO) over a glass-filter.Dry the pre-
cipitatesin a desiccatorcontainingconcentratedH2S04 for 2 d and weigh the
products.
Reducethe concentrationof alcoholin the 80% ethanolsolublehumic frac-
tion by distillation in a water bath, precipitate,wash and dry this fraction as
describedabovefor the otherfractions.
Comments-Use of Other Organic Solvents
Generally,extractingfractions using water-immisciblesolventsgives very
low yields. However, Rice and MacCarthy(1989) have achievedsomesuccess
using a different approachwith the polar water-immiscible solvent, methyl
isobutyl ketone [CH3COCH2CH(CH3)2] (MIBK). They initially separatedthe
aqueousfulvic acid fraction from the nonaqueoushuminlhumicacid fraction at
low pH using HCI to acidify the solution. Following this the pH of the nonaque-
ous phasewas madealkaline using NaOH allowing the separationof the humic
acid in the aqueousphasefrom the humin associatedwith the MIBK nonaqueous
phase.Rice and MacCarthy(1989) were then able to fractionatethe humin resi-
due into an acid solubleform and aqueousand nonaqueousphaseproducts.The
study of the humin fraction of the soil is a neglectedbut importantaspectof soil
research,and this techniqueprovidesan opportunityto study the humin fraction.
Adsorption.The desorptionbehaviourof humic substances sorbedto a gel
(Swinceret al., 1968), resin (Yonebayashi& Hattori, 1990), charcoal(Forsyth,
1947)or alumina(Dragunov& Murzakov, 1970),canbe usedasa meansof frac-
tionation. The purification of the fulvic acid fraction using resinssuchas XAD-
8 or PVP during the extractionof humic substances("Extraction with Aqueous
Solutions")is essentiallyan adsorption/desorption fractionationprocedure.
Materials
1. Sodiumhydroxide,0.1 M, 10 M.
2. Ethanol,50% (v/v).
3. Universalbuffer, 0.02M.
4. Sulfuric acid,SM.
5. XAD-8 resin.
6. Amberlite IR-120 resin (Rohm & HaasCo., Philadelphia,PA).
Method
Dissolve the humic acid samplein 0.1 M NaOH and treat with Amberlite
IR-120 resinto convertit to the H+-exchangedform. Dissolve5 mg of the H+-sat-
uratedhumic acid in 2 mL of aqueoussolution and load onto a column packed
with XAD-8 resin. Elute the humic acid in a stepwisefashion with universal
buffer solutionsadjustedto pH =7 and pH =11, distilled waterand 50% ethanol.
1028 SWIFf
Determinethe elution profile by first adjustingthe pH of the effluent to pH = 12

using 10 M NaOH and then measuringthe optical density at 400 nm. Precipitate
the humic acid containedin eacheffluent using sulfuric acid and then dissolveit
in 0.1 M NaOH. If the humic acid fraction doesnot precipitateon acidification,
absorbit onto a small column of XAD-8 at pH = 3 and elute with NaOH solution.
Dialyze eachof the eluatesagainstdistilled water and freezedry.
Comments
It is possibleto refine this methodby usinga greaterrangeof buffers to sep-
arate the humic acid into a larger numberof fractions. Yonebayashi andHattori
(1990) extendedthe methodby usingboth a pH gradientand a water-ethanolgra-
dient to allow for the ability of obtaininga greaternumberof fractionsif required.
MacCarthyet al. (1979) useda similar methodto fractionatea sampleof humic
substance.Insteadof using buffers a pH gradientsolution was generatedby elut-
ing simultaneouslyfrom two pumps,one containing0.1 M H3P04 and the other
0.1 M NaOH. MacCarthy et al. (1979) separatedout only two fractions, but it
would be possibleto adaptthe methodto obtain more fractions,dependingon the
characteristicsof the humic substanceand the natureof the pH gradientused.
FractionationBasedon Molecular Size

The extremerangein molecular weightsassociatedwith humic substances
extractedfrom soils should theoretically allow the separationof the sampleinto
many specific fractions. In reality the complexintermolecularassociationsmake
it difficult to obtain fractions with insignificant overlap in molecular content.
Nevertheless,fractionationof humic substances on the basisof molecularweight
is a powerful and attractiveprocedure.Gel permeationchromatographyandultra-
filtration are techniqueswhich havebeenusedsuccessfullywith proteinsand car-
bohydratesfor fractionationsbasedon molecular size. Reasonablesuccesshas
beenachievedin using theserapid and reliable techniquesto purify and fraction-
ate humic substances.
Gel PermeationChromatography.The gels used for molecular size
chromatographyhave structuresconsisting of a system of pores, the sizes of
which are determinedby the degreeof cross-linkingin the polymer. When a solu-
tion of humic substanceis appliedto the top of a gel column and elutedwith sol-
vent, large moleculeswhich are unableto enterthe poresin the beadswill bypass
the beadsand will be eluted first from the column. Smallermoleculeswhich are
able to enter the poreswill have their passagethrough the column retarded,the
extentof which will dependon the actualsize and shapeof the molecule.The net
result is that the solutemoleculesare elutedfrom the column in orderof decreas-
ing molecular size, and, for a given family of macromolecules,in order of
decreasingmolecularweight also.
The rangeof molecularsizesover which the gel is able to differentiatethe
moleculeswill depend onthe type of gel chosen,but gels are availablewhich are
able to producefractions in the rangeof severalthousandsto millions of daltons.
Gels are selectedon the basisof the exclusionlimit of the gel and the molecular
weight rangeover which the gel is suitable.Given the extremepolydispersityof
humic substances,it is generallynecessaryto use a rangeof gels to achievesat-
isfactory fractionation.For discussionon the calibration of gels, see "Gel Chro-

matography."
It is important to selectgels that are inert to the solute moleculesso that
there are no chemicalor physical interactionsbetweengel and solute. Otherwise
the resulting separationcannot be entirely attributed to molecular size and/or
weight differences.Swift and Posner(1971) discussedtheseproblemsfully and
show that they can be largely overcomeby careful selectionof the gel matrix and
by the use of appropriatebuffer solutions.By successivelyusing gels of various
exclusionlimits and reapplyingthe excludedor includedportion of a given gel to
anotherwith a higher or lower exclusionlimit it is possibleto successfullysepa-
rate out fractions (Schnitzer & Skinner 1968, p. 41-SS). The method outlined
below is that usedby Swift et ai. (1992).
Materials
1. Sodiumtetraborate,(Na2B407)1% w/v, pH = 8.S.
2. SephadexG-7S, G-1S0.
3. Sephrose6B (Pharmacia,Uppsala,Sweden).
Method
Dissolve the sampleof humic acid in a boratebuffer (1 % w/v, pH = 8.S)
and run SO mL through Sephadex G-1S0in a preparative(S-cm diam.) column to
fractionatethe samplewith respectto molecularweight. Collect the eluateas 10-
mL fractions and read the absorbanceof theseat 400 nm.
Repeatthe aboveprocedurefour times and divide the eluateinto four parts
correspondingto four regionsof the elution curve, namely the excludedpeak(A)
and threesegmentsof the broadpeakof lower molecularweights(E, C, D) (Fig.
3S-2). Dialyze and freezedry eachof the four combinedeluates.
Redissolvethe four fractions using boratebuffer and then fractionatethem
using Sepharose6B, SephadexG-1S0or G-7S gels for fractions high to low mol-
ecularweight, respectively.
Refine the fractions by taking central cuts of the resulting four individual
peaks (shaded section, Fig. 3S-2) and dialyze these against distilled water
exhaustivelybefore freezedrying.
Comments
The volume of the excludedfraction (void volume) can be determinedby
using the nonadsorbingchemical,Blue Dextran2000 (Pharmacia,Uppsala,Swe-
den). To determinethe total effectivecolumnvolume of SephadexgelsuseN-2,4-
dinitrophenylasparticacid (a yellow compound),sucrose(C12H220 11 ), or glucose
(C6H 120 6) as the low molecular weightmarker. Except for gels to which it is
reversibly adsorbed(e.g., Sephadexgels) the sodium salt of 2,6-dichlorophenol
indo-phenol(a blue dye) can also be usedto determinethe total effective volume
of the column.
Typical elution patternsobtainedby proper applicationof gel permeation
chromatography(e.g., Dubachet aI., 1964; Swift & Posner,1971)shouldbe con-
sulted.The elution pattern shown in Fig. 3S-2, using borateor tris buffer is an
exampleof the type of curve that shouldbe obtained.
1030 SWIFf
Primary fractionation (Sephadex G-150)
A B c
:, SecondarY,!ractionations".
, , ,
Sepharose 66 : Sephadex G-150: Sephadex G-150 : Sephadex G-75
Vt Vo Vt Va
Elution volume
Fig. 35-2. Compositediagramshowinginitial elution pattern(absorbanceat 400 nm vs. volume elut-

ed)for whole humicacid (uppersectionof diagram)andpatternsfor the subsequent runsof the sep-
aratefractions, FI, F2, F3 and F4 (lower sectionof diagram).The shadedareasof the secondary
elution patternsrepresentthe "centralcuts" takento obtain the final secondaryfractions.
Ultrafiltration. In recentyearsa rangeof polymer-basedmembranefilters

have beendevelopedwith pore sizesrangingfrom severalmicrometersto a few
nanometers_The nominal molecularweight cut-off values of theserange from
700 to 1 000 000 daltonsand, theoretically,thesemembranefilters shouldbe able
to be usedto fractionatethe polydispersehumic substancesaccordingto molec-
ular weight. However, it has been observed that charge-chargeinteractions
betweenthe solute and the membranesurface may interfere with the filtration
processand humic acidsare highly charged.This fact togetherwith the difficul-
ties encounteredduring the manufactureof the filters and the unresolveddoubts
about the relationshipbetweenmolecularsize and molecularweight for humic
substances, createsomeproblemsconcerningthe overall usefulnessof the results
when studyinghumic substances.
Ultrafiltration is commonly usedas a purification procedurein which the
membraneis usedto separatethe wanted fraction from the unwanted fraction. It
is also usedin conjunctionwith other methodsof fractionation,such as gel per-
meationandultracentrifugationto increasethe flexibility of the fractionationpro-
cedure(Cameronet aI., 1972a).The method below has beenused to study the
complexingcapacityof humic substancesin natural waters(Smith, 1976). It is
possibleto alter the method bychangingthe membranetypes to alter the molec-
ular size rangeof the fractions, and by incorporatinga greaternumberof mem-
branesto increasethe numberof fractions. The membraneselectionwill have to
include somewith higher molecularweight cut-off valuesfor usewith soil humic

substances. It is advisableto begin the ultrafiltration using the membraneof high-
est cut-off value and work downwardsin size in order to limit clogging of the
filters.
Materials
1. Ultrafiltration membranes,XM-l00 (Amicon, Beverly, MA), XM-50
(Amicon, Beverly, MA), PM-lO (Amicon, Beverly, MA), YM-2 (Ami-
con, Beverly, MA), with molecularweight cut-offs 100 x 103, 50 X 103,
10 X 103, 1 X 103 daltonsrespectively.
2. Ultrafiltration cell.
Method
Flush the ultrafiltration cell and membranewith doubledistilled water until
the absorbanceof the filtrate solution recordszero magnitude[using ultraviolet
(UV)-visible spectroanalysis].Introduce a sample of known volume (e.g., 400
mL) into the cell and pressurizethe systemwith N2. Collect the ultrafiltrate and
retain in a preweighedflask. When the samplehas beenconcentratedto approx-
imately 20 mL, depressurizethe system and pour the concentrateinto a pre-
weighedflask. Determinethe volume of this fraction by gravimetryassumingthe
specific gravity of 1.0.
Sequentiallyfractionate the ultrafiltrate from the above step using mem-
braneswith the next lower molecularweight cut-off until that having the small-
est weight cut-off hasbeenused.Determinethe volume of the ultrafiltrate.
The abovefractions could be further refined by repeatingthe ultrafiltration
using the membraneassociatedwith their retention.Dialyze the fractions against
distilled water and freeze dry. Computethe massper volume of the initial ultra-
filtrate to determinethe relative fraction of the original massin the sample.
FractionationBasedon ChargeCharacteristics
The chargecharacteristicsof humic substancesin solution, originate pre-
dominatelyfrom the presenceof carboxylic acid, phenolic and enolic functional
groups.The amountof chargerelatesto the degreeof dissociationof thesefunc-
tional groupswhich is a function of the pH of the solution and the identity of the
counterionsin the surroundingmedium. By taking advantageof the differences
in chargedensity within a sampleof humic substancesit is possibleto fraction-
ate them accordingto their chargecharacteristics.
Electrophoresis.Electrophoresisis the movementof chargedspeciesin a
solution in response to an applied electrical potential. Traditionally, elec-
trophoresiswas carried out in free solution so that movementof the specieswas
able to be related to both chargeand size of a moleculeor ion. By carrying out
the processin a gel medium the movementof the chargedspeciescan be retard-
ed accordingto their size and the size of the poresin the gel. In this way the frac-
tionationof humic substances can be relatedto both the chargeand molecularsize
of the species.Experimentalconditionscan be modified by altering the buffer or
the characteristicsof the gel, to optimize the fractionationobtained.
1032 SWIFf
Gel electrophoresiscan be used as both a preparativefractionation tech-

nique, and as a meansof characterizinghumic substances.A preparativeelec-
trophoretic method(De Nobili et al., 1990a)which produces fractionatedhumic
material able to be usedin other analyseshasbeen outlinedbelow.
Materials
1. Potassiumdihydrogen phosphate
buffer (KH 2P04) 0.02 M pH = 7.0.
2. Glycerol (C3Hg03).
3. Polyacrylamidegel.
4. Sodium pyrophosphate,0.1 M.
5. Poly(vinylpyrrolidone).
Method
Preparethe gel rods by castingthree different overlayedgels of decreasing
acrylamideconcentration(9, 7, and 5%). Preparea samplecontaining15 mglmL
organic C, mix it with glycerol, and then filter the solution through a 0.2-llm
membranefilter beforeapplicationto avoid blocking the poresat the entranceto
the gel. Perform the electrophoresisin 0.02 M phosphatebuffer (PH = 7.0) using
15 rnA per rod (care must be taken to ensurethat the gel in the rods does not
becomeoverheated.
Stop the run when the front of the migrating bandreachesthe bottom of the
9% gel segment.Immediatelyafter the run take 2-cm long slicesof the front frac-
tion, the centerfraction and the tail fraction of the migrating band of humic sub-
stances.Discard the intermediatesections.If doing thefractionationin duplicate,
mince the correspondingslicesand extractthem threetimes with 0.1 MNa4PZ07.
Concentratethe fractionson PVP columns.Thesefractionsare suitableto be used
in the following electrofocusinganalysisor can be dialyzed and freezedried and
preparedfor someother analyses.
Comments
The equipment used by De Nobili et al. (1990a)consistedof 13 x 115 mm
polyacrylamidegel rods and a Bio-Rad 155 Electrophoresiscell, poweredwith
an LKB 2117 power supply (LKB, Bromma,Sweden)and cooledwith water cir-
culating at 4°C. Although presentedas a preparativemethod,it should be noted
that the amountsof materialobtainedby this proceduresare very small.
The techniqueof electrophoresishas been refined by incorporatinga pH
gradient into the gel system.The chargedspeciesmigrate until they reach the
position correspondingto their isoelectricpoint, a pH at which they ceaseto be
charged,in a processcalled electrofocusing.In this method,the gel has relative-
ly large poresso that molecularsize is not involved in the fractionation.
In electrofocusing
t as describedby De Nobili (1988), a pH gradient is set
up by the use of a mixture of compoundswhich are good buffers and able to act
as ampholytesin the separationmedium. The range in pI (isoelectricpoint) val-
uesof the ampholytesin the gel must be betweenthe pH of the electrolytesin the
anodeand cathodecompartments.The most acidic ampholyte(lowest pI) moves
to occupya position closestto the anodeand correspondinglythe most basicam-
pholyte occupiesthe spaceclosestto the cathode(highestpI), with intermediate
ampholytesmoving to occupy spacesarrangedin order of their decreasingacid-

ity in betweenthesetwo.
Electrofocusingis a potentially useful techniquefor characterizinghumic
substances.However, a numberof operationalaspectsindicate that any conclu-
sionsformed from using this techniqueshouldonly be tentative(Duxbury, 1989).
Duxbury believesthat the processis not electrofocusingbut is electrophoresisin
a pH gradient.Someof the reasonsfor this are that humic moleculesare not gen-
erally amphotericand once neartheir isoelectric point tend to lose their charge
and may not evenremainin solution.Thosethat diffuse pasttheir pI are no longer
chargedand so are not focusedback to that point, i.e., true electrofocusingdoes
not occur.
Other problemswhich potentially may be associatedwith the processare
the interactionsof humic moleculeswith the ampholinesand the aggregationten-
denciesof humic substances(Duxbury, 1989).
Fractionationof Polysaccharides
The polysaccharidefraction of soil organic matter will be made up of a

mixture of componentsof varying molecularweight, chargedensity and mono-
saccharidecomposition. Becauseof the similarity in some of the physical and
chemicalcharacteristics between polysaccharidesand humic substancesmany of
the inethodsused forfractionationof humic substances can be appliedto the frac-
tionation of polysaccharides,such as fractional precipitation(Parsons& Tinsley,
1961), electrophoresis(Mortensen& Schwendinger,1963; Barker et aI., 1965),
density fractionation(Strickland& Sollins, 1987), ion-exchangechromatography
(Finch et aI., 1966; Thomaset aI., 1967) and gel filtration (Barker et aI., 1965,
1967; Swinceret aI., 1968).The last two methodsare thosemost commonlyused.
lon-ExchangeChromatography
The method below is basedon that of Barker et ai. (1967). Initially, a pH
gradientelution is carried out to determinethe elution characteristicsof the par-
ticular polysaccharidesample.From this information, it shouldbe possibleto dif-
ferentiatelogical fractions within the sampleand the salt concentrationassociat-
ed with eachof thesefractions. The elution is then repeatedusing a fresh sample
of polysaccharide,and solutionsof specific salt concentrationsin a buffer to sep-
aratethe fractions. In this way the sampleis fractionatedon the basisof increas-
ing chargedensity.
Materials
1. Sodiumchloride, 2 M
2. Phosphatebuffer (0.0371 M KH 2P04, 0.0043M Na2HP04),pH = 6
3. DEAE-cellulose
Method
Preparea column (3.4 by 45 cm) of DEAE-celluloseand carry out a gradi-
ent elution with 0 to 2 M NaCl in phosphatebuffer. Monitor the elution profileby
collecting lO-mL fractions and analyzing thesefor sugars(Dubois et aI., 1956),
1034 SWIFT
and absorbanceat 400 nm. Determinethe numberof fractions to be eluted and

the NaCI concentrationat which their elution is maximized. For each fraction
selected,prepare600-mL portionsof buffer containingthe maximum NaCl con-
centrationassociatedwith that fraction. This preliminary run can be carried out
using a smalleramountof polysaccharidethan that usedin the main preparative
experimentthat follows.
Equilibratethe columnwith the phosphatebuffer. Dissolve200 mg of poly-
saccharidein 50 mL of phosphatebuffer, apply it to the column and elute it suc-
cessivelywith the 600-mL portionsof buffer preparedas above,using increasing
concentrationsof NaCl. Collect lO-mL fractions of the eluate and analyze as
above. Combine the polysaccharide-containing fractions for eachconcentration
of the salt, dialyze againstdistilled water and freezedry.
Gel Filtration
This methodis similar to the previousmethod,exceptthat the separationis
on the basisof size, ratherthan chargedensity.
Materials
1. Sodiumchloride (NaCl), 1 M.
2. SephadexG seriesgels.
3. Biogel P seriesgels.
Method
Pretreatthe gel and equilibratethe column using 1 M NaCI to eliminateion-
ic and adsorptioneffects. Load the column (54 by 3.1 cm) with 3 mL of solution
containing200 to 300 Ilg polysaccharide/mL.Elute the column with 1 M NaCl,
collect 5-mL fractions and analyzetheseusing absorptionmeasurements. Deter-
mine the amount of polysaccharide(Dubois et aI., 1956) as a function of the
elution volume and separateand combine fractions as outlined in "Gel Perme-
ation Chromatography."Suitablegels for polysaccharideanalysisare the Sepha-
dex G series(Pharmacia,Uppsala,Sweden)and Biogel P series(Calbiochem,
Los Angeles).
Density Fractionationof Soil Organic Matter
When soil organic matter becomesintimately associatedwith soil mineral

particles,the densityof the resultingcombinationis greaterthan that of the organ-
ic matteralone. By fractionatingthe soil on the basisof the density of the com-
ponentparticlesit is possibleto separatethe organicmatterinto fractions of dif-
ferent physical characteristicsand almost certainly different chemicalbehavior,
basedon their degreeof decompositionand associationwith mineral particles.
Density fractionation avoids the need for solvent extraction and decreasesthe
possibility of artifact formation. It also is believedthat this type of fractionation
may partially separatethe soil organicmatteraccordingto age,with the youngest
fraction being the lightest.
ORGANIC MA'ITER CHARACTERIZATION 1035
A numberof methodshave been developedto achievethis type of frac-

tionation. Theseinvolve the separationof the soil particleson the basisof their
density in water (Turchenek& Oades1979), and in organic liquids or salt solu-
tions of high specific gravity (Greenland& Ford 1964; Sollins et aI., 1984;
Turchenek& Oades,1979; Dalal & Mayer, 1986, Roth et aI., 1992; Baldock et
aI., 1990). By combiningparticle size fractionation(e.g., sieving, sedimentation,
and continuousflow centrifugation)with density fractionation, it is possibleto
separatethe organic matter in the soil into multiple fractions (Turchenek &
Oades,1979; Oadeset aI., 1987; Ducaroir et aI., 1990).
Initially, the sampleis ultrasonifiedto break up all the aggregatesand to
thoroughly disperseall the colloidal particles.The sampleis then sieved,and/or
separatedon the basisof settling time in water, before fractionatingtheseresul-
tant fractions on the basisof their density in a high specific gravity solution. The
methodbelow is basedon that of Baldock et ai. (1990).
Materials
1. Polytungstatesolution (Na3W04.9W03.H20).
Method
Add approximately20 g of soil to 50 mL of deionizedwater in a 150-mL
beakerand sonicateat constanttemperatureand at a medium output for 5 min.
Passthe dispersedsamplethrough a 53-J..lm sieve and then separatethe sieved
fraction into >2-J..lm and ~2-J..lm fractions using gravitationalseparationin deion-
ized water. Dialyze the ~2-J..lm fraction againstdeionizedwater and freezedry.
Combine the >53-J..lm and the >2-J..lm fractions and separatethese into a
light and heavy fraction by centrifugationin a sodium polytungstatesolution of
density 2.0 g cm-3. Separatethe light fraction from the supernatantby filtration
using a screenwith 5-llm mesh.
Resuspendthe sediment in the polytungstate solution, centrifuge and
remove and filter the supernatantas before. Repeatthis processuntil the super-
natantis clear, then washthe combinedlight fractions and dry at 45°C. Washthe
heavy fraction five times with deionizedwater and then dry it at 45°C.
This method gives three fractions; clay fraction (~2-J..lm diam. particles),
light fraction (>5-J..lm diam. particleswith a density of ~2.0 g cm-3) and heavy
fraction (>2-J..lm diam. particleswith a density >2.0 g cm-3). The methodcould
be extendedto achievea greaternumberof fractions by using different density
solutions(e.g., 1.6 g cm-3).
Comment
The earlier methodsinvolving the use of denseorganicliquids (Greenland
& Ford, 1964) are not generally used nowadays,due to the health, safety and
environmentalproblems associatedwith the use of halogenatedhydrocarbons.
Freezedrying, which is unlikely to alter the compositionof the fractions, may be
preferableto heatdrying.
1036 SWIFf
CHARACTERIZATION OF HUMIC SUBSTANCES
Many of the chemical, physical, and spectroscopicmethodsalready suc-

cessfullyusedin generalorganicchemistry,and in the study of naturally occur-
ring macromoleculeshave beenapplied to the study of soil humic substancesto
try to determinethe compositionand generalstructureof the componentmacro-
molecules.However, becauseof the heterogeneousand polydispersenature of
humic substances,and the complexity of the inter- and intramolecularreactions,
it is often difficult to interpretthe resultsof thesestudies.
Becauseof the diversenatureof humic substances andthe lack of any obvi-
ous meansof referencingdata,it is difficult to quantify the characteristicsof these
samples.A set of standard and reference sampleshave been preparedby the
IHSS to assistin the comparisonof humic substanceswithin and betweenlabo-
ratories,and theseare availablefor purchasefrom IHSS.
Characterizationby ChemicalMethods
The three generalcharacteristicsof a chemicalcompoundare the elemen-

tal composition,the arrangementof theseelementsin the chemicalstructure,and
the typesand locationsof the functional groupsin the structure.
Proceduresto determinethe type and relative abundanceof the elements
within an organic compoundare well defined and measurableexperimentally.
Such proceduresare not dealt with in this chapter. Becauseof their relatively
greateractivity, it is usually far easierto determinethe presenceof the various
functional groupsin large organiccompoundsthan it is to determinethe arrange-
mentsof the elementswithin the structures.However, once the elementalcom-
position and the functional group characteristicsare determined,and an estima-
tion of the molecularweight hasbeenmade(see"Molecular Size and Shape"),it
should be possible to make an assessmentof the type of structuresof which
humic substancesare composed.
Using generalmethodsof organicanalysis,it has beenestablishedthat the
major functional groupsin humic substancesare the carboxylic acids (andother
groups containing the C=O functionality), phenolsand alcohols, with Nand S
located in the minor functional groups.To some extent, the accuracyof these
analysesare open to question,but so long as they are reproduciblethey allow
comparisonsto be madeof humic substancesfrom different sources.
Acidic FunctionalGroup Concentrations

The acidic behaviourof humic substancesis such that they are considered
to be a mixture of stronger(mostly carboxylic acids) and weakerorganic acids
(mostly phenolic acids). The pKa of thesetwo types of acidic groupsare around
pH = 4.5 and pH = 10 respectively(Christensenet aI., 1976; Martell & Smith,
1977). Identical functional groups in a humic substancemay not have identical
pKa values(Perdue,1985).The actualpKa of an acid is not only dependenton the
natureof the acidic functional group, but also is dependenton the chemicalenvi-
ronmentin which it exists, as well as the natureof the compoundto which it is
bound. There would be considerableoverlap in the range of pKa valuesof the
acidic groupsin a solution of humic substances,and the overall rangeof all of

thesevaluescould spanthe pH rangefrom 0 to 13 (Perdue,1985). On the basis
of pKa measurements, it would not be possibleto distinguishunequivocallycar-
boxylic acidsfrom other typesof acids in humic substances (Perdue,1985).
A numberof different typesof methodsfor the analysisof acidshavebeen
usedto determinethe acidic functional groupsof humic acids(Stevensen,1994;
Perdue,1978; Schnitzer& Khan, 1972). Theseinclude direct and indirect titra-
tions, as well as nonaqueousand thermometrictitrations.
Total Acidity. The total acidity of a sampleshould include all the acidic
hydrogenspresent.Thus determinationof total acidity would requirethe useof a
basicreagentat high pH so that eventhe weakestacidspresentwould be account-
ed for. This type of methodhasseveralsourcesof error. First, the volume of base
requiredto reachequilibrium athigh pH valuesis relatively large comparedwith
the amountrequiredfor reactionwith the humic acid. Second,it is necessaryto
carry out the titration rapidly to limit the possibility of errors arising from any
base-catalysed side reactions(Perdue,1985). The resultsof the analysisare also
dependenton the conditionsin solution(e.g.,propertiesof the pH electrode,com-
position of the backgroundelectrolyte,etc.). Becauseof the foregoing consider-
ations, the determinationof the total acidity of a humic substanceis a good ap-
proximation only.
Direct TItrations. Thereis a tendencyfor the pH of the solution to decrease
with time underalkaline conditions,and this effect can be considerablyreduced
by carryingout the titration in the absenceof O2, This drift in pH is thoughtto be
due to the productionof acidsfrom side reactionsassociatedwith the presenceof
the alkali (Swift & Posner,1972). The titration can be carried either in the for-
ward direction (i.e., againsta dilute base)or in the reversedirection (i.e., against
a dilute acid). As humic substancestend to be insoluble in acid solution, it may
be difficult to achievedissolutionof the solid at moderatelyacidic pH values.For
this reasonthe reversetitration, which involves dissolving a known amount \ ,f
humic acid in KOH andtitrating with dilute acid, is morerapid but the error asso-
ciated with titrating in the high pH region (see above) is carried through the
whole titration.
The methodbelow is for the forward titration (Swift & Posner,1972) but
can easily be adaptedto the reversetitration.
Materials
1. W -Dowex 50W resin.
2. Potassiumchloride (KCI), 0.01 M.
3. Potassiumhydroxide,0.1 M.
Method
Preparea solution of humic substancefor titration by treating it with an
excessof H+-Dowex 50W resin. Preparea solutioncontaining5 mg of the humic
substances in 20 mL of 0.01 M KCI andtitrate it againststandardized0.1 M KOH
to pH =7.0 (arbitrary end-point).Add the KOH stepwise (e.g.,50 steps)as rapid-
ly as possiblebut still allowing for the solution to reachequilibrium beforeeach
1038 SWIFr
pH reading.Repeatthe procedureusing a blank solution. Constructa plot of pH

vs. titre volume for eachsolution to determinethe acidity of the humic acid as a
function of pH. All solutionsshouldbe CO2 free and the titration shouldbe car-
ried out underN2 at constanttemperature.
Total acidity (mmolc g-l) =(Vsample- VblanJ X MbaselWsample(g)
whereV blank and Vsample arethe titre volumes(mL) associatedwith the blank and
sampletitrations,respectively,and Wsample (g) is the massof the humic substance
usedin the experiment.(Note, mmolc designatedmillimole of charge.)
Comment
During the titration, the solution may not reachequilibrium in a suitably
shortperiodof time. In this event,choosea setperiodof time betweensuccessive
additionsof alkali or a set time over which the pH is relatively constant,as the
time at which the pH readingis taken.
Indirect titrations. The humic substanceis allowed to equilibrate in an
excessof Ba(OH)2' beforethe unusedalkali is titrated with a standardacid. The
total acidity of the humic substanceis determinedby calculatingthe differencein
titre volume with respectto pH betweenan analysisincluding the humic sub-
stanceanda blank analysisunderthe sameconditions.This methodhasbeenreg-
ularly usedas a meansof determiningthe total acidity of a humic substanceand
asit involvesa solutionof pH > 13, it is consideredthat the resultwould be a rea-
sonableestimationof the total acidity of the sample.
Traditionally, the methodusedhas not specifiedthe type of filter paperto
removethe bariumsalts,andit hasbeenfound (Davis, 1982)that their occurrence
in the titration solution would give an underestimationof the total acidity value.
The methodbelow is basedon that of Schnitzerand Khan (1972) with the
modificationsas suggestedby Perdue(1985).
Materials
1. Barium hydroxide(Ba(OH)2), 0.1 M.
2. Hydrochloric acid, 0.5 M.
Method
Placebetween50 to 100 mg of humic substancein the W-form in a 125-
mL ground-glassstopperedErlenmeyerflask. Add 20 mL of 0.1 M Ba(OH)2 to
the flask containingthe humic substanceand the sameamountto a similar flask
in which thereis no humic substance,to act as a blank. Displacethe air in each
of the flasks with N2, stopper,and then shakefor 24 h at room temperature.Fil-
ter the solutionsthrough0.45-Jlmmembrane,thenwashthe residuesthoroughly
with CO2-free distilled water. Using standardized0.5 M HCI and with the aid of
a pH meter, titrate the filtrate plus washingsof both the sampleand blank solu-
tions to pH =8.4. Determinethe total acidity as describedabovefor direct titra-
tions, using the following equation.
Total acidity (mmolc g-l) =(Vblank - Vsample) X MaciJWsample(g)

StrongerOrganicAcids (Carboxyl Groups).As there are no distinctive

equivalencepoints in the titration curvesof humic substances,the determination
of the pH at which a specifiedgroup of acidsis neutralizedis somewhatarbitrary.
Becauseweak acidsby definition must have a pKa abovepH = 7.0 this value has
sometimesbeenusedasmeansof differentiatingthe acidity associatedwith weak
and strongorganicacids(Gamble,1972; Burch et aI., 1978).Anothermore defin-
itive methodof delineationis to constructa first derivativecurve of titre volume
vs. pH to determinethe maximumchangein pH with changein titre volume. The
volume below this point is then taken to be that associatedwith carboxyl groups
and that abovedue to the phenolicgroupsand other weak acids.
Direct Titrations. The methoddetailedbelow is that of Oliver et al. (1983)
and is carried out on the microscale.It is possibleto scale all the apparatusup-
ward in order to carry out the procedureusing normal-sizedmeasuresand glass-
ware. The results of this analysisgive an operationalestimation of the acidity
associatedwith strongorganic acids and hencecarboxylic acids (Perdue,1985).
As for total acidity titrations, this methodcan be adaptedto be carriedout in the
reversedirection.
Materials
1. Sodiumhydroxide,0.1 M.
Method
Suspend10 mg of humic acid (H± form) in 2 mL of distilled water and
titrate the solution to pH = 7.0 with 0.1 M NaOH in 50-ilL increments.Carry out
a similar titration in the absenceof humic substances.
COOH acidity (mmolc g-l) - (Vsample- Vblank) X MbasJWsample(g)

Indirect Titrations. The most commonmethodusedto determinethe acidi-
ty associatedwith the strongerorganic acids is the calcium acetatemethod in
which the amountof acetic acid generatedfrom the reactionof calcium acetate
with humic substanceis determinedby titration with standardsodium hydroxide
solution.
There are a numberof operationalproblemsassociatedwith this method.
Becauseof poor buffering in the solution, the equilibrium pH of the solution is
determinedby the amountof humic acid added.Complexationof Ca2+ with the
humic acid increasesthe releaseof protons to solution increasingthe apparent
acidity (Perdueet aI., 1980). As in the barium hydroxide methodabove,it is also
importantthat the filtration step removesall solids from solution beforetitration.
This separationcan be achievedby using fine grade filtration (Davis, 1982) or
ultrafiltration (Perdueet aI., 1980).The methodbelow is basedon that of Wright
and Schnitzer (1959) with the alteration to filtration as suggestedby Perdue
(1985).
Materials
1. Calcium acetate[Ca(OAc)z], 0.5 M.
2. Sodiumhydroxide,O. 1 M.
1040 SWIFT
Method
Placebetween50 to 100 mg of humic substancein a 125-mL ground-glass
stopperedErlenmeyerflask. Add 10 mL of 0.5 M Ca(OAc)2 and 40 mL of COz-
free distilled water to the flask containingthe humic substanceand to a similar
flask in which there is no humic substance,to act as a blank. Shakethe flasks for
24 h at room temperature,filter the solutions through 0.45-l1m membrane,and
then wash the residuesthoroughly with CO2-free distilled water. With the aid of
a pH meter, titrate the filtrate plus washingsof the sampleand blank solutionsto
pH = 9.8 using standardized0.1 M NaOH.
COOH acidity (mmole g-l) = (Vsample- Vb1ank) X MbasJWsample(g)
Comments
De Nobili et al. (1990b) hassuggestedan alternativemethodfor the deter-
mination of the carboxylic acid contentof humic substancebasedon their solu-
bility in the presenceof cetyltrimethylammonium(CfA +). The CfA+ is very effi-
cient at quantitativelyprecipitatingthe humic substancesin solutions,thus mini-
mizing someof the problemsin the calcium acetatemethodoutlined by Perdue
(1985). The methodhas severalother advantagesincluding flexibility of pH for
the analysisand no apparentinterferencefrom the phenolicgroups.However,the
procedurehas not beenwidely testedand it is not possibleto recommendit as a
generalmethodat this stage.
Weaker Organic Acids (phenolic groups). The difference betweenthe
total acidity of a humic substanceand the acidity associatedwith the stronger
organic acids(carboxylic acids)is usually attributedto the phenolicgroups,even
thoughotheracidssuchas weak carboxyl groups,alcoholic groupsand enolsare
also probably involved. Before this assumptionis acceptedthe structuralimpli-
cationsof the valuesshould be investigatedfor consistency(see"GeneralCom-
ments"below). (Note, mmolf designatedmillimole of functional groups.)
Phenolic-OHgroups(mmolf g-l)
= total acidity (mmole g-l) - COOH acidity (mmole g-l)
Comment
It is also possibleto determinethe acidity characteristicsof the humic sub-
stance using nonaqueous.titration methods (Stevenson,1994) which tends to
enhancethe strengthsof the weak acids. A method using DMSO is detailed in
Yonebayashi and Hattori (1985)but othersolventswhich can alsobe usedinclude
pyridine (CsHsN) and dimethyl formamide.Thesemethodswarrant more inves-
tigation than they haveso far received.
Hydroxyl (OH) Groups

Total Hydroxyl (OH) Groups.The humic substancesare acetylatedwith
acetic anhydride in pyridine. Acetyl groups are then hydrolyzed to acetic acid
(C2H40 2), which is distilled and titrated with standardbase(Brooks et aI., 1958;
Schnitzer& Skinner, 1965).
Materials
1. Pyridine, 95% purity.
2. Acetic anhydride[(CH3CO)20] 97% purity.
3. Sodiumhydroxide,3 M, 0.1 M.
4. Sulfuric acid, 3 M.
5. Phenolphthalein,0.5% in 95% ethyl alcohol.
Method
Using 5 mL of equalpartsof pyridine and aceticanhydridereflux 50 to 100
mg of humic substancefor 2 to 3 h underan atmosphereof N2. After cooling the
mixture, pour it into distilled water and collect the precipitateby filtration. Wash
the precipitate thoroughly with distilled water and dry under a vacuum in the
presenceofP20 s. Reflux this acetylatedsample(50 mg) with 25 mL of 3 M aque-
ous NaOH solutionfor 2 h underN2. Cool the mixture, add 25 mL of 3 M H2S04
and 25 mL of distilled water and distill through a splashhead.
Collect 25 mL of distillate and titrate with standardized0.1 M NaOH using
phenolphthaleinasthe indicator. Add25 mL portionsof distilled waterto the dis-
tillation mixture and continuethe distillation. Repeatthe collection of distillate,
titration and addition of 25-mL portionsof the distillation mixture until the sam-
ple and reagentblanks titrate equally. Calculatethe acetyl and hydroxyl contents
as follows:
Acetyl (mmolf g-l) = (Vsample - Vblank) X MbasJWsample(g)
Hydroxyl (mmolf g-l) = acetyl content/(1 - 0.042 x acetyl content)
Comments
The factor of 0.042 aboveis a consequence
of the differencein molecular
weight of 42 betweenthe ROH and CH3COOR.The carboxylicand phenolicOH
groupscan be determinedusing a methylationprocedureusing methyl iodide as
outlined by Schnitzerand Skinner(1965)
Phenolic (OH) Groups. The phenolic content of a humic substanceis
assumedto be equivalentto the concentrationof weakerorganicacidsin the sam-
ple [see "WeakerOrganicAcids (PhenolicGroups)"], i.e.,
PhenolicOH (mmolf g-l)
= total acidity (mmolc g-l) - COOH acidity (mmolc g-l)
Alcoholic (OH) Groups.The alcoholic hydroxyl contentof a humic sub-

stanceis assumedto be the differencebetweenthe total hydroxyl contentand the
phenoliccontent.
Alcoholic OH (mmolf g-l)
=total hydroxyl (mmolf g-l) - phenolichydroxyl (mmolf g-l)

1042 SWIFT
Carbonyl (>C=O) Groups

Total Carbonyl (>C=O) Groups. The humic substancesare allowed to
react with an excessof hydroxylamine hydrochloride in methanoV2-propanoi.
The unreactedhydroxylamine hydrochloride is titrated with standardHCI04
solution (Fritz et aI., 1959)
Materials
1. Dimethylaminoethanol[(CH3)2NCH2CH20H]99% purity, 0.25 M (22.5
g in 2-propanol).
2. Hydroxylaminehydrochloride(NH 20H . HCI), 0.4 M (27.8 g in 300 mL
absolutemethanoland dilute to 1 L with 2-propanol).
3. Perchloricacid (HCI04), 0.2 M.
Method
Place50 mg of humic substancein a 50-mL ground-glass stoppered Erlen-
meyerflask. Add 5 mL of 0.25 M 2-diethylaminoethanolsolution and 6.3 mL of
0.4 M hydroxylaminehydrochloridesolution to the flask containingthe humic
substanceand to a similar flask in which there is no humic substance,to act as a
blank. Heat the flasks on a steambath for 15 min. Cool the solutionsand back
titrate potentiometricallythe excessof hydroxylaminehydrochloridewith stan-
dard perchloric acid solution. Determinethe end-pointby plotting milliunits vs.
milliliters of acid.
QuinoneGroups.In the naturalenvironmentalhumic substancesundergo

redox-typeinteractionswith a numberof soil componentsincluding iron (Waite
& Morel, 1984). This type of interactioncan be usedto determinethe concentra-
tion of quinoid groups presentin a sample. In the method outlined below the
humic substances are reducedin alkaline triethanolamine(TEA) solutionby Fe2+.
The excessreductantis back-titratedamperometricallywith standarddichromate
solution (Glebko et aI., 1970)
0= ( d = 0 + 2Fe2+-triethanolamine~ HO-<Q>-OH + 2Fe3+
Materials
1. Sodiumhydroxide,2 M.
2. Triethanolamine(TEA) 97% purity, 2 M.
3. Ferrousammoniumsulfatehexahydrate[FeS04(NH4hS04.6H20],
0.05
M.
4. Potassiumdichromate(K2Cr207), 0.004M.
Method
Dissolve 20 mg of humic substancein a solution of 45 mL distilled water,
25 mL of 2 M NaOH and 25 mL of 2 M TEA in a 200-ml tall form titration cell,
fit the lid and flush with N2 continually during the analysis.Stir the solutionmag-
netically for 30 min before adding 5 mL of 0.05 M ferrous ammoniumsulfate
hexahydratesolution and leave for 30 min. Using standardized0.004M potassi-
um dichromatesolution, backtitrate the excessreductantin solution at a constant
potential of -80mV, determinedusing a platinum foil (2.0 by 0.5 cm)/platinum
wire electrodesystem connectedto a polarograph.Carry out a blank titration
underthe sameconditions.
Quinoid C=O (mmolf g-l) = (Vblank - Vsample) X 6 x M(K2Cr207)/Wsample(g)
Ketonic C=O Groups

Ketonic C=O groups(mmolf g-l)
= total C=O groups(mmolf g-l) - quinonegroups(mmolf g-l)
GeneralComments
Typical values of functional group contentof soil humic substancesfrom
various origins are shown in Table 35-2. Becauseof the complexity of humic
substances,it is not easy to assessthe degreeof accuracyof analytical results.
However, it is possibleto determinethe consistencyof the data with respectto
the logical consequences of the relationshipsbetweeneach set of values. For a
detaileddiscussionof theseusesthe readeris referredto Perdue(1985).
With the determinationof the elementalcompositionand the numberaver-
age molecular weight, theoreticalvalues can be determinedfor the amount of
Table 35-2. Distribution of oxygen-containingfunctional groupsin humic and fulvic acids isolated
from soils of widely different climatic zones(in cmollkg)t (Stevenson,1994).
Climatic zone
Cool, Cool,
temperate temperate
Functionalgroup Arctic acid soils neutral soils Subtropical Tropical Range Average
Humic acids
Total acidity 560 57~90 62()"'{)60 63~770 62~750 56~90 670
COOH 320 15~570 390-450 42~520 380-450 15~570 360
AcidicOH 240 32~570 21~250 21~250 22~300 21~570 390
Weaky acidic + 490 27~350 24~320 290 2~160 20-490 260
alcoholic OH
QuinoneC=O 230
{1~180 {45~560 {8~150 {3~140 {1~560 {290
KetonicC=O 170
OCH3 40 40 30 3~50 6~0 3~0 60
Fulvic acids
Total acidity 1100 89~1420 64~1230 82~1030 64~1420 1030
COOH 880 61~50 52~960 72~1120 52~1120 820
AcidicOH 220 28~570 12~270 3~250 3~570 300
Weakly acidic + 380 340-460 69~950 26~520 26~950 610
alcoholic OH
QuinoneC=O 200 {17~31O {12~260
3~150
{120-420 {270
KetonicC=O 200 16~270
OCH3 60 30-40 8~90 9~120 3~120 80
t From Schnitzer,1977.
1044 SWIFf
unsaturationin the systemand henceupper limits of the main functional groups

suchas carboxylic acids, phenolsalcoholsand enols.
Characterizationby SpectroscopicMethods
The use of spectroscopicmethodsof analysisis importantin the character-

ization of humic substances.The individual techniquesmay yield only small
fragmentsof information but the importanceof this is magnifiedwhen combined
with information from other spectroscopictechniquesand/or other methodsof
characterization.
The aim of this sectionis to outline briefly the typesof spectroscopictech-
niquesgenerallyusedin the studyof humic substances, and how they haveassist-
ed in increasingthe knowledgeof thesesubstances.For more detailedinforma-
tion about the techniquesthe readershould consulta recenttext concernedprin-
cipally with their use. For a comprehensivereview of the applicationsof the
methodsto studieson humic substancesthe text Humic Substances II: In Search
of Structure (Hayeset aI., 1989) is recommended.
Ultraviolet-Visible Spectroscopy
The absorptionof electromagneticradiation in the UV (200-400 nm) and
visible region (400-800nm) is associatedwith the electronictransitionsof the
bondingelectrons.The absorptionof UV -visible radiationby organiccompounds
is due to the presenceof specific segmentsor functional groups (chromophores)
which contain unbondedelectrons(e.g., carbonyl groups, S, N or 0 atoms,and
conjugatedC-C multiple bonds). The electronic transition within a molecular
orbital is termedlocal excitation and the electronictransitioninvolving the trans-
fer of an electronfrom one chromophoreto another(e.g., from an aromaticring
to an OH group) is termedelectron transfer.
Generally,measurementof the absorbanceof a substanceis carriedout by
dissolving it in a solvent and determiningthe difference in absorbanceof that
solutionfrom that of a solutioncontainingonly the solvent, eitherin sequence(in
single beaminstruments)or directly (double beaminstruments).The absorbance
(A) is relatedto the concentrationof the samplein solution (c) accordingto the
Beer-Lambertlaw where
A = €/c
where I is the path length of the cell and € is the absorptivity

Applications of Humic Substances
The absorbancespectrumof a compoundis a characteristicwhich can be
usedin its identification. However, becausethe peaksof absorbancespectraare
relatively broad, it is difficult to identify a particularcompoundin a mixture of
evensimple moleculesand certainly not possiblein a sampleof humic substance.
The absorptionspectraof humic substancesare generallyfeatureless,consisting
of a relatively smoothcurve, with increasingabsorptionwith decreasingwave-
length. The curve representsthe summationof the absorbances of the component
chromophores.Even though it is not known what proportion of moleculescon-
tribute to the spectra,the smoothnessindicatesthat thereare a very large number

of different chromophoresin the sample.
Despite the apparentlack of detailed information in the spectra,different
samplesand fractions of humic substancesdo show slight variationswhich can
be measuredin a numberof ways to allow comparisonsto be made. For exam-
ple, the EJE6 value (the ratio of the absorbanceat 465 nm and 665 nm, of a dilute
aqueoussolution of a substance)is commonly used to characterizehumic sub-
stances(Konova, 1966; Chen et aI., 1977; Stevenson,1994) with the ratio for
humic acids beinggenerallylessthan five and that for fulvic acid more than five.
Becauseof the known differencesbetweenhumic acidsand fulvic acidsthe ratio
would appearto be a measureof the degreeof humification of a sampleof humic
substance.
The magnitudeof the absorbanceat a given wavelength variesslightly with
pH (MacCarthy & O'Cinneide,1974b; Baes& Bloom, 1990) which is probably
due to the ionisation of carboxylic and phenolic functional groups.This change
in absorbancewith pH is dependenton the wavelengthso that the EJE6 ratios of
humic acid and fulvic acid also vary with pH (Chen et aI., 1977; Ghosh &
Schnitzer,1979). In alkaline solution the ratio decreaseswith increasingpH but
below pH =5.5 the EJE6 decreases with decreasingpH (Chenet aI., 1977;Ghosh
& Schnitzer,1979).
Both absorbanceand the EJE6 ratio may be affected by variations in the
salt concentrationof the solution. It has thereforebeensuggestedthat the EJE6
ratio be determinedin 0.05 M NaHC03 (pH = 8) (Chen et aI., 1977).
A more detailedstudy of the dependenceof the absorbanceon pH can be
madeby referencingthe absorbancereadingagainsta spectrumof the sampleat
a different pH (Tsutsuki& Kuwatsuka,1979). Thesedifferencespectraare char-
acterizedby strongerpeaksbut the origin and causeof these peaks is not yet
understoodwith any certainty.
The UV -visible spectraof humic substancesoffer very little assistancein
the identification of their structure.One peculiarity is the observedabsorbanceat
wavelengthsabove500 nm when it has beenfound that compoundsdo not nor-
mally absorblight at thesewavelengths.It hasbeensuggestedthat this is due to
complex unsaturatedstructures(Tsutsuki & Kuwatsuka, 1979) or to electron
donor-acceptorcomplexes(Lindqvist, 1972, 1973).
Infrared Spectroscopy
Infrared spectroscopyis the study of the molecularvibrations of bonded
atoms.The frequencyof the absorptionis characteristicof the atomsin the bond
and the type of motion associatedwith the vibration. The observedfrequencycan
be usedto distinguishthe componentatomsas well as the bondingcharacteristics
of thoseatoms.Hydrogenbondingmay also be apparentas it causesgreatersep-
aration of the bond betweenH and the other atom in the covalentbond, thereby
decreasingthe frequencyof the absorptionand broadeningthe bands.
Detailed interpretationof the spectrato determinethe structureis possible
in simple molecules,but it is generallynot so in complex moleculesor mixtures
of molecules.However,useful informationcan be gainedby comparingthe spec-
tra of different samplesand noting changesin the spectraafter chemically alter-
1046 SWIFT
ing a sample.Using a variety of approachesit is possibleto gain informationon

the functional groupssuch as aromatic,aliphatic and quinonegroupsassociated
with the sample.
Infrared spectraare usually determinedover the frequencyrange4000 to
400 cm-l . Absorptionbandsin the frequencyregion 4000 to 1250cm-l are rela-
tively unaffectedby the remainderof the moleculeand this region is called the
"characteristicgroup frequencyregion." Absorptionbandsbelow 1250em-I are
affectedstrongly by the molecularstructureand so this region is called the "fin-
gerprint region."
The spectralanalysisis usually carried out on a solid sample,but as the
techniquedependson the absorptionof radiation, the sampleis dispersedin a
medium,which is transparentto infrared radiation.The pressed pellet methodof
samplepreparationrequiresa mixture of about 1 mg of humic m~terial per 100
mg of an alkali halide (KBr) both dried and pregroundvery finely. The mixture
is pressed(~7500 kg em-2) into a small disc about100 mm in diameterand 1 to
2 mm thick (Stevenson,1994). Potassiumbromide is transparentover the con-
ventionalsamplerange4000-400cm-l and so doesnot interferewith the spectral
analysis.
Alternatively the fmely ground and dry samplecan be mixed with a low
vapor pressure,medium molecular weight hydrocarbon [commonly "Nujol"
(Harry Holland & Sons,Burr Ridge, IL)]. This substanceis lesssatisfactoryas it
adsorbsradiationat 2900, 1460and 1375cm-l and so imposesits spectrumover
that of the samplebeinganalyzed,and is not very suitablefor the analysisof soil
humic substances.
A commonproblem is the incorporationof water into the samplematrix,
especiallywhen using KBr discs.The presenceof watercausesbandsat 3300to
3000 cm-l and 1720to 1500 cm-l regions.This can be reducedby heatingand
evacuatingthe die prior to pressingor using a nonhygroscopicpelleting matrix
(Stevenson,1994). The problemsassociatedwith water in the samplecan be
avoidedby using castfilms or solutionsin a cell (Bloom & Leenheer,1989).
Applications to Humic Substances
The infrared (IR) spectraof humic substancesare relatively simple with
only a few broadbandsand no well-defmed,sharppeaks,typical of the spectra
of singlecompounds.As for UV -visible spectra,theseundefinedbroadbandsare
evidencethat the functional groupsin the sampleof humic substances exist in a
wide variety of chemicalenvironments.Thereare often similarities in the gener-
al appearance of spectraof different samplesof humic substances.This doesnot
necessarilymeanthat the samplesarecomposedof moleculeswith similar struc-
ture, but doesindicatethat the overall functional groupcontentis similar.
Derivatization techniquesare frequently used to study the attribution of
observedspectralbandsto a particularfunctional group. An excellentoutline of
the bandsassociatedwith the main functional groupsand the type of derivatiza-
tion procedureswhich may assistin the assignmentof the bandsfor a particular
sample,is given in Bloom and Leenheer(1989).
In situationswherethe drying-outprocessmay affect the chemicalequilib-
rium in the samplebeing studied,it may be necessaryto analyzethe samplein
4000 3000 2000 1000

Wavenumber (cm- 1)
Fig. 35-3. Diffuse reflectanceFourier-transformedinfrared spectraof four InternationalHumic Sub-

stancesSociety humic acids(Neimeyeret aI., 1992). Note: the Kubelka-Munk units arise from the
application of the Kubelka-Munk transformation(Kortum, 1969) of the reflectancespectraand
result in spectrasimilar in appearanceto absorbancespectraobtainedfrom transmissionmeasure-
ments(Baes& Bloom, 1989).
Table 35-3. Fourier-transformedinfrared (FTIR) bandsof peathumic and fulvic acids(adaptedfrom

Baes& Bloom, 1989.
Band Assignment
3330-3380 OH stretchof phenolicOH (contribution from aliphatic OH, H20 and possibly NH)
3030 Aromatic CH stretch
2930 AsymmetricCH stretchof -CH2-
2840 SymmetricCH stretchof -CHz-
2600 OH stretchof H-bonded-COOH
1720 -C=O stretchof -COOH
1610 Aromatic C=C stretchand/orasymmetric-COO- stretch
1525 Aromatic C=C stretch
1450 -CH deformationof -CH3 and -CH bendingof -CHz
1350 Symmetric-COO' stretchand/or-CH bendingof aliphatics
1270 -C-OH stretchof phenolicOH
1225 -C-O stretchand OH deformationof -COOH
1170 -C-OH stretchof aliphatic OH
1070 C-C stretchof aliphatic groups
830 Aromatic CH out of plane bending
775 Aromatic CH out of planebending
1048 SWIFr
the aqueousstate. This is difficult in infrared spectroscopyas the absorption

bandsof water are large and centeredaroundthe major areaof interest,3400 to
1640 cm-I , and so interfere with the analysis.This may be overcomeby using
D20 insteadofHzO to shift the positionof the bands(MacCarthy& Mark, 1975).
More recently, Fourier Transform Infrared Spectroscopy(FTIR) and an adapta-
tion to this method using diffuse reflectance rather than transmitted light
(DRIFT), have been applied to the study of humic substances(Baes & Bloom,
1989; Niemeyer et ai., 1992; Ristori et ai., 1992). The Fourier transform tech-
nique eliminatesproblemsassociatedwith the presenceof water and improves
the resolution.The diffuse reflectance adaptationpermits the analysisof opaque
samplesso that whole soil samplescan be studied(Ristori et ai., 1992; Nguyen
et ai., 1991). Reflectancemethodscan be usedwith dispersiveinstruments,and
KBr pellets can be used with FTIR. The combination of FTIR with diffuse
reflectance,however,seemsto offer the best methodfor analysisof humic sub-
stances.
The small amountof samplerequired and the simplicity of the procedure
make IR spectroscopyone of the most commonly used methodsof analysisfor
humic substances.The information gainedfrom the spectrais in many casesnot
definitive asthe assignmentof bandsto particulargroupsis only tentativein com-
plex mixtures of compounds.The resultsshould be confirmed with other meth-
ods of analysis.
Examplesof typical FTIR spectraof humic acids are shown in Fig. 35-3.
The assignmentof structure generally given to the absorptionbands in humic
substances is well documentedin Stevenson(1994) and a summaryof the assign-
mentsis shown in Table 35-3.
NuclearMagneticResonance
When certainnuclei with a particulartype of spin are placedin a magnetic
field, they align themselvessuch thatsomehavetheir spin magneticmomentvec-
tors parallel to the field vector and are at slightly lower energyto othersthat are
antiparallelin the field.
If an oscillating magnetic field is superimposedon the steady magnetic
field with a perpendicularmagneticvector, then for a particularsteadymagnetic
field strength,absorptionof radiationwill occur at certainfrequenciesof oscilla-
tion, allowing transitions betweendifferent energy spin states.This resonance
condition can be met by varying the magnitudeof either the steadystatefield or
the oscillating field. In Fourier transform nuclear magnetic resonance(NMR)
spectroscopythe sampleis subjectedto a pulseof radio frequencyradiationcom-
prising all frequencies.
In a molecular environment, the atoms surroundinga nucleus partially
shield it from the magneticfield so that the frequencyof the oscillating field re-
quired to achieve resonanceis changed.As this shielding is a function of the
chemicalenvironmentassociatedwith the nuclei the resonancefrequenciesof the
atomsin the compoundare able to give information aboutthe chemicalstructure
of the compound.Commonnuclei usedfor the studieswith NMR are 1Hand 13e.
The NMR frequencyof a given nucleus(v sample)is measuredrelative to
that of a standardcompound{typically tetramethylsilane[(CH3)4Si] (TMS)) (v
reference).The frequencyfor resonanceis then given as the difference,or, chem-

ical shift (0) betweenthesetwo frequencies,expressedin partsper million (ppm).
o=(Vsample- Vreference) X 106/vreference

Commonsolvents usedin liquid state13C-NMR are NaOH and NaOO and
theseappearto display no absorptions.The OMSO-d6 provides a sharperspec-
trum but the 30 to 40 ppm region is obscuredunlessthe solventis depletedin 13e.
In IH-NMR the aqueousH is H20, 0 20 and aqueousNaOH obscuresthe 3 to 5
ppm region.
There are a numberof problemsassociatedwith liquid stateNMR such as
the amountof samplerequired(100-200mg), insolubility of some compounds in
suitablesolvents,interferencefrom water in IH-NMR and the long analysistimes
required (2-12 h are common for IH-NMR and up to 1 wk for 13C NMR). In
somesituationsthe dissolutionprocessinterfereswith the analysis.The develop-
ment of solid stateNMR hasavoidedtheseproblemsas well as achievinga high-
er signal to noise ratio, generallygiving greatersensitivity in the spectra.How-
ever, the technicalproblemsin getting good spectraare much greater.

Nuclearmagneticresonancespectroscopyis generallyusedto comparethe
difference in concentrationof the main functional groups betweensamplesof
humic substances.Both IH-NMR and 13C-NMR are able to be usedfor the char-
acterizationof humic substances,although the relatively low abundanceof 13C
(about 1.1%) in the latter techniqueplacesconsiderabledemandson instrumen-
tation and methodology.Nevertheless,thesetwo methodsprovide powerful tools
for studying humic substances(Wilson et aI., 1983; Wilson & Goh, 1983).
The mostcommontype of NMR now usedin the studyof humic substances
is called solid state CPMAS-NMR (cross-polarization,magic angle spinning-
nuclearmagneticresonance)(Wilson et aI., 1987; Krosshavenet aI., 1990). An
exampleof CPMAS 13C-NMR spectraof a soil humic substanceis shownin Fig.
Chemical shift, S(ppm)
Fig. 35-4. Typical CPMAS 13C-NMR spectrumof a soil humic substance.

1050 SWIFr
Table 35-4. Major proton resonanceof humic materialst(Wershaw,1985).

Chemicalshift, 1)
(relative to TMS as0) Assignment
ppm
13.0 Carboxylicacid protons
10.0 Hydroxyl protons
6.0-7.5 Aromatic protons
4.0-5.5 Lactoneprotons
3.7 Methoxyl protons
2.6 Aliphatic protonsattachedC atom IX to a benzenering:j:
1.3 Aliphatic protons~ to a benzenering:j:
0.9 Aliphatic protonsy to a benzenering:j:
t Spectrameasuredin deuterodimethylsulfoxide
[(CD3hSOj.
:j:--CaH;z-CpHrCrH2·
35-4. It is now possibleto comparethe NMR spectraof humic substancesin

whole soil sampleswith those of the associatedextractedfractions. Using this
type of approach,Krosshavenet al. (1992) concludedthat conventionalhumus
fractionationdoesnot significantly changethe contentof the different functional
groups in a sample.Assignmentsof chemicalshifts for IH-NMR and CPMAS
13C NMR are shownin Tables35-4 and 35-5, respectively.
Characterizationof humic substancesusing NMR spectroscopyis a rela-
tively new but widespreadtechnique,both in the number of researchgroups
involved and the type of researchbeing undertaken.Nuclearmagneticresonance
spectroscopyis proving to be important,both as a sourceof information relating
to the structureof humic substances,as well as supportinginformation gained
from other typesof analyses.However,there is a needto standardizethe proce-
duresand techniquesused,especiallyin solid stateNMR, to take full advantage
of this type of researchwith respectto humic substances.
Detailed information on the theoretical aspects
of NMR spectroscopy,and
the applicationof this theory to the studiesof humic substances,
is given in Hayes
et al. (1989).
ElectronSpin Resonance
When moleculescontainingunpaired electrons in a magneticfield are irra-
diated with electromagneticradiation, the electronscan be excited to a higher
energystateandthosewhich are alreadyat the higherenergylevel canbe induced
to move to the lower energystate.For a systeminitially in a stateof thermalequi-
librium there will be slightly more electronsin the lower stategiving a net ab-
sorption of energy. The absorptionof this energy is the basis of Electron Spin
Resonance(ESR) spectroscopywhich involves the study of molecules with
unpaired electrons (free radicals).
The energydifferencebetweenthe two spin states(aE) is
where h is Plank'sconstant,v is the frequencyof the electromagneticradiation,

g is the spectroscopicsplitting factor, ~ is the Bohr magneton(a constant)andHo
is the appliedfield.
Table 35-5. Chemical shift assignmentsin the CPMAS DC NMR spectraof fulvic and humic acids
(adaptedfrom Malcolm, 1989).
Shift range Possibleassignments
ppm
0-50 Unsubstitutedsaturatedaliphatic C atoms
10-20 Terminal methyl groups
15-50 Methylenegroupsin alkyl chains
25-50 Methine groupsin alkyl chains
29-33 MethyleneC (l,~, 0, E from terminal methyl group
35-50 MethyleneC atomsof branchedalkyl chains
41-42 (l-C in aliphatic acids
45-46 R2 NCH3
50-95 Aliphatic C singly bondedto one 0 or N atom
51-61 Aliphatic estersand ethers;methoxy,ethoxy
57-65 Carbon in CH20H groups;C6 in polysaccharides
65-85 Carbonin CH(OH) groups;ring C atomsof polysaccharides;ether-bondedaliphatic C
90-110 Carbonsingly bondedto two 0 atoms;C, anomericin polysaccharides,acetalor ketal
110-160 Aromatic and unsaturatedC
110-120 ProtonatedaromaticC, aryl H
118-122 Aromatic C ortho to O-substitutedaromaticC
120-140 Unsubstitutedand alkyl-substitutedaromaticC.
140-160 Aromatic C substitutedby 0 and N; aromaticether,phenol,aromaticamines
160-230 Carbonyl,carboxyl, amide,esterC atoms
160-190 Largely carboxyl C atoms
190-230 CarbonylC atoms
It is possibleto determinethe concentrationof unpairedelectrons(spins)

by comparingthe integratedabsorptionwith that of a standardunder the same
conditions, althoughthe accuracyof this methodis believedto be quite low. Mea-
surementof the field strengthand the frequencyof the electromagneticradiation
at resonancedeterminesthe g-value, the magnitudeof which is associatedwith
the molecularstructureof the molecule.The hyperfinestructureof the spectracan
be used to determinethe number and types of nuclei interacting with the free
electron,and hencethe structureof the molecule.The concentrationof free radi-
cals is determinedby the width of the absorptionline.
Application to Humic Substances

The ESRspectraof humic substances are generallyonly a single line which
limits the amountof information that canbe gainedfrom the analysis.The hyper-
fine structurein ESR spectra,typical of simple systems,is absentalthougha few
instanceshave beenreportedwith splitting (Atherton et aI., 1967; Senesiet aI.,
1977). An exampleof an ESR spectrumof a humic substanceis shown in Fig.
35-5 with sometypical valuesof the measurableparametersfor soil humic sub-
stancesin Table 35-6. Thesevaluesare only meantto be indicativeof thosecom-
monly found as the magnitudeis dependenton solution pH, metal ion contentand
solventeffects.
Resultsobtainedfrom this type of researchhave led to the realizationthat
the free radicalsin humic substancesare remarkablystablewith respectto time
and chemical attack. This stability has been attributed to the stabilizationof an
unpairedelectron over an aromatic system(Theng & Posner,1967) suggesting
that humic substancescontain free radicals of the semi-quinonetype (Steelink,
1052 SWIFT
-5 o +5
Fig. 35-5.ESRspectraof a podzol FA in NaOD-D20: (A) immediatelyafter mixing fulvic acid with
NaOH; (B) 48 h after mixing. g =2.0040;line width =5.0 x 10-4 T; frequency=9.451 GHz; field
center=3342.0(Steelinket aI., 1983).
1964). Basic solutionsof humic substances containexceptionallylong-lived free

radicals.It hasalso beenfound that the spin content generallyincreasesmarked-
ly with increasingpH.
The concentrationof free radicals in humic substancesis relatively small
(Schnitzer& Skinner,1969),and as the concentrationvarieswith the type of sol-
vent usedin the extraction procedure it may evenbe an artifact of extractiontech-
nique used(Hayeset aI., 1975). Other techniquessuchas NMR and infrared (IR)
are more useful than ESR in characterizingthe functionality of soil humic sub-
stances.However, ESR is a source of additional information and should be
included in the generalresearchprocedure.If the techniquewas to becomemore
reliable and informative the potentialof its use would increaseas it is concerned
with one of the more reactivefractions of humic substances, the free radical con-
tent.
Pyrolysis-MassSpectroscopy
Pyrolysis(Py) is the degradationof a substancethrough the action of heat.
This processis usually carried out in a vacuum,or in a rapid streamof inert gas
to restrict the formation of secondaryproducts.The absorptionof thermalenergy
causesexcitation of the bond vibrational modesresulting in the cleavageof the
weakerbonds.The numberandvariety of the productsformed by using thistech-
Table 35-6. Electron spin resonanceparametersfor various humic and fulvic acids (Steelink et aI.,
1983).
Free-radical Spectroscopic
Sample State concentration Line width splitting factor Reference
spinsg-1x T (g value)
10-17
Soil humic Solid 5-10 4.8 x 10-4-5.2x 10-4 2.0032-2.0047 Chenet al. (1978)
acid
Soil fulvic Solid 1-2 4.5 x 10-4-7.5x 10-4 2.0037-2.0050 Chen et al. (1978)
acid
Soil humic Aqueous 2.1 2.5 x 10-4 2.0037 Varadachariet al.
acid solution (1983)
Soil fulvic Aqueous 1.3 2.5 x 10-4 2.0038 Varadachariet al.
acid solution (1983)
ORGANIC MAlTER CHARACTERIZATION 1053
nique to study humic substancesis very large. It is thereforeessentialto incorpo-

rate massspectrometry(Py-MS) or gaschromatography(Py-GC) into the system,
to separateand identify the reaction products.A further, and highly desirable,
improvementon the techniqueis to use a combinationof both gas chromatogra-
phy and massspp.ctrometry(Py-GC-MS).This systemallows the volatile reaction
productsto be separatedprior to the analysisusing the massspectrometer.Alter-
natively, soft-ionizationtechniquessuchas field ionization(FI) and field-desorp-
tion combinedwith pyrolysis massspectrometry,(Py-FIMS, Py-FDMS) extend
the rangein size of molecularions which can be observedby Py-MS by includ-
ing the small and larger molecularions, respectively.
The datagainedfrom eachanalysiscan be usedas ajingerprint of that sam-
ple under the particular pyrolysis conditions used. This fingerprint can then be
comparedwith the resultsof analysison othersamplesof humic substances or on
known chemicalcompounds.Becauseof the large amountof datacollectedusing
these analytical techniques,multivariate analysiswith the aid of computersis
usedto collate the results(Howarth& Sinding-Larsen,1983).The theoreticaland
technicalaspectsof thesepyrolysis techniquesand the methodsof treatmentof
the accumulateddata are describedby Bracewellet ai. (1989).

Pyrolysis techniquesare used to study humic substancesin an attempt to
correlatesamplesof humic substances with their parentbiopolymers(Meuzelaar
et aI., 1982). Evidencefrom the studiesusing pyrolysis indicate that the humic
macromoleculeshave polysaccharides,polypeptides and lignins incorporated
into their structures(Greenland& Oades,1975; Halma et aI., 1978; Bracewellet
aI., 1980). Pyrolysis techniqueshave also been used to determine differences
betweenthe various fractions of humic substances(Saiz-Himinez et aI., 1978,
1979). The studiescarried out so far indicate that there is very little variation in
the pyrolysatesof humic acidsfrom different soils anddifferent soil types,where-
as the pyrolysatesof a fulvic acid fraction are dependenton the origin of the soil
and the methodof extraction(Bracewellet aI., 1989).
One of the most importantaspectsof pyrolysis with respectto applications
to humic substances is the ability of this techniqueto analyzewhole soil samples.
This permits a relatively rapid analysis of soil organic matter in general and
hence,comparisonof organicmatterassociatedwith soils from different origins
(Baldock et aI., 1991) or as a result of different managementpractices(Schulten
& Hempfling, 1992). Figure 35-6 showsPy-FI massspectra[intensity of the ion
vs. the massof m/z (massto chargeratio of the ion)] of the pyrolysatesobtained
from a soil as a result of different intensitiesof crop rotation and manuretreat-
ment. Thesetwo soil samplesexhibitedalmostidentical total concentrationsof C
and N. Schulten and Hempfling determined that there was little difference
betweenthe pyrolysatesof the soil samplesin the massrangem/z 50 to 200, but
in the higher molecular weight products of the pyrolysis the intensities were
greater for the soil that had less intense cropping and had been treated with
manure.
Comparisonof Py-FIMS spectraof whole soil sampleswith those of the
extractedhumic fractions permits the verification of consistencyof resultswhen
1054 SWIFf
5.0 96
4.0 58 r--xl1f - (a)
3.0 82 110
i 2.0
l1.0
67
1
208
,!;' 0
Cf)
c: 50 100 150 200 250 300 350 400
~ 5.058-_96
(b)
c:
.£ 4.0
«i
~ 3.0 110
2.0
1.0
340
0
50 100 150 200 250 300 350 400
m/z-
Fig. 35-6. AveragedPy-FI massspectraof soil samplesfrom the samesite and with the sameNand
C content, but under different managementsystemsand (a) with manure addition, (b) without
manureaddition.
characterizingthe fractions(Hempfling & Schulten,1991).It hasbeenfound that

humic acid extractsfrom soils are enrichedin polypeptidesrelative to the whole
soil pyrolysate, and the fulvic acid extracts are deficient in polypeptidesbut
enrichedin polysaccharides or pseudo-polysaccharides (Haider et aI., 1977). The
"humin" extracthad a similar fingerprint to that of the humic acid indicating that
the nonextractedmaterialof a soil humic substancehassimilar chemicalcontent
to that of the extractedmaterial.The effectsof different methodsof extractionof
fractionsof humic substances havealso beenobservedusing Py-FIMS by Haider
& Schulten(1985).
Pyrolysis techniquesalso provide supportingdata for the resultsobtained
when using other types of chemicalanalysisand instrumentalanalysis.Zech et
al. (1990) used chemical analysiscombinedwith IR, 13C-NMR and Py-FIMS
instrumentalmethodsto characterizeand comparethe organic matter in soils
from different soil horizons. Beyer et al. (1992) concludedthat the Py-FIMS
resultsconfirmed andextendedthe resultsobtained fromwet chemistrymethods
and NMR methods.
Pyrolysistechniquesusedto study whole soil samplesallow the character-
ization of the organic matter without going through the potentially destructive
and time-consumingprocessesof extraction and purification of the humic sub-
stances.It is for both of thesereasonsthat this approachto organicmatterchar-
acterizationis receivingincreasinginterestand holds considerablepromise.
FluorescenceSpectroscopy
The UV -visible radiationabsorbedby a moleculemay be dissipatedas heat
and/or electromagneticradiation at a longer wavelengththan the incident radia-
tion. This emitted radiation is known as fluorescenceand it is a characteristicof
humic substances.
There are three types of spectragenerally associatedwith this technique.
Excitation spectraare obtainedby scanningthe incident radiation and determin-
ing the intensity of radiation emittedat a fIxed wavelength.Emission spectraare

obtainedby fixing the incident radiation and determiningthe intensity of radia-
tion emitted over a wavelength range. Synchronous excitation spectra are
obtainedby settingthe differencebetweenthe emitted and absorbedradiation to
be a constant(LlA) and determiningthe intensity of the emitted radiation for a
rangeof wavelengths.
In general,only a small fraction of moleculesare known to fluoresce,and
so the proportionof moleculesin a sampleof humic substancecontributingto the
observedfluorescenceis relatively small. Additionally, this observedfluores-
cenceis not the total fluorescenceas humic substancesabsorbradiation over a
wide rangeof wavelengthsand someof the fluorescenceradiation will be reab-
sorbedby the sample.
Fluorescencespectroscopyhasbeenusedto characterizesamplesof humic
substances but for a numberof reasonsit hasnot gainedgeneralacceptance(Sen-
esi, 1990). Becauseof the reabsorbanceof the fluorescedradiation the relative
intensity of the observedradiation at a particularwavelengthis a function of the
concentrationof humic substancesin solution (Spark & Swift, 1994). When
using UV -visible absorbancespectroscopyincreasingthe solution concentration
of dilute solutionsincreasesthe absorbancein a regular manner.In fluorescence
spectroscopyincreasingthe concentrationof humic substancesin dilute solution
doesinitially increasethe observedintensity of fluorescence,but eventuallythis
reachesa maximumand then beginsto decrease.The concentrationat which the
fluorescencereachesa maximum is dependenton the wavelengthof excitation
and emissionas well as on the natureof the humic substance.As yet it has not
beenpossibleto correlatethe detail in the observedfluorescencespectraof humic
substanceswith known characteristicsof thesesubstancesso the usefulnessof
this techniqueis limited.
Measurementsof PhysicalProperties
The physical propertiesgenerally determinedfor humic substancesare

molecularsize, shapeand weight, and chargecharacteristics.The principles of
thesedeterminationsare theoretically soundly based,but in practice the results
are somewhatlimited in value by the large variation in rangeof the propertiesin
eventhe most highly fractionatedsamples.
Molecular Size and Shape

Molecular size and shapeof humic moleculesare two of the most elusive
propertiesof humic substances.Considerableresearchon humic substanceshas
beendirectedtowardsthe determinationof theseproperties,but as knowledgeof
humic substanceshas increased,so has the realizationof the complexitiesasso-
ciatedwith assigningvaluesto the size and shapeof humic molecules.It is prob-
ably becauseof these problems, as well as the rapid developmentof spectro-
scopicinstrumentation,that this areaof humic substance research
hasmadelittle
progressin recentyearswith respectto other areas.
1056 SWIFT
The techniquesgenerally used to study the molecular size and shapeof

humic substancesare gel permeationchromatographyand viscosimetry.A vari-
ety of other techniquessuch as light scattering(see "Light Scattering"), Flow
Field Flow Fractionation(FFFF) (Becketet aI., 1987), ultracentifugation(Posner
& Creeth, 1972; Ritchie & Posner, 1982), and colligative data calculations
(Reuter& Perdue,1981) have also beenused.
Gel Chromatography.The generalprinciplesrelating to gel chromatogra-
phy have previously been discussedwith respectto the fractionation of humic
substances(see "Ion-ExchangeChromatography").For moleculesof the same
shapeand structureit is possibleto calibratethe gel column with respectto elu-
tion volume vs. molecularweight using known compounds,and hence determine
the molecularweight of an unknowncompound.As elution time is dependenton
the molecularshapeand structure,the elution time for two moleculesof the same
molecularweight, one of which is sphericaland the other a long chain molecule,
may be quite different.
The elution volume for a given solute is dependenton the geometryof the
column, the length of the gel bed and differencesin the packing density, and so
cannotbe used to characterizea solute. The distribution coefficient (Kd) can be
usedto comparethe migration ratesof different solutesin different experiments.
where Vc is the elution volume, Vo is the void (excluded)volume Vi is the inner

volume (i.e., the solvent volume inside the gel beads).Becauseof the difficulty
in measuringthe magnitudeof Vi> the total volume of the column (VI) is usedto
determinean approximationof Kav accordingto the equation

A variety of media are now used in this type of work including dextran
(C6H lO OS)m polyacrylamide {[-CH 2C(CONH2)O-]}, agarosegels and porous
glassbeads.The elution characteristicsthat can affect the elution time are inter-
actionsbetweenthe gel and solute,ionic strengthand pH of the solution (Tsutsu-
ki & Kuwatsuki, 1984), concentrationof the sampleand the type of buffer used
(Swift & Posner,1971). Swift and Posner(1971) found that two buffer systems
tris (2-hydroxy-2-aminopropane-1,3-diol) and boratebuffers at pH 9, are partic-
ularly well suitedfor use with humic substances.
Calibrationsof the gels are usually madeusing polysaccharides or proteins
of known molecularweights.It hasbeenfound (Cameronet al., 1972b)that there
is a bettercorrelationbetweenhumic substanceswith polysaccharidesthan with
proteins.This may be becausethe more diffuse or open molecularconfiguration
of polysaccharidesis closer to that of the humic substancesin solution, than are
the condensedprotein configurations.
Calibrationof the column is carriedout by determiningthe elution charac-
teristics of humic substancesof which the molecularweight has beenestimated
using someother technique,such as ultracentifugation(Cameronet aI., 1972b).

Dawsonet al. (1981) had somesuccess calibrating the column using chemicals
thought to be similar to thoseof humic substances.
Ultracentrifugation.Ultracentrifugationis the centrifugationof solutions
at high speed(100 0()(}-600 000 g) under a vacuum to causesedimentationof
solute species.The velocity of sedimentationcan be used to separatemolecules
and to determinemolecularshapeand size.
Sedimentationof a macromoleculerequiresits movementthroughsolution
which may be hinderedby a number of factors including frictional resistance
from the solvent, molecularenlargementdue to solvation, and interactionswith
other sedimentingmoleculesand/orsolutecomponents.In addition to thesefac-
tors if the sedimentingmolecule is chargedits sedimentationvelocity will be
decreaseddue to the accumulationof small counterions behindit. The sedimen-
tation velocity may also be affected by the sedimentationcharacteristicsof the
buffer or backgroundelectrolyte.
A numberof different techniquesare usedin ultracentrifugation.Sedimen-
tation velocity involves the useof high rotor speedand requiresthe development
of well-formed boundaries,which is not possiblein polydispersesystems.Sedi-
mentation equilibrium useslower rotor speedsfor long periodsof time, until the
macromoleculesin solution are in equilibrium. The number average (Mn),
weight-average(Mw) and z-average(Mz) molecularweightsof the samplecan be
determinedusing this methodfor sampleswith a limited amountof poydispersi-
ty without the need forany other measurements.
Both of the abovetechniquesrely on the useof analyticalcentrifugeswhich
haveoptical detectionsystemsattached.When preparativecentrifuges,which do
not haveassociatedoptical systems,are used,it is necessaryto carry out the sep-
arationusing the density gradient technique to overcomeproblemsof convection
and mechanicaldisturbance.The densitygradientis formed in the centrifugetube
by using increasingconcentrationsof a solute such as sucrose,an inorganic salt
or mixturesof H20 and D20.
Applications of Humic Substances
Thereare a numberof problemsassociatedwith applying this techniqueto
the study of humic substances.The color of humic substancesrestrictsthe trans-
missionof light and hencelimits the accuracyof the refractive index determina-
tion. The uncertaintyof the molecularstructureand conformationof humic sub-
stancesmakesit difficult to nominatesuitablecompoundsto be usedas internal
standards.However,the methodis still valuablein being able to determinerela-
tive values of molecular weight betweendifferent fractions, or changesin the
molecularweight of a fraction associatedwith certain conditionsor treatments.
It is necessaryto use a relatively high concentrationof backgroundelectrolyte
concentration(0.1-0.2 M) when analyzing humic substancesto limit interfer-
ences associated with the highly chargednature of the moleculesin thesesam-
ples. This concentrationof electrolytewill also influence the configuration and
degreeof expansionof the humic substancein solution.
The three types of techniques,sedimentationvelocity (Stevensonet aI.,
1953; Cameronet aI., 1972b),equilibrium ultracentrifugation(Posner& Creeth,
lOS8 SWIFf
1972) and densitygradientmethods(Ritchie & Posner,1982) have beenapplied

to samplesof humic substances.In the latter two techniques,carried out using
fractionatedsamplesof humic substances,the large variation in the values of
(Mn), (Mw) and (MJ are strong evidencefor the polydispersityof humic sub-
stances(Swift, 1989). Becauseof the highly polydispersenatureof humic sub-
stancesthe densitygradienttechniqueis potentially the most applicable,particu-
larly for preparativeprocedures.
Light Scattering.The scatteringof light by particles in solution can be
used to determinetheir molecularweight, a techniquecommonly used for the
study of polymers.The molecularweight (M) of the particlesin a monodisperse
systemat a densityp canbe determinedby the usingthe relationshipM =PV. The
volume of the particle (V) is determinedusing the Raleigh equationwhere the
ratio of the intensity of the scatteredlight [/(9)] to the intensity of the incident
light (/0) (Hunter, 1989) is given by
1(9) = 91t2W(n2 - 1)2(1 + cosZS)

10 2,.z~..4(n2 + 2)
whereN, and r are the numberand radiusof the of particlesrespectively,n is the

refractiveindex of the light scatteringparticlesrelative to that of solvent,A. is the
wavelengthof the light and9 is the anglebetweenthe incidentbeamand the scat-
teredbeam.It is assumedthat the particlesare uniform in size and sphericaland
that the particle size is small relative to the wavelengthof the scatteredlight.

Very little use has beenmadeof this techniqueto study humic substances.
Theoretically,light scatteringtechniquesmay be extended tostudy the changes
in solution in responseto changein pH (Wershaw,1989) and to study the com-
plexation of humic particlesin the presenceof heavy metal ions (MacCarthy&
Mark, 1976; Ryan & Weber,1982).
In practiceproblemsarisefrom the absorptionand fluorescenceof light by
humic substances which would significantly interferewith the analysisat wave-
lengthsbelow 500 nm (Wershaw,1989).Reid et aI., (1991) usedlaserlight at 633
nm to study the aggregationcharacteristicsof humic moleculesin solution.Using
laserlights with the new techniqueof quasi-elasticlight scatteringis also a pos-
sibility (Wershaw,1989).
Viscosity. Information about the size and shapeof humic and fulvic acids,
as well as their weight, polyelectrolytic characteristicsand interactionbehavior
with other macromoleculescan be determinedfrom measurements of viscosity.
The equipmentrequiredfor this type of analysisis relatively inexpensiveand the
experimentalproceduresare easierto conductthan other methodsavailable for
determiningthe characteristicsof humic substances.However, it must be noted
that the valuesobtainedfrom the measurements tend to be relative valuesonly.
Both the theory behind this technique,as well as the experimentalinformation,
are well documentedby Clapp et al. (1989).

The viscosity of humic substances in solution hasbeenfound to dependon
both the pH and the salt concentrationof the solution (Chen& Schnitzer,1976;
Stevenson,1994).The results obtainedfrom studiesusing different concentra-
tions of salts (Ghosh & Schnitzer,1980) support the theory that humic macro-
moleculesin solution are polyelectrolytic in characterwith the degreeof coiling
dependenton the concentrationof the salt. It hasbeenfound that for a given mol-
ecularweight, humic substancesfrom different sourcesexhibit varying degrees
of expansion(Visser, 1985).
Besidesbeingusedsimply as a methodof characterizinga humic substance
(Tomar et aI., 1992), viscosity measurementshave also been used to study the
interactionsbetweencontaminantsand humic substances(Chenet aI., 1992) and
betweensoil mineralsand humic substances(Zhanget aI., 1991).
OtherTechniques
A numberof other techniquesare used to characterizehumic substances.

Two of theseare chemicaldegradationstudiesand electronmicroscopy.Neither
of these techniquesare discussedin detail, and the reader is referred to other
sourcesof information.
ChemicalDegradation
The macromolecularsize,the diversearrangementof functional groupsand
the multitude of interactionspossiblebetweenthesegroupsmake the task of de-
termining the structure of humic substancesextremely difficult. The aim of
chemical degradationstudiesof humic substancesis usually to break down the
large macromoleculesinto smaller, more recognizableunits. From theseunits it
may then be theoretically possibleto determinethe origin of the parentmolecules
and, hence,build up a picture of the structureof humic substances.
The processof rebuilding the molecularstructureof humic substances from
the componentmoleculesis complex in itself, but this work is mademuch more
difficult by the inability to control the degradativeprocessto achievea satisfac-
tory set of simpler molecules.The ideal method should yield high amountsof
degradativeproductsof moderatemolecularcomplexity(Stevenson,1994).If the
degradationproduceris too mild, the total amountof eachtype of recognizable
molecules is too small to be determinedanalytically. And, if the degradationpro-
cedureis too extreme,the types of recognizablemoleculesproducedare so sim-
ple that little information is gainedaboutthe natureof the parentmolecule.Some
of thoseproblemsmay be overcomeby using a combinationof different proce-
duresand a rangeof severity (Stevenson,1994; Hayeset aI., 1989).
The types of chemicalproceduresusedin degradativestudiesinclude oxi-
dation, reduction,hydrolysisand depolymerization.
The compoundsproducedwhen applying theseproceduresto humic sub-
stancesare numerous,including complexphenolicand benzenecarboxylicacids,
hydroxycarboxylicacids, nitrophenolsand polycyclic ring compounds(Steven-
son, 1994). The text Humic Substances II. In Search of Structure (Hayeset aI.,
1989) providesa detailedreferencefor this type of research.
1060 SWIFf
ElectronMicroscopy
Electron microscopy has been used to study the physical appearanceof
humic substances.This technique requires that the sample is dried which,
undoubtedly,alterssomeof the physicalcharacteristicsof the humic substances;
It is known that factors,suchas pH, ionic strength,metal complexationand con-
centration,affect the size and shapeof the dried particles of humic substances
(Chen & Schnitzer, 1989).The method of drying the sample also affects the
resultantappearanceof the particle.
It is becauseof these types of problems that the results from electron
microscopyare thought to accentuateor distort the characteristicsof humic sub-
stancesin the natural environment,but do provide an insight into the possible
physicalappearance of thosesubstances.Recentadvancesin electronmicroscopy
allow samplesto be studiedunder moist conditions,and it is important that this
techniquebe applied to the study of humic substances.
CHARACTERISTICSOF SOIL POLYSACCHARIDES
Many of the chemicaland physical methodsfor characterizing humicsub-

stancescan similarly be used to characterizesoil polysaccharides(Cheshire,
1979; Cheshire& Hayes,1990). This sectionwill concentrateon thosemethods
peculiarto polysaccharides.
Determinationof MonosaccharideComposition
Characterizationof a sampleof soil polysaccharideshas similar problems
to humic substances with respectto the complexity of the samplehinderingmost
attemptsat detailed analysis.However, becauseof the polymeric nature of this
soil fraction it is theoretically possible to convert the larger macromolecules
using hydrolysisprocedures,into smallerand perhapsmore recognizableunits.
Hydrolysis of polysaccharidesusually involves the reactionwith hot acid
to the sample to break the glycosidic bondsjoining the different sugars.This
methodis generallyusedto carry out an estimationof total sugars.Becauseof the
different naturesof the monosaccharide units, the natureof the hydrolyzing acid
and the analytical conditionsneededto effect link breakageswithout destruction
of the monosaccharideunits dependson the type of sugar units present.Side-
reactionssuchas acid-reversion[Acid-catalyzeddisaccharideor oligosaccharide
formation (Overendet aI., 1962)] are possible.
Another approachto the characterizationof polysaccharidesis the methyl-
ation technique(Cheshireet aI., 1979, 1983).By methylatingthe polysaccharides
prior to hydrolysis, it is theoretically possible to identify the monosaccharide
linkages(unmethylatedsites on the monosaccharideproducts)within the poly-
saccharidemacromolecule.Completemethylation is difficult even when using
extractedsampleswhich may be due to steric hindrancesarising from the associ-
ation of hydroxyl groupswith nonsugarcomponents(Cheshire& Hayes,1990).
No ideal conditionsexist wherebythe hydrolysisconditionsare optimal for
all the generalgroups of monosaccharides. Thereforea variety of analysis are
usually performed for each of the main groups of monosaccharides.These
include hexosamines,hexoses,deoxyhexoses,pentosesand uronic acids. The

conditionsgenerally used are: hexosamines,HCI (4-8 M); hexoses,H2S04 (0.5
or 1 M); deoxyhexoses,HCI or H2S04 (0.1 M) (Swinceret aI., 1969). It is more
difficult to achievea satisfactorydegreeof recoveryfor the uronic acids and the
pentoses.
Hydrolysis of Polysaccharides
The methodbelow basedon that of Oadeset ai. (1970), uses2.5 M H2S04
to extractonefraction of polysaccharides
prior to a more vigorousextractionpro-
cedurewith 0.5 and 12 M H2S04,
Materials
1. Sulfuric acid, 12 M (72% w/w, sp. gr = 1.634), 2.5 M
2. Sodiumhydroxide,concentrated.
Method
Combine2 g of finely groundair dried soil with 25 mL of 2.5 M H2S04 in
a 50-mL round bottomflask and reflux the suspensionfor 20 min. Filter the mix-
ture through a sinteredglassfilter, wash the residuewith distilled water and dry
over P20S' Retain the filtrate and washingsfor analysisor to be combinedwith
the hydrolysatefrom the next part of the method.
Soakthe residuein 12 M H2S04 for 16 h and then dilute the suspensionto
0.5 M H2S04, Stopperthe flask with a capillary air leak and heat at 100°C for 5
h. Cool the mixture and filter through a sinteredglassfilter (porosity x 3). Com-
bine the filtrate with that from the aboveprocedure,neutralizethis resultingsolu-
tion to pH = 7 using NaOH, make the volume up to 100 mL and then filter again
(Whatmanno. 1 paper).This solution is then ready for analysisof hydrolysates.
Analysis of Hydrolysates
The analysisof the sugarmixture releasedby the hydrolysis procedureis
often carried out using an alkaline ferricyanide and anthronemethod(Cheshire,
1979). An alternativeto this involves reductionand acetylationof the sugarsfol-
lowed by analysisof the solution by gas-liquidchromatography(glc) or high per-
formance liquid chromatography(hplc). For a simple monosaccharidemixture
hplc is probably the most suitabletechnique.However,monosaccharide mixtures
derivedfrom samplesof humic substances are relatively complex.It has beenrec-
ommendedthat glc is a more suitable method for quantitative and qualitative
analysisof thesesamples(Chaplin & Kennedy, 1986).
The methodbelow, basedon that used by Oadeset ai. (1970), is designed
for analysisof the sugarsusing Gc.
Materials
1. Sodiumborohydride(NaBH4)
2. Acetic anhydride(CH3CO)zO
3. Chloroform (CHCI3)
4. Methylenechloride (CH2Cl 2)
1062 SWIFf
Method
Place a solution (water or methanol solvent) of the sugars(1-10 mg),
including internal standards,in a 12.4 by 1.6-cmscrewtop test tube, add 1 mg of
solid sodium borohydrideand leave to standovernight. Placethis reducedsugar
solution in a warm bath at 60°C and blow to drynessusing a streamof air.
Add 1 to 2 mL of methanolcontaining10% (v/v) glacial aceticacid to each
tube in the 60°C water bath and blow it to drynessagainusing a jet of air. Repeat
this procedurefour times. Add 1 to 2 mL of aceticanhydride,then sealthe tubes
using screwcapsfitted with Teflon insertsand heatat 130 to 135°C for 2 h. Dry
the reactionmixture using the water bath and streamof air as before.
Extract the alditol acetatesusing chloroform and filter the solution under
pressurethrough a small column (5 by 10 mm) of Silica Gel G (Bio-Rad, Rich-
mond, CA) usingchloroform as the eluantto removethe fine black solids present
when analysingsoil hydrolysates.Evaporatethe chloroform filtrates to dryness
then dissolvethe alditol acetatesin methylenechloride beforeinjection into a gas
chromatograph.
Comment
Two chemicalssuitablefor useas an internalstandardfor the abovemethod
are myoinositol (C6H 120 6) and quebrachitol(C7H 140 6) (Oadeset aI., 1970). A
suitable internal standardto be used for plant-derivedcarbohydrateanalysisis
pentaerythritol(CSH 120 4) (Chaplin & Kennedy,1986).
FUTURE DEVELOPMENTS
It must be obvious to the readerthat there is still much to learn about the
natureof soil organicmatter.As eachnew techniqueis appliedto this areaof sci-
entific researchit becomesmore apparentthat, in the short term, the chancesof
a major breakthrough' are unlikely. The amountof informationgainedfrom each
individual studyor from the applicationof a single techniqueis usually small, but
the significanceof that information may then be magnifiedwhen combinedwith
that from otherstudiesor results.In this context,it is importantthat asmany tech-
niques as possibleare used to study the samematerial and the information ob-
tained integratedto maximize our knowledgeof this material.The standardand
referencesamplespreparedby the InternationalHumic SubstancesSociety will
prove to be invaluablefor this purpose.
It is anticipatedthat the type of analysisassociatedwith soil organicmatter
researchwill tend towards the desirablesituation in which the analytical proce-
dures arecapableof studyingorganicmatterin situ in the soil matrix. This is like-
ly to involve instrumentationof the complexityandcostlinessof the type of solid-
state NMR, with other nondestructiveinstrumental techniquesalso playing a
major role. In the long term the future directionsof soil organic matter research
will be governedby the requirementsof our society. As the needsfor increased
agricultural productionand decreasedenvironmentalpollution grow, so will the
need for a better understandingof the chemical and physical characteristicsof
soil chemistryand, hence,soil organicmatter.
ACKNOWLEDGMENT
The authoris very grateful to Dr. Kaye Sparkfor invaluableassistancein

the preparationof this chapter.
REFERENCES
Aiken, O.R, D.M. McKnight, R.L. Wershaw,and P. MacCarthy. 1985. Humic substancesin soil, sed-
iment and water. Geochemistry,isolation and characterization.Wiley-Intersci., New York.
Atherton, N.M., PA Cranwell, AJ. Floyd, and RD. Haworth. 1967. Humic acid I. ESR spectraof
humic acids. Tetrahedron23:1653-1667.
Baes,AV., and P.R. Bloom. 1989. Diffuse reflectanceand transmissionfourier transform infrared
(DRIFI) spectroscopyof humic and fulvic acids. Soil Sci. Soc. Am. J. 53:695-700.
Baes, A.V., and P.R Bloom. 1990. Fulvic acid ultraviolet-visible spectra:Influence of solvent and
pH. Soil Sci. Soc. Am. 1. 54:1248-1254.
Baldock, JA, 1.M. Oades,A.M. Vassallo,and MA Wilson. 1990. Solid stateCP/MAS 13C N.M.R.
Analysis of particle size and density fractionsof a soil incubatedwith uniformly labelled 13C_
glucose.Aust. J. Soil Res. 28:193-212.
Baldock,I.A., G.J. Currie, and J.M. Oades.1991. Organicmatteras seenby solid state13C NMR and
pyrolysis tandemmassspectrometry.p. 45-{i0. In W.S. Wilson (ed.) Advancesin soil organic
matterresearch:The impact on agricultureand the environment.Proc. Jt. Symp., Univ. Essex,
3-4 Sept. 1990. R. Soc. Chern.,Cambridge,England.
Barker, S.A, P. Finch, M.H.B. Hayes, R.G. Simmonds,and M. Stacey.1965. Isolation and prelimi-
nary characterizationof soil polysaccharides.Nature (London) 205:68-{i9.
Barker, S.A, M.H.B. Hayes,R.G. Simmonds,and M. Stacey.1967. Studieson soil polysaccharides.
I. Carbohyd.Res.5:13-24.
Becket,R, Z. Jue, and J.C. Giddings. 1987. Determinationof molecularweight distributionsof ful-
vic and humic acids using flow field-flow fractionation. Environ. Sci. Technol. 21:289-295.
Beyer, L., H.-R. Schulten,and R. Friind. 1992. Propertiesand compositionof soil organic matter in
forest and arablesoils of Schleswig-Holstein:1. Comparisonof morphologyand resultsof wet
chemistry,CPMAS-13C-NMRspectroscopyand pyrolysis-field ionisation massspectrometry.
Z. Pflanzenerniihr.Bobenk. 155:345-354.
Bloom, P.R., and J.A Leenheer.1989. Vibrational, electronic,and high-energyspectroscopicmeth-
ods for characterizinghumic substances.p. 409-446./n M.H.B. Hayeset al. (ed.) Humic sub-
stancesII. In searchof structure.Wiley-Intersci., Chichester,England.
Bracewell,I.M., K. Haider, S.R. Larter, and H.-R. Schulten.1989. p. 180-222.Thermal degradation
relevant to structural studiesof humic substances.In M.H.B. Hayes et al. (ed.) Humic sub-
stancesII. In searchof structure.Wiley-Intersci., Chichester,England.
Bracewell,1.M., G.W. Robertson,and B.L. Williams. 1980. Pyrolysis-massspectrometrystudiesof
humification in a peat and a peaty podzol. J. Anal. Appl. Pyrol. 2:239-248.
Brooks, J.D., R.A Durie, and S. Sternhell. 1958. Chemistry of brown coals. Aust. J. Appl. Sci.
9:303-320.
Burch, R.D., C.H. Langford, and D.S. Gamble.1978. Methodsfor the comparisonof fulvic acid sam-
ples: The effects of origin and concentration on acidic properties. Can. J. Chern.
56:11%-1201.
Cameron,R.S., B.K. Thornton, R.S. Swift, and AM. Posner.1972a. Molecular weight and shapeof
humic acid from sedimentationand diffusion measurements on fractionatedextracts.J. Soil
Sci. 23:394-408.
Cameron,RS., R.S. Swift, B.K. Thornton, and A.M. Posner.1972b. Calibration of gel permeation
chromatographymaterialsfor use with humic acid. 1. Soil Sci. 23:342-349.
Chaplin, M.E, and I.E Kennedy. 1986. Carbohydrateanalysis.IRL Press,Oxford.
Chen,S., w.P. Inskeep,S.A. Williams, and P.R Callis. 1992. Complexationof I-Naphthol by humic
and fulvic acids. Soil Sci. Soc. Am. J. 56:67-73.
Chen, Y., and M. Schnitzer.1976. Viscosity measurements on soil humic substances.Soil Sci. Soc.
Am. J. 40:866-872.
Chen, Y., and M. Schnitzer. 1989. Sizesand shapesof humic substancesby electronmicroscopy.p.
62140. In M.H.B. Hayeset al. (ed.) Humic substancesII. In searchof structure.Wiley-Inter-
sci., Chichester,England.
1064 SWIFT
Chen, Y., N. Senesi,and M. Schnitzer. 1977. Information provided on humic substancesby EJE6
ratios. Soil Sci. Soc. Am. J. 41:352-358.
Chen,Y., N. Senesi,and M. Schnitzer.1978. Chemicaland physicalcharacteristicsof humic and ful-
vic acid extractedfrom soils of the mediterraneanregion. Geoderma20:87-104.
Cheshire,M.V. 1979. Nature and origin of carbohydratesin soils. Acad. Press,London.
Cheshire,M.V., and Hayes, M.H.B. 1990. Composition,origin, structures,and reactivities of soil
polysaccharides.p. 307-336.In M.E De Boodt et al. (ed.) Soil colloids and their association
in aggregates.PlenumPress,New York.
Cheshire,M.V., J.M. Bracewell,e.M. Mundie, G.w. Robertson,J.D. Russell,and AR. Fraser.1979.
Structuralstudieson soil polysaccharides.J. Soil Sci. 30:315-326.
Cheshire,M.V., e.M. Mundie, J.M. Bracewell,G.W. Robertson,J.D. Russell,and AR. Fraser.1983.
The extractionand characterizationof soil polysaccharideby whole soil methylation.J. Soil
Sci. 34:539--554.
Christensen,J.J.,L.D. Hansen,and R.M. Izatt. 1976. Handbookof proton ionization heatsand relat-
ed thermodynamicquantities.J. Wiley & Sons,Inc., New York.
Clapp, C.E., W.W. Emerson,and AE. Olness.1989. Sizes and shapesof humic substancesby vis-
cosity measurements. p. 497-514.In M.H.B. Hayeset al. (ed.) Humic substancesII. In search
of structure.Wiley-Intersci., Chichester,England.
Dalal, R.e., and R.J. Mayer. 1986. Long-term trendsin fertility of soils undercontinuouscultivation
and cerealcropping insouthernQueensland.IV. Loss of organiccarbonfrom different densi-
ty fractions.Aust. J. Soil Res.24:293-300.
Davis, J.A 1982. Adsorption of natural dissolved organic matter at the oxide/water interface.
Dawson,H.J., B.E Hrutfiord, R.J. Zasoski,and Ee. Ugolini. 1981.The molecularweight and origin
of yellow organicacids. Soil Sci. 132:191-199.
De Nobili, M. 1988.Electrophoreticevidenceof the integrity of humic substances separatedby means
of electrofocusing.J. Soil Sci. 39:437-445.
De Nobili, M., G. Bragato,J.M. Alcaniz, A Puigbo, and L. Comellas. 199Oa. Characterizationof
electrophoreticfractions of humic substanceswith different electrofocusingbehaviour.Soil
Sci. 150:763-770.
De Nobili, M., M. Contin, and L. Leita. 1990b.Alternative methodfor carboxyl group determination
in humic substances.Can. J. Soil Sci. 70:531-536.
Dragunov,S.S.,and B.G. Murzakov. 1970. Heterogeneityof fulvic acidsof a chernozem.Prochvove-
denie 3:115-121.
Drijber, R.A, and L.E. Lowe. 1990. Nature of humus in andosolsunder differing vegetationin the
SierraNevada,Mexico. Geoderma47:221-231.
Dubach,P., N.e. Mehta, T. Jakab,E Martin, and N. Roulet. 1964. Chemical investigationson soil
humic substances.Geochim.Cosmochim.Acta 28:1567-1578.
Dubois, M., KA Gilles, J.K Hamilton, P.A Rebers,and E Smith. 1956. Colorimetric method for
determinationof sugarsand relatedsubstances.Anal. Chern. 28:350--356.
Ducaroir, J., P. Cambier,J.-P. Leydecker,and R. Prost. 1990.Application of soil fractionation meth-
ods to the study of the distribution of pollutant metals. Z. Pflanzenerniihr. Bodenk.
153:349--358.
Duxbury, J.M. 1989. Studiesof the molecularsize and chargeof humic substancesby electrophore-
sis. p. 593-620. In M.H.B. Hayes et al. (ed.) Humic substancesII. In searchin structure.
Wiley-Intersci., Chichester,England.
Finch, P., M.H.B. Hayes,and M. Stacey.1966. p. 19--32.In G.v. Jacks(ed.) Soil chemistryand fer-
tility. Trans.Jt. Meet. Comm. 2, 4, Int. Soc. Soil Sci. 1966. Univ. Press,Aberdeen,Scotland.
Flaig, w., H. Beutelspacher,and E. Rietz. 1975. Chemical compositionand physical propertiesof
humic substances. p. 28. In J.E. Gieseking(ed.) Soil components.Vol. 1. Organiccomponents.
Forsyth,W.G.e. 1947. Studieson the more soluble complexesof soil organic matter.I. A methodof
fractionation.Biochem.J. 41:176--181.
Fritz, J.S., S.S. Yamamura,and E.C. Bradford. 1959. Determinationof carbonyl compounds.Anal.
Chern. 31:260--263.
Gamble,D.S. 1972. Potentiometrictitration of fulvic acid: Equivalencepoint calculationsand acidic
functional groups.Can. J. Chern. 50:2680--2690.
Ghosh,K, and M. Schnitzer.1979.UV and visible absorptionspectroscopicinvestigationsin relation
to macromolecularcharacteristicsof humic substances.J. Soil Sci. 30:735-745.
Ghosh, K, and M. Schnitzer. 1980. Macromolecularstructuresof humic substances.Soil Sci.
129:266--276.
Glebko, L.I., J.U. U1kina, and O.B. Maximov. 1970. A semi-micro-methodfor the determinationof
quinoid groupsin humic acids. Mikrochim. Acta. 1247-1254.
Greenland,0.1., and G.w. Ford. 1964. Separationof partially humified organic materialsfrom soils
by ultrasonicdispersion.p. 137-148.In Trans. 8th Int. Congr. Soil Sci. 1964. Acad. Socialist
Rep. Romania,Bucharest,Romania.
Greenland,OJ., G.P. Lindstrom, and J.P. Quirk. 1962. Organic materials which stabalisenatural
aggregates.Soil Sci. Soc. Am. Proc. 26:366-371.
Greenland,O.G., and J.M. Oades.1975. Sacharrides.p. 213-261.In 1.E. Gieseking(ed.) Soil com-
ponents.Vol. 1. Organiccompounds.Springer,New York.
Haider, K, and H.-R. Schuten.1985. Pyrolysis-field ionisation massspectrometryof lignins, soil
humic compoundsand whole soil. 1. Anal. Appl. Pyro.. 8:317-331.
Haider, K, B.R. Nagar, H.L.e. Meuzelaar,e. Saiz-Jimenez,and J.P. Martin. 1977. Studieson soil
humic compounds,fungal melaninsand model polymersby pyrolysis-massspectrometry.p.
213-220. In Soil organic matter studies. Proc. Symp. Soil Organic Matter Studies,Braun-
schueis,Germany.6-10 Sept. 1976. Int. At. Energy Agency, Vienna.
Halma, G., M.A. Posthumus,R. Miedema,W. van de Westeringh,and H.L.C. Meuzelaar.1978. Char-
acteriationof soil typesby pyrolysis-massspectrometry.Agrochimica 22:372-381.
Hayes,M.H.B. 1985. Extractionof humic substances from soil. p. 329-361.In G.R. Aiken et al. (ed.)
Humic substancesin soil, sedimentand water. Geochemistry,isolation and characterization.
Wiley Intersci., New York.
Hayes, M.H.B., and R.S. Swift. 1978. The chemistry of soil organic colloids. p. 179-320. In OJ.
Greenlandand M.H.B. Hayes(ed.) The chemistryof soil constituents.Wiley-Intersci., New
York.
Hayes, M.H.B., R.S. Swift, R.E. Wardle, and J.K Brown. 1975. Humic materialsfrom an organic
soil: A comparisonof extractantsand of propertiesof extracts.Geoderma13:231-245.
Hayes,M.H.B., P. MacCarthy,R. Malcolm, and RS. Swift. 1989. Humic substancesII. In searchof
structure.Wiley-Intersci., Chichester,England.
Haynes,RJ., and R.S .Swift. 1990. Stability of soil aggregatesin relation to organicconstituentsand
soil water content.J. Soil Sci. 41:73-83.
Hempfling, R., and H-R. Schulten.1991. Pyrolysis-(gaschromatography!)massspectrometryof agri-
cultural soils and their humic fractions. Z. Pflanzenernahr.Bodenk. 154:425-430.
Howarth, RJ., and R Sinding-Larsen.1983. Multivariate analysis.p. 207-289.In R.J. Howarth (ed.)
Statisticsand dataanalysisin geochemicalprospecting.Elsevier,Amsterdam.
Hunter, R.J. 1989. Foundationsof colloid science.Vol. 1. Oxford Press,Oxford, England.
Konova, M.M. 1966. Soil organic matter. Permagon,Elmsford, New York.
Kortum, G. 1969. Reflectancespectroscopy.Springer-Verlag,New York.
Krosshaven,M., J.O. Bj0rgum, J. Krane, and E. Steinnes.1990. Chemical structure of terrestial
humusmaterialsformed from different vegetationcharacterizedby solid-state\3C NMR with
CP-MAS techniques.1. Soil Sci. 41:371-377.
Krosshaven,M., I. Kiigel-Knabner,T.E. Southon,and E. Steinnes.1992.The influenceof humucfrac-
tionation on the chemicalcompositionof soil organicmatterstudiedby solid-state\3C NMR
J. Soil Sci. 43:473-483.
Kumada,K, and Y. Kawamura.1968. On the fractionation of humic acids by a fractional precipita-
tion technique.Soil Sci. Plant Nutr. 14:198-200.
Kyuma, K 1964.A fractional precipitationtechniqueappliedto soil humic substances. Soil Sci. Plant
Nutr. 10:33-35.
Lindqvist, I. 1972. Chargetransferinteractionof humic acidswith donor moleculesin aqueoussolu-
tions. Swed. J. Agric. Res. 12:105-109.
Lindqvist, I. 1973. Partial reductionof a humic acid. Swed. J. Agric. Res. 13:69-73.
MacCarthy,P., and H.B. Mark, Jr. 1975. Infrared studieson humic acid in deuteriumoxide. I. Evalu-
ation and potentialitiesof the technique.Soil Sci. Soc. Am. Proc. 39:663-668.
MacCarthy, P., and H.B. Mark, Jr. 1976. An evaluationof Job'smethodof continuousvariationsas
applied to soil organicmatter-metalion interactions.Soil Sci. Soc. Am. J. 40:267-276.
MacCarthy, P., and S. O'Cinneide. 1974a. Fulvic acid. II. Interaction with metal ions. J. Soil Sci.
25:429-437.
MacCarthy,P., and S. O'Cinneide.1974b.Fulvic acid. I. Partial fractionation.J. Soil Sci. 25:420-428.
MacCarthy, P., M.J. Peterson,R.L. Malcolm, and E.M. Thurman. 1979. Separationof humic sub-
stancesby pH gradientdesorptionfrom a hydrophobicresin. Anal. Chern. 51:2041-2043.
Malcolm, R.L. 1989. Application of solid-state 13C-NMR spectroscopyto geochemicalstudiesof
humic substances.p. 339-371.In M.H.B. Hayeset al. (ed.) Humic substances.Vol. 2. Wiley
Intersci., Chichester,England.
1066 SWIFf
Martell, A.E.; and RM. Smith. 1977. Critical stability constants.Vol. 3. Other organic ligands.
Plenum,New York.
Ma'shum, M.E., M.E. Tate, G.P. Jones,and J.M. Oades. 1988. Extraction and characterizationof
water-repellentmaterialsfrom Australian soils. J. Soil Sci. 39:99-109.
Meuzelaar,H.L.C., J. Haverkamp,and F.D. Hileman. 1982. Pyrolysis-massspectrometryof recent
and fossil biomaterial~ompendium and atlas. Elsevier,Amsterdam.
Mortensen,J.L., and R.B. Schwendinger.1963. Electrophoreticand spectroscopiccharacterizationof
high molecular weight componentsof soil organic matter. Geochim.Cosmochim.Acta
27:201-208.
Niemeyer,1., Y. Chen,and J.-M. Bollag. 1992. Characterizationof humic acids,composts, andpeat
by diffuse reflectance Fourier TransformInfrared Spectroscopy.Soil Sci. Soc. Am. J.
56:135-140.
Nguyen, T.T., L.J. Janik, and M. Raupach. 1991. Diffuse reflectance infrared fourier transform
(DRIFI) spectroscopyin soil studies.Aust. J. Soil Res.29:49-67.
Oades,1.M.1967. Carbohydratesin someAustraliansoils. Aust. J. Soil Res.5:103-115.
Oades,1.M., MA Kirkman, and G.H. Wagner. 1970. The use of gas-liquid chromatographyfor the
determination of sugars extracted from soils by sulfuric acid. Soil Sci. Soc. Am. Proc.
34:230-235.
Oades,1.M., A.M. Vassallo,A.G. Waters,and M.A. Wilson. 1987. Characterizationof orf,anic mat-
ter in particle size and density fractions from a red-brown earth by solid-state1 C N.M.R.
Aust. J. Soil Res. 25:71-82.
Oliver, B.G., E.M. Thurman,and RL. Malcolm. 1983. The contribution of humic substancesto the
acidity of coloredwaters.Geochim.Cosmochim.Acta 47:2031-2035.
Overend,w.G., C.W. Rees,and J.S. Sequira.1962. No. 675 Reactionsat position 1 of carbohydrates.
Part III. The acid-catalyzeshydrolysisof glycosides.J. Chern.Soc. 3:3429-3440.
Parsons,J.W., and 1. Tinsley. 1961. Chemical studiesof polysaccharidematerial in soil composts
basedon extractionwith anhydrousformic acid. Soil Sci. 92:46-53.
Perdue,E.M. 1978.Solution thermochemistry of humic substances-I.Acid-baseequilibria of humic
acid. Geochim.Cosmochim.Acta 42:1351-1358.
Perdue,E.M. 1985. Acidic functional groupsof humic substances.p. 493-526.In G.R Aiken et al.
(ed.) Humic substancesin soil, sedimentand water. Geochemistry,isolation and characetriza-
tion. Wiley-Intersci., New York.
Perdue,E.M., J.H. Reuter,and M. Ghosal. 1980.The operationalnature of acidic functional group
analysesand its impact on mathematicaldescriptionsof acid-baseequilibria in humic sub-
stances.Geochim.Cosmochim.Acta 44:1841-1851.
Piccolo, A. 1988. Characteristicsof soil humic extractsobtainedby someorganic and inorganicsol-
ventsand purified by HCI-HF treatment.Soil Sci. 146:41~26.
Piccolo,A., and Mirabella, A. 1987. Molecular weight distribution of peat humic substances extract-
ed with different inorganicand organicsolutions.Sci. Total Environ. 62:39-46.
Posner,A.M. 1966.The humic acidsextractedby variousreagentsfrom a soil. Part I. Yield, inorgan-
ic components,and titration curves.J. Soil Sci. 17:65-78.
Posner,A.M., and1.M. Creeth.1972.A studyof humic acid by equilibrium ultracentrifugation.1. Soil
Sci. 23:333-341.
Posner,A.M., B.K.G. Theng,and1.RH. Wake. 1968.The extractionof soil organicmatterin relation
to humification. p. 153-162.In 1.W. Holmes (ed.) Trans. 9th Int. Congr. Soil Sci., 3. 1968.
August Robertson,Sydney,Australia.
Reid, P.M., A.E. Wilkinson, E. Tipping, and M.N. Jones.1991. Aggregationof humic substancesin
aqueousmediaas determinedby light-scatteringmethods.1. Soil Sci. 259-270.
Reuter,J.H., and E.M. Perdue.1981. Calculationof molecularweightsof humic substances from col-
ligative data: Application to aquatichumusand its molecularsize fractions. Geochim. Cos-
mochim. Acta 45:2017-2022.
Rice, 1., and P. MacCarthy.1989. Isolation of humin by liquid-liquid partitioning. Sci. Total Environ.
81/82:61~9.
Ristori, G.G., E. Sparvoli, M. De Nobili, and L.P. D' Acqui. 1992. Characterizationof organicmatter
in particle-sizefractions of vertisols. Geoderma54:295-305.
Ritchie, G.S.P.,and A.M. Posner.1982. The effect of pH and metal binding on the transportproper-
ties of humic acids.1. Soil Sci.33:233-247.
Roth, C.H., W. Wilczynski, and C. de CastroFilho. 1992. Effect oftiIIage and liming on organicmat-
ter composition in a Rhodic Ferralsol from Southern Brazil. Z. Pflanzenerniihr. Bodenk.
155:175-179.
Ryan, D.K., and1.H. Weber. 1982. Fluorescencequenchingtitration for determinationof complexing

capacitiesand stability constantsof fulvic acid. Anal. Chern. 54:986--990.
Saiz-limenez,e., K. Haider, and H.L.e. Meuzelaar.1979. Comparisonsof soil organicmatterand its
fractions by pyrolysis-mass spectrometry. Geoderma22:25-37.
Saiz-limenez,C., e. Martin, K. Haider, and H.L.e. Meuzelaar.1978. Comparisonof humic and ful-
vic acids from different soils by pyrolysis-massspectrometry.Agrochimica 22:353-359.
Sanderson,G.W., and B.P.M. Perera.1966. Removalof polyphenoliccompoundsinterfering with car-
bohydratedeterminationsin plant extractswith an insoluble polyphenol adsorbent.Analyst
91:335-336.
Schlesinger,W.H. 1984. Soil organic matter: A source of atmosphericCO2, p. 111-127. In G.M.
Woodwell (ed.) The role of terrestrialvegetationin the global carboncycle: Measurementby
remotesensing.SCOPE,Wiley, Chichester,England.
Schnitzer,M. 1977. Recentfindings on the characterisationof humic substancesextractedfrom soils
from widely differing climatic zones.p. 117-131.In Proc. Symp. Soil Organic Matter Stud-
ies, Braunschweig,Germany.6--10 Sept. 1976. Int. At. Energy Agency, Vienna.
Schnitzer,M. 1990. Selectedmethodsfor characterizationof soil humic substances.p. 65-90. In P.
MacCarthy et al. (ed.) Humic substancesin soil and crop sciences:Selectedreadings.ASA
and SSSA, Madison,WI.
Schnitzer,M., e.A. Hindle, and M. Megliz. 1986.Supercritical gasextractionof alkanesand alkonoic
acids from soils and humic materials.Soil Sci. Soc. Am. 1. 50:913-919.
Schnitzer,M., and S.U. Khan. 1972. Humic substancesin the environment.Marcel Dekker Inc., New
York.
Schnitzer, M., and S.I.M. Skinner. 1965. Organo-metallicinteractions in soils: 4. carboxyl and
hydroxyl groupsin organic matterand metal retention.Soil Sci. 99:278-284.
Schnitzer,M., and S.I.M. Skinner. 1968. Gel filtration of fulvic acid, a soil humic compound.Isotopes
and radiation in soil organic matterstudies.Int. At. Energy Agency, Vienna.
Schnitzer, M., and S.I.M. Skinner. 1969. Free radicals in soil humic compounds. Soil Sci.
108:383-390.
Schulten,H.-R., and R. Hempfling. 1992. Influenceof agricultural and managementon humuscom-
positon and dynamics:Classicaland modemanalytical techniques.Plant Soil 142:259-271.
Senesi,N. 1990.Molecularand quantitativeaspectsof the chemistryof fulvic acid and its interactions
with metal ions and organicchemicals.PartII. The fluorescencespectroscopyapproach.Anal.
Chim. Acta 232:77-106.
Senesi,N., Y. Chen, and M. Scnitzer. 1977. Hyperfine splitting in electronspin resonancespectraof
fulvic acid. Soil BioI. Biochem. 9:371-372.
Skjemstad,1.1992. Genesisof podzolson coastaldunesin southernQueensland.III. The role of alu-
minum-organiccomplexesin profile development.Aust. J. Soil Res. 30:645-{)65.
Smith, R.G., Jr. 1976. Evaluationof combined applications of ultrafiltration and complexationcapac-
ity techniquesto natural waters.Anal. Chern.48:74-76.
Soliins, P., G. Sprycher,and e.A. Glassman.1984.Net nitrogenmineralizationfrom light- and heavy-
fraction forest soil organicmatter. BioI. Biochem. 16:31-37.
Spark,K.M., and R.S. Swift. 1994. Investigationof someof the parametersaffectingthe fluorescence
spectraof humic substances.p. 153-160.In N. SenesiandT.M. Miano (ed.) Humic substances
in the global environmentand implications on human health.Elsevier,Amsterdam.
Steelink, e.A 1964. Free radical studiesof lignin, lignin degradationproducts,and soil humic acid.
Steelink,C.A., M.A Mikita, and K.A Thorn. 1983. Magnetic studiesof humatesand related model
compounds.p. 83-105. In R.E Christman and E.J. Gjessinger(ed.) Aquatic and terrestrial
humic materials.Ann Arbor, Sci., Ann Arbor, MI.
Stevenson,FJ. 1994. Humuschemistry.Genesis,composition,reactions.2nd ed. John Wiley & Sons,
New York.
Stevenson,FJ., Q. van Winkle, and w.P. Martin. 1953. Physiochemicalinvestigationsof clay-
adsorbedorganiccolloids II. Soil Sci. Soc. Am. Proc. 17:31-34.
Strickland, T.e., and P. Sollins. 1987. Improved method for separatinglight- and heavy-fraction
organic material from soil. Soil Sci. Soc. Am. I. 51:139(}"1393.
Swift, R.S. 1985.Fractionationof soil humic substances. p. 387-408.In G.R. Aiken et al. (ed.) Humic
substancesin soils, sediment,and water. Wiley-Intersci., New York.
Swift, R.S. 1989. Molecularweight, size,shape,andchargecharacteristicsof humic substances:some
basicconsiderations.p. 449-466.In M.H.B. Hayeset al. (ed.) Humic substancesII. In search
of structure.Wiley-Intersci., Chichester,England.
Swift, R.S., and AM. Posner.1971. Gel chromatographyof humic acid. J. Soil Sci. 22:237-249.
1068 SWIFT
Swift, R.S.,andA.M. Posner.1972.Autoxidationof humic acid underalkalineconditions.J. Soil Sci.

23:381-393.
Swift, R.S., R.L. Leonard,R.H. Newman,andB.K.G. Theng.1992.Changesin humic acid composi-
tion with molecularweight asdetectedby \3C-nuclearmagneticresonancespectroscopy.Sci.
Total Environ. 117/118:53-61.
Swincer,G.D., I.M. Oades,and D.J. Greenland.1968. Studieson soil polysaccharides.I. The isola-
tion of polysaccharidesfrom soil. Aust. 1. Soil Res.6:211-224.
Swincer,G.D., I.M. Oades,and OJ. Greenland.1969. The extraction,characterization,and signifi-
canceof soil poysaccharides. Adv. Agron. 21:195-235.
Theng,B.K.G., and A.M. Posner.1967. Natureof the carbonyl groupsin soil humic acid. Soil Sci.
104:191-201.
Theng,B.K.G., K.R. Tate,andP. Becker-Heidmann.1992.Towardsestablishingthe age,location,and
identity of the inert soil organic matter of a spodosol. Z. Pflanzenerniihr. Bodenk.
155:181-184.
Theng,B.K.G., 1.R.H. Wake, and A.M. Posner.1968.The fractional precipitationof soil humic acid
by ammoniumsulphate.Plant Soil 29:305-316.
Thomas,R.L., J.L. Mortenson,and EL. Himes. 1967. Fractionationand characterizationof a soil
polysaccharideextract.Soil Sci. Soc.Am. Proc. 31:568-570.
Thurman,E.M., and R.L. Malcolm. 1981.Preparativeisolationof aquatichumic substances. Environ.
Sci. Technol. 15:463-466.
Tomar, N.K., R.P. Yadav, and P.S. Relan. 1992. Characterisationof humic and fulvic acidsextracted
with NaOH and NaOH-Na-pyrophosphate mixture from soils of arid and subhumidregions1.
Analytical characteristics.Arid Soil Res. Rehab.6:177-185.
Tsutsuki, K., and S. Kuwatsuka. 1979. Chemical studieson soil humic acids. VII. pH dependent
natureof the ultra-violet and visible absorptionspectraof humic acids.Soil Sci. Plant. Nutr.
25:373-384.
Tsutsuki,K., and S. Kuwatsuka.1984. Molecular size distribution of humic acid as affectedby the
ionic strengthand degreeof humification.Soil Sci. Plant Nutr. 30:151-162.
Turchenek,L.W., and I.M. Oades.1979.Fractionationof organo-mineralcomplexesby sedimenta-
tion and density techniques.Geoderma21:311-343.
Varadachari, C., A.K. Barman, and K. Ghosh. 1983. Electron spin resonanceand scanning
microscopyof humic sbustances. J. Indian Soc. Soil Sci. 31:28-30.
Visser, S.A. 1985. Viscosimetricstudieson molecularweight fractionsof fulvic and humic acidsof
aquatic,terrestrialand microbial origin. Plant Soil 87:209-221.
Waite, T.D., and M.M. Morel. 1984.Coulometricstudy of the redox dynamicsof iron in seawater.
Anal. Chern.56:787-792.
Watanabe,A., and S. Kuwatsuka.1991.Fractionationof soil fulvic acidsusing polyvinyl-pyrrolidone
and their ionization differencespectra.Soil Sci. Plant Nutr. 37:611-617.
Wershaw,R.L. 1985.Application of nuclearmagneticresonancespectroscopyfor determiningfunc-
tionality in humic substances.p .561-584.In G.R. Aiken et al. (ed.) Humicsubstances in soils,
sedimentand water. Wiley-Intersci.,New York.
Wershaw,R.L. 1989.Sizesand shapesof humic substances by scatteringtechniques.p. 545-559.In
M.H.B. Hayeset al. (ed.) Humic substances II. In searchof structure.Wiley-Intersci.,Chich-
ester,England.
Whitehead,D.C., and J. TInsley. 1964. Extraction of soil organic matter with dimethylformamide.
Soil Sci. 97:34--42.
Williams, B.G., OJ. Greenland,andJ.P. Quirk. 1967.The effect of polyvinyl alcohol on the nitrogen
surfaceareaand pore structureof soils. Aust. J. Soil Res.5:77-92.
Wilson, M.A., andK.M. Goh. 1983.NMR spectroscopyof soils. Structureof organicmaterialin sodi-
um deuteroxideextractsfrom PatuaLoam, New Zealand.J. Soil Sci. 34:305-313.
Wilson, M.A., A.H. Gillam, and PJ. Collins. 1983.Analysisof the structureof dissolvedhumic sub-
stancesand their phytoplanktonprecursorsby lH and \3C nuclearmagneticresonance.Chern.
Geol. 40:187-201.
Wilson, M.A., A.M. Vassallo, E.M. Perdue,and I.H. Reuter. 1987. Compositionaland solid-state
nuclearmagneticresonancestudy of humic and fulvic acid fractions of soil organic matter.
Anal. Chern.59:551-558.
Wright, I.R., and M. Schnitzer.1959.Oxygen-containingfunctional groupsin the organicmatterof a
podzol soil. Nature(London) 184:1462-1463.
Yonebayashi,K., andT. Hattori. 1985.Non-aqueoustitration of functional groupsin humic acid. Org.
Geochem.8:47-54.
Yonebayashi,K., and T. Hattori. 1990. A new fractionationof soil humic acids by adsorptionchro-
matography.Geoderma47:327-336.
Zech, W., R. Hempfling, L. Haumaier,H.-R. Schulten,and K. Haider. 1990. Humification in sub-
alpine rendzinas:Chemicalanalyses,IR and 13C NMR spectroscopyand pyrolysis-field ioni-
sationmassspectometry.Geoderma47:123-138.
Zhang,T., H. Gan,and P.F. Low. 1991.Effect of sodium-humateon the rheologicalcharacteristicsof
montmorillonitesuspensions. Soil Sci. Soc.Am. I. 55:989-993.
Published 1996
Chapter 36
Extraction of Organic Chemicals
B. L. SAWHNEY, The Connecticut Agricultural Experiment Station,

New Haven, Connecticut
Agricultural and industrial operationsare the major sourcesof organicchemicals

enteringthe soil. Agricultural chemicals,as pesticides,havebeenapplied to soils
for decadesand continueto be appliedto enhanceproductionand quality of food
and fiber, and to control pestsin the environment.Industrial chemicals,including
solvents,can enterthe soil throughstoragein lagoons,spills and inadequatedis-
posal methods. In addition, military operationsinvolving defenseand energy
sourcesare being recognizedas sourcesof soil contamination.Some of these
chemicalsare hazardous to humanand environmentalhealth. Determinationof
the organic chemicalsin soils is essentialbecauseof potential contaminationof
food cropsfrom their useandbecauseof their potentialerosionor leachingto sur-
face or groundwater leadingto their entry into the food chain. Methodsfor quan-
titative determinationof organic compoundshave been developed.However,
before undertakingtheir analysis,the organic chemicalsneedto be quantitative-
ly extractedfrom the various matricessuchas water, soil, sludge,waste,etc.
For analysesof organic contaminantsin soils, sedimentsor wastes,it is
essentialto procurea representativesample,and also, the integrity of the sample,
i.e., its chemicaland physical propertiesat the time of collection must be main-
tained. The organic chemicalthen has to be extractedfrom the sample into an
organicsolventfor analysis.For efficient extractionof the organicchemical,the
extractantmust solubilize the chemicaland must favorably competefor the sites
on which the organic chemicalmay be sorbed.To achievethis, a variety of sol-
ventsand combinations have beenused.Theseinclude aqueoussolutionsof ace-
tone (C3H60), methanol(CH30H) or acetonitrile(C2H3N), salt solutions,acids,
bases,and metal complexing agents as citrate and ethylenediaminetetraacetic
acid (EDTA). Basedon the available scientific information, USEPA (1986) has
compiled a Laboratory Manual SW-846, Test Methods for Evaluating Solid
Wastes, which includes extraction proceduresfor volatile (purgeable), semi-
volatile and nonvolatile organic compoundsfrom environmentalsamplesof dif-
ferent matrices.The convenientformat for categorizingorganic chemicalsinto
purgeableand semi- and nonvolatile organics outlined in the manual will be
adopted here. Furthermore,becausesoil scientists are increasingly being in-
Copyright © 1996 Soil ScienceSociety of America and American Society of Agronomy. 677 S.
Book Seriesno. 5.
1071
1072 SAWHNEY
volved in use and analysisof environmentalsamples,including sewagesludge,

yard and sewagesludgecompost,and MSW (municipal solid waste),as well as
wastewater, landfillleachatesand groundwaterbelow the soil receivingagricul-
tural chemicalsand wastes,extraction of organic compoundsfrom both liquid
and solid matriceswill be consideredhere.
SAMPLING AND PRESERVATION
Liquid Samples
For the analysis of volatile organic compounds(VOCs, boiling point

<200°C), USEPA(1986) recommendsthat the samplebe taken in a 40-mL glass
vial with Teflon-lined septumcap and for semivolatileand nonvolatile organics,
0.5 to 1 gal (1.89-3.78L) sampleshould be obtained.The former can be pre-
servedfor up to 2 wk at 4°C and the latter shouldbe extractedwithin 1 wk. The
containersshouldbe filled gently to capacityand placedin separateplastic bags,
preferablycontainingsome activatedcarbonto avoid contamination,especially
thoseto be analyzedfor VOCs. Such samplescan be contaminatedby diffusion
of VOCs through the screwcap septum.To checkfor suchcontamination,a field
blank of reagentwater shouldbe carried through samplingand subsequenthan-
dling process.
Solid Samples
For the analysisofVOCs in solid or semisolid(sludges)matrices,the sam-

ple shouldbe mixed well and taken in duplicatein a 4-oz (120-mL) wide mouth
glasscontainerwith Teflon-lined screw-captop. The containersshould be filled
to capacitywithout introducingany air bubbles.The samplecan be preservedat
4°C for up to 2 wk beforeanalysis.For the analysisof semivolatileor nonvolatile
organics,a 8-oz (240 mL) wide mouth glass containeris suggested.
EXTRACTION PROCEDURES
Volatile organic compoundsare generallypurgedout of the liquid or sus-

pensionby an inert gas,asHe or Nz, and trappedon asorbentcolumn(purge-and-
trap extraction).The liquid samplealso can be injected directly into the analyti-
cal instrument(suchas gaschromatograph).Semivolatileand nonvolatile chem-
icals are extractedfrom the sample by an appropriatesolvent using Soxhlet,
Sonicationor SupercriticalFluid Extraction(SFE) techniquedescribedbelow.
Purgeand Trap
When VOCs in solution are to be analyzed,they are purgedfrom the solu-

tion by bubbling an inert gas. A solid sample,however,has to be first dispersed
(generally5 g soil/sediment)into water or methanol,and purging is carried out
EXTRACTION OF ORGANIC CHEMICALS 1073
Liquid samples Solid samples
Proceed as In liquid
samples above
Sorb on GC column
Fig. 36-1. Extraction of volatile organic contaminantsfrom liquid and solid samplesby purge and
trap technique.
on a heated(about 85°C) sample.The VOCs purged into the vapor phaseare

sweptthrougha sorbentcolumn wherethey are adsorbed.The sorbentcolumn is
then heatedand backflushedwith the inert gas to desorbthe VOCs onto a gas
chromatographic(GC) column for analysis.The purge-and-trapmethod is out-
lined in Fig. 36-1. The method is suitable for compoundsthat elute as sharp
peaksfrom a packedGC column, including low molecularweight halogenated
hydrocarbons,aromatics,ketones,nitriles, acetates,acrylates,ethers, and sul-
fides. While the methodis satisfactoryfor determiningVOCs in liquid samples
(Sawhney& Kozloski, 1984), it gives low recoveriesin solid matrices(Hiatt,
1981).
Several purge-and-trapdevices are commercially available. They essen-
tially consist of the purging chamber,the trap and the desorber.Method 5030
(USEPA, 1986) for VOCs analysisby purge-and-trapprocedure,recommends
the following specificationsto comply with ResourceConservationand Recov-
ery Act (RCRA). The recommendedpurging chamberis designedto accept5-
mL sampleswith at least 3-cmdeepliquid column. The gaseousheadspacevol-
ume betweenthe liquid column and the trap should be less than 15 mL. The
purgegasshouldbe introducedat lessthan 5 mm from the baseof the liquid col-
umn. The trap must be at least 25- by 0.267-cmi.d. containingTenax GC (2,6-
Diphenyleneoxide polymer) as the packing material. If fluorocarbonsare to be
determined,one-third of the volume of trap must contain charcoaland, if com-
poundswith boiling point <35°C are to be analyzed,anotherone-thirdof the trap
volume shouldcontain silica gel.
Before use, the trap should be conditionedovernight at 180°C by back-
flushing with the inert purgegasat a flow of at least20 mLimin, venting the trap
effluent to the hood. Before daily use, the trap shouldbe conditionedfor 10 min
at 180°C. The desorbershould be capableof rapidly heatingthe trap, the poly-
1074 SAWHNEY
mer sectionof the trap shouldnot be heatedabove180°Cand the remainingsec-

tions not above220°C.
Calibrationof Purge-and-Thap
System
Add 5 mL reagentwaterinto the purgingchamberthroughthe sampleinlet
using a glass syringe. Reagentwater is purified water, preparedby filtration
througha carbonfilter bed or otherpurification systemsuchas Millipore Super-
Q or equivalent,in which compoundsof interestare not detectedby the analyti-
cal methodused.To the water in the chamber,add aliquotsof a calibrationstan-
dard solution using a syringewith needlebelow the surfaceof the reagentwater.
Similarly addthe internalstandardsolutionto the water. Carry out purgeandtrap
analysisusing a purge gas flow rate of 40 or 20 mUmin with a corresponding
purgetime of 12 or 15 min. The trappedchemicalis then desorbedfor 2 to 4 min
at a desorbtiontemperatureof 180°Conto a GC column for analysis.Identifica-
tion and quantitationof the compoundis obtainedfrom its retention time and
peakareain comparisonto the calibrationstandard.To avoid contaminationof
the purge-and-trapsystemwith high concentrationsof analytesit is recommend-
ed that the samplessuspectedto be contaminatedshouldbe diluted beforecarry-
ing out purgeand trap analysis.
SolventExtraction
Prior to the analysis,semivolatile and nonvolatile organic chemicalsin-

cluding base/neutral,acid and organochlorinepesticides,have to be extracted
with an appropriate solvent.Prior to extraction, the sampleis ground to pass
througha I-rom standardsieveto makeit more homogeneous.Commonlyused
solventsand solventmixturesinclude, toluene(~H8)/methanol, 10:1 (v/v), ace-
tone/hexane(CJI14), 1:1 (v/v) and methylenechloride (CH2CI2). All solvents
shouldbe pesticidegradeor equivalent.To removemoisturefrom the extract,it
is then filtered througha drying agent,suchas Na2S04and concentratedusing a
concentratortube as discussedbelow. When necessary,the organicanalyteis ex-
changedinto a solvent which is compatiblewith the analytical determinative
method.For example,methylenechlorideextractwill haveto be exchangedinto
anothersolvent, such as hexane,for analysis using electron capture detector
(ECD), becausea halogenatedsolventasmethylenechloridewould captureelec-
trons in the detectorand result in large backgroundresponse.
AqueousSamples
1\vo proceduresthat havebeencommonlyusedfor the extractionof semi-
volatile and nonvolatileorganiccontaminantsfrom liquid samplesare: separato-
ry funnel techniqueand continuousliquid-liquid extraction.The former is gener-
ally usedfor semivolatileheat-labilepesticidesandotherorganicsin liquid sam-
ples.The latter is usedfor thermally stableorganicsand for matriceswhich may
tend to form emulsionsin the separatoryfunnel procedure.A third more recent
technique,solid phaseextraction,is usefulfor traceorganicsin liquids. It alsohas
the advantagein that while the othertwo methodsarenonspecific,the solid phase
extraction can be tailored to the extraction of individual organic molecule by

using a cartridgewith a specific absorbate(see below). A wide variety of car-
tridgeswith different absorbatesare now available.
1. SeparatoryFunnel Technique.Transfer a 200-mL liquid sample in a
500-mLseparatoryfunnel. Add 100-mLmethylenechloride to the sample,shake
for 2 min. and let standuntil the two phases,aqueousand organic,haveseparat-
ed. Drain methylenechloride phasethrough a funnel containingglass woolwith
20 g anhydrousNaZS04 on top into a 500-mL round bottom evaporationflask
attachedto a Kudema-Danish(K-D) concentratortube. Repeatmethylenechlo-
ride extractionof the aqueousphasetwo more times and passextractsthrough
the NaZS04 drying column. Then washNaZS04two times with 25 mL eachwith
methylenechloride.If the aqueoussampleand the solventform an emulsion,the
emulsion can either be broken by adding about 5 mL of 10% NaCL or 20%
NaZS04 solution or the extraction with the organic solvent can be carried out
using a continuousliquid-liquid extractor(seebelow). Add 1 to 2 boiling chips
to the concentratortube, attach a three-ball Snydercolumn to the evaporation
flask and evaporatethe extract to near dryness(about 1 mL) on a water bath
(80-90°C), with concentratortube partially immersedin hot water. If solvent
exchangeis requiredadd 50 mL hexaneto the flask and againconcentrateto near
dryness.Repeatthe hexanetreatmenttwo more times to ensureremoval of all
tracesof methyleI).echloride, concentratingthe last extractto 10 mL. Then con-
centratethe extractfurther to 1 to 2 mL using Nz gas.
2. ContinuousLiquid-Liquid Extractor. This method involves simultane-
ous distillation and extraction.A numberof different typesof continuousliquid-
liquid extractorsare commerciallyavailable.The organicchemicalscontainedin
aqueousphaseand the organicsolvent,such as methylenechloride or isooctane
(2,2,4-trimethylpentane)(commonly used solvents), are distilled together,
digesting the sample during the boiling pr.ocess.The mixed condensatethen
flows into a capillarytube down to the bottom of the apparatuswhereseparation
occurs and the solvent with dissolvedorganicsflows to the solvent flask and
water to the sampleflask. Distillation is run overnightand the organicchemicals
are separatedinto solventflask (Pylypiw et al.,1985).The extract is then passed
througha drying agentand concentratedas describedabovebeforeanalysis.The
procedureis outlined in Fig. 36-2.
Solid PhaseExtraction.An aliquot (200 mL) of aqueoussampleis filtered
under vacuumthrough a specially modified silica cartridge,such as a common-
ly used C-18 cartridge where modification is achievedby coating silica with
octadecylsilane.Before using, the cartridge is washedwith 5 mL hexane to
removeany contaminants,followed by 10 mL of methanolto wet the cartridge
surfaceand then with 20 mL of reagentwater. The aqueoussample(200 mL) is
passedthrough the cartridgewhich takesabout 10 min and effectively removes
the organicchemicalsfrom the sample.The cartridgeis washedtwice with 5-mL
aliquotsof reagentwater. Air is then drawn through the cartridgeto removeany
water. The organicchemicalssorbedon the cartridgeare extractedwith about 3
mL hexane,collecting the first 2 mL in a volumetric flask for analysis.The pro-
cedureis outlined in Fig. 36-3. The methodis fast, convenientand conservative
in the use of solventvolumesand is frequently usedfor routine analysis.
1076 SAWHNEY
Liquid· Liquid Extraction
Transfer 200 mL sample into 500 mL separatory

funnel, add 100 mL methylene chloride
Shake 2 minutes, let stand, drain methylene

chloride phase through Na"SO. column
Repeat methylene chloride extraction twice
Combine extracts and concentrate using

K·O apparatus
Solvent exchange if needed,

concentrate extract
Fig, 36-2. Extraction of semivolatile and nonvolatile Clean-up extract, inject into
organicsfrom liquid samplesby liquid-liquid extrac- GC column
tion,
Solid Samples
Solid samplessuch as a soil or sediment,should be mixed thoroughly,
especiallycompositesamples,and ground to passthrougha I-mm sieve. Waste
samplesconsistingof multiphasesshouldbe phase-separated and extractedsep-
arately when possible.Among the three extractiontechniquesdescribedbelow,
the Soxhlet extraction is the earliest developedprocedure.Although it is more
time consuming,it hasthe advantageover the sonicationtechniquein that it min-
imizes the use of extractionsolvent. Application of theseproceduresin extrac-
tion of pesticidesfrom field contaminatedsoils has beendiscussedby Sawhney
Solid Phase Extraction
*
Filter under vacuum 200 mL sample through
a modified silica cartridge
Wash cartridge twice with 5 mL reagent water
Draw air through cartridge to remove water
Extract organic chemicals sorbed on cartridge

with 3 mL of hexane
Fig. 36-3. Extraction of semivolatile and nonvolatile

organicsfrom liquid samplesby solid phaseextrac-
Inject extract into GC column
tion.
Soxhlet Extraction
Mix 109 sample with 109 anhydrous Nat SO,
Place mixture in extraction thimble, or in Soxhlet

extraction column between two glass wool plugs
Add 300 mL extraction solvent and 2-3 boiling

chips in extraction flask and extract 16-24 hours
Filter extract through Na501t column and

concentrate uSing K-O apparatus
Solvent exchange If needed, concentrate
Clean-up extract, Inject Into GC for analysIs Fig. 3~. Extraction of organic contaminantsfrom
solid samplesby Soxhletextraction.
et al. (1988), amongothers.The supercriticalfluid extractionmethodhas been

developedmore recently and eliminatesthe use of organic solventsfor extrac-
tion. A procedureto removeorganic liquids form spill sites by their adsorption
on porouspolyethylenehas recently been suggested(Cary et aI., 1991) but its
efficiency and applicationto different matriceshave not yet beentested.
1. Soxhlet Extraction. Mix 10 g of ground samplewith 10 g anhydrous
Na2S04in an extractionthimble or betweentwo plugsof pyrex glasswool in the
Soxhletextractorcolumn. Add 300 mL of extractionsolventandoneor two boil-
ing chips in the 500-mL round bottom flask. Attach the flask to the extractorand
extractthe samplefor 16 to 24 h, adjustingthe boiling rate to distill aboutfive to
six volumes perhour. Dry the extract by passingthrough a lO-cm column of
anhydrousNa2S04into K-D flask attachedto the concentratortube and concen-
trate the extract and conduct solvent exchange,when necessary,as described
aboveunder aqueoussamples.The procedureis outlined in Fig. 36-4.
2. SonicationExtraction. In contrastto the Soxhlet extraction,this proce-
dure is carriedout at room temperature,but it insuresintimate contactof the sam-
ple matrix with the extractionsolvent,and is thus quite effective.However,care
mustbe exercisednot to overheatthe sampleto avoid any loss of the solventand/
or the analyte. For samplescontaininglow concentrations(less than 20 mg/kg)
of individual organicchemicals,a larger sampleand a more rigorous extraction
procedureis requiredbecauselow levelsare more difficult to extract(seebelow).
The procedureis outlined in Fig. 36-5.
Low concentrationextraction. Mix 30 g samplewith enough anhydrous
Na2S04 into a 20-mL vial and mix well. Add appropriatespiking standards.
Bring volume to 10 mL with the extraction solvent. Sonicatefor 2 min using
0.32-cm taperedmicrotip ultrasonic probe. Filter extract through a 2- to 3-cm
pyrex glass woolpackedin a concentratortube for further concentration.Con-
1078 SAWHNEY
SOnication
Low concentration sample:

l
High concentration sample:
Less than 20 mg/kg contaminant More than 20 mglkg .contaminant
Mix 30 9 sample with Na.-:;O.. to lonn Mix 2g sample with 2g Na..SO.,.

lree flowing powder
Add 100 mL 1:1 Methylenechloride:acetone Add 10 mL 1:1 Methylene chloride:acetone
Sonicate for 3 min. placing probe tip 1 em Sonicate for 2 min. using micratip
below solvent surface. but above sediment layer ultrasonic probe
Filter extract using vacuum filtration Filter extract through 2-3 em pyrex
glass wool
Repeat solvent extraction twice more, Concentrate extract using K-O appa-atus 'I
concentrate extract using K·D apparatus
Clean-up extract. inject into GC Clean-up extract, inject into GC

for analysis for analysis
Fig. 36-5. Extractionof organiccontaminantsfrom solid samplesby sonication.
centratethe extractand solventexchange,if required,following proceduresout-

lined above.
3. Supercritical Fluid Extraction (SFE). While solvent extraction tech-
niquesdescribedaboveare widely usedin the analysisof routine as well as reg-
ulatory samples, recent developmentsis SFE methods are leading to their
increaseduse. Becauseof various advantagesof SFE(outlined below) for ex-
tracting organic chemicalsfrom soils and environmentalsamples,this method
may soonemergeas a methodof choicein this field.
The methodinvolves a pump to supply a supercriticalextractionfluid (a
substanceabove its critical temperatureand pressure),such as CO2, into an
extractionvesselcontainingthe sampleto be analyzed.To maintainthe temper-
atureof the supercriticalfluid abovethe critical temperature,the extractionves-
sel is placedin a heatedoven.The analytesextractedinto the supercriticalfluid
are sweptthrougha flow constrictorinto the collectionvesseldevice,which is at
ambient pressure.The depressurizedsupercriticalfluid, which is now gas, is
vented and the extractedanalytes,which remain as liquids (Fig. 36-6), can be
analyzedby chromatographicor other appropriatetechniques.
The SFE methodhasseveraladvantagesover the liquid-solventextraction
methods(Hawthorne,1990). One, SFE is rapid; extractionsby this methodare
generally completein 10 to 60 min, whereasliquid solvent extractionsrange
Extraction Vessel
Pump
/'
Flow
tJ
Restrictor -,
J~I
Collection
Device
Supercritical
Fluid Fig. 36--6. Schematicof supercriticalfluid extrac-
tion system.
from hours to days. This is becausesupercriticalfluids have much better mass

transfercharacteristicsdue to their higher solute diffusivities and lower viscosi-
ties; hence,a higher rate of extraction.Two, the solventstrengthof supercritical
fluids can be controlled easily by controlling pressure and temperature.
Consequently,a single samplecan be extractedat different pressures(or temper-
atures)to permit selectiveextractionsof different organics.Three,becausesuper-
critical fluids are gasesat ambienttemperatureand are easily separatedfrom the
analytes,concentrationstepsafter extractionare reduced.Four, since CO2 (com-
monly usedfluid) is ventedinto the atmosphere,large volumesof liquid solvents
used in other extraction proceduresare eliminated, reducing disposalproblems
and exposureof laboratorypersonnelto potentially toxic solvents.Finally, direct
coupling of SFE with chromatographysystems(Levy & Rosselli, 1989; Anton et
aI., 1988) permit greater sensitivity and less handling betweenextraction and
analysis.
Despiteits relatively recentdevelopment,supercriticalfluid extractionhas
been used in investigationsinvolving large number of semivolatile and non-
volatile organicchemicalsin a wide rangeof samplematrices.Carbondioxide is
the most commonly usedextractionfluid becauseof its low critical temperature
(31.3°C) and pressure(7.39 MPa) and nontoxicity, as well as its availability in
pure form. Other less frequently usedfluids include ammonia(NH4), nitrous ox-
ide (N20), methanol,ethane(C2H6), ethylene(C2H4) and dichlorofluorome-thane
(CHCI2F). Richards and Campbell (1990) compared SFE with conventional
Soxhlet and sonication extraction for 16 significant semivolatile organic com-
pounds,using soil matrix as the test substrate.They found that using CO2 modi-
fied with 2% methanolwas more efficient for all, but three,compounds.The con-
ditions found for optimum extraction were: 80°C temperature,39.5 MPa pres-
sure, and 30 to 40 min extraction time. The SFE techniquehas been used for
extractingpolychlorinatedbiphenyls(PCBs) from sediments(Shantz& Chesler,
1986); polycyclic hydrocarbons(PAHs) from sedimentandfly ash(Hawthorne&
Miller, 1987); PAHs and alkanesfrom soot and sediments(Levy & Rosselli,
1989) and hazardousorganic chemicalsfrom aqueouswastesfrom steel, pesti-
cide, and acrylonitrile (C2H3N) manufacturingplants (Rice et aI., 1987). The
technique also has been used for extracting bound pesticide residues in soil
(Capriel et aI., 1986; Khan, 1991). McNally and Wheeler (1988) have demon-
1080 SAWHNEY
stratedits usefor extractingherbicidesand metabolitesin soil, plant, andcell cul-

tures.
EXTRACT CLEANUP
Soil and waste extractsoften require cleanuptreatmentsbefore injection

into a gas or liquid chromatographfor analysisof organicchemicals.Untreated
extractscan deterioratecolumn efficiency, detectorsensitivity, and shortenthe
life of expensivecolumnsin addition to causingextraneouspeaksand prevent-
ing peakresolution.A numberof procedureshavebeendevelopedto cleanupthe
extract to overcometheseproblems.In general,the methodsinvolve separation
of different compoundcategoriesalong a chromatographiccolumn, basedupon
their molecularsize or affinities for the column sorbent.Methodsadoptedby the
USEPA (1986) for cleanupfor different analytesare given below.
Gel PermeationChromatography
This is the most common cleanup techniqueused for a broad range of

semivolatile and nonvolatile organicsand pesticides.It has been used success-
fully for all the semivolatilebase,neutral and acid compoundsassociatedwith
the USEPA Priority Pollutantandthe SuperfundHazardousSubstanceLists. The
Gel PermeationChromatography(GPC) is a size exclusionprocedurein which a
60- to 70-cmby 2.5-cmi.d. glasschromatographiccolumnfitted for upwardflow
operationis filled with 70 g of Bio-BeadsS-X3 and calibratedas outlined in Test
MethodsManual (USEPA, 1986).
Add a 5-mL aliquot of the sampleextract in methylenechloride onto the
column. Any othersolventextractshouldbe concentratedto 1 mL and diluted to
10 mL with methylenechloride to avoid severe shrinkingor swelling of the GPC
packing.Elute the column with methylenechloride, collect eluate containing
analytesof interest,as determinedfrom elution patternsof calibrationstandards.
Concentrateeluateand analyzeusing appropriateanalyticalprocedure.Choiceof
the column packing material and the elution solvent can vary dependingon the
type of organic compoundsto be analyzed.It is important to purge the sample
loading tube thoroughly with the solvent and to run a GPC blank to check for
carry-over.
OtherCleanupProcedures
Adsorption column chromatography,using alumina, florisil and silica gel

columns;acid-basepartitioning to separateacidic and basicorganics;and sulfur
cleanupto remove S from the extracts,are used for specific amalyte groups.
Acid-base partitioning and sulfur cleanup are briefly described below and
adsorptioncolumn chromatographicproceduresare outlined in Table 36-l.
For acid-baseseparationof the extract,add 20 mL methlyene chlorideand
20 mL reagentwater to a lO-mL aliquot of the sampleextract. Adjust pH to 12
to 13 with NaOH, shakefor 1 to 2 min in a separatoryfunnel. Let standfor 10
Table 36-1. Extractcleanupfor specificanalytesusingcolumn chromatography.

Method Analyte Elution Solvent
Alumina column Phathalateesters Hexane
Nitrosoamines Ethyl etherin pentane
F10risil column Phathalateesters Ethyl etherin hexane
Nitrosoamines Acetone,ethyl ether
Organochlorineandorganophosphorus Hexane
pesticidesand haloethers
Chlorinatedhydrocarbons Petroleumether
Nitroaromaticsand isophosone
Silica Gel column Polynuclear aromatic
Methylenechloride/pentane
Hydrocarbons
Derivatizedphenols Hexane
Alumina column (following Base-neutralaliphatics Hexane
acid baseseparation) Base-neutralaromatics Methylenechloride
Base-neutralpolars Methanol
min, separateaqueouslayer which containsphenolsand organic acids. Repeat

extractiontwo more times. Transferto a separatoryfunnel, adjustaqueouslayer
phaseto pH 1 to 2 with sulfuric acid, add 20 mL methylenechloride, shakefor
2 min, separatesolventphase.Repeatextraction twice,concentrateextract.This
separationis generallycarriedout for petroleumwastesprior to further cleanup
throughaluminacolumn (see Table36-1).
Sulfur is generally encounteredin sedimentsamplesand interfereswith
determinationof organochlorineand organophosphoruspesticides,especially
when electroncapturedetectoris used.For sulfur cleanup,the sampleis mixed
with either Cu, Hg or tetrabutylammoniam[(C4H9)W] (TBA)-sulfite reagent.
The mixture is shakenand the extractis separatedfrom sulfur cleanupreagentas
describedin Test Methodsfor EvaluatingSolid Waste(USEPA, 1986).
"BOUND" PESTICIDERESIDUES
Numerousinvestigationshaveshownthat when pesticideresiduesremain

in the soil for extended periodsof time, they becomemore resistantto solvent
extraction (Hamaker et aI., 1966; Khan et aI., 1975; Smith, 1981; Rao &
Davidson,1980; Pavlostathis& Mathavan,1992; Scribneret aI., 1992). Capriel
et ai. (1986) using radioisotopesas tracersin pesticidemoleculesobservedthat
a considerableportion of pesticideresidues remained "bound" or nonextractable.
Steinberget ai. (1987) found that 1,2-dibromoethane(CHzBrChzBr) (EDB)-a
volatile, watersoluble,weakly sorbingorganiccompoundusedas a fumigant-
persistsin soils for a numberof yearsand only a fraction of this EDB is extract-
ed by the commonly usedextractionmethods,such as purge-and-trap,Soxhlet,
and sonication(Sawhneyet aI., 1988). Sawhneyet ai. (1988) describeda proce-
dure involving extractionusing a hot solvent,such as methanol,for a 24-h peri-
od. A hot solvent technique,similar to the one usedabove,has beenemployed
for completerecoveryof slowly reversiblefractions of severalhalogenatedsol-
ventsand soil fumigants(Pignatello,1990)and of herbicides,atrazine(6-chloro-
1082 SAWHNEY
N-ethyl-N'-1,3,5-triazine-2,4-diamine)and metolachlor [2-chloro-N-(2-ethyl-6-

methylphenyl)-N-(2-methoxy-1-methylethyl)-acetamide] in field soil samples
(Huang & Pignatello,1990). The details of the methodare given below.
Weigh 5 g soil in a 40-mL glassscrew-capvial, add 25 mL methanol,and
cap firmly with an unused Teflon-backed silicone-rubberseptum liner after
ensuringthat the septumface andvial lip are free from particles.Acetoneor ace-
tonitrile (~H3N) can be substitutedfor methanolas extractionsolvents.Invert
the vial and mark the level of liquid as referencefor determiningany leak during
heating.Place the vial inverted in an incubator at 75°C for 16 to 24 h. Gentle
orbital shaking(60 rpm or 62.8 rad/s)of the incubatorfacilitates maintainingthe
soil in suspension.As a precautionagainstbreakagethat could result in injury, the
vials should be placedinside anothercontainerand not handleduntil cool. After
cooling, checkthe level of liquid and discardvials where loss in volume is indi-
cated.Centrifugeat 1000 rpm (1047 rad/s) for 10 min, transfersupernatantinto
200-mL flask. Repeatextraction, combine supernatantsand phasetransfer into
hexane,concentrateand analyzeextract using an appropriatetechnique.
Supercriticalfluid extractionprocedureswhich are more efficient than the
solvent extraction of organic chemicalsfrom solid matrices have not yet been
testedfor their effectivity in extractingthe "bound" residuesin field soils. How-
ever, it is likely that under appropriatepressureand temperature,SFE would
effect completeremoval of the "bound" residues.
MATRIX AND SURROGATESPIKING SOLUTIONS
A solution in methanolor acetoneof an organic chemicalfrom an analyte

group (matrix spiking solution) is addedto the sampleand the blank to estimate
recoveryof the organicchemicalscorrespondingto that analytegroup. For water
and soiUsedimentsamples,the concentrationsof the base/neutraland acid com-
poundsshould be 100 and 200 flglmL respectively.Concentrationsof organo-
chlorine pesticidematrix spiking solutionsshould rangefrom 0.2 to 0.5 flglmL.
Concentrationsof the matrix spikesshould be five times higher for waste sam-
ples.
A solution of a surrogatestandard,chemicallyinert substancenot expected
to occur in the sample,also is addedto eachsampleand blank prior to extraction
in order to monitor any unusualmatrix effects or sampleprocessingerrors. An
acceptablerecovery of the surrogatestandardassumesabsenceof errors due to
matrix effectsand sampleprocessing.A list of matrix and surrogatestandardsfor
different analytegroups(USEPA, 1986) is given in Table 36-2.
CONCLUSIONS
Organiccontaminantsincluding solvents,pesticides,PCBsand othersused

in industrial, agricultural and military operationscan enter soil, sedimentand
groundwater. Thus, a variety of methodsdependingon the contaminantcatego-
ry and the natureof the contaminatedsample,whetherliquid, suspensionor solid,
EXTRACflON OF ORGANIC CHEMICALS 1083
Table 36-2. Matrix and surrogatespiking standardsfor different analytegroups.

Purgeableorganics Base/neutrals Acids Pesticide
Matrix standards
l,l-dichloroethene 1,2,4-trichlorobenzenePentachlorophenol Lindane
Trichloroethene Acenaphthene Phenol Heptachlor
Chlorobenzene 2,4-dinitrotoluene 2-chlorophenol Aldrin
Toluene Pyrene 4-chloro-3-methylphenol Dieldrin
4,4'-DDT
Benzene 1,4-dichlorobenzene
Surrogatestandards
p-Bromofluorobenzene2-fluorobiphenyl 2-fluorophenol Ditubylchlorendate
(DBC)
1,2-dichloroethane-d
4 Nitrobenzene-d
s 2,4,6-tribromophenol 2,4,6-tetrachloro-meta-
xylene (TCMAX)
Toluene-ds
solid, have beendevelopedfor the extractionand determinationof theseconta-

minants.Proceduresusedfor their extractionfrom different matrices,including
field-contaminatedsoils, and techniquesfor the cleanupof the extractsprior to
their analysisby determinativemethodsare described.
ACKNOWLEDGMENTS
The authorthanksDr. H.M. Pylypiw for many helpful suggestions.
REFERENCES
Anton, K., R. Menes,and H.M. Widmer. 1988. Direct coupling of CO2 fluid extractionwith capil-
lary fluid chromatography.Chromatographia26:221-223.
Capriel,P., A. Haisch,and S.Y. Khan. 1986.Supercriticalmethanol:An efficaceoustechniquefor the
extraction of bound pesticide residuesfrom soil and plant samples.l. Agric. Food Chern.
34:7(}"'73.
Cary, l.W., l.R. McBride, and C.S. Simmons. 1991. Essayof organic liquid contentsin predomi-
nantly water-wetunconsolidatedporousmedia.l. Contam.Hydrol. 8:135-142.
Hamaker,l.W., c.A.1. Goring, and C.R. Youngson.1966.Organicpesticidesin the environment.Am.
Chern.Soc.,Washington,DC.
Hawthorne,S.B. 1990.Analytical-scalesupercriticalfluid extraction.Anal. Chern. 62:633A-642A.
Hawthorne,S.B., and DJ. Miller. 1987. Extracti~n and recovery of polycyclic aromatichydrocar-
bons from environmentalsolids using supercriticalfluids. Anal. Chern. 59:1705-1708.
Hiatt, M.H. 1981. Analysis of fish and sedimentfor volatile priority pollutants. Anal. Chern.
53:1541-1543.
Huang,L.Q., and1.1. Pignatello.1990. Improvedextractionof atrazineand metolachlorin field soil
samples.l. Assoc. Off. Anal. Chern.73:443-446.
Khan, S.U. 1991. Bound residues.p. 265-279.In R. Grover and AJ. Cessna(ed.) Environmental
chemistryof herbicides.Vol. 2. CRC PressInc., Boca Raton,FL.
Khan, S.U., R. Greenhalgh,andW.P. Cockrane.1975.Determinationof Linuron residuein soil. Bull.
Environ. Contam.Toxicol. 13:602-610.
Levy, L.M., and A.C. Rosselli. 1989. Quantitativesupercriticalfluid extractioncoupledto capillary
gaschromatography.Chromatographia28:613-616.
McNally, M.E.P., and l.R. Wheeler. 1988. Supercriticalfluid extraction coupledwith supercritical
fluid chromato-graphyfor the separationof sulfonylureaherbicidesand their metabolites
from complex matrices.l. Chromatogr.435:63-71.
1084 SAWHN~Y
Pavlostathis,S.G., and G.N. Mathavan.1992.DesorptionKinetics of selectedvolatile organiccom-

poundsfrom field contaminatedsoils. Environ. Sci. Technol.26:532-538.
Pignatello,1.1. 1990.Slowly reversiblesorption of aliphatic hal0carb0nsin soils. 1. Formation of
residualfractions.Environ. Toxicol. Chern.9:1107-1115.
Pylypiw, H.M. Ir., F. Zimmerman,and G.W. Harrington.1985.Apparatusfor tracedeterminationof
volatile N-Nitrosaminesin small samples.Anal. Chern.57:2996-2997.
Rao, P.S.C.,andI.M. Davidson.1980.Estimationof pesticideretentionand transformationparame-
ters required in nonpoint sourcepollution models. p. 234J7.In M.R. Overcashand I.M.
Davidson(ed.)Environmentalimpactof nonpointsourcepollution. Ann Arbor Sci. Publ.,Ann
Arbor, MI.
Rice, P.N., W.E. McGovern, and G.S. Kingsley. 1987. Near critical CO2 extractionof hazardous
organicsfrom acrylonitrile, pesticideandsteelmill wastes.Proj. Summ.EPN600/S2-87!OO5.
USEPA,Cincinnati,OH.
Richards,M., and R.M. Campbell. 1990.Comparisonof supercriticalfluid extraction.Soxhletand
sonicationmethodsfor the determinationof priority pollutantsin soil. LC-GC 9:358-364.
Sawhney,B.L, and R.P. Kozloski. 1984. Organic pollutants in leachatesfrom landfill sites. 1.
Sawhney,B.L, 1.1. Pignatello,andS.M. Steinberg.1988.Determinationof 1,2-dibromoethane (EDB)
in field soils: Implicationsfor volatile organiccompounds.1. Environ. Qual. 17:149-152.
Scribner,S.L, T.R. Benzing,S. Sun, and SA Boyd. 1992. Desorptionand bioavailability of aged
simazineresiduesin soil from a continuouscom field. 1. Environ. Qual. 21:115-120.
Shantz,M.M., and S.N. Chesler.1986. Supercriticalfluid extractionprocedurefor the removal of
traceorganicspeciesfrom solid samples.1. Chromatogr.363:397-401.
Smith, AE. 1981. Comparisonof solventsystemsfor the extractionof atrazine,benzoylprop,flam-
prop, andtrifluralin from weatheredfield soils. 1. Agric. Food. Chern.29:111-115.
Steinberg,S.M., JJ.Pignatello,andB.L Sawhney.1987.Persistenceof 1,2-dibromomethane in soils:
Entrapmentin intrapartic\emicropores.Environ. Sci. Technol.21:1201-1208.
U.S. EnvironmentalProtectionAgency. 1986Testmethodsfor evaluatingsolid waste.SW-846.U.S.
Gov. Print. Office, Washington,DC.
Published 1996
Chapter37
Nitrogen-Total
J. M. BREMNER, Iowa State University, Ames, IA
The determinationof total N in soils and other complexheterogeneous materials

containingseveralforms of N presentsmany difficulties. Total N analysisof soils
is further complicatedby the inadequacyof knowledgeconcerningthe forms of
N presentand by the low N contentof the material under analysis.The total N
contentof soils rangesfrom <0.02% in subsoilsto >2.5% in peats.The surface
layer of most cultivatedsoils containsbetween0.06 and 0.5% N.
Two methodshavegained generalacceptancefor determinationof total N:
the Kjeldahl (1883) method,which is essentiallya wet oxidation procedure,and
the Dumas(1831) method,which is fundamentallya dry oxidation (i.e., combus-
tion) procedure.
In the Kjeldahl method,organic N in the sampleunder analysisis convert-
ed to NHt -N by digestion with concentratedH2S04 containingsubstancesthat
promotethis conversion,and the NHt-N is determinedfrom the amountof NH3
liberatedby distillation of the digest with alkali. In the original Kjeldahl proce-
dure, H2S04 alone was employedfor digestion, and KMn04 was used to com-
plete the oxidation of organic matter. However, numerousinvestigationshave
shown that both the speedand the completenessof conversionof organic N to
NHt -N by digestion with H2S04 can be increasedby adding salts to raise the
temperatureof digestionor by adding catalyststo promoteoxidation of organic
matter. In practically all Kjeldahl methodsnow employed,K2S04 or Na2S04is
usedto raisethe temperatureof digestion,and catalystssuchas Se, Hg, or Cu are
usedto promoteoxidation of organicmatter.
In the classicalDumasmethodof determiningtotal N, the sampleis heat-
ed with CuO at a high temperature(usually above600°C) in a streamof purified
COz, and the gasesliberatedare led over hot Cu to reducenitrogenoxides(main-
ly N20) to N2, and then over CuO to convert CO to CO2, The Nz-C0 2 mixture
thus obtainedis collected in a nitrometercontaining concentratedalkali, which
absorbsthe CO2, and the volume of N2 gas is measured.
The customaryDumasand Kjeldahl methodsfail to recoversomeforms of
N, particularly N in certain heterocycliccompoundsand compoundscontaining
N-N and N-O linkages.Before 1960there was goodreasonto questionthe relia-
bility of conventionalKjeldahl methodswhen they were appliedto soils because,
Book Seriesno. 5.
1085
1086 BREMNER
apart from the fact that the identity of a considerableproportion of soil N was
(and still is) obscure,the literatureon determinationof total N in soils indicated
that thesemethodswere not satisfactory.For example,Alves and Alves (1952)
studiedthe effect of varying the period of digestionin Kjeldahl analysisof soils
using different catalystsand found that the maximal total N value obtainedand
the period of digestionrequiredto attain this value dependedgreatly on the cata-
lyst used.Apart from the wide variation observedin the resultsobtainedwith dif-
ferent catalysts,the most disturbingfinding in this work was that N was lost with
each of the catalysts tested when digestion was continued beyond the time
requiredto achievethe maximal value. However,work by Bremner(1960) pro-
vided goodevidencethat conventionaltypesof Kjeldahl methodscanbe usedsat-
isfactorily for determinationof total N in soils provided certain precautionsare
observed.This evidencewasobtainedby taking advantageof the fact that numer-
ous modifications of customary Kjeldahl methods have been developedthat
greatlyextendthe scopeof the Kjeldahl procedureandpermit inclusionof almost
all combinedforms of N. Application of a selectionof thesemodified Kjeldahl
methodsto a variety of soils containingfrom 0.03 to 2.7% N showedthat with
eachof the soils tested,the resultsby the variousmodified methodswere identi-
cal and in close agreementwith the results obtainedby conventionalmethods.
This work showed that several Kjeldahl methodsthat had been proposedfor
determinationof total N in soils, including the procedurerecommendedby the
Association of Official Agricultural Chemists(1955), were unsatisfactorybut
that good resultscould be obtainedwith thesemethodsby increasing therecom-
mendedperiod of digestion.It also indicatedthat very little of the N in the soils
examinedwas in the form of highly refractoryorganicN compoundsor of com-
poundscontainingN-N or N-O linkages. This work left little reasonabledoubt
that the Kjeldahl methodrecommendedhere("RegularKjeldahl Method") is sat-
isfactory for total N analysisof most soils, becausenone of the other methods
testedgavehighervalues,and thereare few, if any, naturally occurringcombined
forms of N that cannot be determinedby one or anotherof these methods.It
shouldbe emphasized,however,that this work was designedto test the accuracy
of conventionalKjeldahl methodsin analysisof soils containingthe usual low
levelsofNOj" and NOi. Suchmethodsdo not completelyrecoverNOj"-N or NOi
-N, andthey must be modified to include theseforms of N if the soil underanaly-
sis containssignificant amountsof NOj" or NOi, or if, as in sometracerinvesti-
gationsusing 15N-labeledcompounds, it is essentialto recoverNOj"-N and NOi
-N quantitativelyin Kjeldahl analysis fordeterminationof total 15N. Many inves-
tigationsof N transformationsin soils, particularlystudiesof denitrification, have
beenvitiated by the use of Kjeldahl methodsthat fail to quantitatively recover
NOj"-N or NOi-N.
No comprehensivereview of the extensive literature on the Kjeldahl
methodis available,but much of the importantliteratureand information on this
methodis discussedin a monographby Bradstreet(1965) and in articlesby Kirk
(1950), Lake et a1. (1951), Middleton and Stuckey (1951), McKenzie and Wal-
lace(1954),andBaker(1961). Literatureon useofthe Kjeldahl methodfor deter-
mination of total N in soils has beenreviewedby Bremner(1960, 1965), Hesse
(1971), Nelson and Sommers(1980), Bremnerand Hauck (1982), Bremnerand
NITROGEN·TOTAL 1087
Mulvaney (1982), and Bremnerand Yeomans(1988). Literatureon use of auto-

mated Dumas combustion methods for total N analysis of soils has been
reviewed by Bremner and Tabatabai (1971), Bremner and Hauck (1982),
Tabatabaiand Bremner(1982, 1991), and Bremnerand Yeomans(1988).
DUMAS METHODS
Dyck and McKibbin (1935) analyzed26 organicsoils by Dumasand Kjel-

dahl methodsand found that with every sampletested,the Dumasmethodgave
a considerablyhigher N value. Similar resultswere obtainedwith organic soils
in comparativework reportedby Bremnerand Shaw(1958), but they found that
Dumas and Kjeldahl methodsgave practically identical results with mineral
soils. Thesefindings were confirmedby Stewartet ai. (1963),who found that the
ColemanModel 29 N analyzer(an automatedinstrumentfor Dumas analysis;
ColemanInstruments,Inc., Maywood, IL) gave higher valuesthan the Kjeldahl
method with soils containing >0.3% N but gave similar valueswith soils con-
taining <0.3% N (seealso Bremner,1965; Morris et aI., 1968).
Stewartet ai. (1963, 1964) showedthat the higher valuesobtainedby the
Dumas procedurewith soils of high organic-mattercontent were causedby
incompletecombustion,which resultedin methane(CH4) being formed (instead
of CO2) and measuredas N2• Stewart et ai. (1964) and Keeney and Bremner
(1967) subsequentlydemonstratedthat when the Coleman Model 29 or 29A
nitrogen analyzerwas modified to effect completecombustionof soil samples
and eliminate interferenceby CH4, the total N valuesobtainedby theseDumas
analyzerswith both organicand mineral soils agreedclosely with thoseobtained
by Kjeldahl methods.
Stewartet ai. (1964)found that the ColemanModel 29 analyzergavegood
recoveryof NO)-N addedto soil, and Keeneyand Bremner(1967)found that the
Model29A analyzergavenearly quantitative(96-98%)recoveryof both NO)-N
and NOi-N, evenwhen the sampleanalyzedcontainedan appreciableamountof
water. However, Keeney and Bremner (1967) also found that, like customary
Kjeldahl methodsof analysis, Dumas analysisusing the Coleman Model 29A
analyzerdid not give complete recovery of indigenousfixed NHt -N in some
soils containinghigh levels of fixed N.
Use of the ColemanModel 29 analyzerfor soil analysisis complicatedby
sample-sizelimitations that make it essentialto grind soil samplesvery finely
before samplingto reducesamplingerror. The ColemanModel 29A analyzeris
much more suited for soil analysisthan the ColemanModel 29 instrumentbe-
causeit permits analysisof up to 3 g of mineral soil and 1 g of organicsoil, and
the sampleanalyzedneednot be finely ground (Keeney& Bremner,1967).
The Coleman Model 29 and 29A nitrogen analyzerswere developedby
ColemanInstruments,Inc. They did not gain acceptancefor soil analysisbecause
althoughthey requiredvery little laboratoryspacecomparedwith the equipment
neededfor total N analysisby the Kjeldahl method,they were difficult and cost-
ly to operateand allowed only 30 to 40 analysesper day. OtherautomatedN ana-
lyzers have beendevelopedin the USA, Italy, Britain, and Japan,but most have
1088 BREMNER
beendesignedand usedfor total N analysisof purecompoundswith high N con-

tents(Colombo & Giazzi, 1982; Kirsten & Hesselius,1983; Kirsten & Jansson,
1986) and do not appearsuitedfor total N analysisof soils, which usually con-
tain lessthan 0.5% N.
Recentwork indicatesthat the Leco CHN-600 ElementalAnalyzer manu-
factured by LaboratoryEquipmentCorporation,St. Joseph,MI, and the Carlo-
Erba NA-1500 Nitrogen Analyzermanufacturedby Carlo-ErbaStrumentazione,
Milan, Italy, can be usedfor total N analysisof soils. The Leco CHN-600 Ana-
lyzer is an automatedinstrumentin which the sampleunderanalysisis combust-
ed by heatingit to a high temperature(950°C) by a resistancefurnacein a stream
of purified O2, The gasesproducedby combustionare analyzedfor CO2 by a
selectiveinfrared analyzer,and an aliquot is carriedby He to a thermalconduc-
tivity cell for measurementof N2. The time of analysisis 4 min. Yeomansand
Bremner(1991) recently evaluatedthis instrumentfor soil analysisby applying
it to 20 soils selectedto obtain a wide rangein total C and total N contents.They
found that with all 20 soils analyzed,the resultsof total C analysisby the CHN-
600 instrument agreed closely with those obtained by the wet-combustion
methodof Allison (1960), and the resultsof total N analysisagreedclosely with
thoseobtainedby the Kjeldahl methoddescribedin "RegularKjeldahl Method."
They concludedthat this analyzeris well suitedfor routine analysisof soils for
total C and total N (also see Sheldrick, 1986; McGeegan& Naylor, 1988).It
should be noted, however, that the precision of total N analysisby the Leco
instrumentwaslower than that observedwith the Kjeldahl methodandthat there
is a needfor further work to evaluatethis instrumentfor accurateand precise
determinationof total N in soils.
Other automatedinstrumentsthat merit considerationfor total N analysis
of soils are the Leco UP-14SNitrogen Determinator,the Leco 1N-15 Nitrogen
Determinator, the Leco TC-36 Nitrogen/OxygenAnalyzer, the Leco FP-228
Analyzer, the Leco FP-428Nitrogen Determinator,and the Carlo-ErbaNA-1500
Analyzer. The Leco FP-228instrumentis similar in principle to the Leco CHN-
600 Analyzer, but it determinesonly total N and permits more rapid analysis
(160 s) than the Leco CHN-600 instrument.Total N analysisby the Leco1N-15
and the Leco TC-36 Analyzersinvolvesconversionof total N to N2 by fusing the
samplein a graphitecrucibleat high temperature(2500-3500°C)in an inert (He)
atmosphereand subsequentdeterminationof N2 by a gas chromatographicpro-
cedure.The Carlo-ErbaNA-1500 instrumentalso utilizes gas chromatography
for determinationof N2, but it involves flash combustionof the samplein O2 at
1800 to 2OOO°C.
Vittori Antisari and Sequi(1988) recently reportedthat the resultsof total
N analysisof soils by the Carlo-ErbaModel 1102 CHN Analyzer were consid-
erably higher than thoseobtainedby conventionalKjeldahl proceduresand that
a procedureinvolving microwavedigestionof soil with HF-HCI, H 3B03 (boric
acid), and H20 2 also gave significantly higher valuesthan a conventionalKjel-
dahl method. But He et al. (1990) found that this microwave procedure gave
much lower valuesthan conventionalKjeldahl methodswhen it was applied to
soil or plant samples,and otherworkers(e.g., Yeomans& Bremner,1991) have
NITROGEN-TOTAL 1089
found that the resultsof total N analysisof soils by Kjeldahl and automatedcom-
bustionmethodsare in closeagreement.
The mostsignificantrecentdevelopmentin regardto methodologyfor 15N
tracer researchon N transformationsin soils and other biological systemshas
beenthe introductionof automatedsystemsthat permit simultaneousdetermina-
tion of 15N and total N using an automatedN analyzerinterfacedto a continu-
ous-flow isotope-ratiomassspectrometer(ANA-MS system).In the ANA-MS
methoddescribedby PrestonandOwens(1983),the sampleN is convertedto N2
by combustionin a pulseof pure 02. Water is trappedand N2 and CO2 are sepa-
ratedin a gaschromatographiccolumnbeforegasis introducedto the massspec-
trometer.Resultsfrom useof this methodfor 15N andtotal N analysishavebeen
reportedby Barrie andWorkman(1984),Marshall and Whiteway(1985),Schep-
ers et al. (1989), Harris and Paul (1989), Egsgaardet al. (1989), and Jensen
(1991). TheseANA-MS systemsutilize a Carlo-ErbaNA-1500 or Roboprep
Analyzer(EuropaScientific, Crewe,England)for C and N analyses.
Apart from their high cost, the main disadvantageof most automated
Dumascombustioninstrumentsis that they do not permit analysisof more than
about 50 mg of soil or 15 mg of plant material,which meansthat the material
underanalysismust be dried and very finely groundbeforeanalysisin order to
reducesamplingerror. This commentdoesnot apply to the Leco CHN-6ooAna-
lyzer and other Leco instrumentsbecausethey permit analysisof much larger
samplesof soil and plant material.Scheperset al. (1989) studiedthe sampleho-
mogeneityproblem in N analysisof soil and plant samplesby the Carlo Erba
Model NA 1500 Analyzer and concludedthat these samplesshould be ball
milled beforeanalysis.Harris andPaul(1989)evaluatedthe RoboprepCoN Ana-
lyzer for N analysisof soil and plant samplesand recommendedthat samples
should be dried at 60°C and finely ground «250 J.lm) before analysisby this
instrument.
Antek Instruments,Inc. (6005 N. Freeway,Houston, TX 77076), devel-
opedautomatedN analyzersin which the sampleunderanalysisis pyrolyzedin
an oxygen-argonatmosphereto convertN in the sampleto NO, which is deter-
mined by a procedureinvolving useof a chemiluminescentdetectorto measure
the light emitted by metastableN02 producedby mixing NO with 03. These
instrumentshave been used to determineN in petroleumfractions (Drushel,
1977)and to measuregaseousloss of N from plantsduring transpiration(Stutte
& Weiland, 1978),but have not beenevaluatedfor total N analysisof soils.
KJELDAHL METHODS
General
The Kjeldahl proceduresgenerallyemployedfor determinationof total N

involves two steps:(i) digestionof the sampleto convert organicN to NH.t-N
and(ii) determinationof NH.t-N in the digest.The digestionis usuallyperformed
by heatingthe samplewith H2S04 containingsubstances that promoteoxidation
1090 BREMNER
of organic matter and conversionof organic N to NHt-N-the substancesgen-

erally favored are salts such as K ZS04 or NaZS04' which increasethe tempera-
ture of digestion,and catalystssuch as Hg, Cu, or Se,which increasethe rate of
oxidation of organicmatterby HzS04. The NHt-N in the digestis usually deter-
mined by collection of the NH3 liberatedby distillation of the digestwith alkali
[or when the digest contains Hg, with alkali containing sodium thiosulfate
(NaZSZ03),sodiumsulfide (NazS),or zinc dust] and analysisof the distillate for
NHt-N, usually by a titrimetric procedure.
The two-step Kjeldahl proceduredescribedhas proved satisfactory for
total N analysisof most of the nitrogenouscompoundsknown to occur in soils
and plant materials,but it does not give accurateresultswith compoundscon-
taining N-N and N-O linkages (e.g., azo, nitroso, and nitro compounds,
hydrazines,hydrazones,oximes, pyrazolones,isooxazoles,1,2-diazines,1,2,3-
triazines,nitrites, nitrates).However,most of thesecompoundscan be analyzed
successfullyif the customarytwo-stepKjeldahl procedureis modified by inclu-
sion of an appropriatepretreatment.The pretreatments found mostgenerallysuc-
cessfulin Kjeldahl analysisof organiccompoundscontainingN-N and N-O link-
ages are the HI pretreatmentintroduced by Friedrich and his coworkers
(Friedrich, 1933;Friedrichet aI., 1933)andthe zinc-iron pretreatmentintroduced
by Steyermarket al. (1958). Bremner(1960) found that inclusion of thesepre-
treatmentsdid not affect the resultsobtainedin total N analysisof mineral and
organic soils by the Kjeldahl methoddescribedin "Regular Kjeldahl Method,"
which indicatesthat very little, if any, of the N in soils is in the form of organic
compoundscontainingN-N or N-O linkages.However,soils sometimescontain
z,
significant amountsof inorganic-N in the form of NO) or NO which contain
N-O linkages,and this N is not recoveredquantitativelyby the two-stepKjeldahl
proceduresgenerallyemployedfor determinationof total N. The pretreatments
z,
that have been employedfor inclusion of NO), NO and other forms of N in
Kjeldahl analysisof soils are discussedin the next section.
Most of the Kjeldahl methodsusedfor total N analysisof soils and other
materialsbefore about 1960 were macro methodsinvolving use of 350- to 800-
mL Kjeldahl digestionflasks, but semimicroversionsof thesemethodsinvolv-
ing use of 30- to 50-mL flasks gainedpopularity during the 1960sand are now
usedmuch more extensivelythan macro methods.For this reason,only semimi-
cro-Kjeldahl methodsare describedand discussedhere. Descriptionsand dis-
cussionsof macro-Kjeldahlmethodscan be found in the chapteron total N by
Bremner (1965) in the first edition of this monograph. Semimicro methods
gained favor over macro methods because,besidesrequiring less expensive
equipmentand considerablylesslaboratoryspacethan macro methods,they are
safer and require much smaller amountsof soil and reagents.However, when
semimicro-Kjeldahlmethodsare usedfor total N analysisof soils, the soil sam-
ple under analysismust be more finely ground and thoroughly mixed than for
macro-Kjeldahlanalysisto minimize samplingerror.
Pretreatmentof Sample
Soil samplesto be analyzedfor total N are usually dried, ground, and
sieved before analysis,and they are often stored for analysisin paperbags or
NITROGEN-TOTAL 1091
other containersthat are not airtight. Although the effects of thesesoil pretreat-
mentson the resultsof total N havereceived littleattention,thereis evidence that
air or oven drying of soil samplescan lead to substantialloss of NHt -N and
NOz-N (Bremner& Shaw, 1958; Bremner,1965).It seemsunlikely that air dry-
ing of soil sampleswith the customarylow percentage(1-3%) of total N in the
form of inorganicN will significantly affect the resultsof total N analysis.How-
ever, air or oven drying of soil samplescontainingunusually large amountsof
inorganic N may lead to significant loss of N, and drying should be avoided if,
as in many 15N-tracerinvestigations,the soil sampleunderanalysiscontains15N_
labeled NHt-N, NOz-N, or NO:)-N, and it is essentialto obtain quantitative
recoveryof theseforms of N in total N analysisfor determinationof 15N.
Jackson(1958) recommendedthat soil samplesfor macro-Kjeldahlanaly-
sis be ground to passthrough a 0.15-mm(100-mesh)screento ensurecomplete
oxidation of organic matter within small aggregates.But Bremner and Shaw
(1958) found that the resultsof macro-Kjeldahlanalysisof 14 mineral soils con-
taining from 0.07 to 0.54% N using samplesground to passthrough a 0.5-mm
(32-mesh)screen were highly consistentand were not significantly different
from those obtained using samplesground to passthrough a 100-mesh(0.15-
mm) screen.Bremner(1965) recommendedthat beforemacro-Kjeldahlanalysis,
soils containing<0.5% N be groundto passthrougha 32-mesh(0.5-mm)screen,
soils containingfrom 0.5% to 1.0% N be groundto passthrougha 60-mesh(250-
J..lm) screen,and soils containing>1 % N be ground to passthrough a 150-mesh
(150-J..lm)screen.He also recommendedthat before semimicro-Kjeldahlanaly-
sis, soils containing <1% be ground to pass through a 100-mesh(0.15-mm)
screenandsoils containing>1% Nbe groundto passthrougha 150-mesh(0.105-
mm) screen.
Bal (1925) found that the total N valuesobtainedby Kjeldahl analysisof
some Indian soils containing a high percentageof clay were increasedsignifi-
cantly when the soils weretreatedwith waterbeforedigestion.He also notedthat
the residuesfrom digestion of these soils with H2S04 were much coarserand
darkerin color when the pretreatmentwith waterwas omitted,and that the effect
of this treatmentwas markedly greaterwith the clay fractions of the soils than
with the silt or fine silt fractions. From these and other observations,he con-
cluded that the soils containedsome material that cementedthe soil particles
together and protectedorganic matter inside the particles from the action of
H2S04, and that this cementingmaterial was not readily soluble in concentrated
H2S04, but was easily dissolvedby dilute H2S04•
Several workers (Srinivasan, 1932; Walkley, 1935; Ashton, 1936) con-
firmed Bal's observationsregardingthe effect of treating clay soils with water
before Kjeldahl analysis. Walkley (1935) found that ball-milling of such soils
before analysisfor total N had an even greatereffect than treatmentwith water
and deducedthat the lower N valuesobtainedwith thesesoils when the treatment
with water was omitted were not due to the presenceof cementingmaterials
insoluble in concentratedH2S04 but to failure of the soil crumbsto dispersein
this acid.
Sincework during 1956 to 1960 showedthat a considerableportion of the
N in some soils (particularly clay soils and subsoils) is in the form of NHt
1092 BREMNER
trappedin the latticesof clay minerals,the observationsby Bal and Walkley sug-
gestedthat their clay soils containedNHi-N (and possiblyorganicN) within the
latticesof clay mineralsandthat this N wasnot recoveredby the Kjeldahl method
unlessthe soils were first ball-milled to destroyclay latticesor were treatedwith
waterto permit expansionof theselatticesduring digestionwith H2S04• Howev-
er, Bremner(1959, 1960) and Bremnerand Harada(1959) found that a pretreat-
ment with water had no effect on the total N valuesobtainedwhen soils contain-
ing substantialamountsof clay and clay-fixed NHi-N were analyzedby a macro
version of the Kjeldahl method describedin "Regular Kjeldahl Method," and
later work by Bremnerand Mulvaney (1982) showedthat a pretreatmentwith
waterdid not increasethe total N valuesobtainedin Kjeldahl analysisof diverse
Iowa soils by the semimicro-Kjeldahlmethod describedin "Regular Kjeldahl
Method," andtendedto slightly decreasethe valuesobtainedby this methodwith
soils containingmore than about3% of their total N as NO)". But Moraghanet al.
(1983) found that a pretreatmentwith water led to a significant increasein the
value obtainedin total N analysisof an Indian Vertisol soil (51% clay) by macro-
and semimicro-Kjeldahlmethods.They also found that the effect of the water
pretreatmentwas greatly reducedwhen the soil was ground to passa 1oo-mesh
(0.15-mm)screeninsteadof a 16-mesh(0.25-mm)screenbeforeKjeldahl analy-
sis and that it appearedto be associatedwith incompletedigestionof organicN
ratherthan incompleterecoveryof fixed NHt -N.
Bremner(1959, 1960) and Bremnerand Harada(1959) studiedthe possi-
bility that clay-fixed NHt-N may not be recoveredquantitativelyby customary
Kjeldahl methodseven if a pretreatmentwith water and a prolongedperiod of
digestionareemployed.They found thatNH.t-N fixed on additionofNHt to soils
and clay mineralswas recoveredquantitativelyby a macro version of the Kjel-
dahl method describedin "Regular Kjeldahl Method," and that the results ob-
tained by this methodwith soil samplescontainingappreciableamountsof clay
and indigenousclay-fixed NHi-N were not increasedby pretreatmentof the sam-
ples with HF to destroyclay mineralsbeforeKjeldahl digestion(also seeNelson
& Bremner,1966). Stewartand Porter(1963), however,found that this pretreat-
ment led to a significant increasein the total N valuesobtainedin Kjeldahl analy-
sis of profile samplesof two soils containing11 to 55% of their N as indigenous
clay-fIXed NHi-N (see also Meints & Peterson,1972). This led Keeney and
Bremner(1967) to further investigatethe effect of pretreatmentwith HF in Kjel-
dahl analysisof soils containingfixed NHi-N. They found that the Kjeldahl and
HF-Kjeldahl methodsdescribedin "Regular KjeldahlMethod" and "Hydrofluo-
ric Acid Modification of Kjeldahl Method to Include Fixed Ammonium" gave
practically identical resultswith 25 surfacesoils and 7 subsoilscontainingup to
23% of their N as fIXed NHi, but that the HF-Kjeldahl methodgave higher val-
ueswith two subsoilscontaining29 and 66% of their N as fixed NHi-N. These
fmdings supportedthe conclusionthat fixed NHi -N in most soils is determined
satisfactorilyby the Kjeldahl proceduredescribedin "RegularKjeldahl Method."
Further support for this conclusionwas provided by work by Ahlrichs (1965)
using 11 soils containing20 to 32% of their N asfixed NHi andby studiesreport-
ed by Mogilevkina (1970). However, since the work of Stewart and Porter
(1963), Keeneyand Bremner(1967), and Meints and Peterson(1972) leavesno
doubt that the Kjeldahl methoddescribedin "RegularKjeldahl Method" doesnot

completelyrecoverfixed NHt-N in somesoils, an HF-Kjeldahl methodfor total
N analysisof suchsoils is describedin "Hydrofluoric Acid Modification of Kjel-
dahl Method to Include Fixed Ammonium."
Soils presentunusualproblems in total N analysis,becausebesidescon-
taining organic N in a variety of combinationsand NHt -N trappedin the lattices
of minerals,they sometimescontain significant amountsof NO)"-N and NOz-N.
The Kjeldahl methodsusually employedfor determinationof total N do not effect
quantitativerecovery of NO)"-N or NOz-N, but they normally include some of
this N. Therefore,total N in soils containingNO)" and NOz cannotbe calculated
by assumingthat the N recoveredby thesemethodsrepresents(organic + NHt)-
N, and adding(NO)" + NOz)-N as determined byseparateprocedures.The recov-
eries of NO)"-N and NOz-N in Kjeldahl analysisof soils by customarymethods
are highly variable, and they appearto dependlargely on the amountsof water
and organic matter in the sample.High recoveriescan be obtainedwith organic
soils, and loss of NO)"-N during digestioncan be reducedby addition of organic
matter in the form of glucose(Bremner& Shaw, 1958; Goh, 1972).
Three modifications of the Kjeldahl method have been used to recover
NO)"-N and NOz-N in total N analysisof soils: the salicylic acid modification
introducedby Cope (1916), the alkaline reductionmodification of Davissonand
Parsons (1919), and the permanganate-reduced iron modification of Olsen
(1929).
In the salicylic acid modification of the Kjeldahl method, the sample is
treatedwith salicylic acid (C7H60 3) dissolved in concentratedH2S04, and the
nitro compoundsformed by reactionof salicylic acid with NO)" in acid medium
are reducedto the correspondingamino compoundsby heatingthe mixture with
sodiumthiosulfateor zinc dustbeforeKjeldahl digestion.The identity of the nitro
compoundsformed in this procedurehas not beenfully established,but work by
Stalcupand Williams (1955) indicatedthat the main productof nitration is 5-ni-
trosalicylic acid (C7HsNOs) and that small amountsof 3-nitrosalicylic acid also
are formed. Although this methodhas beenusedextensivelyfor total N analysis
z,
of soils containingNO)" and NO seriousdoubtshaveexistedabout its ability to
recover NOz-N quantitatively and its applicability to undried soils (Bremner,
1965). However, work discussedin "Salicylic Acid-ThiosulfateModification of
Kjeldahl Method to Include Nitrate and Nitrite" leavesvery little doubt that the
salicylic acid proceduredescribedin that section gives good recovery of both
NO)"-N and NOz-N and that it is not essentialto dry soil samples beforeanalysis
to eliminate interferenceby water as indicated by Piper (1947). Drying of soil
samplesto eliminate interferenceby water involves the risk of loss of N, and ex-
tensive loss of NHt-N and NOz-N can occur if drying is performed at 105°C
(Bremner& Shaw, 1958; Bremner,1965).
In the alkaline reduction modification of the Kjeldahl method,the sample
is heatedin a Kjeldahl flask with Devardaalloy and alkali to reduce NO)" and
NO z to NH 3, and the NH3 liberatedis collectedin H2S04 in an absorptiontower
connectedto the Kjeldahl flask. When reduction is judged complete,the H2S04
in the absorptiontower is transferredto the Kjeldahl flask and usedfor Kjeldahl
digestion of the sample.This is a tedious and complicatedprocedure,and its
1094 BREMNER
applicationto soils hasbeenvery limited. It wasoriginally designedfor inclusion

of NO)"-N, and its accuracywhen usedto include NOz-N doesnot appearto have
beentested.
In the permanganate-reduced iron modification of the Kjeldahl method,the
sampleis treatedbefore Kjeldahl digestion with KMn04 and H 2S04 to oxidize
NOz to NO)", and then with reducedFe to reduce NO)" to NHt, the reduction
being effectedby the nascenthydrogengeneratedby reactionof the Fe with the
H 2S04 used in the permanganatepretreatment.This method receivedvery little
attentionuntil work by Bremnerand Shaw (1958) showedthat it gave quantita-
tive recovery of NOz-N and NO)"-N added to soils and was applicableto soil
samplescontaininglarge amountsof water. It is the preferredmethodfor total N
analysisin 15N-tracerresearchrequiring quantitativerecovery of small amounts
of 15N-IabeledNOz-N or NO)"-N.
Liao (1981) hasdescribeda modified Kjeldahl methodfor soil analysisthat
involves the useof Devardaalloy and H 2S04 for reductionof NO)" to NHt before
Kjeldahl digestion,and Prudenet al. (1985) have describeda modified Kjeldahl
methodin which Zn is usedto reduceCr(IlI) to Cr(II), which in turn reducesNO)"
to NHt. In the latter method,the sampleis mixed with zinc powder and treated
with an acidified (H2S04) solution of Cr(III)K(S04h. and the mixture is allowed
to stand at room temperaturefor 2 h. The usual Kjeldahl digestion reagentsare
then addedand digestionis performedin a block digesterat 360°C. This method
has the attraction that the pretreatmentdoes not involve heating, and it gives
quantitativerecoveryof NO)" addedto soils or plant materials.However,like the
methodof Liao (1981), it doesnot give quantitativerecoveryof NOi-N (recov-
z
ery of NO is about 80%).
Digestionof SamiJle
In the literature on the Kjeldahl method, the term digestion is used to

describethe processof heatingwith H 2S04 to convertorganic N to NHt-N, the
adjectiverefractory is appliedto organicN compoundsthat aredifficult to digest,
and the term clearing is used to describethe stage in digestion at which the
digestedmaterial ceasesto lose color and is free from char or other suspended
material.Soil digests do not clear in the usual senseof the term becausethey
always containsomesuspendedmaterial.
Studiesby Bremner(1960) showedthat the most important factor in Kjel-
dahl digestion of soils is the temperatureof the treatmentwith H 2S04, which is
controlled largely by the amount of K 2S04 (or Na2S04)employed.If the con-
centrationof K 2S04 is low (e.g., 0.3 glmL of H 2S04), it is necessaryto digestthe
samplefor severalhours to ensureaccurateresults; if it is high (e.g., 1.0 glmL of
H2S04), short periodsof digestion are adequate.Catalystshave a markedeffect
on the rate of digestionwhen the salt concentrationis low but havepractically no
effect when the salt concentrationis high, and satisfactoryKjeldahl analysisof
soil can be performedwithout the use of catalystsby digestion for a compara-
tively short time with H2S04 containinga high concentrationof K 2S04 (e.g., 1 g
of K 2SOJmLof H 2S04).
Kjeldahl methodsthat involve short periodsof digestionusing a high con·

centrationof K 2S04 would appearto have advantages,becausespeedof analysis
is usually an important consideration,and most of the difficulties and dangers
associatedwith Kjeldahl digestion (bumping, spattering,etc.) increasewith the
period of digestion. However, the use of a high concentrationof K 2S04 has the
complicationthat the digestis likely to solidify on cooling, which meansthat con-
siderabletime may be requiredto take up the digestwith waterbeforedistillation
with alkali. It is often stated(or implied) that solidification of a digeston cooling
indicatesthat loss of N occurredduring digestion (Bradstreet,1965), but this is
not true. When Kjeldahl digestionis performedby the methodof McKenzie and
Wallace (1954) using 1 g of K 2SOJmLof H2S04, the digestalwayssolidifies on
cooling, yet this is one of the best methodsavailable for Kjeldahl analysis of
refractory N compounds.Digestsfrom Kjeldahl methodsusing more than about
0.8 g of K 2SOJmLof H 2S04 usually solidify on cooling, and when solidification
occurs,it is often necessaryto reheatthe Kjeldahl flask after addition of water to
disintegratethe solid cake of digest. Bumping usually occursduring this reheat-
ing even if the flask is swirled continuously,and it is sometimesso violent that it
causesfracture of the Kjeldahl flask and loss of the digest.Another disadvantage
of methods using high concentrationsof K 2S04 is that considerablefrothing
occursduring digestionof organicsoils, so that althoughthe period of digestion
requiredafter clearing is considerablyshorterthan with methodsusing low con-
centrationsof K 2S04, the clearingtime is much longer. A further disadvantageof
thesemethodsis that seriousfrothing also occursduring removal of water when
the pretreatmentwith water recommendedby Bal (1925) for Kjeldahl analysisof
clay soilsis used.However,both of thesedifficulties can be minimized by adding
the K 2S04 after a preliminary digestionwith H 2S04•
The main defectof methodsusing highconcentrationsof K 2S04 is that they
will standless abusethan methodsusing low concentrations,becausethe risk of
loss of N during Kjeldahldigestionis more seriouswhen the initial concentration
of K 2S04 is high. Loss of N occurs when the temperatureof digestion exceeds
about400°C, and this temperatureis attainedwhen the concentrationof K 2S04 is
about 1.3 to 1.4 glmL of H 2S04. This is much higher than the concentrationusu-
ally employed for Kjeldahl digestion of soil (0.22-0.33 g of K 2SOJmL of
H2S04) and is appreciablyabove the concentrationrequired for rapid Kjeldahl
digestionof refractory N compoundssuchas tryptophan(C u H 12N20 2) and nico-
tinic acid (C6HsN02) (1.0 g of K 2SOJmLof H2S04). But loss of H2S04 occurs
during Kjeldahl digestion, and this leads to an increasein the salt concentration
and the temperatureof digestion. Under the conditions normally employedfor
Kjeldahl digestion, loss of H2S04 by volatilization and decompositionis very
small if a properly designeddigestion standis used,but a significant amountof
H2S04 is consumedduring Kjeldahl digestionof soil in oxidation of organicmat-
ter and reactionswith the mineral constituents.Determinationsof the amountsof
H2S04 consumedduring Kjeldahl digestion of mineral and organic soils have
shown that there is practically no dangerof loss of N due to rise in temperature
causedby consumptionof H 2S04 when the low salt concentrationKjeldahl meth-
ods recommendedhere are used,even if the amountof soil taken for analysisis
1096 BREMNER
twice the amount specified (Bremner, 1960). This does not hold for Kjeldahl
methodsinvolving high concentrationsof K 2S04, and if suchmethodsare adopt-
ed, consumptionof H2S04 must be taken into considerationand allowed for if
necessary,particularly if the methodsare modified to include N03" and N02.
If sufficient informationis availableregardingthe soils to be analyzed,acid
consumptionduring Kjeldahl digestioncan be estimatedfrom the data in Table
37-1, which showsthe amountsof concentratedH2S04 consumedduring Kjel-
dahl digestionby varioussoil constituentsand by reagentsusedin modifications
of the Kjeldahl methodto include N03" and N02. The acid consumptionvaluefor
the organicfraction of soils is much higher than the correspondingvaluesfor the
inorganic soil constituents,and acid consumptionduring Kjeldahl digestion of
organicsoils can exceed6 mUg of soil. Thefact that ferric and aluminumoxides
have higher valuesthan other inorganicsoil constituentssuggeststhat soils rich
in sesquioxidesmay require specialattention in Kjeldahl analysis.Piper (1947)
apparentlyencountereddifficulties in Kjeldahl analysisof ferruginous and lat-
eritic soils, becausehe recommendedthat the amountof H2S04 normally em-
ployed for Kjeldahl digestionbe increasedfor analysisof suchsoils.
Consumptionof H2S04 is not the only factor to be consideredin assessing
the risk of loss of N during Kjeldahl digestionof soils, becausesoils differ from
mostmaterialsanalyzedby the Kjeldahl methodin havinga high mineralcontent.
The reactionof H2S04 with the mineral constituentsof soils leadsto the forma-
tion of salts,which increasethe temperatureof digestion. But most of the salts
formed during Kjeldahl digestionof soils [CaS04,Fe2(S04)3,Al2(S04)3,etc.] are
not very soluble in concentratedH2S04, and their effect on the temperatureof
digestionis probablyvery small comparedwith the effectsof the saltsformed by
Kjeldahl digestion of the reagentsemployed in modifications of the Kjeldahl
methodto include N03" and N02. The influenceof thesereagentson the temper-
ature of digestion does not appearto have been studied, but there seemslittle
doubt that the sodium thiosulfate usedin the salicylic acid modification of the
Kjeldahl methodhasa markedeffect on the temperatureof digestion.
Table 37-1. Amountsof H2S04 consumedby variousmaterialsduring Kjeldahl digestion(Bremner,

1960).
Material Acid consumption
mL of IBM H2SOJgof material
Soil organicC 10.0
Soil organicmatter 5.8t
Ai 20 3 1.63
Fe203 1.04+
Clay 0.60
CaC03 0.55
Silt 0.30
Sand o
Salicylic acid 6.76
Na2S203• 5H20 0.58
ReducedFe 1.50+
t Calculatedfrom organicC value on the assumptionthat soil organicmattercontains58% C.
+Calculatedon the assumptionthat the digestionproductis Fe2(S04n.
NITROGEN-TOTAL 1097
Sodiumsulfate hasbeenused insteadof K 2S04 to raise the temperatureof

digestion,but this usually increasesspatteringduring Kjeldahl digestionof soils.
The catalystsmost frequently employedfor Kjeldahl digestion are those
containingSe, Hg, and Cu. Their efficiency in producingrapid clearing in Kjel-
dahl digestionof soil decreasesin the order Se > Hg > Cu. Seleniumis not sig-
nificantly more effective than Hg when catalysisof both clearingand conversion
of organicN to NHt-N are considered,but both Seand Hg areconsiderablymore
effective than Cu. Considerablecontroversyexists concerningreports that Se
causesloss of N when usedas a catalystin Kjeldahl digestion,but a critical eval-
uation of the literatureon this subjectindicatesthat Se is a safeand effective cat-
alyst when properly usedand that loss of N occursonly if Se is usedin excessive
quantity, or with large amountsof K 2S04 or Na2S04,and digestionis fairly pro-
longed(Bradstreet,1965). Work by Bremner(1960) supportedtheseconclusions
and showedthat no loss of N occurredduring Kjeldahl digestionof soils by the
selenium-catalyzedmethod recommendedhere ("Regular Kjeldahl Method")
evenwhen digestionis continuedfor as long as 12 h.
It is difficult to evaluatethe extensiveliteratureon the effectivenessof dif-
ferent catalystsin Kjeldahl digestionof organic N compounds,but if effects on
both rate of clearingand conversionof organicN to NHt -N are considered,there
is considerableevidencethat Hg is the most effective single catalyst.However,
the useof Hg hasbeendiscouragedby the practicaldifficulties encounteredin the
determinationof NHt in digestscontainingHg. When a digest containingHg2+
is treatedwith alkali, someof the NHt in the digest reacts withthe HgO precip-
itated by the alkali to form a Hg-NHt complex(Clark, 1943),and the NHt in this
complexis not readily liberatedby distillation with alkali. To avoid low resultsin
the determinationof NHt, it is necessaryto add sodiumsulfide or thiosulfateto
precipitateHg2+ as HgS, or zinc dust to reducethe HgO to metallic Hg. Numer-
ous difficulties have beenexperiencedwith the techniquesusedto preventinter-
ferenceby HgO in the determinationof NH.t. For example,it hasbeenfound that
they can lead to the evolutionof H2S, to the appearance of metallic Hg in the dis-
tillate, to seriousbumping when the Hg is precipitatedas the sulfide, and to the
formation of a black deposit in the condenserof the distillation apparatus.
McKenzie and Wallace (1954) found that when sodium thiosulfatepentahydrate
was usedto eliminateinterferenceby HgO, the successof the methodwas depen-
dent on the ratio of thiosulfateto HgO. Somered HgO was precipitatedwhen the
Na2S203• 5H20-HgO ratio was three, whereasblack HgS was always precipi-
tatedwhen the ratio was five. Theseobservationshave beenconfirmed by other
workers,and it has beenfurther observedthat if a considerableexcessof alkali is
usedfor distillation after precipitation of Hg as HgS, the mixture turns yellow,
and metallic Hg distills into the flask usedto collect the NH3 (Clark, 1943; Brem-
ner, 1960). The precisereasonsfor someof the difficulties experiencedin distil-
lation of NHt from digestscontainingHg are still obscure.In view of this and of
the toxicity of Hg, use of mercury-catalyzedmethodsfor Kjeldahl digestion of
soils is not recommended.
Kjeldahl methodsin which digestion is performedusing 0.6 to 0.7 g of
K 2SOJmL of H2S04 have distinct advantagesover methodsemploying lower
concentrationsof K 2S04 in respectto both speedand completenessof digestion
1098 BREMNER
of organic N, and they appearto be free of some of the defects of methods

employinghigherconcentrationsof K 2S04 (e.g.,frothing, solidification of digest,
risk of loss of N). But experiencewith thesemethodshasbeentoo limited to per-
mit an evaluationof their suitability for soil analysis,and their accuracywhen
modified to include N03-N and N02'-N in Kjeldahl analysisof soils hasnot been
studied.In contrast,the Kjeldahl methodrecommendedhere("RegularKjeldahl
Method") hasbeenfound to give highly reproducibleresultswith a wide variety
of soils andto give excellentrecoveriesof N03-N and N02'-N when modified to
include theseforms of N as describedin "Permanganate-Reduced Iron Modifica-
tion of Kjeldahl Methodto IncludeNitrate and Nitrite" and"Salicylic Acid-Thio-
sulfate Modification of Kjeldahl Method to Include Nitrate and Nitrite."
Severalinvestigators(White & Long, 1951; Grunbaumet aI., 1952) have
found that the useof sealedtubespermitsdigestionat temperatures above400°C
without loss of N and allows completedigestionof heterocyclicN compounds
without addition of the substancesusually employedto promote conversionof
organicN to NUt -N in digestionwith H 2S04, The use of sealedtubesfor diges-
tion also eliminateslossesof N through bumping and preventscontamination
with NH3 from laboratory air. Stevenson(1960) found that these advantages
madethe sealedtube techniquevaluablefor Kjeldahl analysisof rocks, silicate
minerals, and subsurfacesoils containing small amountsof N. However, this
techniqueis too complicatedand hazardousfor routine use in Kjeldahl analysis
of soils.
Honda(1962) describeda Kjeldahl methodof determiningtotal N in soils
involving digestionof the soil samplewith a mixture of H2S04 and H3P04 con-
taining K 2S04 and CUS04 • 5H20. This method appearsattractive for routine
total N analysisof soils, becausethe time of clearing(2-7 min) andtime of diges-
tion after clearing (10 min) are much shorter than the correspondingtimes in
other methods.Work reportedby Skjemstadand Reeve(1976) indicatedthat a
modification of Honda'smethodgavesatisfactoryresultsandcould be adapted to
include N03. However,Nelsonand Sommers(1980) found that seriousbumping
and spatteringoccurredduring digestionof soil samplesby Honda'smethodand
that the clearingtime requiredin this methodwas much longer than 7 min.
Early workers attemptedto use H20 2 to accelerateKjeldahl digestion,but
found that use of this oxidant can lead to low results.There is evidence,howev-
er, that H20 2 can be usedsatisfactorily for Kjeldahl analysis.For example,the
Kjel-FossAutomaticAnalyzer developedby FossElectric of Hillerud, Denmark,
for rapid Kjeldahl analysisusesH20 2 to reducethe time requiredfor conversion
of total N to NHt by the Kjeldahl digestiontechnique,and it has proved satis-
factory for determinationof total N in plants,animals,feeds,and fertilizers (Wall
et aI., 1975; Noel, 1976; Larson & Peterson,1979; Bjamo, 1980). Another Kjel-
dahl method involving the use of H20 2 that merits attentionis the method pro-
posedby Hach et ai. (1985, 1987), which involves digestionwith H2S04 and
H20 2 in the absenceof K 2S04 and catalyst.Watkins et ai. (1987) found that this
methodwas satisfactoryfor routine determinationof total N in variousfeedstuffs
andbiological products,andChristiansonandHolt (1989)found that it gavegood
recovery(average,98.2%)of total N in six soils analyzed.For useofthis method,
NITROGEN-TOTAL 1099
it is necessaryto purchasethe DigesdahlDigestionApparatusmarketedby Hach

Co., Loveland, CO.
Sharmaand Sud (1980) describeda rapid method for total N analysisof
soils and reportedthat the total N valuesobtainedby this methodwere usually
significantly higher than thoseobtainedby conventionalKjeldahl procedures.In
their method, the soil sample is digestedwith a mixture of chromium trioxide
(Cr03) and concentratedH2S04 at 300 ± 10°C for 15 min, and the NHt produced
by this digestionis determinedby estimationof the NH3 liberatedby distillation
of the digestwith sodiumhydroxide.Flowersand Bremner(1991a)recentlyeval-
uated this method by comparing its results with 12 diverse surface soilswith
those obtainedby the Kjeldahl methoddescribedin "Regular Kjeldahl Method
Using Block Digester."They found that the chromic acid methodrecoveredonly
87.5 to 94.1% (average,90.5%) of the soil N recoveredby the Kjeldahl method
and that this was at least partly due to oxidation of ammoniumto nitrate and/or
nitrite by chromicacid and subsequent gaseousloss of theseoxidized forms of N.
The most significant developmentsince 1965 in regardto Kjeldahl diges-
tion has been the introduction of techniquesin which digestion is performedin
Pyrex tubes placed in holes drilled in an aluminum block that is heatedby an
electric hot plate or by internal electric heatersconnectedto a temperaturecon-
trol. Block digestershave significant advantagesover the customarydigestion
stands,becausebesidespermitting better temperaturecontrol during digestion,
they requirelessattentionand fume hood spaceand allow simultaneousdigestion
of 40 to 100 samples.Nelson and Sommers(1972), Schumanet al. (1973), and
Gallaher et al. (1976) were the first to describe the use of aluminum block
digestersfor Kjeldahl digestion of soils. The digestion techniquesadoptedby
theseworkers are outlined in Table 37-2. Aluminum block digesterswith inter-
Table 37-2. Methodsproposedfor Kjeldahl digestionof soils using AI block digesters.

Reference
Details of Nelson & Sommers Schumanet al. Gallaheret al.
method (1972) (1973) (1976)
Pyrex digestion 50 ml,t with funnel in 7S ml, with constricted 75 ml, with funnel in
tube neck neck neck
Soil sample,g 0.2 0.5 1.0
Soil meshsize <100 «0.15 mm) Not stated <60
H2S04, mL 3 10 4
K 2S04, g 1.0 3.4 1.1
Catalyst CUS04 • 5H20 (100 mg) CUS04 (105 mg) CUS04 (33 mg)
Se (IOmg)
Pumice,mg None 35 11
AI block digester Fabricatedblock heatedby TecatorAB digester with Fabricatedblock with
hot plate temperaturecontrol internal heaterand
temperaturecontrol
Temperatureof >350 375 375
digestion,°C
Time of digestion 3.0 2.5 1.5
after clearing,
hours
t Folin-Wu nonproteinN tube.
1100 BREMNER
nal heatersand temperaturecontrols are available commercially from Tecator,

Inc. (Herndon,VA 22070), and from TechniconInstrumentsCorp. (Tarrytown,
NY 10591).Schumanet al. (1973) found thatthe Tecatordigesterwas satisfac-
tory for routine Kjeldahl digestionof soil samples,andwork by NelsonandSom-
mers (1980) indicated that the Technicon digester also is satisfactoryfor soil
analysis(seealso Bremner& Breitenbeck,1983). It is unfortunatethat the high
cost of the block digestersavailablecommerciallyhaslimited their use,because
Bremnerand Mulvaney (1982) found that severalelectrically heateddigestion
standsmarketedin the USA were poorly designedand requiredmodification for
satisfactoryuse. For example,they found that standssuppliedby one manufac-
turer did not provide sufficient heat for satisfactoryKjeldahl digestion of soil
samplesevenwhen the highestheatsettingwas used.They also found that stands
purchasedafter 1970from anothermanufacturerprovidedtoo much heatfor sat-
isfactory digestionevenat the lowest heatsetting,and that evenwhen the lowest
heatsettingwas used,the total N valuesobtainedusing thesestandsfor Kjeldahl
analysisof soils were 1 to 3% lower thanthoseobtainedusing standspurchased
from the same manufacturerduring the 1960s1• This may explain reports that
Kjeldahl methodsof soil analysisinvolving use of block digestersgave slightly
higher valuesthan similar methodsinvolving use of customarydigestionstands
(Nelson & Sommers,1972, 1980; Schumanet al., 1973; Gallaheret al., 1976).
Determinationof Ammonium in Digest
Although severalmethodshave beenproposedfor direct determinationof

NHt in Kjeldahl digests,this determinationis usuallyperformedby estimationof
the NH3 liberatedby distillation of the digestwith strongalkali. In the distillation
methodoriginally employed bymost workers, the NH3 liberatedby distillation
was collectedin an accuratelymeasuredvolume of standardmineral acid (usual-
ly H2S04) and determinedby titration of the excessacid with standardalkali
(usually NaOH) using indicators such as methyl red, bromocresolgreen, and
methyl red-methyleneblue mixtures. This method gives excellent results, but
Winkler's (1913) modification in which the NH3 is distilled into H3B03 and
titrated with standardH2S04 is equally accurate,and it has the advantagethat it
is a direct methodrequiring only one standard reagent, whereasthe original pro-
cedureis a differencemethodrequiring two standardreagents.Neither the vol-
ume nor the strengthof the H3B03 solution usedto collect the distillate needsto
be known accuratelybecausethe ammoniumborate formed by the reaction of
NH3 with H3B03 is titrated back to H3B03 when the distillate is titrated with
H 2S04
NH3 + H3B03 =NHi + H2BOj"

H+ + H2BOj" =H3B03.
1 There appearslittle doubt that the lower valuesobtainedwith the standspurchasedafter 1970
were due to the loss of N throughoverheating,becausewhen thesestandswere modified to prevent
overheating,the total N valuesobtainedin Kjeldahl analysisof soils using the modified standswere
identical to thoseobtainedusing standspurchasedfrom the samemanufacturerduring the 1960s.
A further advantageof the Winkler modification is that a sufficiently large

excessof H3B03 can be usedto ensurecompleteabsorptionof NH3 even if the
amount of NH3 liberated is larger than anticipated.Theuse of H3B03 solution
containing the titration indicator has additional advantages,becausethe color
changesthat occur when the distillate is collected in H3B03-indicator solution
serveto indicateboth the presenceof sufficient alkali in the digestfor distillation
of NH3 to occur and the completion of the NH3 distillation. Severalindicators,
including methyl orange (Winkler, 1913), Congo red (Winkler, 1913), bro-
mophenolblue (Scales& Harrison, 1920), and methyl red (Meeker & Wagner,
1933), have been employed for the titration with standardmineral acid using
Winkler's modification, but the general experiencehas been that the sharpest
end-pointsare obtainedusing mixed indicatorssuch as tetrabromophenolblue-
methyl red (Stover& Sandin,1931),methyleneblue-methylred (Meeker& Wag-
ner, 1933),bromocresolgreen-methylred (Ma & Zuazaga,1942), and bromocre-
sol green-p-nitrophenol-new coccine (Sher, 1955). Automatic titrators with pH-
sensingelectrodesare now commonly used for determinationof NH4 in distil-
latescollectedin H3BOr indicator solution.
When the NH3 liberatedby distillation of Kjeldahl digestsis collected in
H3BOr indicator solution, it is usually determinedby titration of the distillate
with H2S04 or HCI. Titration alsocanbe performedwith sulfamicacid (H3N03S)
(Wagner et aI., 1952) or potassiumbiodate (KHI0 3) (McKenzie & Wallace,
1954), and thesereagentshave the advantagesthat they can be usedas primary
standards.
Tecator,Inc., has developeddistillation systemsthat permit rapid determi-
nation of NH4 in Kjeldahl digests whendigestionis performedwith their 40-tube
block digester. These systemshave the important advantagethat they do not
require transfer of the digestfor NH4 analysis,but their use hasbeen limitedby
their high cost. However, Bremnerand Breitenbeck(1983) developeda simple
and inexpensivesteam distillation apparatusthat permits direct distillation of
NH4 from the Pyrex tubes used for Kjeldahl digestion in commercial 40-tube
block digesters,and they showed that this apparatusallows rapid and precise
determinationof NH4+ in semimicro-Kjeldahlanalysisof soils and plant materi-
als using thesedigesters.Use of this distillation apparatusand a 40-tube block
digesterfor total N analysisof soils is describedin "Regular Kjeldahl Method
Using a Block Digester."
Severalworkers(e.g., Henzell et aI., 1968; Holz & Kremers, 1971; Skjem-
stad& Reeve,1976) have usedautoanalyzersystemsfor colorimetric determina-
tion of NH4 in Kjeldahl digestsof soils. In thesemethods,NH4 in the digest is
separatedfrom otherdigestconstituentsby distillation or dialysis to preventinter-
ference in colorimetric analysis. The NH4 thus separatedis treated with the
reagentsneeded to produce a colored indophenol complex by the Berthelot
(1859) reaction, and the color formed is measuredat absorptionwavelengths
rangingfrom 620 to 660 nm. The autoanalyzersmost frequently employedhave
been those manufacturedby Technicon InstrumentsCorporation.These instru-
ments are currently marketed by Bran & Luebbe, Hamburg, Germany (U.S.
office is in Elmsford, NY). An automatedflow injection analysissystemdevel-
opedby LachatInstruments,Milwaukee, WI, hasbeenproposedfor colorimetric
1102 BREMNER
determinationof NHt in Kjeldahl digestsof soils, but no work to evaluatethis

systemfor NHt analysisof Kjeldahl digestshasbeenreported.
Baethgenand Alley (1989) havedescribeda colorimetric methodfor direct
determinationof ammoniumin Kjeldahl digestsof soils and plant materials.This
methodis a manualmodification of an automatedcolorimetricprocedureusedby
Wall et al. (1975) for determinationof ammoniumin Kjeldahl digestsof proteins.
It is basedon the color reactionbetweenammoniumand a weakly alkaline mix-
ture of sodium salicylate(C7HsNa03)and sodium hypochlorite(NaOCI), and it
involves the use of sodium nitroprusside{-Na2[Fe(CN)sNO]-] to increasethe
intensity and rate of formation of the greencolor producedby this reactionand
the use of sodium tartrate(C4H4Na206)to preventprecipitationof hydroxidesof
heavy metals in the digests. Baethgenand Alley (1989) found that the results
obtainedwhen this methodwas usedto determineammoniumin Kjeldahl digests
of soils and plant materialswere similar to thoseobtainedby the customarydis-
tillation-titration method.
As notedin "Digestion of Sample,"Hach et al. (1985,1987)haveproposed
a Kjeldahl methodthat involves digestionwith H2S04 and H20 2 in the absence
of K 2S04 and catalyst.An attractivefeatureof this methodis that it permitsdirect
determinationof NHt in the digest by a colorimetric procedurethat utilizes
Nessler'sreagent.This colorimetric procedurehas proved satisfactoryfor NHt
analysisof digestsobtainedby heatingvarious feedstuffsand biological materi-
als with H2S04 and H 20 2 as describedby Hach et al. (1985, 1987), but it has not
beenevaluatedfor NHt analysisof soil digests.
A significant developmentsince 1965 in regardto determinationof NHt in
Kjeldahl digestsof soils has beenthe use of NH3 gas-sensingelectrodesfor this
analysis.Theseelectrodespermit rapid, direct determinationof NHt in Kjeldahl
digestsand eliminatethe needfor distillation of the digest(Bremner& Tabatabai,
1972; Gallaher et aI., 1976). Determination of NHt using an NH3 electrode
involves dilution of the Kjeldahl digest to standardvolume and addition of alka-
li to convert NHt to NH 3. The NH3 electroderespondsto the partial pressureof
dissolvedNH3 gas in the samplesolution, which is relatedto the NH3 concentra-
tion. The electrode is calibrated using solutions of known NHt concentration
treatedwith alkali.
Six semimicro-Kjeldahlmethodsfor soil analysisare describedin the fol-
lowing sections.Five involve useof the customarytype ofmicro-Kjeldahl diges-
tion stand.The first methodis referredto as the regularmethodto indicate that it
is applicablein the usual situation in which the soil samplecontainsonly a small
quantity of NOj"-N and NOz-N, and it is not necessaryto include this N. The sec-
ond is a modification of the regular methodin which NHt in the digest is deter-
mined directly by an NH3 electrode.The third methodis a HF modification of the
regularmethodto include fixed NHt. The fourth and fifth methodsare modifica-
tions of the regularmethodto include NOj" and NOz. The sixth methodis a mod-
ification of the regular method in which digestion is performed with a block
digesterinsteadof a digestion stand and the ammoniumformed by digestion is
determinedby a steamdistillation procedurethat allows useof the block digester
tubes as distillation chambersand thereby eliminates the need to transfer the
digestbeforedistillation.
NITROGEN-TOTAL 1103
DISTILLED
o 5 10 WATER
cm. ~ RESERVOIR
TEFLON
<>- NEEDLE
VALVE
! 34/4 5
I
ADJUSTABLE TO TO WATER JET TO VARIABLE
SUPPORT WASTE VACUUM PUMP TRANSFORMER
Fig. 37-1. Modified Hoskinssteamdistillation apparatus.
REGULAR KJELDAHL METHOD2

SpecialApparatus
1. Micro-Kjeldahl digestionflasks, 30 or 50 mL.

2. Micro-Kjeldahl digestionstand.
3. Microburette(or automatictitrator).
4. Steamdistillation apparatus:The apparatusrecommendedis illustrated
in Fig. 37-1. It is essentiallythe Hoskins(1944) steamdistillation appa-
ratus modified to make it suitable for distillation and determinationof
ammonium in 15N-tracerresearch.The main modificationsof the Hos-
kins designin this apparatusare as follows: (i) the baseof the funnel of
the apparatusis fitted with a standard-taper(to/30) ground joint and
funnel plug insteadof a stopcock,(ii) the condenserand spray trap are
connectedby a standard-taper(12/30) groundjoint, (iii) the baseof the
water jacket on the condenseris fitted with a condensatetrap, (iv) the
tube at the baseof the steamjacket of the distillation chamberis con-
nectedby Teflon tubing to a three-waystopcockthat is connectedto a
water-jet vacuum pump, (v) steamis generatedby electrical heatingof
distilled water in a 5-L flask that is connectedto a reservoirof distilled
water and containsa small amountof H2S04 (to trap any ammoniumin
the distilled water) and chips or stones that promote smooth boiling
(e.g., Teflon boiling stones supplied by Berghof/America, Concord,
CA). Before use, the distillation apparatusshould be steamedout to
2 Regular Kjeldahl methoddescribedby Bremner(1960).

1104 BREMNER
removetracesof NH 3, and the rate of steamgenerationshouldbe adjust-

ed so that about6 mL of distillate are collectedper minute. The flow of
cold water through the condenserof the apparatusshould be such that
the temperatureof the distillate obtainedusing this rate of distillation
doesnot exceed25°C.
Reagents
1. Potassiumsulfate-catalystmixture: Preparean intimate mixture of 200
g of K2S04, 20 g of cupric sulfate pentahydrate(CUS04• 5H20), and 2
g of Se. Powder the reagentsseparatelybefore mixing, and grind the
mixture in a mortar to powderthe cake that forms during mixing.
2. Sulfuric acid (H 2S04), concentrated(18 M).
3. Sodium hydroxide (NaOH) solution, approximately10 M: Place3.2 kg
of reagent-gradeNaOH in a heavy-walled10-L Pyrex bottle markedto
indicate a volumeof 8 L. Add 4 L of C0z-freewater, and swirl the bot-
tle until the alkali is dissolved.Cool the solution with a rubberstopper
in the neck of the bottle, and make it to 8 L by the addition of C0z-free
water. Then swirl the bottle vigorously to mix the contents,and fit the
neckwith somearrangementthat permitsthe alkali to be storedand dis-
pensedwith protection from atmosphericCO2 (Piper, 1947; Jackson,
1958).
4. Boric-acid-indicatorsolution: Place 80 g of pure boric acid in a 5-L
flask markedto indicate a volume of 4 L, add about 3800 mL of water,
and heat and swirl until the H3B03 is dissolved. (Dissolution of the
H3B03 also can be effectedby vigorousstirring with a high-speedlabo-
ratory stirrer.) Cool the solution,and add80 mL of mixed indicatorsolu-
tion preparedby dissolving0.099g of bromocresolgreenand 0.066g of
methyl red in 100 mL of ethanol(CH3CH20H). Then add 0.1 M NaOH
cautiouslyuntil the solution assumesa reddishpurple tint (pH approxi-
mately 5.0), and make the solution to 4 L by addition of water. Mix the
solution thoroughly before use.
5. Sulfuric acid (H2S04) or hydrochloricacid (HCI), 0.005M standard.
Procedure
Place a sampleof soil containing about 1 mg of N in a micro-Kjeldahl
digestion flask, add 1.1 g of K 2S04-catalyst mixture and 3 mL of concentrated
H2S04, and heat the flask cautiously on the digestion stand. When frothing has
ceased,increasethe heat until the digest clears, and thereafterboil the mixture
gently for 5 h. Regulatethe heating during this boiling so that the H2S04 con-
densesaboutone-thirdof the way up the neck of the digestionflask.
After completionof digestion,allow the flask to cool, and add about20 mL
of water (slowly, and with shaking).Then swirl the flask to bring any insoluble
material into suspension,and transferthe contentsto the distillation chamberof
the Hoskins apparatusvia the funnel of the apparatus.Rinse the Kjeldahl flask
three times with a total of about 9 mL of water to completethe transfer. Add
enoughwater to the distillation chamberto bring the level of the solution to a
NITROGEN-TOTAL 1105
mark madepreviouslyto indicatea volume of 50 mL, and closethe stopcockcon-

necting the funnel and distillation chamber.Add 5 mL of H3B03-indicator solu-
tion to a 50-mL Erlenmeyerflask marked to indicate a volume of 35 mL, and
placethe flask underthe condenserof the distillation apparatusso that the end of
the condenseris about4 cm abovethe surfaceof the H3B03. Then add 20 mL of
10 M NaOH to the funnel of the apparatus,and run the alkali slowly into the dis-
tillation chamberby opening the funnel stopcock.When about 1 mL of alkali
remainsin the funnel, rinse the funnel rapidly with about 15 mL of water. Allow
enoughof this water to run into the distillation chamberto bring the level of the
solution to a mark made previously to indicate a volume of 80 mL. Then close
the funnel stopcock,and immediatelycommence distillationby closing the steam
bypasstube at the baseof the steamjacket of the distillation chamber.(It is not
necessaryto interrupt to flow of steam to the steamjacket of the distillation
chamberbeforeaddition of the digestand alkali, but the bypasstube of the steam
jacket must be kept openduring theseadditions.)When the distillate reachesthe
35-mL mark on the receiver flask, stop the distillation by opening the steam
bypasstube, rinse the end of the condenser,and determineNHt-N in the distil-
late by titration with 0.005M H2S04 using a lO-mL burettegraduatedat O.OI-mL
intervals(1 mL of 0.005M H2S04 = 0.14 mg ofNHt-N). The color changeat the
end-pointis from greento pink.
Comments
Sincea variety of gas-heatedand electrically heatedmicro-Kjeldahl diges-

tion standsare available commercially, commentson their suitability for soil
analysismust be limited to the following generalstatements:
1. The gas or electric heatersshould have individual controls that permit
rapid adjustmentof the heat, and they should provide sufficient heat to
causebrisk boiling of a mixture of 3 mL of concentratedH2S04 and 3 g
of K 2S04. At full heat the heaterson a micro-Kjeldahl digestion stand
found satisfactoryfor soil analysisbrought 25 mL of water in a 50-mL
Pyrex Kjeldahl flask to a rolling boil in approximately2.5 min.
2. The design should be such that the Kjeldahl flask is supportedat an
angle <45 from the horizontal and is exposedto direct heatonly in the
0
region occupiedby liquid during digestion.Seriousloss of N can occur

during digestion if the flask is exposedto the heaterabove the level of
the liquid.
3. The insulation and venting should be such that a mixture of 3 mL of
H2S04 and 1 g of K2S04 can be boiled continuouslyfor severalhours,
with the H2S04 condensingabout one-third of the way up the neck of
the digestionflask.
Whatever type of digestion stand is used, it is essentialto have some
arrangementfor effective disposal of the acid fumes formed during digestion.
Most of the micro-Kjeldahl digestion standsavailable commercially are fitted
with glassmanifolds that effectively disposeof acid fumes when connectedto a
water-jet vacuumpump.
1106 BREMNER
During the processof digestion, the Kjeldahl flasks should be swirled at

intervals to dislodgeany material adheringto the walls and to bring it into con-
tact with the acid. This is particularly importantwith clay soils becauseclay pro-
motes spattering,and the spatteredmaterial may adhereso tenaciouslyto the
walls of a flask that it is not washeddown into the digestionmixture by the con-
densingH2S04.
Bumping is commonlyencounteredduring Kjeldahl digestionof soils (par-
ticularly sandy soils), but this problem can usually be eliminatedby addition of
glass beadsor other materials that promote smooth boiling. Kjeldahl methods
involving short periods of digestion using a high concentrationof K 2S04 may
prove useful for analysisof sandysoils, becausethe severityof bumpingusually
increaseswith the period of digestion.The dangerof loss of N due to spattering
and bumpingduring digestionis considerablyreducedif the Kjeldahl flaskis sup-
ported at an angle <45 0 from the horizontal as previously recommended.
The 5-hr period of digestion after clearing specified in the method de-
scribedneednot be adoptedif it is not essentialto obtain highlyaccurateresults.
The NHt-N producedby digestionof soils for 2 h after clearing is rarely <98%
of the amount formed by digestion for 5 h, and with most soils, the results of
analysesusing2- and5-h periodsof digestionare identical.A 1-h periodof diges-
tion after clearing is adequatefor routine analysisof soils and for other analyti-
cal work that doesnot require high accuracy.
Severaltypesof steamdistillation units for determinationof NHt are avail-
able commercially,and most of thesecan be employedsuccessfullyto determine
NUt to soil digests.The macro version of the Hoskins unit is recommended
becauseit has significant advantagesover other commercialsteamdistillation
units, and detailedinformation is availableconcerningits use for determination
of NHt (Bremner,1960). Its main advantagesover the Markham (1942) appara-
tus are that it has a large distillation chamber(80 mL of liquid can be safely dis-
tilled) and a very efficient spray trap. The steamdistillation apparatusof Bremn-
er and Breitenbeck(1983) describedin "RegularKjeldahl Method Using a Block
Digester" has similar advantages,and it is recommendedthat this apparatusbe
usedif the Hoskins apparatusis not available.
Considerabletime can be savedby employingtwo Hoskinsunits with their
steambypasstubesconnectedto a water-jetpump as shownin Fig. 37-1, because
the chambersof the units can then be emptiedand washedvery rapidly by suc-
tion. With this arrangementit is not necessaryto tum off the supply of steam
betweensuccessiveanalysesto obtain the vacuum required to empty and wash
the distillation chamber,and it is easily possibleto perform 80 distillations in a
normal working day.
It is convenientto replacethe 500-mL steamgeneratorflask suppliedwith
the Hoskinsapparatusby a 5-L flask andto usean electricheatingmantlefor gen-
eration of steam(Fig. 37-1). With this arrangement,the desiredrate of distilla-
tion is readily obtainedusing a variabletransformerto adjustthe input to the heat-
ing mantle,and oncethis adjustmenthasbeenmade,it is rarely necessaryto alter
it during a seriesof distillations to maintain the desired rate of distillation. A
small amountof H2S04 should be addedto the steamgeneratorflask to trap any
NHt-N in the distilled water usedto generatesteam.It also is advisableto add
NITROGEN-TOTAL 1107
pumiceor glassbeadsto eliminatebumpingin the steamgeneratorflask, because

bumping cancausethe liquid in the distillation chamberto siphoninto the steam
jacket.
Unsatisfactoryresultsare sometimesobtainedif water condensingon the
externalsurfaceof the condenserof the steamdistillation apparatusentersthe
NH3 receiverflask, and a trap for this condensate(Fig. 37-1) or a collar of filter
papershouldbe attachedto the baseof the waterjacket on the condenserto pre-
vent this. It is not necessaryto distill with the endof the condenserunderthe sur-
face of the H3B03 (Bremner,1960).
The sharpnessof the end-pointobtainedin the titration with standardacid
to determineNat in distillatescollectedin H3B03 dependson the quality of the
H3B03, the strengthof the H3B03solution,andthe indicator.The betterthe qual-
ity of the H3B03, and the more dilute the H3B03 solution consistentwith com-
plete retentionof NH3, the sharperis the end-point(Yuen & Pollard, 1953). Var-
ious indicatorshavebeenusedfor the titration ("Determinationof Ammonium in
Digest"), but most workers favorthe bromo-cresolgreen-methylred indicator
mixture recommendedby Ma and Zuazaga(1942). The sharpnessof the end-
point obtainedwith bromocresolgreen-methylred mixture dependson the source
of the indicatorsemployed,and it can vary with different batchesof indicator
from the samesource.It may be necessary,therefore,to alter the recommended
proportionsof the constituentsof mixed indicatorsto obtain a satisfactoryend-
point.
The 5 mL of 2% H3B03 solution used in the method describedshould
effectively absorbabout5 mg of NH3-N (Scales& Harrison,1920; Yuen & Pol-
lard, 1953).This is aboutfive timesthe amountthat will be liberatedif the recom-
mendationsconcerningsamplesize in analysisare followed.
The H2S04 (or HCI) solution employedfor determinationof NUt in the
methoddescribedcanbe standardizedsatisfactorilyusingborax,anhydroussodi-
um carbonate,and other alkaline reagents(Vogel, 1961). Tris(hydroxymethyl)a-
minomethane[2-amino-2(hydroxymethyl)-1,3-propanediol] is recommendedfor
this standardizationbecauseit is availablecommercially(underthe tradenameof
THAM) as a highly purified reagentthan can be usedas a primary acidimetric
standard.This reagenthas a high equivalentweight, is not appreciablyhygro-
scopic,andis readily solublein water.Aqueoussolutionsof this substanceabsorb
CO2 at a slow rate and shouldbe protectedfrom atmosphericCO2 (Bates& Het-
zer, 1961).
Controlsshouldbe performedin eachseriesof analysesto allow for N in
the reagentsused.
It is recommendedthat acetanilide(CgH9NO) or THAM be usedto check
recoveryof organicN by the methoddescribed.Acetanilidespeciallypurified for
useasa standardin N analysiscanbe obtainedfrom the National Bureauof Stan-
dards,Washington,DC, and highly purified THAM suitablefor use as a N stan-
dard (Baker & Chesterfield,1977) is readily availablecommercially.
The method describedgives consistentresults with most mineral soils
when appliedto samplesgroundto passthrough a 60-mesh(250-11m)sieve,but
it is usually necessaryto usemore finely groundmaterialto obtain highly repro-
ducible resultswith organicsoils or with mineralsoils containinga high percent-
1108 BREMNER
age of sand. To ensurepreciseresults, it is recommendedthat soils containing

<1 % N be ground to passthrough a 100-mesh(150-)lm) screen,that soils con-
taining >1% N be groundto passthrougha ISO-mesh(105-)lm) screen,and that
the sievedmaterial be thoroughly mixed before analysis.
The Kjeldahl proceduredescribeddoes not include the pretreatmentwith
water recommendedby Bal (1925) for Kjeldahl analysisof clay soils because
repeatedtestsin the author'slaboratoryusing a wide variety of soils have failed
to reveala single casein which this pretreatmenthasaffectedthe resultsobtained
by this procedure(Bremner,1965; Bremner& Mulvaney, 1982).However,inclu-
sion of this pretreatmentdoesnot add materially to the time requiredfor total N
analysisby the proceduredescribed,and it may prove to be a safeguardagainst
low results in analysisof certain clay soils (see"Pretreatmentof Sample").To
include the pretreatment,treat the soil sample in the micro-Kjeldahl digestion
flask with 2 mL of water before addition of K 2S04-catalystmixture and concen-
tratedH2S04, and swirl the flask for a few minutesand allow it to standfor a fur-
ther 30 min beforeaddition of thesereagents.
AMMONIA ELECTRODEMODIFICATION
OF KJELDAHL METHOD3
SpecialApparatus
1. Micro-Kjeldahl digestionflasks, 30 or 50 mL.
2. Micro-Kjeldahl digestionstand.
3. Orion Model 95-10 NH3 electrode(Orion Research,Inc., Boston, MA)
connectedto an Orion specific ion meter (Orion Model 401, 404, or
407).
4. Magneticstirrer and Teflon-coatedstirring bars.
Reagents
1. Reagents1 and 2 describedin the previous"Reagents."
2. Sodium hydroxide solution, approximately2.5 M: Dissolve 200 g of
reagent-grade NaOH in about1 L of water,anddilute the cooledsolution
with water to 2000 mL. Store in a tightly stopperedbottle.
3. Standardammonium-nitrogen(NHt-N) solution: Dissolve 0.382 g of
pure, dry ammoniumchloride (NH4CI) in about 500 mL of water in a
1000-mLvolumetric flask, and dilute with water to 1000 mL. This solu-
tion contains100 )lg of NHt -N/mL.
Procedure
Place a sample of soil containing about 1 mg of N in a micro-Kjeldahl
digestion flask, and digest it with 3 mL of concentratedH2S04 and 1.1 g of
K2S04-catalystmixture as describedin the previous"Procedure."After comple-
3 Ammonia electrode modification of Kjeldahl method describedby Bremner and Tabatabai

(1972).
NITROGEN-TOTAL 1109
tion of digestion,allow the flask to cool, and add about30 mL of water (slowly,
and with shaking).Then swirl the flask to bring any insoluble material into sus-
pension,and transferthe contentsto a 100-mL volumetric flask. Rinse the Kjel-
dahl flask three times with about 20 mL of water to completethe transfer, and
makethe diluted digestto volume (100 mL) after allowing it to cool to room tem-
perature.
To determineNUt -N in the diluted digest, mix the contentsof the volu-
metric flask thoroughly, and transfera 10-mL aliquot to a 150-mL beakercon-
taining a Teflon-coatedstirring bar. Placethe beakeron a magneticstirrer, add 80
mL of water and 10 mL of 2.5 M NaOH, and insert into the solution an Orion
Model 95-10 NH3 electrodeconnectedto an Orion specific ion metercalibrated
for NHt analysis(the selectorswitch of the meter should be in the monovalent
anion position). The tip of the NH3 electrodeshouldbe about2 cm below the sur-
face of the solution, and the electrodeshould be held at a 20° angle with respect
to the vertical to preventair entrapmentunderthe electl;ode(as recommendedin
the instructionmanualfor the electrode).Stir the solution magneticallyfor 1 min,
record the readingon the logarithmic scaleof the meter,and calculateNHt -N in
the digestfrom this reading.
To calibratethe specific ion meterfor NHt analysis,follow the procedure
describedfor determinationof NHt -N in the diluted digest, but use lO-mL ali-
quots of NHt standardscontaining Illg and 10 llg of NHt-N/mL insteadof 10
mL of the diluted digestand adjustthe meter,as describedin the instructionman-
ual for the specific ion meter, so that a scalereadingof 100 is obtainedwith the
aliquot containing 100 llg of NHt -N and a scale readingof 10 is obtainedwith
the aliquot containing10 llg of NHt -N. To preparestandardscontainingl11g and
10 llg of NHt-N/mL, dilute 10- and 100-mL aliquots of the standardNHt-N
solution with water to 1000 mL in volumetric flasks, and mix the diluted solu-
tions thoroughly.
Comments
For commentsregarding the sample digestion, see the previous "Com-
ments."
All water usedin the proceduredescribedshould be deionized(NHt-free)
distilled water, and controls should be performed in each seriesof analysesto
allow for NHt in the reagentsused. To perform controls, follow the procedure
describedfor determinationof NHt in the diluted sample·digest, but use 10-mL
aliquotsof the diluted control digest,and treat thesealiquotswith 75 mL of water
and 5 mL of the NHt standardcontaining10 llg of NHt-N/mL insteadof 80 mL
of water (allow for the 50 llg of NHt-N thus addedwhen calculatingNHt-N in
the control from the meter readingobtainedwith the NHt -treatedaliquot of the
diluted control digest).
The electrodeshouldbe checkedbeforeuse,rinsedand dried betweenmea-
surements,and storedwhen not in use as recommendedin the instruction manu-
al for the NH3 electrode.
Ammonium analysisof digestsand NHt standardsby the electrodeproce-
dure describedshould be performed immediately after the addition of alkali,
becauseNHt is lost as NH3 gas if digestsor standardsare allowed to standafter
1110 BREMNER
this addition (the pH after the addition of alkali is 13.0 to 13.2). The digestsand
standardsanalyzedin eachseriesof analysesshouldbe at the sametemperature,
and the rate of stirring during their analysisshouldnot be so rapid as to causefor-
mation of a vortex.
Gallaheret al. (1976) have describeda semiautomatedprocedurefor total
N analysisof soil and plant samplesthat involves Kjeldahl digestionof samples
using an aluminumblock digesterand direct determinationof NH.t in the digests
using an Orion Model 95-10 NH3 electrodeconnectedto a Coming Model 101
digital electrometer.The NH3 electrodeis placedin a reactionvesselthat permits
rapid, semiautomaticintroductionof the sampleand alkali, measurement of NH.t-
N concentration,sampleremoval,and a rinse of the electrodeand apparatus.This
semiautomatedprocedurepermitsanalysisof more than 100 samplesin a normal
working day and appearswell suitedfor routine analysisof soils.
The methoddescribedwas developedusing the Orion NH3 electrodespec-
ified, but other NH3 electrodesshould prove satisfactory(e.g., the Orion Model
95-12 NH3 electrodeand the NH3 electrodessuppliedby Coming, Inc., Coming,
NY (Model 476130) and by Unicam Ltd., Cambridge,England(Model 80003),
and any sensitivepH/millivolt metercanbe usedinsteadof the Orion specificion
meterspecified.
HYDROFLUORIC ACID MODIFICATION OF KJELDAHL METIIOD

TO INCLUDE FIXED AMMONIUM4
SpecialApparatus
1. Sameas in "SpecialApparatus"under"RegularKjeldahl Method."
Reagents
1. Reagents1, 3, 4, and5 describedin "Reagents"under"RegularKjeldahl
Method."
2. Hydrofluoric acid-hydrochloricacid solution (approximately5 M HF-l
M HCI): Place about 1500 mL of water in a 2.5-L polyethylene or
polypropylenebottle marked to indicate a volume of 2 L, and add167
mL of concentratedHCI (specific gravity of 1.19) and 325 mL of 52%
(wt/wt) HF (approximately31 M). Then dilute the solution to the 2-L
mark, and mix it thoroughly.
3. Sulfuric acid solution, approximately 4.5 M: Add 510 mL of conc
H2S04 to about 1.5 L of water in a 3-L Pyrex flask markedto indicatea
volume of 2 L. Dilute the cooled solution with water to 2000 mL, and
mix the solution thoroughly.
Procedure
Placea sampleof soil containingabout 1 mg of N in a 25-mL polyethylene
bottle, and add5 mL of 5 M HF-l M HCI solution. Stopperthe bottle tight! y with
a rubberstopperor polyethylenecap, and shakeit on a mechanicalshakerfor 24
4 The hydrofluoric acid modification of Kjeldahl methodto include fixed ammoniumdescribedby

Keeneyand Bremner(1967).
NITROGEN-TOTAL Ull
h. Transferthe resulting suspensionto a micro-Kjeldahl digestionflask contain-

ing 12 mL of 4.5 M H2S04, Perform this transfervia a polyethylenefunnel with
the end of its stem below the surfaceof the 4.5 M H2S04 in the flask, and swirl
the flask during the transfer.After completingthis operation,heat the flask gen-
tly on a digestionstanduntil the water is removed.Then cool the soil-acid mix-
ture, add 1.1 g of K2S04-catalyst mixture,andproceedas describedin "Proce-
dure" under "Regular Kjeldahl Method" for digestion and NHt analysis after
treatmentof the soil samplewith H2S04 and K 2S04-catalystmixture.
Comments
The mixture of soil and HF-HCI solution is shakenin a polyethylenebottle
and is transferredto the digestion flask via a polyethylenefunnel becauseHF
attacksand dissolvesglass. For this reason,micro-Kjeldahl flasks used for the
proceduredescribedshould be inspectedafter use and discardedif they are so
damagedthat they have thin walls and appearfragile.
It is essentialthat the transferof the mixture of soil and HF-HCI solution to
the micro-Kjeldahl digestionflask be performedas specified(i.e., via a polyeth-
ylene funnel with the end of its stembelow the surfaceof the 4.5 M H2S04 in the
flask), becauselow resultsare obtainedif this procedureis not followed.
For other comments,see"Comments"under"Regular Kjeldahl Method."
PERMANGANATE-REDUCEDIRON MODIFICATION
OF KJELDAHL METHOD TO INCLUDE NITRATE AND NITRITES
SpecialApparatus
Reagents
1. Reagents1 to 5 described in "Reagents" under "Regular Kjeldahl
Method."
2. Potassiumpermanganate(KMn04) solution: Dissolve 25 g of KMn04
in 500 mL of water, and store the solution in an amberbottle.
3. Dilute sulfuric acid: To 500 mL of water in a 2-L Pyrex flask, add slow-
ly, and with continuousstirring, 500 mL of concentratedH2S04,
4. ReducedFe: Use a finely divided product and sieve it to remove any
materialthat doesnot passthrougha lOa-mesh(0.15-mm)screen.A sat-
isfactory product is suppliedby FisherScientific Co.
5. n-Octyl alcohol.
Procedure
Placea samplecontainingabout 1 mg of N in a micro-Kjeldahl digestion
flask, add 1 mL of KMn04 solution, and swirl the flask for about30 s. Then hold
the flask at a 45° angle, and pipette 2 mL of dilute H2S04 down the neck. Use a
5 Semimicromodificationof methoddescribedby Bremnerand Shaw (1958).

1112 BREMNER
pipettewith a fine tip to ensureslow addition of the acid, andswirl the flask con-
tinuously during this addition. After addition of acid, allow the flask to standfor
5 min, and add one drop of octyl alcohol. Then add 0.50 ± 0.01 g of reducedFe
througha funnel with a long stemthat reachesdown into the bulb of the Kjeldahl
flask. Swirl the flask to bring the Fe in contactwith the acid, allow it to standuntil
the initial strong effervescencehas ceased(approximately15 min), and heat it
gently on the digestionstandfor 45 min. (The heatingshouldbe suchthat it does
not lead to significant loss of water.) Then allow the flask to cool, add 1.1 g of
K 2S04-catalystmixture and 3 mL of concentratedH2S04, and heatthe flask cau-
tiously on the digestion stand. When the water is removed and frothing has
ceased,increasethe heat until the mixture assumesa yellowish greencolor, and
completethe digestionby boiling the mixture gently for 5 h.
Mter completion of digestion, allow the Kjeldahl flask to cool, and add
about 15 mL of water, slowly, and with shaking.If the digest has solidified on
cooling and the solid cake of digest does not disintegrateduring the addition of
water, heatthe flask cautiouslywith vigorousswirling until the digestis in a fluid
condition. Then cool the flask under a cold-water tap, and transfer the diluted
digest to the distillation chamberof the Hoskins apparatusvia the funnel of the
apparatus.Rinse the Kjeldahl flask four times with about7 mL of water to com-
plete the transfer.Then proceedas describedin "Procedure"under"RegularKjel-
dahl Method" for distillation and determinationof NHt -N after transfer of the
digest to the distillation apparatus.
Comments
Analysesof reduced'Fe from different sourceshaveshownthat someprod-
ucts contain substantialamountsof N, and appreciableamountsof N have been
detected in some batchesof the product recommended.However, this N is
allowed for in control analyses,and highly consistentresults are obtained in
analysesusing 0.5-g samplesof the finely divided material specified.
The methoddescribedis a semimicromodificationof a macro-Kjeldahlpro-
cedurefound to give quantitativerecovery of NOj"-N and N02"-N (Bremner &
Shaw, 1958). It gives quantitative recovery of KNOrN or NaNOz-N addedto
soils.
It is recommendedthat the recoveryof NOj"-Nand N02"-N by the method
describedbe checkedusing purified KN03 and NaN02 supplied by J.T. Baker
Inc., Phillipsburg,NJ ("Baker-Analyzed"reagents)as standards.
For other comments,see"Comments"under"Regular Kjeldahl Method."
SALICYLIC ACID-THIOSULFATE MODIFICATION OF KJELDAHL

METHOD TO INCLUDE NITRATE AND NITRITE6
SpecialApparatus
6 Semimicromodification of methoddescribedby Bremner(1965).

NITROGEN·TOTAL 1I13
Reagents
1. Reagents1, 3, 4, and 5 describedin "Reagents"under"RegularKjeldahl

Method."
2. Salicylic acid-sulfuricacid mixture: Dissolve25 g of salicylic acid in 1
L of concentratedH2S04,
3. Sodium thiosulfatepentahydrate(Na2S203• 5H20): Powdercrystalsof
sodium thiosulfate pentahydrateto pass through a 60-mesh(250-llm)
screen.
Procedure
Place a sample of soil containing about 1 mg of N in a micro-Kjeldahl

digestionflask. Add 4 mL of salicylic acid-sulfuric acid mixture, and swirl the
flask until the acid is thoroughly mixed with the soil. Allow the mixture to stand
for several hours (or overnight), add 0.5 g of sodium thiosulfate pentahydrate
througha dry funnel with a long stemthat reachesdown into the bulb of the Kjel-
dahl flask, and heat the mixture cautiouslyon the digestionstand until frothing
hasceased.Thencool the flask, add 1.f g of K 2S04-catalystmixture, andproceed
as describedin "Procedure"under"Regular Kjeldahl Method" for digestionand
NHt analysisafter treatmentof the soil samplewith H2S04 and K2S04-catalyst
mixture.
Comments
As noted in "Pretreatmentof Sample,"seriousdoubt has existedconcern-

ing the ability of the salicylic acid modification of the Kjeldahl methodto recov-
z
er NO -N quantitativelyandthe applicability of this methodto undriedsoils. Goh
(1972) increaseddoubt aboutthe reliability of this procedure,becausehe report·
ed that it gave poor recovery of N03-N addedto a Muscatinesoil and that the
total N valuesobtainedby this procedurein analysisof soils containing>40 mg
kg-1 of N03-N were significantly lower than those obtainedby a reducediron
modification of the Kjeldahl method to include N03-N. According to Piper
(1947), nitration of salicylic acid doesnot occur if more than a trace of water is
present,which indicatesthat the salicylic acid procedurecannotbe applied suc-
cessfully to undried soil samples.However, Cheng and Bremner(1964) found
that the proceduredescribedgave goodrecoveryof N03-N andNOz-N evenwith
soil samplescontainingas muchas 0.6 mL of water/gramof soil. Also, Bremner
and Mulvaney (1982) found that the total N values obtainedby the procedure
describedwith soils treatedwith KN03 or NaN02 so that they contained74 to
z
89% of their N as NO)" or 70 to 91% of their N as NO were identical to those
obtained by the permanganate-reduced iron proceduredescribedin "Perman-
ganate-Reduced Iron Modification of Kjeldahl Method to Include Nitrate and
Nitrite."
Work by Chengand Bremner(1964) indicatedthat it may not be necessary
to add salicylic acid to obtain quantitativerecoveryof N03-N and NOz-N by the
salicylic acid-thiosulfatemodification of the Kjeldahl method.Dalal et al. (1984)
1114 BREMNER
TmON TUBING
NO 4 tlEOI'RENE 51OPf'ER
WITH 15MM HOLE
TEfLON TUBING
10 1,5
Fig. 37-2. Steamdistillation apparatus(Bremner& Breitenbeck,1983).
have reportedwork supportingthis conclusion andhavedescribeda modification

of the Kjeldahl method in which only NaZSZ03 is used to obtain quantitative
recoveryof NOz-N and N03'-N.
It is recommendedthat the recoveryof N03'-N and NOz-N by the proce-
dure describedbe checkedby analyzingthe purified KN03 and NaNOz supplied
by J. T. Baker Inc.
For othercomments,see"Comments"under"RegularKjeldahl Method."
REGULAR KJELDAHL METHOD USING A BLOCK DIGESTER'
SpecialApparatus
1. Block digestertubes,100 mL.

2. Tecatoror Technicon40-tubeblock digester.
3. Steamdistillation apparatus.The apparatusused(Fig. 37~2) is designed
so that the Pyrex tubes used for Kjeldahl digestion in the Tecator and
Technicon40-tubeblock digesterscan be usedas distillation chambers.
The steamrequiredfor distillation is generatedby heatingdistilled water
in a 5-L flask that containsa small amountof HzS04 (to trap any ammo-
nium in the distilled water) and chips or stonesthat promote smooth
7 The regular Kjeldahl method using a block digester describedby Bremner and Breitenbeck
(1983).
NITROGEN-TOTAL 1115
boiling (e.g., Teflon boiling stonessuppliedby Berghof/America,Con-

cord, CA). Before use, the distillation apparatusshould be steamedout
for about 10 min to removetracesof NH3, and the rate of steamgener-
ation shouldbe adjustedso that about5 mL of distillate are collectedper
minute. The flow of cold water through the condensershould be such
that the temperatureof the distillate obtainedusing this rate of distilla-
tion does not exceed22°C. The desired rate of distillation is readily
obtained if the steam generatorflask is heatedby an electric heating
mantle and the power supply is controlled by a variable transformeras
in the steam generatorunit describedin "Special Apparatus" under
"Regular Kjeldahl Method." The trap at the baseof the condenseris to
preventwater condensing on the externalsurfaceof the condenserfrom
enteringthe flask usedto collect the distillate.
4. Microburette(or automatictitrator).
Reagents
Reagents1 to 5 describedin "Reagents"under"RegularKjeldahl Method."
Procedure
Placea sampleof soil containingabout1 mg of N in a 100-mL Pyrex block

digester tube, add 1.1 g of K 2S04-catalyst mixture and 3 mL of concentrated
H2S04, and place the tube in a 40-tube Tecator or Techniconblock digesterat
375°C for 3 h. Then allow the digestto cool, treat the cooled digestwith 15 mL
of distilled water, swirl the tube to mix the contents,and allow the diluted digest
to cool to room temperature.
To determineammoniumN in the diluted digest, add 5 mL of boric acid-
indicator solution to a 50-mL Erlenmeyerflask markedto indicate a volume of
20 mL, and place the flask under the condenserof the distillation apparatusso
that the end of the condenseris below the surfaceof the H3B03 solution. Then
add 15 mL of 10 M NaOH (slowly, and without shaking) to the cooled diluted
digest, connectthe digestion tube to the distillation apparatusas shown in Fig.
37-2 and immediately commencesteamdistillation by closing the stopcockon
the steam-bypass tube of the distillation apparatus.When thedistillate reaches the
20-mL mark on the receiverflask, lower the flask until the tip of the condenseris
abovethe surfaceof the distillate, stop the distillation by openingthe stopcockon
the steam-bypass tube, rinse the end of the condenser,and determineNHt-N in
the distillate by titration with 0.005 M H2S04 from a lO-mL burettegraduatedat
0.01-mL intervals (1 mL of 0.005 M H2S04 = 0.14 mg of NHt-N). The color
changeat the end-pointis from greento pink.
Comments
Bremnerand Breitenbeck(1983) found that the resultsobtainedin ammo-

nium N analysisof Kjeldahl digestsof soils and plant materialsby the direct dis-
tillation method describedagreedclosely with, and were slightly more precise
1116 BREMNER
than, those obtainedby the customarymethod involving transfer of the digest

beforedistillation. They also found that it was not necessaryto collect more than
15 mL of distillate to achievequantitative(99.8-100.2%)recoveryof ammonium
N by the distillation methoddescribed,but thatit wasessentialto conductthe dis-
tillation with the end of the condenserbelow the surfaceof the boric acid solu-
tion as specifiedin the proceduredescribed(approximately3% of ammoniumN
distilled was not recoveredwhen this precautionwas not observed).
The digestiontube illustrated in Fig. 37-2 is the 30- by 2.5-cm tube used
for Kjeldahl digestion in the Tecator40-tubeblock digester.The tubes used for
digestionin the Technicon40-tubeblock digesterhave the samediameteras the
tube illustratedin Fig. 37-2 but are 5 cm shorter,so the Teflon tubing attachedto
the end of the glasstube that deliverssteamto the digestionflask must be short-
enedby 5 cm when thesetubes are used. The distancebetweenthe end of the
Teflon tubing and the bottom of the digestiontube should not exceed1 cm.
TecatorInc. marketsa manifoldexhaustsystemthat effectively disposesof
acid fumes when it is placedover a Tecator40-tubeblock digesterand connect-
ed to a water-jetvacuumpump in the fume hood usedfor Kjeldahl digestion.It
is essentialthat this manifold or a similar type of fume disposal system be
employedwhen a block digesteris usedfor Kjeldahl digestionof soils because
use of block digesterswithout such systemscan lead to seriousdamageto fume
hoodsusedfor Kjeldahl digestion.
Bremnerand Breitenbeck(1983) found that the steamdistillation proce-
dure in the methoddescribedwas satisfactoryfor determinationof ammoniumin
digestsobtainedby the semimicro-Kjeldahl methods describedin "RegularKjel-
dahl Method," "Permanganate-Reduced Iron Modification of Kjeldahl Method to
Include Nitrate and Nitrate," and "Salicylic Acid-Thiosulfate Modification of
Kjeldahl Method to Include Nitrate and Nitrite." They also found that the results
obtainedwith Iowa soils by the Kjeldahl methoddescribedusing 3-and5-h peri-
ods of digestionwere identical and that a I-h period of digestionwas adequate
for routine analysisof soils for total N.
The methoddescribedis simplerand more rapid than the methoddescribed
in "Regular KjeldahlMethod" and is the methodof choice in the author'slabo-
ratory in the usualsituationin which the soil sampleunderanalysiscontainsonly
a small quantity of N03"-Nor NOi -N and it is not necessaryto include this N.
OTHER METHODS
Dalal and Henry (1986) have proposedthe use of near infrared diffuse
reflectance(NIR) spectrophotometryfor simultaneousestimation of total N,
moisture, and organic C in air-dried soils. This techniqueallows rapid, nonde-
structive estimationof total N in soil, but it seemsuseful only for routine analy-
sis of finely groundsoils that havea limited rangein color and containmoderate
amountsof organicmatter(0.3-2.5%C).
Two methodsinvolving the use of persulfatefor oxidation of organicN to
nitrate have been proposedfor total N analysisof soils. In one, the sample is
digestedwith potassiumpersulfatesolution in an autoclaveat 1.5 x UP Pa for 2
NITROGEN-TOTAL 1117
h, and the digest is treatedwith Devardaalloy to reducenitrate to ammonium,

which is determinedby a colorimetric procedure(Raveh& Avnimelech, 1979).
In the other, the soil sampleis digestedwith potassiumpersulfate(K20SS2) and
sodiumhydroxidesolution in an autoclaveat 125°Cfor 30 min, and the digest is
filtered, acidified, and analyzedfor nitrate by a spectrophotometricprocedure
(Huang & Yang, 1987).Thesemethodshave not beenevaluatedby comparison
with othermethodsusinga variety of soils, andthey havereceivedlittle attention.
Flowers and Bremner (1991b) recently describeda simple procedurefor
routine estimationof total N in soils. It involves (i) digestionof the soil sample
with an acidified dichromate(K2Cr20TH2S04)solution for 45 min in a Pyrex
digestion tube in a 40-tube block digester preheatedto 170°C, (ii) titrimetric
determinationof the NH3-N liberated by direct steamdistillation of the digest
with alkali (NaOH), and (iii) calculationof the total-N value by applicationof a
correction factor (1.1) to the measuredNH3-N value. Studies using 14 soils
selectedto obtaina wide rangein total N contentandotherpropertiesshowedthat
the uncorrectedrecovery of N as Nat by this procedurerangedfrom 88.5 to
92.3% and averaged90.5%. When a correctionfactor of 1.1 was applied to the
results,the recoveryofN rangedfrom 97.4 to 101.5%and averaged99.6%.This
dichromateprocedureis rapid and precise,and it permitsat least80 analysesin a
normal working day. The only equipmentrequiredfor its use is equipmentcom-
monly usedfor total N analysisof soils by the Kjeldahl method(40-tubeblock
digesterandsteamdistillation unit), and the acidified dichromatesolutionusedin
the procedurefor digestionof soil is the reagentcommonly usedfor determina-
tion of organicC in soil by a modified Mebiusprocedure(Yeomans& Bremner,
1988). Besidesbeing simpler and more rapid than the Kjeldahl method, this
dichromateprocedurehas the advantagethat it does not require the expensive
fume disposalsystemneededfor Kjeldahl digestion. It is clearly not suitable,
however,for soil investigationsthat require high accuracyin total N analysis.
ACKNOWLEDGMENTS
Contribution from the Departmentof Agronomy, College of Agriculture,

Iowa StateUniversity, Ames, IA 50011.
REFERENCES
Ahlrichs, L.E. 1965.Ammoniumfixation andreleaseon someMinnesotasoils. Ph.D.diss.Univ. Min-
nesota(Diss. Abstr. 66-8853).
Allison, L.E. 1960.Wet-combustionapparatusandprocedurefor organicand inorganiccarbonin soil.
Soil Sci. Soc.Am. Proc. 24:36-40.
Alves, J.A., and E.L.N. Alves. 1952. Sobrea determinacaodo azoto em sole pelo macrometodode
Kjeldahl. Melhoramento5:77-83.
Ashton, F.L. 1936. Seleniumas a catalystin the Kjeldahl methodas applied to soil and grassanaly-
sis. 1. Agric. Sci. 26:239-248.
Associationof Official Agricultural Chemists.1955. Official methodsof analysis.8th ed. AOAC,
Washington,DC.
Baethgen,W.E., and M.M. Alley. 1989.A manualcolorimetric procedurefor measuringammonium
nitrogenin soil and plant Kjeldahl digests.Commun.Soil Sci. PlantAnal. 20:961-969.
1118 BREMNER
Baker,HJ., and R.S. Chesterfield.1977.Tris-(hydroxymethyl)-aminomethane: A dual purposestan-

dard for Kjeldahl nitrogendeterminations.Food Technol.Aust. 29:497-499.
Baker, P.R.W. 1961. The micro-Kjeldahl determinatonof nitrogen.An investigationof the effectsof
addedsaltsand catalysts.Talanta8:57-71.
Bal, D.V. 1925. The determinationof nitrogen in heavy clay soils. 1. Agric. Sci. 15:454-459.
Barrie, A, and C.T. Workman. 1984. An automatedanalytical systemfor nutritional investigations
using 15N tracers.Int. 1. Spectrosc.3:439-447.
Bates,R.G., and H.B. Hetzer.1961.Absorptionof carbondioxide by solutionsof 2-amino-2-(hydrox-
ymethyl)-I,3-propanediol.Anal. Chern.33:1285.
Bjamo, O-C. 1980. Kjel-Foss automaticanalysisusing an antimony-basedcatalyst:A collaborative
study.1. Assoc.Off. Anal. Chern. 63:657-663.
Berthelot,M.P. 1859. Violet de'aniline.Chim. Appl. 1:284.
Bradstreet,R.B. 1965.The Kjeldahl methodfor organicnitrogen.Acad. Press,Inc., New York.
Bremner,I.M. 1959. Determinationof fixed ammoniumin soil. 1. Agric. Sci. 52:147-160.
Bremner, I.M. 1960. Determination of nitrogen in soil by the Kjeldahl method. 1. Agric. Sci.
55:11-33.
Bremner,I.M. 1965. Total nitrogen. p. 1149-1178.In C.A. Black et al. (ed.) Methodsof soil analy-
sis. Part 2. Agron. Monogr. 9.
Bremner,J.M., and G.A Breitenbeck.1983. A simple method for determinationof ammoniumin
semimicro-Kjeldahlanalysisof soils andplant materialsusing a block digester.Commun.Soil
Bremner,J.M., and T. Harada.1959. Releaseof ammoniumand organic matter from soil by hydro-
fluoric acid and effect of hydrofluoric acid treatmenton extractionof soil organic matterby
neutraland alkaline reagents.1. Agric. Sci. 52:137-144.
Bremner,I.M., and R.D. Hauck. 1982. Advancesin methodologyfor researchon nitrogen transfor-
mationsin soils. p. 467-502.In FJ. Stevenson(ed.) Nitrogen in agriculturalsoils. ASA, Madi-
son, WI.
Bremner,I.M., and C.S. Mulvaney. 1982.Nitrogen-Total.p. 595-624.In AL Pageet al. (ed.) Meth-
ods of soil analysis.Part 2. 2nd ed. Agron. Monogr. 9.
Bremner,I.M., and K. Shaw. 1958. Denitrification in soil. I. Methodsof investigation.1. Agric. Sci.
51:22-39.
Bremner,I.M., and M.A. Tabatabai.1971. Use of automatedcombustiontechniquesfor total carbon,
total nitrogen, and total sulfur analysisof soils. p. 1-15. In LM. Walsh (ed.) Instrumental
methodsfor analysisof soils and plant tissue.SSSA,Madison,WI.
Bremner,I.M., and M.A Tabatabai.1972. Use of an ammoniaelectrodefor determinationof ammo-
nia in Kjeldahl analysisof soils. Commun.Soil Sci. Plant Anal. 3:159-165.
Bremner,I.M., and I.C. Yeomans.1988. Laboratorytechniquesfor determinationof different forms
of nitrogen. p. 399-414.In I.R. Wilson (ed.) Advancesin nitrogen cycling in agricultural
ecosystems.C.AB. Int., Wallingford, England.
Cheng,H.H., and I.M. Bremner.1964. Use of the salicylic acid-thiosulfatemodification of the Kjel-
dahl methodfor determinationof total nitrogen in soils. p. 21. In Agronomy abstracts,ASA,
Madison,WI.
Christianson,C.B., and LS. Holt. 1989. Rapid digestion procedurefor the determinationof total
nitrogen and nitrogen-15contentof soils. Soil Sci. Soc. Am. 1. 53:1917-1919.
Clark, E.P. 1943. Semimicroquantitativeorganicanalysis.Acad. Press,Inc., New York.
Colombo, B., and G. Giazzi. 1982. Total automaticnitrogen determination.American Laboratory,
luly, p. 38-45.
Cope, W.C. 1916. Kjeldahl modification for determinationof nitrogen in nitro substitution com-
pounds.1. Ind. Eng. Chern.8:592-593.
Dalal, R.C., and R.I. Henry. 1986. Simultaneous determinationof moisture, organiccarbon,and total
nitrogenby nearinfrared reflectancespectrophotometry. Soil Sci. Soc.Am. 1. 50:120-123.
Dalal, R.C., K.L. Sahrawat,and RJ.K. Myers. 1984. Inclusion of nitrate and nitrite in the Kjeldahl
nitrogen determinationof soils and plant materialsusing sodium thiosulfate.Commun. Soil
Davisson,B.S., and I.T. Parsons.1919.The determinationof total nitrogen including nitric nitrogen.
1. Ind. Eng. Chern. 11:306-311.
Drushel, H.V. 1977. Determinationof nitrogen in petroleumfractions by combustionwith chemilu-
minescentdetectionof nitric oxide. Anal. Chern.49:932-939.
Dumas,I.B.A.1831. Procedesde I'analyseorganique.Ann. Chim. Phys.247:198-213.
Dyck, A W.l., and R.R. McKibbin. 1935. The non-proteinnatureof a fraction of soil organic nitro-
gen. Can. 1. Res. 13B:264-268.
Egsgaard,H., E. Larsen,and E.S. Jensen.1989. Evaluationof automateddeterminationof nitrogen-

15 by on-line combustion.Anal. Chim. Acta 226:345-349.
Flowers,T.H., and J.M. Bremner. 1991a.Evaluationof a rapid chromic acid methodfor determina-
tion of total nitrogen in soils. Commun.Soil Sci. Plant Anal. 22:1417-1422.
Flowers,T.H., and lM. Bremner.1991b.A rapid dichromateprocedurefor routine estimationof total
nitrogen in soils. Commun.Soil Sci. Plant Anal. 22:1409-1416.
Friedrich,A. 1933.Die Praxisder quantitativenorganischenMikroanalyse.E Deuticke,Leipzig, Ger-
many.
Friedrich,A., E. Kuhaas, and R. Schnurch. 1933. Uber die generelleAnwendung der MikroKjel-
dahlbestimmung.Z. Physiol. Chern. 216:68--76.
Gallaher,R.N., C.O. Weldon, and Ee. Boswell. 1976. A semiautomatedprocedurefor total nitrogen
in plant and soil samples.Soil Sci. Soc. Am. J. 40:887--889.
Goh, K.M. 1972. Comparisonand evaluationof methodsfor including nitrate in the total nitrogen
determinationof soils. J. Sci. Food Agric. 23:275-284.
Grunbaum,B.w., EL. Schaffer,and P.L. Kirk. 1952. Kjeldahl determinationof nitrogen with sealed
tube digestion.Anal. Chern.24:1487-1490.
Hach, e.e.,S.Y. Brayton,and A.B. Kopelove.1985.A powerful Kjeldahl nitrogen methodusingper-
oxymonosulfuricacid. l Agric. Food Chern. 33:1117-1123.
Hach, e.e.,B.K. Bowden,A.B. Kopelove,and S.Y. Brayton. 1987. More powerful peroxideKjeldahl
digestion.J. Assoc. Off. Anal. Chern. 70:783-787.
Harris, D., and E.A. Paul. 1989. Automatedanalysisof 15N and 14C in biological samples.Commun.
He, X.T., R.L. Mulvaney, and w.L. Banwart.1990.A rapid methodfor total nitrogen analysisusing
microwavedigestion.Soil Sci. Soc. Am. J. 54:1625-1629.
Henzell, E.E, 1. Vallis, and lE. Lindquist. 1968. Automatic colorimetric methodsfor the determina-
tion of nitrogen in digestsand extractsof soils. p. 513-520.In Trans. 9th Int. Congr. Soil Sci.
3, Int. Soil Sci. 1968.
Hesse,P.R. 1971. A textbookof soil chemicalanalysis.Chern. Publ. Co., New York.
Holz, E, and H. Kremers.1971. Die automatischeBestimmungdesStickstoffsals Indophenolgrunin
Boden und Pflanzen.Landwirtsch.Forsch.Sonderh.26:177-199.
Honda, e. 1962. Rapid determinationof nitrogen in soil by Kjeldahl method.(In Japanese.)1. Sci.
Soil Manure 33:195-200.
Hoskins,J.L. 1944. An interchangeablemicro and macrosteamdistillation apparatus.Analyst (Lon-
don) 69:271.
Huang,e., and L. Yang. 1987. New methodfor determiningtotal nitrogenin soils and fertilizers. (In
Chinese.)Fenxi Huaxue15:384.
Jensen,E.S. 1991.Evaluationof automatedanalysisof 15N and total N in plant materialand soil. Plant
Soil 133:83-92.
Keeney, D.R., and I.M. Bremner.1967. Use of the ColemanModel 29A Analyzer for total nitrogen
analysisof soils. Soil Sci. 104:358--363.
Kirk, P.L. 1950. Kjeldahl methodfor total nitrogen.Anal. Chern. 22:354-358.
Kirsten, W.J., and G.U. Hesselius.1983. Rapid, automatic, high-capacityDumas determinationof
nitrogen. Microchem.J. 28:529-547.
Kirsten, W.J., and K.H. Jansson.1986. Rapid and automaticdeterminationof nitrogen using flash
combustionof large samples.Anal. Chern. 58:2109-2112.
Kjeldahl, 1. 1883. Neue Methodezur Bestimmungdes Stickstoffs in organischenKorpem. Z. Anal.
Chern. 22:36&-382.
Lake, G.R., P. McCutchan,R. Van Meter, and J.e. Neel. 1951. Effects of digestion temperatureon
Kjeldahl analyses.Anal. Chern. 23:1634-1638.
Larson,S.L., and H.P. Peterson.1979. Semiautomated determinationof nitrogen in nitrate-containing
fertilizers. Anal. Chern. 51:2414-2415.
Liao, C.EH. 1981. Devarda'salloy method for total nitrogen determination.Soil Sci. Soc. Am. J.
45:852--855.
Ma, T.S., and G. Zuazaga.1942. Micro-Kjeldahl determinationof nitrogen.A new indicator and an
improved rapid method.Ind. Eng. Chern.Anal. Ed. 14:280-282.
Markham, R. 1942. A steamdistillation apparatussuitablefor micro-Kjeldahl analysis.Biochem. J.
36:790-791.
Marshall, R.B., andJ.N. Whiteway. 1985.Automationof an interfacebetweena nitrogenanalyserand
an isotoperatio massspectrometer.Analyst 110:867--871.
1120 BREMNER
McGeegan,S.L., and D.Y. Naylor. 1988. Automatedinstrumentalanalysisof carbonand nitrogen in

plant and soil samples.Commun.Soil Sci. Plant Anal. 19:493-505.
McKenzie, H.A, and H.S. Wallace. 1954.The Kjeldahl determinationof nitrogen: A critical study of
digestionconditions-temperature, catalyst,and oxidizing agent.Aust. J. Chern. 7:55-70.
Meeker, E.W., and E.C. Wagner. 1933. Titration of ammoniain presenceof boric acid. Macro- and
micro-Kjeldahl procedures.Ind. Eng. Chern.Anal. Ed. 5:396-398.
Meints, V.w., and G.A Peterson.1972. Furtherevidencefor the inability of the Kjeldahl total nitro-
gen methodto fully measureindigenousfixed ammoniumnitrogen in subsoils.Soil Sci. Soc.
Am. Proc. 36:434-436.
Middleton, G., and R.E. Stuckey. 1951. The standardizationof the digestion processin the Kjeldahl
determinationof nitrogen.J. Pharm.Pharmacol.3:829-841.
Mogilevkina, LA 1970.Testingthe applicability of the Kjeldahl methodfor the determinationof total
nitrogen in soils with a high contentof fixed ammonium.(In Russian.)Pochvovedenie1970:
139-143.
Moraghan,1.T., T.I. Rego, and KL. Sahrawat.1983. Effect of water pretreatmenton total nitrogen
analysisof soils by the Kjeldahl method.Soil Sci. Soc. Am. 1. 47:213-217.
Morris, G.F., R.B. Carson,A Shearer,and W.T. Jopkiewicz. 1968. Comparisonof the automatic
Dumas(ColemanModel 29A Nitrogen Analyzer II) and Kjeldahl methodsfor the determina-
tion of total nitrogen in agricultural materials.J. Assoc. Off. Anal. Chern. 51:216-219.
Nelson; D.W., and J.M. Bremner.1966.An evaluationof Mogilevkina'smethodof determiningfixed
ammoniumin soils. Soil Sci. Soc.Am. Proc. 30:409-410.
Nelson,D.W., and L.E. Sommers.1972.A simple digestionprocedurefor estimationof total nitrogen
in soils and sediments.1. Environ. Qual. 1:423-425.
Nelson, D.W., and L.E. Sommers.1980. Total nitrogen analysisfor soil and plant tissues.J. Assoc.
Off. Anal. Chern.63:770-778.
Noel, R.I. 1976. Collaborativestudy of an automatedmethodfor the determinationof crude protein
in animal feeds.1. Assoc. Off. Anal. Chern.59:141-147.
Olsen, C. 1929. On the determinationof nitrogen in soils. With special referenceto tire presenceof
nitratesand nitrites. C.R. Lab. Carlsberg.Vol. 17.
Piper, C.S. 1947. Soil and plant analysis.Univ. Adelaide,Adelaide.
Preston,T., and N.l.P. Owens.1983.Interfacingan automaticelementalanalyserwith an isotope-ratio
massspectrometer:The potential for fully automatedtotal nitrogen and 15N analysis.Analyst
110:867-871.
Pruden,G., S.J.Kalembasa,and D.S. Jenkinson.1985. Reductionof nitrate prior to Kjeldahl diges-
tion. J. Sci. Food Agric. 36:71-73.
Raveh,A, and Y. Avnimelech.1979. Total nitrogenanalysisin water,soil andplant materialwith per-
sulfateoxidation. Water Res. 13:911-912.
Scales,EM., and AP. Harrison. 1920. Boric acid modification of the Kjeldahl method for crop and
soil analysis.1. Ind. Eng. Chern. 12:350-352.
Schepers,J.S.,D.D. Francis,and M.T. Thompson.1989. Simultaneousdeterminationof total C, total
N, and 15N on soil and plant material. Commun.Soil Sci. Plant. Anal. 20:949-959.
Schuman, G.E.,M.A Stanley,and D. Knudsen.1973. Automatedtotal nitrogen analysisof soil and
plant samples.Soil Sci. Soc. Am. Proc. 37:480-481.
Sharma,R.C., and KC. Sud. 1980.An improved rapid chromic acid method for determiningtotal
nitrogen in surfacesoils. 1. Indian Soc. Soil Sci. 28:232-235.
Sheldrick,B.H. 1986. Test of the Leco CHN-600 Determinatorfor soil carbonand nitrogen analysis.
Can. 1. Soil Sci. 66:543-545. .
Sher,LH. 1955. Two-stepmixed indicator for Kjeldahl nitrogen titration. Anal. Chern. 27:831-832.
Skjemstad,J.O., and R. Reeve.1976. The determinationof nitrogen in soils by rapid high-tempera-
ture Kjeldahl digestionand autoanalysis.Commun.Soil Sci. Plant Anal. 7:229-239.
Srinivasan,A 1932. Determinationof nitrogen in soils. Indian J. Agric. Sci. 2:525-530.
Stalcup,H., and R.w. Williams. 1955. Volumetric determinationof nitrocelluloseand nitroguanidine
by transnitrationof salicylic acid. Anal. Chern.27:543-546.
Stevenson,EJ. 1960. Microdeterminationof nitrogen in rocks and silicate minerals by sealedtube
digestion.Anal. Chern. 32:1704---1706.
Stewart, B.A, and L.K Porter. 1963. Inability of the Kjeldahl method to fully measureindigenous
fixed ammoniumin somesoils. Soil Sci. Soc. Am. Proc. 27:41-43.
Stewart,B.A, L.K Porter,and W.E. Beard. 1964. Determinationof total nitrogen and carbonin soils
by a commercialDumasapparatus.Soil Sci. Soc. Am. Proc. 28:366-368.
Stewart, B.A, L.K Porter, and F.E. Clark. 1963. The reliability of a micro-Dumasprocedurefor
determiningtotal nitrogen in soil. Soil Sci. Soc. Am. Proc. 27:377-380.
NITROGEN-TOTAL 1121
Steyermark,A, B.E. McGee, E.A. Bass,and R.R. Kaup. 1958. Micro-Kjeldahl methodfor nitrogen
in certain organic compoundscontainingnitrogen-nitrogenand nitro-oxygen linkages.Anal.
Chern. 30:1561-1563.
Stover,N.M., and R.B. Sandin. 1931. Use of boric acid in micro-Kjeldahl determinationof nitrogen.
Ind. Eng. Chern.Anal. Ed. 3:240-242.
Stutte, e.A, and R.T. Weiland. 1978. Gaseousnitrogen loss and transpirationof severalcrop and
weedspecies.Crop Sci. 18:887-889.
total nitrogen, and total sulfur in soils by combustiontechniques.p. 171-194.In K.A Smith
(ed.) Modern instrumentaltechniquesin soil analysis.Marcel Dekker, New York.
nitrogen, and sulfur in soils by combustiontechniques.p. 261-289.In K.A Smith (ed.) Soil
analysis:Modem instrumentaltechniques,2nd ed. Marcel Dekker, New York.
Vittori Antisari, L. V., and P. Sequi. 1988. Comparisonof total nitrogen by four proceduresand
sequentialdeterminationof exchangeableammonium,organicnitrogen,and fixed ammonium
in soil. Soil Sci. Soc. Am. J. 52:1020-1023.
Vogel, AI. 1961.A textbookof quantitativeinorganicanalysis.3rd ed. JohnWiley & Sons,Inc., New
York.
Wagner,W.E, J.A Wuellner, and C.E. Feiler. 1952. Sulfamicacid as a standardreagentfor alkalime-
try. Anal. Chern. 24:1491-1492.
Walkley, A 1935. An examinationof methodsfor determiningorganiccarbonand nitrogen in soils.
J. Agric. Sci. 25:598-609.
Wall, L.L., Sr., C.w. Gehrke,T.E. Neuner,R.D. Cathey,and P.R. Rexroad.1975. Total protein nitro-
gen: Evaluation and comparison of four different methods. J. Assoc. Off. Anal. Chern.
58:811-817.
Watkins, K.L., T.L. Veum, and G.E Krause. 1987. Total nitrogen determinationof various sample
types:A comparisonof the Hach, Kjeltec, and Kjeldahl methods.J. Assoc. Anal. Chern.
70:410-412.
White, L.M., and M.e. Long. 1951. Kjeldahl microdigestionsin sealedtubesat 470°e.Anal. Chern.
23:363-365.
Winkler, L.w. 1913. Beitrag zur titrimetrischen Bestimmungdes Ammoniaks. Z. Agnew. Chern.
26:231-232.
Yeomans,J.e., and J.M. Bremner. 1988. A rapid and precise method for routine determinationof
organiccarbonin soil. Commun.Soil Sci. Plant Anal. 19:1467-1476.
Yeomans,J.C., and J.M. Bremner. 1991. Carbon and nitrogen analysisof soils by automatedcom-
bustion techniques.Commun.Soil Sci. Plant Anal. 22:843-850.
Yuen, S.H., and AG. Pollard. 1953. Determinationof nitrogen in soil and plant materials:Use of
boric acid in the micro-Kjeldahl method.J. Sci. Food Agric. 4:490-496.
Published 1996
Chapter 38
Nitrogen-Inorganic Forms
R. L. MULVANEY, University of Illinois, Urbana, Illinois
Most soils contain inorganic nitrogen (N) in the form of ammonium(NHt) and
z)
nitrate (NO)"). Nitrite (NO also may be present,but the amount is usually too
small to warrant its determination,exceptin caseswhere NHt or NHt-forming
fertilizers are applied to neutral or alkaline soils. Severalother forms of inorgan-
ic N have beenproposedas intermediatesduring microbial transformationsof N
in soils, including hydroxylamine (NH20H), hyponitrous acid (H 2N20 2), and
nitramide (NH2N02), but thesecompoundsare thermodynamicallyunstableand
have not beendetectedin soil.
Until the 1950s,inorganic N was believedto accountfor <2% of total soil
N, on the assumptionthat NHt and NO)" are completelyrecoveredby extracting
soil with a neutral salt solution. The validity of this assumptionwas challenged
by the finding that some soils contain NHt in a form that is not extractedby
exchangewith other cations (e.g., Rodrigues, 1954; Dhariwal & Stevenson,
1958; Stevenson& Dhariwal, 1959; Bremner & Harada,1959; Bremner, 1959;
Schachtschabel, 1960, 1961; Young, 1962), and by estimatesthat the proportion
of soil N in this form can exceed50% for some subsurfacesoils (Stevenson&
Dhariwal, 1959; Young, 1962). In such cases,NHt is said to be fixed, and fixed
NHt hassubsequentlybeendefined asthe NHt in soil that cannotbe replacedby
a neutral potassiumsalt solution (SSSA, 1987), such as 1 or 2 M KCI or 0.5 M
K 2S04, in contrastto exchangeable NHt, which is extractableat room tempera-
ture with such a solution. Existing information indicatesthat fixed NHt occurs
largely, if not entirely, betweenthe layers of 2: I-type clay minerals,particularly
vermiculite and illite (hydrous mica),and that fixation resultsfrom entrapmentof
NHt in ditrigonal voids in the exposedsurfacesupon contractionof the clay lat-
tice (Nommik & Vahtras,1982). The term, nonexchangeable NHt, hasbeenused
by Bremner(1965) and Keeney and Nelson (1982) in previous editions of this
publication as a more precisealternativeto fixed NHt. The sameterm is usedin
the presenttreatment,with specific referenceto NHt determinedby the method
describedin "Determinationof Nonexchangeable Ammonium," which involves
digestionwith an HF-HCI solution following treatmentof the soil with alkaline
KOBr to removeexchangeableNHt and labile organic-Ncompounds.
Book Seriesno. 5.
1123
1124 MULVANEY
The determinationof exchangeableNH!, NO)", and N02" in field soils is

complicatedconsiderablyby extensivespatialvariability (Biggar, 1978),andspe-
cial sampling techniquesbased on a knowledge of this variability may be
required(Van Meirvenne& Hofman, 1989).A further complicationis that these
forms of N are subject to microbial transformations,which can lead to rapid
changesin the inorganic-N content of a soil or soil sample.To minimize such
changes,analysesof inorganic N should be carried out immediatelyafter sam-
pling. In most cases,however,somedelay is unavoidablebecauseof the needto
transportthe sampleto the laboratoryand sieveit beforethe analysiscan be per-
formed, and in somecases,the samplemay be storedfor later analysis.Various
chemicalshavebeenemployedto minimize microbial growth during transportor
storage of soil samplesfor inorganic-N analysis (e.g., volatile organic com-
pounds,salts of heavy metals), but such treatmentshave given erratic results
(Storrier, 1966; Robinson,1967), and they can interfere with analysesfor inor-
ganic N (Bremner,1965).
Deep-freezestorageoften hasbeenemployedas a meansof preservingsoil
samplesfor inorganic-N analyses.Severalstudiesto evaluatethis practicehave
beenreported,but the findings are inconsistent.In somecases,storageof soils at
subzerotemperatureshas been found to have little effect on their content of
exchangeableor nonexchangeable NH! or NO)" (Gasser,1958; Nelson & Brem-
ner, 1972; Westfall et aI., 1978). In other cases,deep-freezinghas led to a sub-
stantialincreaseor decreasein analysesfor NH! and/orNO)" (Allen & Grimshaw,
1962; Harding & Ross, 1964; Robinson, 1967; Selmer-Olsenet aI., 1971;
Breimer & Slangen,1981). It is difficult to accountfor such divergentfindings,
but attentionshouldbe drawn to the possibilitiesof contaminationduring storage
and of microbial N transformationswhen frozen soils are allowed to thaw grad-
ually at room temperature(Nelson & Bremner, 1972). To minimize thesediffi-
culties, soils shouldbe transferredto air-tight containers fordeep-freezestorage,
and analysesfor inorganicN shouldbe carriedout as soonas possibleafter rapid
thawing at room temperature(Nelson& Bremner,1972).
To facilitate sieving, soil samplesfor inorganic-Nanalysisare often air- or
oven dried, and this has the advantagethat dried soils are convenientlystoredat
room temperature.However,drying canlead to significantchangesin the NH! or
NO)" content of soil (e.g., Gasser,1958; Allen & Grimshaw, 1962; Nelson &
Bremner, 1972; Breimer & Slangen,1981), particularly if done at an elevated
temperature(e.g., 35-70°C),in which caseseriousloss of exchangeableNH! or
NO)" can occur (Selmer-Olsen etaI., 1971; Nelson & Bremner,1972; Breimer &
Slangen,1981). For this reason,drying of soil for inorganic-N analysisis often
carriedout in the openlaboratoryat room temperature(go°C), but careshould
be taken to minimize exposureto atmosphericNH3, which is readily sorbedby
air-dried soils (Bremner, 1965). Cleaningsolutionsand floor waxesare a com-
mon sourceof such exposure,so drying should be done where thesematerials
will not be used,and the dried soil shouldbe storedin an air-tight glassor plas-
tic container(Nelson & Bremner, 1972).Drying of soils should not be done in
caseswhere a significant amountof N02" may be present,becausethe N02"-N is
subjectto gaseousloss and chemicalfixation by soil organic matter (Bremner,
1965;Nelson& Bremner,1969).In suchcases,the field-moist samplesshouldbe
NITROGEN-INORGANIC FORMS 1125
extractedas soonas possibleafter collection (Nelson & Bremner,1972; Keeney

& Nelson, 1982).
METHODS FOR DETERMINATION OF INORGANIC NITROGEN

IN SOILS
Although methodshave been describedto determineexchangeableNHt,

NO)", or NOz in soils without extraction(e.g., Keeney & Bremner, 1966), most
methodsfor thesedeterminationsinvolve the preparationof extracts,which are
easily stored for later analysis. Numerous methods have been employed for
determinationof inorganicforms of N in soil extracts,including colorimetry, ion-
selectivepotentiometry,steamdistillation, microdiffusion, ion chromatography,
and ultraviolet (VV) spectrophotometry. Table 38-1 briefly evaluatesthe various
methods,with referencesfor additional information.
Colorimetric methods have been used extensively for determinationof
inorganic N in soil extracts. Ammonium-N is readily determinedby several
methodsbasedon the Berthelot reaction involving phenol (Keeney & Nelson,
1982; Dorich & Nelson, 1983) or salicylate(Nelson, 1983; Kempers& Zweers,
1986). Nitrite-N is easily determinedby the Griess-Ilosvaymethod (Bremner,
1965; Keeney& Nelson, 1982), and the samemethodcan be employedto deter-
mine NO)"-N following reductionto NOz-N by passagethrougha columnof cop-
perizedcadmium(Huffman & Barbarick,1981; Keeney& Nelson,1982; Dorich
& Nelson, 1984). Thesemethodsare simple and sensitive,and they have been
widely usedfor automatedanalysesof soil extractsand natural waters. Of par-
ticular importanceis the fact that all three determinationscan be performedby
extractionwith a single reagent(2 M KCI).
Ion-selectiveelectrodesare simpler and more convenientto use than col-
orimetric methods,but they have not gainedwide acceptancefor routine deter-
mination of inorganic N in soils or soil extracts,due in part to the high cost of
these electrodesand their limited operational lifetime. The gas-sensingelec-
trodesusedto measureNHt and NOz are highly sensitiveand specific, but NO)"
electrodesare subjectto seriousinterferencesby other anions,require continual
restandardization,and lack sensitivity.
The steam-distillationmethodsdescribedby Bremner(1965) for determi-
nation of NHt, NO)", and NOz in soil extracts,and subsequentlyevaluatedin
publicationsby Bremnerand Edwards(1965) and Bremnerand Keeney (1965,
1966), havebeenwidely used.Distillation is carriedout with a mild base(MgO)
to convertNHt-N to NHrN, with or without a reducingagent(Devarda'salloy)
z
to convert (NO)" + NOz)-N or N03-N [in caseswhere NO is absentor the
extract has beentreatedwith sulfamic acid (H}NO}S)] to NH3-N, and the NH r
N liberatedis collected in boric-acid indicator solution and determinedby titra-
tion with standardacid. Thesemethodsare basedon the finding that alkali-labile
organic-Ncompoundsdo not undergohydrolysis if distillation is performedfor
a short period with steam using a small amount of MgO (Bremner & Keeney,
1965, 1966). Steam-distillationmethodsare accurateand precise,and they per-
mit isotope-ratioanalysisof the N collectedin the distillate (Hauck, 1982).
....
....
N
="
Table 38-1. Methodsfor determinationof inorganicforms of N in soil extracts.

Method Comments Reference(
s)
Ammonium-Nitrogen
Colorimetric
Salicylate Simple, sensitive,amenableto automation See"Ammonium" under"Colorimetric Methods"
Indophenolblue Like salicylatemethod,but usesphenol,which is toxic Keeney& Nelson (1982)
Nessler Insensitive,requirespreliminary distillation to eliminate interference Am. Public Health Assoc. (1992)
Ion electrode Sensitive,convenient,highly specific See"Ammonium" under"Ion-SelectiveElectrodes"
Steamdistillation Widely used,reliable See"Steam-DistillationMethods"
Microdiffusion Simple but slow See"Microdiffusion Methods"
Nitrate-Nitrogen
Colorimetric
Reductionto NOi by Cd; Sensitive,determines(NO) + NOn-N, error can arise from incomplete See"Nitrate" under"Colorimetric Methods"
Griess-Ilosvaymethod reduction,amenableto automation
Brucine Brucine is carcinogenic Baker (1967)
Chromotropicacid UsesconcentratedH 2S04 Clark & Jennings(1965), Sims & Jackson(1971),
Kowalenko & Lowe (1973), Haby (1989)
Phenoldisulfonicacid Tediousand time-consuming,usesNH40H and concentratedH2S04, Bremner(1965)
Cl- interferes
Transnitrationof salicylic acid UsesconcentratedH 2S04, Cl- interferes Vendrell & Zupancic(1990)
Ion electrode Insensitive,accuracycan be a problem,samplesmust be bufferedto See"Nitrate" under"Ion-SelectiveElectrodes"
commonionic strength,many interferences
2
~
~
~
t'1
~
"!-
z
-
o

Method Comments Reference(
s) ~
-;:5~
Steamdistillation Widely used,reliable, recovers(NO) + N02)-N by reductionwith See"Steam-DistillationMethods"
Devarda'salloy, NOi removedwith sulfamic acid ~
Microdiffusion Simple but slow, utilizes the samechemistry as steamdistillation See"Microdiffusion Methods" r:n
Ion chromatography Sensitive,allows simultaneousdeterminationof NO) and other anions Tabatabai& Frankenberger(1996), seeChapter8
UV difference Sensitiveand precise,multiple measurements are madeto correct for Norman & Stucki (1981), Heinzmannet al. (1984),
UV absorbanceby non-NO) species Norman et al. (1985)
Nitrite-Nitrogen
Colorimetric
Griess-I1osvay Simple, sensitive,amenableto automation See"Nitrite" under"Colorimetric Methods"
Ion electrode MeasuresgaseousNOx in acidified extract, few interferences See"Nitrite" under "Ion-SelectiveElectrodes"
Steamdistillation Widely used,reliable, NOi-N determinedby differencefrom distilla- See"Steam-DistillationMethods"
tions with and without sulfamic acid pretreatment
Microdiffusion Simple but slow, utilizes the samechemistry as steamdistillation See"Microdiffusion Methods"
Ion chromatography Sensitive,allows simultaneousdeterminationof NOi and other anions Tabatabai& Frankenberger(1996), seeChapter8
UV difference Measurementsare madewith and without sulfamic acid treatment Norman & Stucki (1981)
N
-....
1128 MULVANEY
Microdiffusion methods offer several advantages.They are less labor-

intensive than steam-distillation methods and require very little equipment,
which allows analysesto be performedat low cost. When properly conducted,
the results are accurateand precise.However, sensitivity is limited by the vol-
ume of soil extractthat can be accomodated(4-10 mL with most microdiffusion
methods;up to 100 mL with the proceduredescribedin "Methods" for "Microd-
iffusion Methods"). Moreover, the resultscannotbe obtainedimmediately,as a
period of 1 to 2 d is requiredfor completediffusion. Perhapsbecauseof these
limitations, microdiffusion methodshavenot beenwidely usedfor determination
of inorganicforms of N in soil extracts.
Ion chromatography permits simultaneous determination of severaldiffer-
ent anions or cations. It has been used successfullyfor determinationof NO)"
(Dick & Tabatabai,1979; Tabatabai& Dick, 1979; Reynders& Vlassak, 1981;
Hem et aI., 1983; Vilsmeier, 1984; Bradfield & Cooke, 1985; Kuchnicki et aI.,
1985; Nieto & Frankenburger,1985a; Kuchnicki & Webster, 1986; Barak &
z
Chen, 1987; Stewart, 1987; du Preez & Laubscher,1989), NO (Vilsmeier,
1984; Nieto & Frankenberger,1985a),and NHt (Nieto & Frankenberger,1985b)
in soil extracts.The techniqueis rapid and sensitive, butanalysesare subjectto
interferenceby high salt concentrations,so extractionsare usually carried out
usingwateror a dilute salt solution,which precludesdeterminationof exchange-
able NHt. A further limitation is the high costof a dedicatedion chromatograph,
althoughanalysescan be carried out with conventionalhigh-performanceliquid
chromatography(HPLC) equipmentusing a low-capacityion exchangecolumn.
Nitrate and NOzabsorbUV radiation,which suggeststhat the concentra-
tion of theseions in a soil extractcould be determinedby a direct spectrophoto-
metric analysis.However, the measurements are subject to seriousinterference
by organic matter and other constituentsthat absorbat the samewavelengthas
z
NO)" or NO (~21O nm). Attempts have been made to remove the interfering
speciesthrough the use of alumina creamas a clarifying agent (Cawse,1967),
but removal may be incomplete,and the alumina cream can affect absorbance
measurements(Norman & Stucki, 1981). A more satisfactoryalternative is to
measureUV absorbancebeforeand after reductionof NO)" to nonabsorbingspe-
cies, NO)" being determinedfrom the differencein absorbance(Norman & Stuc-
z
ki, 1981; Heinzmannet aI., 1984). If NO is present,the difference represents
z,
NO)" + NO but the two ions can be determinedseparatelyby carrying out an
additional measurementinvolving treatmentwith sulfamic acid to decompose
z
NO (Norman & Stucki, 1981). The only interferenceis from carbonate,which
is easily eliminated by addition of mineral acid (Norman & Stucki, 1981),
althoughacidification will necessarilyvitiate analysesfor NO z.
The UV difference method is sensitiveand precise,and good agreement
has beenobtainedwhen NO)" analysesby this methodare comparedto thoseby
steamdistillation (Norman & Stucki, 1981; Heinzmannet aI., 1984). The analy-
sis time per samplealso is comparable,but this is reducedconsiderablywith a
dual-wavelengthmethod developedby Norman et al. (1985), which eliminates
the needfor a chemicaltreatmentto remove NO)". Instead,correctionfor inter-
ferenceby non-NO)" speciesis made by subtractingan empirically determined
multiple of the absorbanceat 270 nm (due to non-NO)" species) from the
absorbanceat 210 nm (due to NO] + non-NO] species).The dual-wavelength

approachis rapid and convenient,but it is inherently less accurateand precise
than the UV differenceapproachinvolving chemicalreduction,and seriouserror
can arise from variability in extraction of non-NO] species,as can occur with
soils receiving organicamendmentssuchas manureor sewagesludge.
EXTRACTION OF EXCHANGEABLE AMMONIUM

AND NITRATE AND NITRITE
Introduction
As noted by Bremner (1965), methods for extraction of exchangeable
NHt, NO], and NOz from soils must meetthe following requirements:(i) extrac-
tion of the form(s) of N under analysismust be practically quantitative; (ii) no
changemay occur in the amountof this N as a result of biological or nonbiolog-
ical reactions; (iii)the extractmust be compatible withthe method(s)of analysis
to be employedand must not contain any substancethat would interfere with
analysis;and (iv) the extractmust be stableso that it can be safelystoredfor later
analysis. In addition, the method of extraction should be simple and rapid
(Keeney& Nelson, 1982).
To prevent loss of NOz-N through chemical decompositionof HN02
formed under acidic conditions, a neutral or alkaline reagentis normally em-
ployed in extracting soil for determinationof NO)" or NOZ-. Although other
reagentsmay be requiredto avoid interference(seeTable 38-2), a 2 M solution
of KCI is usually preferredbecauseit extractsexchangeable NHt as well as NO)"
and NOz, which has the advantagethat all three forms of N can potentially be
determinedfrom a single extraction.Moreover, the high salt concentrationsim-
plifies filtration by flocculating soil solids, and the extract is stable for several
months if stored in a refrigerator (Bremner & Keeney, 1966; Robinson, 1967;
Selmer-Olsenet aI., 1971; Nelson & Bremner, 1972), with no formation of pre-
cipitate, as occursduring refrigerationof 0.5 M K 2S04 extracts.
Soil extracts fordeterminationof inorganicforms of N may be obtainedby
either leaching or equilibrating the soil with the extractant.With the leaching
Table 38-Z. Extractantsfor use with different methodsof determininginorganic forms of N in soil
extracts.
Method Extractant
Steamdistillation ZMKCI
Microdiffusion ZMKCI
Colorimetric
Salicylate(for NHt-N) ZMKCI
Cd reduction(for NOj"-N) ZMKCI
Griess-I1osvay(for NOi-N) ZMKCI
Ion electrode
NH3 electrode ZMKCI
NO) electrode Water
NO, electrode IMKCI
Ion chromatography O.OlMKCI
1130 MULVANEY
method,the soil sampleis treatedwith the extractantfor a specifiedperiod (with

or without shaking).The suspensionis then filtered, the residueon the filter is
leachedwith the extractant,and the filtrate is brought to an exact volume. With
the equilibrium method,the soil sampleis shakenwith an accuratelymeasured
volume of extractantfor a specified period. The suspensionis then filtered, but
the residueis not leached,and the filtrate is not broughtto volume. In theory, the
leaching method should be more quantitative,but if properly performed,both
methodsgive essentiallythe sameresults. The equilibrium method is generally
preferredbecauseit is simpler and more convenientthan the leachingmethod,
and becauseextractionscan be performedusing a smallervolume of extractant,
which facilitates storage and analysis of the extract. Whichever method is
employed,the cation concentration,the volume of extractant,and the period of
extractionshouldbe adequateto ensurequantitativereplacementof exchangeable
NHt·
The extractionproceduredescribedinvolves equilibrium extractionof soil
with 2 M KCI, using 10 mL of extractantper gram of soil and a 1-h period for
shaking.Studiesto evaluatethis procedurehave shown that it gives quantitative
recoveryof NO.3 and N02" addedto soil and that recoveryof addedNHt also is
quantitative,providedthe soil has no ability to fix NHt or fixation is blockedby
a prior addition of K+ (Bremner& Keeney,1966).
Method
SpecialApparatus
1. Reciprocatingshaker.
2. Buchnerfunnel. A two-piece polypropylenefunnel for 5.5-cm diam.
filter paperis satisfactory.
3. Filtering apparatus.Filtration can be carried out using a heavy-walled
filtering flask, but a much more convenientalternativeis the five-place
filter funnel stand(cat. no. 1300) manufacturedby SoilmoistureEquip-
ment Corp., P.O. Box 30025, Santa Barbara, CA 93105. With this
device, the filtrate is convenientlycollected in 118-mL (4-oz.) screw-
cap bottles (the Qorpak bottles available from Fisher Scientific, 711
ForbesAve., Pittsburgh,PA 15219, as cat. no. 03-326-5Care satisfac-
tory).
Reagents
1. Potassiumchloride (KCI) solution, approximately2 M. Dissolve 1.5 kg
of reagent-gradeKCI in 8 L of deionizedwater, and dilute the solution
to 10 L.
2. Filter paper. Whatman No. 42 (5.5-cm diam.) is satisfactory.Before
use, it shouldbe washedby filtering 20 mL of 2 M KCI and 20 mL of
deionized water in a Buchner funnel, followed by air- or oven
(40-70°C)drying.
Procedure
Place 10 g of soil (oven-dry weight) in a 250-mL widemouth bottle, and
add 100 mL of 2 M KCl. Stopperthe bottle, and shakeit for 1 h with a recipro-
cating shaker.Allow the suspensionto settle until the supernatantis clear (nor-
mally 15-30 min), then decanta small amountof the supernatantto moistenthe
filter paperin the Buchnerfunnel. When the paperis moistenedcompletely,bring
the funnel undervacuum,decantthe suspensioninto the funnel, and carry out fil-
tration. Store the filtrate in a refrigerator.
Comments
The proceduredescribedwas developedfor extractions with 2 M KCI
(Bremner& Keeney, 1966), but the sameprocedurecan be employedwith 1 M
KCI and 0.5 M K 2S04• For extractionsusing water or a dilute salt solution (e.g.,
0.01 M KCI), as required for NO)" determinationsby the NO)" electrodeor ion
chromatography(Table 38-2), the soil suspensionshould be centrifugedbefore
filtration, or the filtrate shouldbe clarified by passingit through a membranefil-
ter (0.45 11m).
Cellulosefilters commonlycontainsignificantamountsofNHt-N, but very
little, if any, NO)"- or N02"-N (Sparrow & Masiak, 1987; Qasim & Flowers,
1989).The NH.t-N is effectively removedby washingwith a KCI or K 2S04 solu-
tion (Sparrow & Masiak, 1987; Qasim & Flowers, 1989), and such washing is
highly recommendedwheneverthe filtrate is to be analyzedfor NHt. An alter-
native is to use a more expensiveglassfiber filter, in which casethe contentof
NHt and NO)" is sufficiently low that washingis unnecessary(Sparrow& Masi-
ak,1987).
If desired,extractionscan be carried out using <10 g of soil, with a pro-
portionatedecreasein the volume of 2 M KCI. Extractions alsocan be performed
with> 10 g of soil, but the volume of KCI shouldbe increasedto maintaina 1:10
ratio of soil to extractant,as a lower ratio may lead to incompleteextraction of
NHt-N (Bremner & Keeney, 1966). Incompleteextraction also can arise if the
soil suspensionis not thoroughly shakenfor at least 1 h (Bremner & Keeney,
1966).
Filtration of the extract can be omitted if analysesare to be performedby
steamdistillation within a few hoursafter extraction.In suchcases,an aliquot of
the supernatantcan be taken for analysisusing a pipette with a wide tip to pre-
vent plugging by suspendedmaterial (Bremner& Keeney,1966).
DETERMINATION OF AMMONIUM, NITRATE, AND NITRITE

IN SOIL EXTRACTS
Steam-DistillationMethods
Introduction
Procedures involving steamdistillation of a 2M KCI soil extractwith MgO
and Devarda'salloy are described.Theseproceduresare summarizedby Table
38-3. The sameprocedureshave beendescribedby Bremner(1965) and Keeney
and Nelson(1982) in previouseditions of this publication, and descriptionsalso
can be found in articles by Bremnerand Keeney (1965, 1966). The latter refer-
1132 MULVANEY
Table 38-3. Steam-distillationmethodsfor determinationof inorganicforms of N in soil extracts.

Form of N Methodt
NH,t Steamdistillation with MgO

NO:! Steamdistillation with MgO and Devarda'salloy after destructionof
NOi with sulfamic acid and removal of NH,t by steamdistillation
with MgO:j:
NHt +NOj Steamdistillation with MgO and Devarda'salloy after destructionof
NOi with sulfamic acid:j:
NOj+ NOi Steamdistillation with MgO and Devarda'salloy after removalof
NHt by steamdistillation with MgO
NHt + NOj + NOi Steamdistillation with MgO and Devarda'salloy
t With eachmethod,NH3 liberatedby steamdistillation is collectedin H3BOr indicatorsolution and

determinedby titration with 0.0025M H2S04 ,
:j: If NOi is absent,the sulfamic acid treatmentis omitted.
encesreport resultsfrom studiesto evaluatethe steam-distillationproceduresby

a variety of methods,including recovery,interference,and specificity tests.The
resultsof thesestudiesindicatethat the proceduresare not subjectto interference
from organic or inorganic soil constituentsand are applicableto neutral, acidic,
and calcareoussoils.
The steam-distillation proceduresdescribed for determination of NHt,
N03", and NOz in soil extractsdevelopedfrom the finding that interferenceby
alkali-labile organic-N compoundsin alkali-distillation methodsof determining
NHt is eliminatedif the distillation is performedwith steamfor only a shortperi-
od using a small amountof MgO. Theseproceduresare accurateand reliable, and
they have advantagesover colorimetric methodsin that the analysesare unaf-
fected by various organic and inorganic substancescommonly present in soil
extracts,and analysesmay be performedon colored or turbid extractswithout
difficulty. A further advantageof steam-distillationproceduresis that they permit
isotope-ratio analysis of exchangeableNHt-N, N03"-N, and NOz-N in tracer
studiesconcerningthe fate of labeledN applied to soil. The only specialappara-
tus required is the steam-distillationapparatusillustrated by Fig. 38-1, and the
sameapparatuscan be used in determiningnonexchangeable NHt (see "Deter-
mination of Nonexchangeable Ammonium"), total N (Bremner,1996, seeChap-
ter 37), and organic forms of N (Stevenson,1996, seeChapter39) in soils. The
only standardreagentrequiredis 0.0025M H2S04, usedin titrating the NHt lib-
eratedby steamdistillation.
In previouseditions of this publication, proceduresare describedwhereby
steamdistillation with MgO and Devarda'salloy may be performeddirectly on
the soil samplefollowing treatmentwith 2 M KCI (Bremner, 1965; Keeney &
Nelson,1982), therebyeliminating the needfor extraction.The direct-distillation
procedureshave beenusedsuccessfullywith a wide variety of soils (Keeney &
Bremner, 1966; du Preezet aI., 1987), but there is clearly greaterpotential for
interferencefrom soil organic-Ncompoundsthan with the extraction-distillation
approach, and substantially higher values have been observed in measuring
exchangeableNHt -N in organic and paddy soils by direct distillation than by
extraction-distillation (Robinson, 1967; Sahrawat & Ponnamperuma,1978).
Moreover, liberation of CO2 during distillation of calcareousor anaerobicsoils
Steam-bypass
assembly
§ 26115
o• 6
em L.LJ
Fig. 38-1. Steam-distillationapparatus

. (Available from O'Brien's Scientific Glass Blowing. 725A
West Bridge Street.Monticello, IL 61856 USA.)
has been found to causenegativeerror in titrimetric determinationof NH.t- or

NO)"-N (Clausenet aI., 1981; Sahrawat,1982), and the analysesfor NO)"-N are
subjectto interferenceby high concentrationsof exchangeableMg2+ (Clausenet
aI., 1980). In view of such difficulties, direct proceduresfor steam distillation
should be usedonly after checkingthe recovery of addedN for the soils under
study.
Principles
z
The methodsfor determining NH.t, NO)", and NO by steam distillation
with MgO and Devarda'salloy (Table 38-3) are basedon the finding that NH.t-N
in solutions containingglutamine and other alkali-labile organic-N compounds
can be determinedquantitatively from the NH3-N liberated by steam-distilling
thesesolutionswith a small amountof MgO for 3 to 4 min and that (NH.t + NO)"
+ N02)-N in suchsolutionscan be determinedquantitativelyby the samemethod
if finely divided Devarda'salloy is addedimmediatelybeforedistillation (Brem-
ner & Keeney, 1965). The methodsfor determinationof NH.t and NO)" in the
z
presenceof NO developedfrom the finding that sulfamicacid decomposes NO z
rapidly and quantitatively at room temperature(HN02 + NH 2S03H ~ N2 +
H2S04 + H20), but doesnot react with NH.t or NO)" and doesnot interfere with
their determinationby steamdistillation with MgO and Devarda'salloy (Brem-
ner & Keeney,1965).
Methodsl
Special Apparatus
1. Steam-distillationapparatus(Fig. 38-1). The unit illustrated is a modi-
fied version of the apparatusdescribedby Bremner(1965) and Keeney
1 After Bremner(1965), and Bremnerand Keeney(1965, 1966).

1134 MULVANEY
and Nelson(1982). The major modificationsare: (i) addition of a stop-

cock to the steam-bypass assemblyto allow completeterminationof the
steamsupply to the distillation head in distillation of Kjeldahl digests
(this stopcockis left openat all times in distillation of soil extracts),and
(ii) addition of a Teflon sleeve(3 cm long, 1O-mm o.d., 8-mm Ld.) to
permit use of interchangeablesteam-inlettubeswith different types of
distillation flasks. The apparatusis equippedwith an O-ring ball joint
(~ 28/15) for attachmentof Pyrex boiling flasks (via a pinch clamp)
used as distillation chambers.Steam for distillation is generatedby
boiling deionizedwater in a 5-L flask containinga few Teflon boiling
chipsand equippedwith an electricheatingmantle controlledby a vari-
able transformerand a needlevalve stopcockto supply deionizedwater.
Before use, the apparatusshould be steamedout for about 10 min to
removetracesof NH3, during which the transformeris adjustedso that
distillate is collected at a rate of 7 to 8 mL min-I. A trap is installed
below the waterjacket of the condenserto preventcondensatethat col-
lects on the outside surfaceof the condenserfrom contaminatingthe
distillate.
2. Distillation flasks. The flasks employedare 100-mL Pyrex round bot-
tom boiling flasks with standard-taper joint (1'24/40)that havebeenfit-
ted with a socketjoint (~28/15) for attachmentto the distillation appa-
ratus. Their dimensionsshould be such that when the flasks are con-
nectedto the steam-distillationapparatus,the distancebetweenthe tip
of the steam-inlettube and the bottom of the flask does not exceed4
mm.
3. Microburette(5-mL, graduatedat O.OI-mL intervals)or automatictitra-
tor.
Reagents
1. Magnesiumoxide (MgO). Heat heavy MgO in an electric muffle fur-
naceat 600 to 700°C for 2 h. Cool the product in a desiccatorcontain-
ing potassiumhydroxide (KOH) pellets, and store it in a tightly stop-
peredbottle.
2. Boric acid-indicatorsolution. Add 400 g of reagent-gradeboric acid
(H3B03) to 18 L of deionizedwater in a 20-L Pyrex bottle marked to
indicate a volume of 20 L, and stir vigorously with a motorizedstirrer
to dissolve the H3B03. Then add 400 mL of indicator solution (pre-
paredby dissolving0.495 g of bromocresolgreenand 0.33 g of methyl
red in 500 mL of ethanol),and bring the volume to 20 L with deionized
water. With continuousstirring, adjust the pH to 4.8 to 5.0, or until the
solution assumesa reddishpurple tint, by cautiouslyadding 1 M sodi-
um hydroxide (NaOH) or single NaOH pellets. If excessNaOH is
added,the pH can be reducedby addingdilute HCI. Dispensethe solu-
tion throughTeflon tubing.
3. Devarda'salloy. A satisfactoryalloy is producedby Merck, Darmstadt,
Germany(cat. no. 5341; availablefrom EM Science,P.O. Box 70, 480
DemocratRoad, Gibbstown,NJ 08027).
4. Sulfamicacid, 0.2 M. Dissolve2 g of sulfamic acid (the certified mate-

rial availablefrom FisherScientific is satisfactory)in 100 mL of deion-
ized water. Storethe solution in a refrigerator.
5. Sulfuric acid (H2S04), 0.0025M standard.
6. Standard(NHt + N03")-N solution. Dissolve 0.2358 g of ammonium
sulfate [(NH4)zS04] and 0.3609 g of potassium nitrate (KN0 3) in
deionizedwater (both reagentsare availablefrom FisherScientific as a
primary-standardgrade),dilute the solution to a volume of 1 L in a vol-
umetric flask, and mix thoroughly. If pure, dry reagentsare used, this
solution contains50 Ilg of NHt-N and 50 Ilg of N03"-N mL-I. Storethe
solution in a refrigerator.
7. Standard (NHt + N03" + NOz)-N solution. Dissolve 0.2358 g of
(NH4)zS04,0.3609g of KN0 3, and0.1232g of sodiumnitrite (NaN02)
in deionizedwater, dilute the solution to a volume of 1 L in a volumet-
ric flask, and mix thoroughly.If pure, dry reagentsare used,this solu-
tion contains50 Ilg of NHt-N, 50 Ilg of N03"-N, and 25 Ilg of NOz-N
mL-I. Store the solution in a refrigerator.
Procedures in Absence of Nitrite
Ammonium-Nitrogen. Add 5 mL of H3BOr indicator solution to a 50-mL
beakermarkedto indicate a volume of 35 mL, and position the beakerunderthe
condenserof the distillation apparatusso that the tip of the condenseris in con-
tact with the side of the beakerand about 1 cm below the top. Pipettean aliquot
(normally 10-20 mL) of the soil extractinto a distillation flask, and add0.2 g of
MgO througha dry powderfunnel having a long stemthat reachesdown into the
bulb of the flask. Immediatelyattachthe flask to the steam-distillationapparatus
as shown in Fig. 38-1, and commencesteam distillation by closing the lower
stopcockon the steam-bypass assembly.When the volume of distillate reaches35
mL, rinse the tip of the condenser,and stop the distillation by openingthe lower
stopcock on the steam-bypassassembly.Titrate the distillate with 0.0025 M
H2S04. At the end-point,the color changesfrom greento a permanent,faint pink.
Nitrate-Nitrogen. Following steam distillation with MgO as describedin
the previoussection,removethe stopperfrom the side-armof the flask, add 0.2
g of Devarda'salloy rapidly through a dry powderfunnel that reachesdown into
the flask about 1 cm below the baseof the ground joint that servesas the side-
arm, and immediatelyreplacethe stopperin the neck of the side-arm.Then carry
out steamdistillation as describedin the previous "Ammonium-Nitrogen" sec-
tion, and titrate the distillate with 0.0025M H2S04.
(Ammonium + Nitrate)-Nitrogen. Proceed as described in the previous
"Ammonium-Nitrogen"section,but add 0.2 g of Devarda'salloy to the distilla-
tion flask immediately after the addition of MgO and before attachingthe flask
to the distillation apparatus.
Procedures in Presence of Nitrite
Ammonium-Nitrogen. Proceedas describedin the previous "Ammonium-
Nitrogen" section.
1136 MULVANEY
(Nitrate + Nitrite)-Nitrogen. Proceedas describedin the previous"Nitrate-

Nitrogen" section.
(Ammonium + Nitrate + Nitrite)-Nitrogen. Proceedas describedin the pre-
vious "(Ammonium + Nitrate)-Nitrogen"section.
(Ammonium + Nitrate)-Nitrogen. Proceedas described in the previous
"(Ammonium + Nitrate)-Nitrogen"section,but treat the samplein the distillation
flask with 1 mL of sulfamic acid, and swirl the flask for a few secondsto decom-
poseNOz , before the addition of MgO and Devarda'salloy.
Nitrate-Nitrogen. Proceedas describedin the previous"Nitrate-Nitrogen"
section,but perform the analysison a samplethat hasbeentreatedwith sulfamic
acid to destroyNOz as describedin the "(Ammonium + Nitrate)-Nitrogen"sec-
tion immediatelyabove.
Calculations. Calculate the amount of N liberated by steam distillation
from the expression,(S - C) x T, whereS is the volume of H 2S04 used in titra-
tion of the sample,C is the volume used in titration of a control (obtainedby
steam-distilling an aliquot of extractant), and T is the titer of the titrant (for
0.0025 M H2S04, T = 70 flg N mL- 1).
Comments.The steam-distillationapparatusillustrated by Fig. 38-1 was
designedso that it can be rapidly disassembledbetweendistillations to prevent
cross-contaminationof samplesduring distillation of NHt -N for 15N analysis
(Hauck, 1982), although this is unnecessarywith the proceduresnormally
employedto prevent cross-contamination(Mulvaney, 1986). A similar type of
apparatusin which the partsare connectedby rubber or plastic tubing insteadof
ground-glassjoints can be usedif 15N analysiswill not be carriedout on NHt-N
in the distillate. To simplify operation,Teflon stopcocksare recommendedfor the
steam-bypassassembly. Ground-glassstopcocks also may be used, but they
should be greasedperiodically. If desired, the lower stopcockon this assembly
can be replacedby a pinchcock. If the apparatuswill not be used for distillation
of Kjeldahl digestsor soil hydrolysates,it can be fabricatedwithout a funnel, and
the upper stopcockcan be omitted from the steam-bypass assembly.The O-ring
joints specified are more convenientthan conventionalground-glassjoints, but
care is requiredin assemblingthe apparatusto ensurea properfit of the O-rings,
which is essentialto avoid leaks.The O-rings andground-glassjoints of the appa-
ratus should not be greased.
As described,steam distillations are performed using flasks with a side-
arm. The main advantageof the side-armis that it permitsaddition of Devarda's
alloy while the flask is attachedto the distillation apparatus,which is necessary
for determinationof N03"-N or (N03" + NOz)-N following removal of NHt by
steamdistillation with MgO. If this determinationis not required,distillation can
be accomplished using flasks without a side-arm.
The MgO employedfor distillation of NHt is ignited before use to remove
any MgC03 present,and the ignited material is storedin an airtight containerto
minimize its contentof this impurity, which interferesin titrimetric determination
of NHt-N in the distillate. The use of heavy MgO is recommended,as the light
variety tendsto creepup the walls of the distillation flask and migratethroughthe
distillation apparatus.
z
The Devarda'salloy employedfor reductionof NO) and NO to NHt must
be finely ground, becausestudieshave shown that both the amountof alloy and
the time required for this reduction are decreasedmarkedly by use of a finely
ground alloy (Bremner& Keeney, 1965). The alloy specified is supplied in the
form of a fine powder,and further grinding is unnecessarybefore use. A coarser
alloy may be used if it is ball-milled with a ceramicvial and ceramicballs until
all of the material will pass through a 150-llm (lOO-mesh) screen.The finely
ground alloy tends to adhereto the distillation flask and steam-inlettube, but
removal is easily accomplishedusing dilute HCI or H2S04.
The 0.2-g additionsof MgO and Devarda'salloy specifiedin the methods
describedneednot be accuratelyweighed,and are more easily made using cali-
bratedglassor metal spoons.
The certified sulfamic acid available commercially contains only trace
amountsof NHt, and further purification (Bremner,1965) is unnecessarybefore
use. Aqueoussolutions of sulfamic acid are not completely stable, as sulfamic
acid is slowly hydrolyzed to ammonium bisulfate (NH2S03H + H20 -?
NH4HS04). However,hydrolysis is very slow at room temperature,and extreme-
ly slow at 5°C, and only trace amountsof NHt havebeendetectedin 0.2 M solu-
tions of purified sulfamic acid that were storedfor 6 mo in a refrigerator(Brem-
ner & Keeney,1965). When usedin the methodsdescribed,such a solution does
not liberate a measurableamountof NHt.
The sharpnessof the end-point obtained during titration with 0.0025 M
H2S04 to determineNHt collectedin H3B03-indicatorsolutiondependsupon the
sourceof the indicatorsusedin preparingthis solution, and, if necessary,the rec-
ommendedproportionsof bromocresolgreenand methyl red may be altered to
obtain a more satisfactoryend-point.The sharpnessof the end-pointalso depends
upon the purity of the H3B03 and the concentrationof the H3B03 solution (Yuen
& Pollard, 1953). A sharperend-pointis obtainedwith a more dilute solution of
H3B03 (Yuen & Pollard, 1953), but the concentrationmust be high enough to
ensurecompleteretention of NH 3. The H3BOr indicator solution recommended
in the methodsdescribedcontains20 g of H3B03 L-l, and 5 mL of such a solu-
tion can adsorbabout5 mg of NHt -N (Yuen & Pollard, 1953).
The H2S04 usedfor titration can be standardizedusing accuratelyknown
amountsof tris(hydroxymethyl)aminomethane, which is available from several
commercial sourcesas a highly purified acidimetric standardunder the trade
name THAM. The THAM can be weighed directly (after drying for 2 h at
105°C), but it is usually more convenientto pipette aliquots from an aqueous
solution. Such a solution is stable for severalmonths if stored in a refrigerator,
but the containershould be stopperedtightly to excludeatmosphericCOz, which
is slowly absorbed.
Titrations with standardH2S04 are convenientlycarriedout using an auto-
matic titrator with potentiometric(pH) end-pointdetection,but titrations also can
be done manually using a microburette.In the latter case,the burette should be
equippedwith a three-waystopcockfor rapid refilling from a reservoir,and titra-
tions should be performedwith continuousstirring, using a variable-speedmag-
netic stirrer and a Teflon- or glass-coatedstirring bar.
1138 MULVANEY
tions shouldbe performedwith continuousstirring, using a variable-speedmag-

netic stirrer and a Teflon- or glass-coatedstirring bar.
Extensivefoaming can occur during steamdistillation of 2 M KCl soil
extractswith MgO and Devarda'salloy, andthe analysiswill be vitiated if any of
the foam contaminatesthe distillate. Foamingcanbe controlledby addinga small
amountof mineraloil to the samplebeforedistillation, but this hasbeenfound to
lead to low or erratic results in analysisfor NO) or N02" (Bremner& Keeney,
1965) and cannotbe recommended.Rather,the distillation apparatusshouldbe
equippedwith larger flasks (150- to 300-mLcapacity)when foaming is a prob-
lem. A longersteam-inlettube also will be required,but the necessarychangeis
easily madeif this tube is interchangeableas in the apparatusillustratedby Fig.
38-l.
Studies with glutamine (CSHlON203) and other alkali-labile organic-N
compoundshaveshownthat someof thesecompoundsyield measurableamounts
of NHS when they aresubjectedto steamdistillation with MgO for more thanthe
3- to 4-min period recommendedfor analysis of NH.t-N by the procedures
described(Bremner& Keeney,1965).Suchfindings suggestthat the greatestrisk
of interferenceby organic-Ncompoundsis in the methodsfor determinationof
NO)-N and(NO) + N02)-N, in which casethe total time of distillation with MgO
is at least7 min. An alternativeis to calculateNO)-N and (NO) + N02")-N from
the resultsobtainedin analysisfor NH.t-N, (NH! + NO)-N, and (NHS + NO) +
N02)-N. However,testsusing a variety of mineral soils haveshownthat identi-
cal results are obtainedby either approach (Bremner & Keeney, 1965), which
supportsother evidencethat the organic-N compoundsin soil extractsdo not
releasesignificantamountsof NRt underthe conditionsof the methodsdescribed
(Bremner& Keeney,1966).
Reductionof N03" and N02" to NHS by Devarda'salloy is subjectto inter-
ferencefrom high concentrationsof variousinorganicanionsandcations,notably
phosphate(Bremner& Keeney,1965; Freney,1971; Bremner& Bundy, 1973),
silicate (Bremner& Keeney,1965; Bremner& Bundy, 1973), and Mg2+ (Brem-
ner & Keeney,1965; Clausenet aI., 1980; Best & Craswell, 1985). The concen-
trationsrequiredfor interferenceare much higher than would normally occur in
2 M KCI soil extractsobtainedwith a 1:10 ratio of soil to extractant.However,
BestandCraswell(1985)reportedseriousinterferenceby Mg2+ during steamdis-
tillations to determineN03" in 2 M KCI extractsobtainedusing a soiVsolution
ratio of 1:2.5.
As described,analysesby steamdistillation are performedon soil extracts
obtainedusing 2 M KCI, but analysesalso can be carriedout when otherextrac-
tantsare used,suchas water, 1 M KCl, or 0.5 M K2S04, and acidic extractscan
be analyzedif they are neutralized(by addition of NaOH) before distillation
(Bremner,1965).
The accuracyand precisionof analysesby the methodsdescribedcan be
checkedby analyzingappropriatealiquotsof Reagents6 or 7 (seethe previous
"Reagents"section)to encompassthe rangeof inorganicN encounteredin analy-
sesof soil extracts.Both reagentsare stable for severalmonths if stored in a
refrigerator.
Controlsshouldbe performedin eachseriesof analysesto allow for NH;t-N

derivedfrom the reagentsemployed(including the reagentsusedfor extractionof
exchangeableNH;t, NO)", and NOil
Microdiffusion Methods
Introduction
The useof microdiffusion to determineinorganicforms of N in soil extracts
offers several advantages.The proceduresare simple and require very little
equipment.Coloredor turbid extractsmay be analyzedwithout difficulty. Diffu-
sion is carriedout at room temperature,and consequentlythe risk of interference
by alkali-labile organic-N compoundsshould be lower than with steam-distilla-
tion methods,provided a mild alkali such as MgO is employed (Bremner &
Shaw,1955). However,diffusion methodshavebeenusedmuch lesswidely than
steam-distillationmethods,presumablybecausea 1- or 2-d period is requiredto
obtain the results.Moreover,useof diffusion is complicatedby the fact that, with
most methods,no more than 4 to 10 mL of extractcan be safely analyzed.
An important considerationin diffusion methodsis the size and design of
the container employed, which necessarilydeterminesthe surface-to-volume
ratio for the sampleand absorbingsolutions.To increasethe rate of diffusion, a
shallow dish is normally usedwith separatechambersfor sampleand absorbent.
Two types of units are availablecommercially.One is of glassconstructionand
follows the design of Conway (1947), in which the top of the dish is sealedby
afftxing to it a flat plate with petroleumjelly or gum arabic. The other is of plas-
tic constructionand follows the three-chamberdesignof Obrink (1955), in which
a flanged cover fits into an annular moat containing the alkaline agent used to
treat the sample.The latter designhas the advantagethat no fixative is required,
but neither design is really satisfactoryfor diffusion of soil extracts.The most
seriousproblemis the volume of the samplechamber,which can accomodateno
more than 4 mL of extract, and with the commercially available three-chamber
unit, this problemis compoundedby the limited height of the wall that separates
the chambersfor sampleand absorbent.Larger versionsof both units have been
described(Bremner & Shaw, 1955; Bremner, 1965; Freney & Wetselaar,1967;
Hauck, 1982), and with these,the maximal volume of sampleis 6 to 20 mL. A
still larger unit, having sufficient capacity to easily accomodate100 mL of
extract,is employedwith the methodsdescribedin the following "Methods" sec-
tion. Although not commercially available, the latter unit is readily fabricated
from inexpensivecomponents.
A survey of microdiffusion methodsfor determinationof inorganic forms
of N in soil extractsrevealsthat the choiceof reagentshas often beenlimited by
the designof the microdiffusion unit employed.As originally proposed(Conway,
1947), microdiffusion of NH3 was carriedout by treatmentof the samplewith 1
mL of a saturatedsolution of K 2C03, which forms KOH. A 3.3 M (45%) solution
of the samereagenthas beenemployedin diffusion of soil extracts(Stanfordet
aI., 1973; Keeney& Nelson, 1982); however,interferencecan arisefrom decom-
position of labile organic-N compoundssuch as glucosamine(C6H 13N05), and
1140 MULVANEY
analysescannot be performed on acidic extracts owing to liberation of CO2,

which interferesin titrimetric determinationofNHrN (Bremner& Shaw, 1955).
To avoid the latter problem, Conway (1947) suggestedthat KOH or potassium
metaborate(K2B20 4) be used insteadof K 2C03 for analysisof acidic samples,
but Bremnerand Shaw (1955) found that thesereagentsalso liberated NHrN
from glucosamineat room temperature.Their work showedMgO to be the best
alternative,and in the procedurethey developed,the soil extractis madealkaline
by treatmentwith 3 mL of an aqueoussuspensionof this reagent(Bremner &
Shaw, 1955; Bremner,1965).
As with steamdistillation, microdiffusion methodsof determiningN03"-N
involve use of a reducing agent to convert N03"-N to NHS-N. In some cases,
Devarda'salloy has been used (Conway, 1947; Stanford et aI., 1973; Hauck,
1982; Keeney& Nelson, 1982), and this material has the advantagethat it quan-
titatively reducesboth N03" and NOz to NHS underalkaline conditions.Howev-
er, use of Devarda'salloy is complicatedconsiderablywhen microdiffusion is
carried out in a shallow dish, owing to the productionof Hz, which generatesan
alkaline spray that may contaminatethe acidic absorbent(Bremner & Shaw,
1955), and the resulting increasein pressuremay causethe lid to leak (Hauck,
1982). A coarselyground alloy may be used to slow the liberation of H2 (Stan-
ford et aI., 1973; Hauck, 1982; Keeney & Nelson, 1982), and in somecases,an
antifoamagenthasbeenrecommendedto control spraying(Stanfordet aI., 1973;
Keeney& Nelson, 1982), but most workers haveresortedto a different reducing
agent,the alternativesbeing Ti2(S04)3 (Bremner& Shaw, 1955; Bremner, 1965;
Freney& Wetselaar,1967) or a mixture of FeS04and Ag2S04 (Premi & Corn-
field, 1967)or zinc powder(Li, 1968).While thesereagentsquantitativelyreduce
NO) to NHS, reductionofNOz is incomplete;therefore,NOz-N cannotbe direct-
ly determined,and a pretreatmentwith sulfamic acid is adviseableto ensureits
absenceduring determinationsof N03"-N (Bremner & Shaw, 1955; Bremner,
1965).
Conway (1947) suggestedthe use of H3BOr indicator solution to absorb
NH3 in microdiffusion analysis, and this practice has been widely adopted in
microdiffusion methodsof determininginorganic N in soil extracts.The reagent
recommendedby Conway (1947)contained10 g ofH3B03 L-l, but Bremnerand
Shaw(1955) obtainedhigher recoveriesofNHS-N using a reagentthat contained
20 g of H3B03 L -1. The boric acid-indicator solution used in the methods
describedin the following "Methods" sectioncontains40 g of H3B03 L-1. The
NHS -N collectedduring diffusion is readily determinedby titration with standard
acid.
Principles
The methods described are exactly analagousto the steam-distillation
methodsdescribedin "Methods" for "Steam-DistillationMethods,"in that NHS,
z
NO), and NO are determinedthrough conversionto NH3, the major difference
being that the NH3 liberatedby theseconversionsis separatedby microdiffusion
at room temperatureinsteadof by distillation with steam.In the absenceof NO z,
NHS-N is determinedfrom the NH3 liberated by treatmentof the samplewith
NITROGEN·INORGANIC FORMS 1141
o Stainless-steel machine screw

eO·ring
6) Stainless-steel nut
e Screw-down mounting base
" Nylon cable tie (releasable)
o Cable clamp
e Nylon machine screw
o Nylon nul
Fig. 38-2. Mason-jarmicrodiffusion unit.
MgO, and N03-N is determinedfrom the NH3 liberatedby subsequentaddition

of Devarda'salloy. In the presenceof N02, N03-N is determinedby the same
procedureafter treatmentof the samplewith sulfamic acid to decomposeN02,
and N02-N is determinedas the differencebetweenresultsobtainedby addition
of MgO and Devarda'salloy, with and without sulfamic acid. In eachcase,the
NH3 liberatedis absorbedin H3BOr indicator solution and determinedby titrat-
ing this solution with standard(0.0025M) H2S04•
Methods2
Special Apparatus
1. Microdiffusion unit (Fig. 38-2). The unit illustrated consistsof a 473-
mL (l-pt) wide-mouth Mason jar (cat. no. 70610-00518,Kerr Glass
ManufacturingCorp., P.O. Box 76961,Los Angeles,CA 90076or Ball
brand cat. no. 14400-66000,Alltrista Corp., Muncie, IN 47305) with
86-mm dome lid (Ball brand only) modified to supportthe bottom for
a 60-mm (diam.) Pyrex Petri dish (55-mm diam., 17 mm high). The lid
is modified by: (i) drilling a 3.6-mm (9/64-in.) hole 12 to 14 mm from
the edge;(ii) attachinga 6-32 by 38- to 64-mm (1.5-2.5 in.) stainless-
steelmachinescrewwith two O-rings(Viton, 2.9-mmi.d., 6.4-mmo.d.)
and two stainless-steelnuts; and (iii) fasteninga screw-downmounting
basefor a releasablenylon cabletie to the stainless-steel screwvia a 3.2
mm (1/8 in.) cable clamp, a 6-32 nylon machinescrew (13 mm long)
and nut, and four stainless-steelnuts. To improve the fit of the lid, the
cable clamp is notchedwith a bench grinder, and the nylon screw is
ground flush with the nut. The cable tie for mounting the Petri dish
should be tightened sufficiently that the dish cannot slip from the
mount, yet can be easily removedand replaced.Double stainless-steel
nuts are tightenedfirmly againsteachother to prevent loosening.The
2 Modified from Saghiret al. (1993).

1142 MULVANEY
cabletie mountingbaseillustratedin Fig. 38-2 is manufacturedby Pan-

duit, Tinley Park, IL (cat. no. TM2S6-C), and is available from elec-
tronic suppliers.The othercomponentsusedin modifying the lid canbe
obtainedfrom McMaster-CarrSupply Co., Chicago,IL.
2. Orbital shaker(optional). A heavy-dutyunit is recommended.A ply-
wood box (91 cm long, 66 cm wide, 30 cm high) may be mountedto
serveas a platform for shaking.Up to 144 samples,in threelayers,may
be accommodatedif the jars are placed ontrays (e.g., 12 jars per tray).
3. Microburette,SmL, graduatedat O.01-mL intervalsor automatictitra-
tor equippedwith a microelectrode.A satisfactorymicroelectrodeis
available from Microelectrodes,Inc., 40 Harvey Road, Bedford, NH
03110(Model MI-411S).
Reagents
1. Magnesiumoxide. Either light or heavypowdermay be used.
2. Boric acid-indicatorsolution. Add 800 g of reagent-grade(H3B03) to
18 L of deionizedwaterin a 20-L Pyrex bottle markedto indicatea vol-
ume of 20 L, and stir vigorously with a motorizedstirrer to dissolvethe
H3B03. Then add 0.099 g of water-solublebromocresolgreen and
0.066 g of methyl red (as a water-solublesodium salt) (satisfactory
indicatorsare availablefrom EM Science,P.O. Box 70, 480 Democrat
Road,Gibbstown,NJ 08027),and bring the volume to 20 L with deion-
ized water. With continuousstirring, adjust the pH of this solution to
4.2-4.3 by cautiously adding 1 M NaOH or single NaOH pellets, and
then check that a pH of 4.8 to 5.0 is obtainedwhen an aliquot of the
boric acid-indicatorsolution is diluted with an equalvolume of deion-
ized water. If excessNaOH is added,the pH can be reducedby adding
dilute HCI. Dispensethe solution throughTeflon tubing.
3. Devarda'salloy. Use the alloy specifiedin the previous"Reagents"sec-
tion.
4. Sulfamicacid solution. Prepareas describedin the previous"Reagents"
section.
5. Sulfuric acid, 0.0025M standard.
6. Standard(NHt + NO]")-N solution. Prepareasdescribedin the previous
"Reagents"section.
7. Standard(NHt + NO]" + NOz)-N solution. Prepareas describedin the
previous"Reagents"section.
Procedures in Absence of Nitrite
Ammonium-Nitrogen. Pipettean aliquot (to 100 mL) of soil extractinto the
Mason jar; if necessary,add 2 M KCI to increasethe volume to at least 10 mL.
Dispense4 mL of boric acid-indicator solution into a Petri dish bottom, and
attach thedish to thejar lid with the cabletie. Then add 0.2 g of MgO using a cal-
ibratedspoon,and swirl the jar to mix the contents.After allowing 15 to 30 s for
any magnesiumoxide dust to settle, place the lid on the jar and seal it with a
screw band. Then place the jar on a laboratorybenchtop(for diffusion without
shaking), or transfer it to an orbital shakerfor continuousagitation (75-85 rev
min-I). After a sufficient period for completediffusion of NH3 into the H3B03
solution (Table 38-4), removethe Petri dish from the jar, add 5 mL of deionized
water to the dish, and titrate the H3B03 solution with 0.0025M H2S04 from a
microburetteor automatictitrator. At the end-point,the color changesfrom green
to a permanent,faint pink.
Nitrate-Nitrogen. Following diffusion with MgO as describedin the previ-
oussection,openthe jar, andremovethe Petri dish from the jar lid. Attach anoth-
er Petri dish containing4 mL of H3B03-indicator solution to the lid. Treat the
samplein the jar with 0.2 g of MgO from a calibratedspoon,followed by 0.2 g
of Devarda'salloy, swirl the jar briefly to mix the contents,and seal the jar by
attachingthe lid with the Petri dish. Carry out diffusion (with or without shaking)
for the sameperiod specifiedpreviously.Titrate the H3B03 solution with 0.0025
MH 2S04•
(Ammonium + Nitrate)-Nitrogen. Proceedas describedin the previous
"Ammonium-Nitrogen"section,but add 0.2 g of Devarda'salloy to the sample
in the jar after the addition of MgO.
Procedures in Presence of Nitrite
Ammonium-Nitrogen. Proceedas describedin the previous"Ammonium-
Nitrogen" section.
(Nitrate + Nitrite)-Nitrogen. Proceedas describedin the previous"Nitrate-
Nitrogen" section.
(Ammonium + Nitrate + Nitrite)-Nitrogen. Proceedasdescribedin the pre-
vious "(Ammonium + Nitrate)-Nitrogen"section.
(Ammonium + Nitrate)-Nitrogen. Proceedas describedin the previous
"(Ammonium + Nitrate)-Nitrogen"section,but treat the samplein the jar with 1
mL of sulfamic acid, and swirl the jar for a few secondsto decomposeN02",
beforethe addition of MgO and Devarda'salloy.
Nitrate-Nitrogen. Proceedas describedin the previous"Nitrate-Nitrogen"
section,but perform the analysison a samplethat hasbeentreatedwith sulfamic
acid to destroyN02" as describedin the "(Ammonium + Nitrate)-Nitrogen"sec-
tion immediatelyabove.
Table 38-4. Diffusion periodsfor different volumesof soil extract.

Minimal period for completediffusion
Volume Without shakingt With shaking:!:
mL
10 26 h 18 h
20 44h 30h
50 5d NR
100 8 d (9 d) NR
*
t Value in parenthesisindicatesthe minimal diffusion period when using the Ball jar specified.
NR = not recommended.
1144 MULVANEY
Calculations.Calculatethe amount of N liberated by diffusion from the

expression,(S - C) x T, whereS is the volume of H2S04 usedin titration of the
sample,C is the volume usedin titration of a control (obtainedby diffusing an
aliquot of extractant),and T is the titer of the titrant (for 0.0025M H2S04, T =70
~gN mL-l).
Comments.The microdiffusion unit describedhasseveraladvantagesover
the units previously employedfor determinationof inorganicforms of N in soil
extracts.The Masonjar specifiedis readily availablefrom a numberof discount
storesand supermarkets,and at very low cost. It is large enoughto easily accom-
modate100 mL of extract, yet diffusion is rapid becauseof the large diameter,
which effects a high surface-to-volumeratio. Diffusion is further hastened
through the use of a Petri dish to contain the H3B03 solution, which providesa
large areafor absorptionof NH3, and the dish is positionedabovethe solution to
maximize the surfaceareaof sampleand avoid contaminationof the absorbing
solution by creepof salt or alkali. The lid sealstightly without the needfor fIXa-
tive agents,and there is no risk of leakagefrom the increasein pressurecaused
by liberation of H2 from Devarda'salloy.
The Kerr brand 473-mL (I-pt) wide-mouthMasonjar is somewhatlarger
in diameterand somewhatshorterthan the correspondingBall jar. The effect in
both casesis to increasethe rate of diffusion, and for this reason,the Kerr jar is
probably a better choice than the Ball jar. No difference in recovery has been
observedwhen diffusions areperformedfrom 10, 20, or 50 mL of soil extractfor
the periodsspecifiedin Table 38-4, but with tOO mL of extract, an extra day is
requiredfor completerecoverywhen using the Ball jar. Becauseof the difference
in dimensions,the Kerr jar is somewhateasierto cleanthan the Ball jar, and this
can be a considerableadvantagein performing a large number of analyses.
Regardlessof which jar is used,a Ball lid is strongly recommended.Experience
hasshownthat the sealingsurfaceof the Kerr lid becomesgroovedfrom a single
use, and thereafterthe lid may not seal reliably, in which casediffused NHt-N
will be underestimated.The sealingsurfaceon the Ball lid is more resilient, and
no suchdifficulty hasbeenobserved,evenafter repeateduse for many analyses.
As described, microdiffusionanalysesfor inorganic N are performedon
samplesobtainedby extractionof soil with 2 M KCI, but with longerperiodsfor
diffusion, the samemethodscan be employedfor analysisof extractsobtained
using other reagents,such as water, 1 M KCI, or 0.5 M K 2S04, and for analysis
of natural waters.Acidic extractscan be analyzedafter they have beenneutral-
ized (by addition of NaOH). Determinationsof NHt in Kjeldahl digestscan be
accomplishedby a simple modification of the methodfor soil extracts,in which
a vial containingan aliquot of digest is overturnedinside a jar containingNaOH
(Stein et aI., 1993).
The microdiffusion units must be cleanedthoroughlybeforeuseto remove
all tracesof alkali and Devarda'salloy, and this is most easily accomplishedif it
is donesoonafter eachuse.Thejars may be cleanedby a procedurein which they
are washedwith tap water usingthe fingers or a small brush to aid removal of
adheringmaterial,and then rinsedthoroughlywith deionizedwater. The lids and
Petri dishesmay be cleanedby thoroughly rinsing them with deionizedwater.
The boric acid-indicator solution used in the microdiffusion methods

described contains twice the concentration of H3B03 used in the reagent
employed with the steam-distillation methods described in "Methods" for
"Steam-DistillationMethods." The higher concentrationof H3B03 dramatically
increasesthe capacityfor absorptionof NH 3, and also reducesthe periodfor com-
plete diffusion, particularly with 100 mL of soil extract. To improve sensitivity,
the H3B03 solution is diluted with deionizedwater beforetitrimetric determina-
tion of the absorbedNHrN. Water-solubleforms of the indicators,bromocresol
green and methyl red, have been specified in "Reagents"for "Microdiffusion
Methods"for their easeof aqueousdissolution.If alcohol-solubleforms of these
indicators are used, they should be dissolved in ethanol as described in
"Reagents" under "Steam-Distillation Methods." The concentrations of
bromocresolgreen and methyl red in the boric acid-indicatorsolution specified
are 75% lower than the concentrationsof theseindicatorsin the H3B03 reagent
recommendedfor steamdistillations (see "Reagents"under "Steam-Distillation
Methods").The reductionwas madeto facilitate isotope-ratioanalysesof the dif-
fused NHt-N. No difficulty hasbeenobservedin detectingthe end-pointduring
acidimetric titrations as described,but if desired,a higher concentrationof indi-
cator can be employedin caseswhere 15N analyseswill not be performed;e.g.,
0.396 g of bromocresolgreenand 0.264 g of methyl red per 20 L of boric acid-
indicator solution.
The other reagentsemployedin the microdiffusion methodsdescribedare
the sameas those used in steamdistillations, and referenceshould be made to
"Reagents"and "Comments" under "Steam-Distillation Methods" for specific
information regardingthesereagents.The MgO may be usedwithout being ignit-
ed to remove MgC03, as no interferencedue to CO2 has been detectedusing
unignited MgO. As noted in "Comments"under "Steam-DistillationMethods,"
high concentrationsof phosphate,silicate, or Mg2+ can interfere with reduction
z
of NO)" and NO to NHt by Devarda'salloy, but such interferencedoesnot nor-
mally occurin analysisof 2 M KCI extractsobtainedusing a soil/solutionratio of
1:10. A secondaddition of MgO has beenfound to increaserecoveryin the pro-
ceduresfor determinationof NO)"-N and (NO)" + N02)-N (Saghiret aI., 1993).
The 0.2-g additionsof MgO and Devarda'salloy specifiedin the methods
describedneednot be accuratelyweighed,and are more easily madeusing cali-
bratedglassor metal spoons.Care should be taken in the use of MgO to elimi-
nate any possibility that this reagentmay come in contactwith Petri dishescon-
taining boric acid-indicatorsolution, becausethis can lead to overestimationof
diffused NHS-N.
When diffusions are performedfor the periodsspecifiedin Table 38-4, the
4 mL of boric acid-indicatorsolution usedin the methodsdescribedquantitative-
ly absorbup to 4 mg of N as NHS or NO)" from 10 or 20 mL of 2 M KCI, and up
to 2 mg of N from 50 or 100 mL of 2 M KCI.
If desired,shakingmay be employedto reducethe periodrequiredfor com-
plete diffusion with 10 or 20 mL of soil extract. To avoid contaminationby
splashing,an orbital shakeris used rather than a reciprocatingunit, and the jars
are placedso that they cannotrock. Shakingshould be done at the speedspeci-
1146 MULVANEY
fied, as the rate of diffusion will be reducedwith a lower speed,whereassplash-

ing is likely to occurwith a higherspeed.Orbital shakersarecommonlyequipped
with a meterto indicatethe speedof shaking,but experiencein the author'slab-
oratorywith two of theseshakershasshownthat the readingcanbe grosslyinac-
curate,so the speedshouldbe checkedmanually using a stopwatch.Shakingis
not recommended with 50 or 100 mL of soil extract,becausethe periodfor com-
pletediffusion is not appreciablyreduced,and thereis considerablerisk of cont-
aminationby splashing.
The diffusion periodsspecifiedfor different volumesof soil extractwere
determinedthroughstudiesin which diffusion wascarriedout at a temperatureof
25°C. Satisfactoryresultsmay be obtainedat lower temperaturesby increasing
the diffusion periodsspecifiedin Table 38-4. For example,a 25% increaseis
requiredfor completerecoveryat 20°C.
Titrationswith standardH2S04 are convenientlyperformedusing an auto-
matic titrator, but a microelectrodeis requiredowing to the shallowdepthof liq-
uid in the Petri dish. Titrations also can be done manually using a microburette
equippedwith a three-waystopcockfor rapid refilling from a reservoir.A vari-
able-speedmagneticstirrer and a small stirring bar can provide continuousstir-
ring during manualtitrations, but mixing is more convenientlyaccomplishedby
swirling the solutionin the dish.
The methodsdescribedcanbe checkedby analyzingappropriatealiquotsof
Reagents6 or 7 (see"Reagents"for "Microdiffusion Methods")to encompass the
rangeof inorganicN encounteredin analysesof soil extracts.Both reagentsare
stablefor severalmonthsif storedin a refrigerator.
Controlsshouldbe includedin eachseriesof analysesto allow for NH4-N
derivedfrom the reagentsemployed(including the reagentsusedfor extractionof
soil).
ColorimetricMethods
Introduction
Colorimetric methodshave beenusedextensivelyto determineinorganic
forms of N in soil extracts.Their main advantagesare speedand convenience,
particularlywhen analysesare to be performedon a large numberof samples.In
such cases,an automatedanalyzersystemmay be employedfor simultaneous
z,
determinationof NHt, NO), andNO at a rateof up to severalhundredsamples
per hour.
Ammonium.The Nesslermethodhasbeenusedto determineNHt in soil
extracts(e.g.,Greweling& Peech,1960); however,sensitivityis limited, and the
measurements are subjectto interferenceby severalsoil constituents,particular-
ly Ca, Mg, Fe, and sulfide. The interferencescanbe eliminatedby steamdistilla-
tion (Am. Public Health Assoc.,1992),but NHt in the distillate is more readily
determinedby acidimetric titration than by colorimetry.Becauseof its limita-
tions, the Nesslermethodhas not beenusedwidely for analysisof soil extracts.
Much bettermethodshavebeendevelopedby utilizing the Berthelotor indophe-
nol reaction.
Berthelot (1859) noted that an intenseblue color was formed by mixing

NH3, phenol (C6H60), and hypochlorite(NaOCI). Van Slyke and Hiller (1933)
utilized this reactionto developa methodfor determiningNH4" in blood, which
becameknown as the indophenolblue, or phenate,method. Thismethodproved
to be more sensitivethan nesslerizationandwaswidely usedin clinicallaborato-
ries. Subsequentwork showedthat sensitivity is enhancedby carrying out the
reaction in the presenceof a catalyst, preferably sodium nitroprusside
(CsFeN,.Na30)(Lubochinsky& Zaita, 1954), and that the samereactionoccurs
with at least65 substitutedphenolshaving an unsubstitutedpara-position (Solo-
way & Santoro,1955). Phenolhas beenused more widely than other phenolic
compoundsin determinationof NH4 by the Berthelotreaction(Searle,1984),but
sodiumsalicylate(NaC7Hs03) also hasbeenusedwidely becauseit is less toxic
than phenol and is a more convenientreagent.Despitenumerousclaims to the
contrary, recent work by Krom (1980) indicates that similar sensitivities are
obtainablewith different phenoliccompounds.
Severalmanualprocedureshavebeendescribedfor analysisof NH4 in soil
extractsby the Berthelot reaction (Kempers, 1974; Gerlach, 1980; Keeney &
Nelson,1982; Dorich & Nelson,1983; Nelson,1983; Kempers& Zweers,1986;
Wang& 0ien, 1986;Qui et ai., 1987).The mostseriousinterferenceis from diva-
lent and trivalent cations(notably Ca2+, Mg2+, and Fe3+), which form insoluble
hydroxidesat the high pH (11-13) requiredfor the analysis.To avoid suchinter-
ference,Kempers(1974) steam-distilledthe extract before analysis,but subse-
quentwork hasled to direct methodsof analysis,in which the soil extractis treat-
ed with a chelatingagent [ethylenediaminetetraacetic acid (EDTA) or a citrate
buffer] to complex polyvalent cationsand therebypreventtheir precipitationat
high pH (Keeney& Nelson,1982; Dorich & Nelson, 1983; Nelson, 1983; Kem-
pers & Zweers, 1986; Qui et ai., 1987). This techniquewas first employedfor
determinationof NHt in seawater (Roskam& de Langen,1964), and has been
usedwidely in automatedmethodsfor Nat analysisof soil extracts(Henzell et
ai., 1968; Selmer-Olsen,1971; Rowland, 1983; Markus et aI., 1985; Gentry &
Willis, 1988).
The Berthelotreactionis not entirely specificfor NH4. In addition to NH4,
a color is obtainedwith a variety of organic compoundshaving a free amino
group, including amino acids, amines,and amides(see Searle,1984). Several
investigationshave shown that amino acids interfere in automatedanalysisof
NH4 in soil extractsby methodsbasedon the Berthelotreaction(White & Gosz,
1981; Rowland, 1983; Adarnsenet ai., 1985; Burton et ai., 1989), and manual
methodsfor this analysisare undoubtedlysubjectto the sameinterference.The
results from these investigationssuggestthat this type of interferenceis most
likely to occurwith soils havinga high contentof organicmatter(White & Gosz,
1981), and with soils that have beenfumigatedor subjectedto air- or oven dry-
ing (Burton et ai., 1989).
The optimum conditions for analysisof NH4" by the Berthelot reaction
dependupon the reagentsemployed.Color developmentis strongly affectedby
pH, and a buffer is commonlyusedto ensurea uniform pH for all analyses.With
phenol,maximumcolor intensity is obtainedat a pH of 11 to 12 (Weatherbum,
1967; Kempers, 1974), whereasa pH of 12 to 13 is required with salicylate
1148 MULVANEY
(Krom, 1980; Nelson, 1983; Kempers& Zweers,1986), presumablybecauseof

its higher pKa value (Krom, 1980). In some casesorganochlorinecompounds
suchas dichloroisocyanurate (C30 3N3Cl2) havebeenusedto generatehypochlo-
rite for analysesof NRt by the Berthelot reaction(e.g., Krom, 1980; Adamsenet
aI., 1985); however, most workers have preferredto use a solution of NaOCI,
which is readily availableas a householdbleach.Color developmentcan be car-
ried out at room temperature(Kempers, 1974; Gerlach, 1980; Kempers &
Zweers, 1986; Wang & 0ien, 1986) or even in a refrigerator(Kempers& Kok,
1989),but gentleheating(e.g., to 37-40°C)is often employedto reduceanalysis
time and increase sensitivity (Weatherburn, 1967; Keeney & Nelson, 1982;
Dorich & Nelson, 1983; Nelson, 1983; Stock, 1983). The color is stable for at
least 1 h after development(Nelson, 1983; Kempers& Zweers,1986), and then
slowly fades.
For convenience,analysesof NHt by the Berthelot reactionmay be carried
out using various combinationsof the reagentsrequiredfor color development.
Weatherburn(1967) describeda procedurethat usesonly two reagents,phenol
plus nitroprusside(CsFeN602-) and alkali plus hypochlorite.Similar procedures,
using either phenol or salicylatewith nitroprussideand a buffered hypochlorite
solution, havebeensuccessfullyemployedin analysesof soil extracts(Kempers,
1974; Keeney& Nelson, 1982; Dorich & Nelson,1983; Nelson, 1983).
Nitrate. The phenoldisulfonic acid (C6H60 7S2) method has been used
widely to estimateNO)" in soil extractsby colorimetry (Bremner,1965); howev-
er, the procedureis complicated,and the analysesare subjectto interferenceby
organicmatter,NOl", and Cl-. The latter interferenceprecludesthe useof KCI as
an extractant,and even the levels of Cl- that occur naturally in some soils can
lead to significant error, requiring a chemicaltreatmentto remove Cl- from the
samplebefore the analysisfor NO)" is carried out (Bremner, 1965; Puttanna&
PrakasaRao, 1981).The limitations of the phenoldisulfonicacid method have
drawn attentionto the needfor a more satisfactoryalternative,and severalother
colorimetricmethodsof determiningN03" havebeenproposedfor analysisof soil
extracts(Barker, 1974; Keeney& Nelson, 1982).
Brucine (Cz3H26N204)reactswith N03" to give a blue color that exhibits
maximal absorbanceat 410 nm. This reactionhas beenutilized for the analysis
of N03" in soil extracts(Wolf, 1944; Fisher et aI., 1958; Robinsonet aI., 1959;
Baker, 1967). While the analysesare simpler and more convenientthan with the
phenoldisulfonicacid method,they are subjectto interferenceby organicmatter
and NOl", and are particularly sensitiveto the presenceof strong oxidizing or
reducing agents. Moreover, the brucine reagent must be preparedfrequently
becauseit is unstable, and this is a significant limitation given the fact that
brucineis extremelytoxic.
Under strongly acidic conditions. N03" reactswith xylenol (dimethylphe-
nol; CSHlOO) to form nitroxylenol (CSH9N02), which may be determinedcolori-
metrically by treatmentwith NaOH after steam distillation or extraction with
toluene(C7HS). Various versionsof the xylenol methodhave beenemployedfor
determinationof NO)" in soil extracts (Balks& Reekers,1955; Buckett et aI.,
1955; Lewis, 1961); however, the proceduresare even more complicatedthan
with the phenoldisulfonicacid method,and the analysesare likewise subjectto

interferencefrom organicmatter, N02", and Cl-.
A simpler approachto determiningNO)" by colorimetry utilizes the reac-
tion with chromotropic acid (4,5-dihydroxy-2,7-naphthalenedisulfonic acid) in
concentratedH2S04. Originally developedfor analysisof naturalwaters(West&
Lyles, 1960),the chromotropicacid methodhas beenemployedby severalwork-
ers for determinationof NO)" in soil extracts(Clarke & Jennings,1965; Sims &
Jackson,1971; Kowalenko & Lowe, 1973; Hadjidemetriou,1982; Haby, 1989).
Analysesby this methodare rapid and convenient,and there are fewer interfer-
ences than with the phenoldisulfonic acid method. However, McNeilly and
Howard (1973) found that the chromotropicacid methodlackedsensitivity when
usedfor analysisof 1 M KCl extracts,and that the methodseriouslyunderesti-
matedNO)" in Ca(OH)2extractsfrom woodlandsoils due to the presenceof sol-
uble organicmatterthat interferedwith color development.
The reaction of NO)" with salicylic acid in concentratedH2S04 to form
nitrosalicylic acid, which can be determinedcolorimetrically after addition of
NaOH, has been utilized for NO)" analysisof wastewater(Scheiner,1974) and
plant extracts(Cataldoet aI., 1975), and recentwork by Vendrell and Zupancic
(1990) indicates that the same reaction may be employed in analysis of soil
extracts.The procedureis simple and convenient.The major limitation appears
to be interferenceby Cl-, which precludesthe use of KCl as an extractant.Fur-
ther evaluationis neededto check for other interferences,particularly organic
matter.
Severalothercompoundshavebeenutilized to developcolorimetric meth-
ods of determining NO)", including strychnine (C21H22N202) (Snell & Snell,
1949),pyrogallol sulfonic acid (CJI606S)(Snell & Snell, 1949), diphenylamine
(C12H ll N) (Snell & Snell, 1949), iron (II) sulfate(Swann& Adams, 1956),crys-
tal violet (~H3oN3Cl) (Shukla& Singh,1968),fluorescein(~OH1205) (Axelrod
et aI., 1970),bianthronyl(C28H 1S0 2) (Nawratil et aI., 1974), resorcinol(C6H60 2)
(Nakamura,1980), and 2-sec-butylphenol(Tanakaet aI., 1982). Most of these
methodshavenot beenevaluatedfor analysisof soil extracts.
Nitrate also may be determinedby reducingit to N02", in which casea col-
orimetric analysis is easily performed by the Griess-Ilosvay method (see
"Nitrite"). Various reducingagentshavebeenemployedfor this purpose,includ-
ing a mixture of zinc and MnS04 • H20, with or without CUS04 • 5H20 (e.g.,
Singh, 1988); hydrazineand CUS04·5H20, (e.g., Kamphakeet aI., 1967; Hen-
zell et aI., 1968; Best, 1976; Markus et aI., 1985; Am. Public Health Assoc.,
1992);crudeor purified preparationsof the enzyme,nitrate-reductase (e.g., Lowe
& Gillespie, 1975; Rice et aI., 1984); powderedzinc (Heanes,1975); NaCl in 18
M H2S04 (Nakamura,1981; Fakhri et aI., 1983); TiCl 3 (Al-Wehaid & Town-
shend,1986);a powderedcadmiumcatalyst(Lambert& DuBois, 1971); amalga-
mated cadmium metal (Gaugush& Heath, 1984); spongy cadmium particles
(Jones,1984); cadmium-silverwire (Willis & Gentry, 1987); a hollow cadmium
coil (Elliott et aI., 1989); and copperizedcadmium metal (e.g., Henriksen &
Selmer-Olsen,1970; Huffman & Barbarick, 1981; Keeney & Nelson, 1982;
Dorich & Nelson, 1984; Adamsenet al., 1985; Am. Public Health Assoc.,1992;
1150 MULVANEY
Tel & Heseltine,1990). The latter reductanthas been used extensivelyin N03"
analysisof soil extractswith autoanalyzersystems(see"Automation"), and man-
ual procedureshave beendescribedthat utilize the samereductant(Huffman &
Barbarick, 1981; Keeney & Nelson, 1982; Dorich & Nelson, 1984). Hydrazine
reductionalso hasbeenemployedfor NO)" analysisof soil extracts(e.g.,Henzell
et aI., 1968; Best, 1976; Markus et aI., 1985),but the high pH requiredfor reduc-
tion by this method(> 11) can lead to interferencefrom polyvalentcations,which
form hydroxideprecipitates(Ananth & Moraghan,1987).
Nitrite. The colorimetricmethodsgenerallyemployedfor determinationof

z z
NO are basedon the reactionof NO with primary aromaticamines(diazotizing
reagents)under acidic conditionsto producediazoniumsalts, which react (cou-
pIe) with aromatic compoundscontaining suitable amino or hydroxyl groups
(coupling reagents)to form coloredazo compoundsthat may be measuredspec-
trophotometrically(Boltz & Taras, 1978; Fox,1985). Thesemethodshave their
origin in work by Griess(1879), who performedthe diazotizationreactionusing
sulfanilic acid (C6H7N03S) in H2S04 solution and utilized 1-naphthylamine
(ClOH9N) as the coupling reagent.Ilosvay (1889) modified the original method
by carrying out the diazotizationand coupling reactionsin acetic acid solution,
and this so-called Griess-Ilosvaymethod, or a modification thereof, has been
z
used almost exclusively for determinationof NO in soils and other biological
materials.In the versionof this methoddescribedby Bremner(1965) and Keeney
and Nelson (1982) for analysisof soil extracts,sulfanilamideis usedinsteadof
sulfanilic acid as the diazotizing reagent,andN-(l-naphthyl)-ethylenediamine is
used insteadof 1-naphthylamineas the coupling reagent.These modifications
have been found to increasethe rateof color development,the stability of the
color, and the sensitivity of the method(Shinn, 1941).
The Griess-Ilosvaymethodof determiningNOz is more sensitivethan the
steam-distillationor microdiffusion methodsfor this determination,and even
greatersensitivity has beenachievedif the azo chromophoreis concentratedby
solventextraction(e.g., Baveja& Gupta, 1982; Chaubeet aI., 1984), or is mea-
suredby fluorometry (e.g., Nakamura,1980; Rubio et aI., 1984),or is measured
by fluorometry (e.g., Nakamura,1980; Rubio et aI., 1984). Thesetechniquesare
up to 100 times more sensitivethan the Griess-Ilosvaymethod; however,they
also are more complicated,and most have a narrow working rangeof NOz con-
centrations.
Puttannaand PrakasaRao (1986) have describeda two-step modification
of the Griess-Ilosvaymethodthat extendsthe working range.In the first step,the
sample is treated with sulfanilic acid and N-(l-naphthyl)-ethylenediamineto
form the azo chromophore.If the color intensity at 550 nm exceedsthat of the
higheststandard(0.12 Ilg of NOz-N mL-l), the solution is treatedwith acetone
(C3H60) andsodiumacetate(CzH3Na02),and the absorbanceis measuredat 480
nm. The latter procedurefollows Beer'slaw at NOz concentrationsup to 0.40 Ilg
of N mL-l, which is equivalentto 100 Ilg of NOz-N g-l of soil when extractions
are performedas describedin "Procedure"under "Extraction of Exchangeable
Ammonium and Nitrate and Nitrite," using a 1:10 ratio of soil to extractant.
Automation. Increasingly,automatedanalyzersystemsare being usedfor

z
colorimetricdeterminationsof NHt, NOj", and NO in soil extracts.Most of these
systemsutilize flow analysis,in that an aliquot of sampleis mixed with reagents
in a flowing carrierstreamto developa color, the intensity of which is measured
before dischargeto waste.This techniquewas originatedby Skeggs(1957), and
subsequentlyled to the developmentby the TechniconInstrumentsCorporation
of the AutoAnalyzer, which was first used for clinical analyses,but was subse-
quently adaptedfor other applications,including the analysisof inorganic forms
of N in water and soil extracts(e.g., Kamphakeet aI., 1967; Henzell et aI., 1968;
Henriksen& Selmer-Olsen,1970; Selmer-Olsen,1971; Brown, 1973; Jacksonet
aI., 1975). An improved version, the AutoAnalyzer II, remains available, and
similar systemsare now offered by manufacturersin severalcountries.All are
modular in design,the basiccomponentsbeing a sampler,peristalticpump, mix-
ing manifold, photometer,and recorderor other output device. Additional com-
ponentsare often employed,such as a heating bath for color developmentor a
dialyzer to eliminate interferencefrom turbidity, and many of the newer models
are equippedwith a microprocessoror microcomputer,which is utilized to oper-
ate the systemand acquiredata.
There are two commonforms of flow analysis.In one form, referredto as
continuous-flowor segmented-flowanalysis,the analytical streamis segmented
by the regular introductionof air bubblesto limit sampledispersion,promotethe
mixing of sampleand reagents,and scrubthe walls of the mixing manifold. In the
otherform, referredto as flow-injection analysis,the sampleis introducedinto an
unsegmentedcarrierstreamfrom an injection valve, forming a zone that dispers-
es becauseof laminar flow in the mixing manifold. The merits of continuous-
flow and flow-injection analysishave beenthe subjectof much discussion,and
further information can be found in a numberof publications,including thoseof
Snyder (1980), Horvai and Pungor (1987), Ruzicka and Hansen (1988), and
Smith and Scott (1991). It suffices here to note that either techniquemay be
employedfor analysisof NHt, NOj", andNO z in soil extractsand that, with some
systems,all three analysescan be carried out simultaneously,at a rate of more
than 100 samplesper hour.
Severalmethodshavebeenemployedfor automatedanalysisof NH! in soil
extracts.Most of thesemethodsare basedon the Berthelotreaction,using either
phenol(Henzell et aI., 1968; Selmer-Olsen,1971; Brown, 1973; Rice et aI., 1984;
Markus et aI., 1985; Tel & Heseltine, 1990) or salicylate (Rowland, 1983;
Adamsenet aI., 1985; Botha & Johnson,1988; Gentry & Willis, 1988), although
the Nessler reactionalso hasbeenemployed(Krug et aI., 1979). To preventpre-
cipitation of cadmiumand magnesiumhydroxidesat the high pH requiredfor the
Berthelot reaction, color developmentis often carried out in the presenceof a
chelatingagentsuch as a tartrateor citrate buffer (Henzell et aI., 1968; Selmer-
Olsen, 1971; Rowland, 1983; Markus et aI., 1985; Gentry and Willis, 1988) or
EDTA (Markus et aI., 1985). Interferencealso can arise from the presenceof
amino acids (White & Gosz, 1981; Rowland, 1983; Adamsenet aI., 1985; Bur-
ton et aI., 1989), someof which form a colored complex or liberate NHt under
the conditions employedto carry out the Berthelot reaction.This type of inter-
1152 MULVANEY
ferencehas beenfound to causeseriouserror in NHt analysisof extractsfrom

forest soils with a high contentof organicmatter(White & Gosz,1981),andseri-
ous error also hasbeenobservedwith soils that were air- or oven dried or fumi-
gated(Burton et aI., 1989). White and Gosz(1981) found that the error observed
in their work was practically eliminatedby reducingthe concentrationof NaOH
in the alkaline phenol reagentand by decreasingthe temperatureof the heating
bath, but subsequentstudiesby Rowland (1983) suggestthesemodificationsto
be ineffective. Steamdistillation hasbeenproposedas a meansof avoiding inter-
ferenceby amino acidsin automated analyses of soil extractsfor NHt (Burton et
aI., 1989). Automatedproceduresinvolving distillation have beendescribedfor
NHt analysiswith the TechniconAutoAnalyzer(Keay & Menage,1969; Skjem-
stad& Reeve,1978).
The Griess-Ilosvaymethodhas beenuniversally employedfor automated
analysesof NOi" in soil extracts(e.g., Henzell et aI., 1968; Markus et aI., 1985),
and the samemethod has been used to determineNO)" plus NOi", from which
NO)" is obtainedby difference,following reductionof NO)" to NOi" by hydrazine
and CUS04• 5HzO (Henzellet aI., 1968; Best, 1976; Markus et aI., 1985),nitrate
reductasefrom a bacterialculture (Lowe & Gillespie, 1975; Rice et aI., 1984),
copperizedcadmium (Henriksen& Selmer-Olsen,1970; Adamsenet aI., 1985;
Botha & Johnson,1988; Tel & Heseltine,1990), or cadmium-silverwire (Willis
& Gentry, 1987).The useof cadmiumis generallypreferred,as hydrazinereduc-
tion requires a high pH, which effects precipitation of Ca and Mg (Ananth &
Moraghan,1987), and also is subject to interferenceby soluble organic matter
(Rowlandet aI., 1984),whereascell-culturetechniquesmustbe employedto pro-
duce nitrate reductase,and the reduction is incompletewith 2 M KCl extracts
(Rice et aI., 1984).
Methods
Ammonium
Principles. In the proceduredescribed,NHt extractedfrom soil with 2 M
KCI is determinedby measuringthe intensity of the emeraldgreen color that
forms upon treatmentof an aliquot of the extractwith salicylateand hypochlorite
at high pH. A catalyst(sodium nitroprusside)increasesthe rate and intensity of
color development,and a chelatingagent(EDTA) preventsthe precipitationof
divalent and trivalent cationsas hydroxides.
Figure 38--3 shows a three-stepmechanism forcolor developmentby the
methoddescribed.In the first step,NH3 reactswith hypochloriteto form mono-
chloramine (NH 2CI). The monochloraminethen reactswith salicylate to form
benzoquinonemonoimine, which coupleswith salicylate to give the colored
indophenoldye.
For maximum sensitivity and accuracyin NHt analysesby the method
described,absorbancemeasurements are madeat 667 nm. When measurements
are performedat this wavelengthusing a light path of 1 cm, the methodobeys
Beer'slaw up to at least20 flg of NHt-N in 25 mL of solution (the volume rec-
ommendedfor color development).Under theseconditions,the limit of detection
is approximately0.2 flg of NHt-N. With a 5-cm light path, the detectionlimit is
about0.04 flg of NHt-N.
(A) NH,CI + OW
(I)
(8) 00. COO·

+ NH,CI
(II)
(C) 00. HNQO - ·ONQO

COO·
+
COO· ·OOC COO·
(II) (III)
Fig. 38-3. Reactionsin methodfor colorimetricdeterminationof NHt: (A) monochloramine(I) for·

mation; (B) oxidative imination of salicylate(II); (C) coupling reactionto form indophenoldye
(III).
Method 3
SPECIAL APPARATUS
1. Spectrophotometer,
equippedto provide a l-cm light path and capable
of absorbancemeasurements
at 667 nm.
REGENTS
1. Sodium salicylate-sodiumnitroprussidereagent.Dissolve 7.813 g of
sodium salicylate (NaC7Hs03) and 0.125 g of sodium nitroprusside
[disodium pentacyanonitrosylferrate, Na2Fe(CN)sNO• 5H20] in ap-
proximately 80 mL of deionizedwater in a 100-mL volumetric flask,
and bring the solution to volume with deionizedwater. Mix thorough-
ly, and then transferthe solution to an amberbottle for protectionfrom
light. Store in a refrigerator.
2. Buffered hypochlorite reagent.Dissolve 2.96 g of sodium hydroxide
(NaOH), and then 9.96 g of sodium monohydrogenphosphatehepta-
hydrate(Na2HP04• 7H20), in approximately60 mL of deionizedwater
in a lOO-mL volumetric flask. Add 10 mL of sodium hypochlorite
(NaOCl) solution(Clorox bleachor equivalent).Adjust the pH to 13.0
with NaOH, and dilute to 100 mL with deionizedwater.
3. Ethylenediaminetetraacetic acid reagent.Dissolve 6 g of the disodium
salt of ethylenediaminetetraacetic acid (Na2EDTA) in 100 mL of deion-
ized water in a volumetric flask.
4. Standardammoniumsolution. Dissolve0.4717g of ammoniumsulfate
[(NH4hS04] in deionizedwater (a primary-standardreagentis avail-
able from FisherScientific), and dilute the solution to a volume of 1 L
in a volumetric flask. If pure, dry (N~hS04 is used,the solution con-
tains 100 Ilg of NJIt-N mL- 1. Store the solution in a refrigerator. To
preparea working standardthat contains2 Ilg of NHt -N mL-1, dilute 4
mL of the concentratedsolution to 200 mL in a volumetric flask with
deionizedwater.
3 After Nelson(1983).
1154 MULVANEY
PROCEDURE.Pipettean aliquot (nonnallyS;3 mL, not to exceed5 mL) ofthe

soil extractinto a 25-mL volumetricflask. Add 1 mL of EDTA reagent,and swirl
the flask to mix the contents.Then add 4 mL of salicylate-nitroprusside reagent,
swirl the flask, and bring the volume to approximately20 mL with deionized
water. Add 2 mL of bufferedhypochloritereagent,immediatelybring the volume
to 25 mL with deionizedwater, and thoroughly mix the contentsof the flask.
Placethe flask in a waterbath at 37°C to developthe color. After 30 min, remove
the flask from the bath, allow the solution to cool to room temperature(approx.
10 min), and measureits absorbanceat 667 nm againsta reagentblank solution.
Calibrateabsorbancemeasurements by analysisof standardscontaining0,
2, 4, 6, 10, and 20 Ilg of NHt -N. To preparethesestandards,pipette into six 25-
mL volumetricflasks the samevolume of 2M KCI (the extractingsolution)asthe
aliquot of soil extract taken for analysis,and add 0, 1, 2, 3, 5, or 10 mL of the
working standard(NH4)2S04 solution (2 Ilg of NHS-N mL- 1). Then carry out
color development,and measurethe absorbanceby the proceduredescribedfor
analysisof the extract.
CALCULATIONS. Detenninethe NHS concentrationof the sampleusing the
equationobtainedby linear regressionof the concentrationsof the standardson
the correspondingabsorbancemeasurements. Alternatively, make this detenni-
nation by referenceto a calibrationcurve preparedfrom analysesof standards.
COMMENTS. The method describedis essentially a modification of the
indophenolblue methoddescribedby Keeneyand Nelson (1982), the major dif-
ference being that sodium salicylate is used to carry out color development
insteadof phenol. Sodium salicylateis neither toxic nor caustic,and the reagent
is suppliedin the fonn of a powderthat is easily weighedand transferred.
The addition of EDTA is necessaryto preventinterferencefrom precipita-
tion of polyvalent cations, particularly Ca2+ and Mg2+. According to Nelson
(1983), the EDTA treatmentemployedin the methoddescribed(1 mL of a solu-
tion containing60 mg of Na2EDTA) will complexat least 8 mg of Ca2+ or 5 mg
of Mg2+, which greatly exceedsthe nonnal content of theseions in 5 mL (the
maximum volume recommendedfor analysis)of a 2 M KCI soil extract(10 mL
of KCI g-l of soil). If a precipitatedevelopsupon addition of the bufferedhypo-
chlorite reagent,the analysisshould be perfonnedusing a smalleraliquot of soil
extract, or 2 mL of the EDTA reagentmay be addedto double the capacityfor
complexation.
A pH of 13 is required for maximal color developmentin the method
described(Nelson,1983).A lower pH will lead to a loss of sensitivity. For exam-
ple, Nelson(1983) found that absorbancewas reducedby up to 41% when color
developmentwas carried out at pH 12 insteadof pH 13. The methoddescribed
may be employedfor analysisof soil extractsobtainedusing acidic extractants,
but the extractshouldfirst be neutralized(by addition of NaOH) to ensureprop-
er color development.
The buffered hypochlorite reagent is unstable and should be prepared
immediatelybeforeuse.The salicylate-nitroprusside reagentis stablein the dark,
but should be preparedweekly to ensuremaximal sensitivity. For proper color
development,the salicylate-nitroprussidereagent must be added before the

bufferedhypochloritereagent.
As described,color developmentis carried out at 37°C for 30 min, which
maximizes molar absorptivity (Nelson, 1983). Alternatively, the color can be
developedat room temperature(23°C); however, maximal color development
requires2 h, and molar absorptivity is somewhatlower than when the color is
developedat 37°C (Nelson, 1983). The color is stablefor at least 4 h (Nelson,
1983).
Like othermethodsbasedon the Berthelotreaction,analysesby the method
describedare subjectto interferenceby organic forms of N, particularly amino
acids. Caution is advisedin the use of this methodfor analysisof extractsthat
may contain appreciableamountsof organic N, such as those from fumigated
soils or forest floor litter samples.In suchcases,analysesmay be performedby
the steam-distillationmethodsdescribedin "Methods" under"Steam-Distillation
Methods," or by the microdiffusion methods described in "Methods" under
"Microdiffusion Methods."
Nitrate
Principles. In the proceduredescribed,N03" extractedfrom soil with 2 M
z
KCI is reducedto NO by passage through a columnof copperizedcadmium,and
z
the NO formed is determinedby a modified Griess-Ilosvaymethod.Quantitative
z
reductionof N03" to NO is accomplishedby carryingout the reactionwith cop-
perizedcadmiumin an NH4Cl matrix, at a pH of 5 to 10. Nitrite is estimatedcol-
orimetrically after treatmentof the column leachatewith a diazotizing reagent
[sulfanilamide(CJISN20 2S)] and a couplingreagent[N-(1-naphthyl)-ethylenedi-
amine] in HCl solution to form an azo chromophore(see"Nitrite"). The intensi-
ty of the reddishpurplecolor that develops isproportionalto the concentrationof
z z
N03" in the soil extract,or to the concentrationof N03" plus NO if NO is pre-
sent.
For maximum sensitivity and accuracy, absorbancemeasurementsare
madeat a wavelengthof 540 nm, although,if necessary,any wavelengthmay be
employedbetween510 and 550 nm. When the measurements are madeusing a
light path of 1 em, the methodobeysBeer'slaw up to at least20 ~g of N03"-N in
100 mL of solution(the volume recommendedfor analysis).The excellentsensi-
tivity of the methodallows low levels of N03" to be measured,and therebyper-
mits dilution of soil extractsto minimize any interferencethat might ariseduring
color development.
Method'
SPECIAL APPARATUS
1. Nitrate reductiontube. A Pyrex tube (1.2-cm o.d., 30 cm long) is fitted

with a fritted glassplate and stopcockat one end to serveas an outlet,
and with a 5-cm length of 5.1-cm o.d. tubing at the other end to serve
as a reservoirfor 75 mL of solution. The outlet end of the stopcockis
4 After Keeneyand Nelson(1982), and Dorich and Nelson(1984).

1156 MULVANEY
connectedvia vinyl tubing to a Pyrex tube (4-mm o.d.) insertedthrough

a two-hole size 00 rubberstopper.A secondtube (4-mm o.d.) through
the stopperconnectsto a flow regulatorand a vacuumsource.
2. Spectrophotometer, equippedto provide a 1-cm light path and capable
of absorbancemeasurements at 540 nm.
REAGENTS
1. Copperizedcadmiumreagent.PlaceSO g of cadmiummetal (coarsely

powderedor granular,0.2S-to 1-mm diam., :::;2 mm long; a satisfacto-
ry material is availableas cat. no. C-3 from FisherScientific) in a SOO-
mL beaker,and treat it with 2S0 mL of 6 M hydrochloric acid (HCI).
After 1 min, decantthe HCI, and thoroughly wash the cadmiummetal
with deionizedwater. Then treat the metal with 2S0 mL of a cupric sul-
fate pentahydrate(CUS04 • SH20) solution (20 g of CUS04 • SH20
L- 1). Allow the mixture to stand for at least 1 min; then decantthe
CUS04 • SH20 solution, and wash the metal with deionizedwater. Re-
treat with 2S0 mL of CUS04 • SH20, decant the solution, and thor-
oughly wash the copperizedcadmium with deionized water until all
traces of blue and light gray color have disappearedfrom the wash
water. Transferthe metal to a NO)" reductioncolumn as describedin
the previous"SpecialApparatus"section.
2. Ammoniumchloride (NH4CI) solution,concentrated.Dissolve100 g of
reagent-gradeNH4CI in SOO mL of deionizedwater. Storein a glassor
plastic bottle.
3. Ammonium chloride solution, diluted. Dilute 50 mL of concentrated
NH4CI solution to 2 L in a volumetric flask with deionizedwater. Mix
thoroughly,and transferto a glassor plastic bottle for storage.
4. Diazotizing reagent.Dissolve 0.5 g sulfanilamidein 100 mL of 2.4 M
HCI in a volumetric flask. Storethe solution in a refrigerator.
5. Coupling reagent. Dissolve 0.3 g of N-(l-naphthyl)-ethylenediamine
dihydrochloridein 100 mL of 0.12 M HCI in a volumetricflask. Trans-
fer the solution to an amberbottle for storagein a refrigerator.
6. Standard nitrate solution. Dissolve 0.3609 g of potassium nitrate
(KN03) in deionized water (a primary-standardreagent is available
from FisherScientific), and dilute to 1 L in a volumetric flask. If pure,
dry KN03 is used,this solution contains50 j..lg of NO)"-N mL-l. Store
the solution in a refrigerator.To preparea working standardthat con-
tains 2 j..lg of NO)"-N mL-l, dilute 20 mL of the concentratedsolution to
500 mL in a volumetric flask with 2 M KCI.
PROCEDURE
PREPARATION OF CADMIUM COLUMN. With the outlet stopcockclosed,fill the

NO)" reduction tube with dilute NH4CI solution (Reagent3) to the baseof the
upper reservoir.Pour in sufficient copperizedcadmiumparticlesto give a depth
of 20 cm, tapping the tube as the cadmiumparticlesare addedto ensurethat all
air bubblesare removed.Drain the excessNH4CI solutionthroughthe outlet stop-
cock, and then wash the cadmium column thoroughly (10 pore-volumes)with
dilute NI4CI solution,usinga flow rateof approximately8 mL min-to Leavesuf-

ficient solution in the tube that the cadmiumcolumn is coveredto a depth of at
least 1 cm.
Immediatelybeforecarrying out NOj" analyses,add 1 mL of concentrated
NI4CI solution (Reagent2) to the cadmium column,and usethe outlet stopcock
to lower the level of liquid until it is evenwith the top of the column. Then add
75 mL of dilute NI4CI solution (Reagent3) to the upper reservoirof the NOj"
reductiontube, attach a 100-mL volumetric flask to the outlet stopcock-...ia the
rubberstopper,and by regulatingthe supplyof vacuumand/orthe position of the
stopcock,allow the NH4CI to flow throughthe column at a rateof 110 mL min-to
ANALYSIS OF EXTRACT. Drain excessNI4CI solution from the cadmiumcol-
umn until the liquid level is evenwith the top of the column.Pipette1 mL of con-
centratedNI4CI solution (Reagent2) onto the top of the column, and then an
aliquot of soil extract(normally 2-5 mL) containingnot more than 20 ~g of NOj"
-N. Attach a lOO-mL volumetric flask to collect the outlet flow from the column,
and open the stopcock to drain the treated extract into the cadmium column.
Immediatelywash the inside of the reduction tube with 2 mL of dilute NH4CI
solution, and add75 mL of dilute NH4CI solution to the upper reservoiron the
reductiontube. Use the stopcockand/or the supply of vacuumto adjust the flow
rate to 110 mL min-to When the level of liquid reachesthe top of the cadmium
column, close the outlet stopcock,and disconnectthe flask used to collect the
flow from the column.Add 2 mL of the diazotizingreagentto the contentsof the
flask, and mix thoroughly. Allow to stand5 min; then add 2 mL of the coupling
reagent,bring to volumewith deionizedwater,and mix thoroughly.After 20 min,
measurethe absorbanceof the solution at 540 nm againsta reagentblank solu-
tion.
2,4,6,10,and 20 ~g of N03"-N. To perform theseanalyses,add 0,1,2,3,5,and
lO mL of the working standardN03" solution (2 ~g of N03"-N mL- 1) to the cad-
mium column, and carry out reduction and color developmentas describedfor
the soil extract.
The method describeddeterminesany NOz initially present in the soil
extract,plus that formed by reductionof N03". If necessary,the amountof N03"
in the extractcan be determinedby treating the aliquot under analysiswith sul-
z
famie acid to decomposeNO as describedin "Methods" under"Steam-Distilla-
tion Methods"or "Methods"under"Mierodiffusion Methods,"or by carryingout
z
a separatedeterminationfor NO by the Griess-Ilosvayproceduredescribedin
the "Nitrite" sectionunder"Colorimetric Methods."
CALCUlATIONS. Determinethe N03" concentrationof the sampleusing the
the correspondingabsorbancemeasurements.Alternatively, make this determi-
nation by referenceto a calibrationcurve prepared fromanalysesof standards.
COMMENTS. The reductionof N03" to NOzis the most likely sourceof dif-
ficulty in the methoddescribed,and care is requiredin the preparationand oper-
ation of the copperizedcadmiumcolumn employedfor this reduction.The cad-
1158 MULVANEY
mium metal usedshouldbe of the size specified,as the efficiency of reduction

dependsupon particle size, and a different flow rate may be requiredif the cad-
mium particlesare largeror smallerthan specified.To ensurequantitativereduc-
tion of NO)", copperizationof the cadmiummetal mustbe complete.The copper-
ized metal must be thoroughlywashed,as excessCu causesNO)" to be reduced
to an oxidation state lower than NOz (Nydahl, 1976). The cadmium column
shouldbe packedto a depthof 20 cm; Dorich and Nelson(1984)obtainedincom-
plete reductionof NO)" with a 10- or IS-em column,and incompleterecoveryof
NOzwith a 30-cm column. Air bubblesin the column seriously interfere with
NO)" reduction,so careshouldbe takento ensurethat all air bubblesareremoved
during packing of the column, and that a layer of liquid is always maintained
abovethe cadmiummetal to excludeair. If thereis any failure to maintainsucha
layer, the columnshouldbe repacked.The flow ratemustbe regulatedat 110 mL
rnin-1 during operationof the column.Dorich andNelson(1984)obtainedincom-
z
plete recoveryof NO)" and NO when elution was carriedout at a rate of 7 to 10
mL min-I, asrecommendedin severalothermanualmethodsfor NO)" analysisby
cadmium reduction (e.g., Huffman & Barbarick, 1981; Am. Public Health
z.
Assoc., 1992), which was attributedto reductionof NO Excessiveflow rates
will lead to incompletereductionof NO)".
A cadmiumcolumn has a finite capacityfor reductionof NO)", and with
use,a declinewill eventuallyoccur in the efficiency of reduction(normally after
severalhundredsamples),as indicatedby a decreasein the absorbancevalues
measuredduring calibration.In suchcases,columnefficiency may be checkedby
z,
comparing analysesfor NO)" and NO using samplesthat contain the same
amountof N (e.g., 10 llg). A difference in absorbancevalues indicatesincom-
plete reductionof NO)", and althoughanalyseswill still be possibleif the amount
of NOzformed is proportionalto the amountof NO)" in the sample,the cadmium
metal should be recopperized(as describedin the previous"Reagents"section)
to restoresensitivity.
Soil extractsobtainedwith 2 M KCI are sometimescolored,but no diffi-
culty has been encounteredin analysis of colored extracts by the method
described.Becauseof the excellentsensitivityof the method,soil extractsmay be
diluted with deionizedwater beforeanalysis,and by this technique,evenhighly
coloredextractsmay be analyzedwithout difficulty.
Although pH is not critical to the reduction of NO)" by the method
described,extractsobtainedusing acidic or alkaline extractantsshould be neu-
tralized (by addition of NaOH or HCI) before analysis,to ensureproper color
developmentby the Griess-Ilosvaymethod.The solutionpH after additionof the
diazotizingreagentshouldbe about1.6, andthe pH after additionof the coupling
reagentshouldbe about1.5 (Bremner,1965).
The methoddescribedis free from interferenceby any commonconstituent
of soil extracts,but the efficiency of N03" reductionis reducedby high concen-
trationsof sulfide (Wood et aI., 1967)or phosphate(Olson, 1980).Sulfide reacts
with Cd to form CdS,whereasphosphateis adsorbedby the cadmiumcolumn.
The standardNO)" solution is stablefor at least6 mo if storedin a refriger-
ator. Owing to the excellentsensitivity of the method for NO)" and NO the z,
(A) RONH Z + NO; + 2H+ - - - - - - R O N. N + 2H zO

(I)
(8) RON.N+ 8 R'
(II)
Fig. 38-4. Diazotizationandcouplingreactionsin methodfor colorimetricdeterminationof N0Z-: (A)

diazotizationreaction-(I)sulfanilamide(R =-S02 • NH2); (B) coupling reaction-(II)N-(l-naph-
thyI)-ethylenediamine(R' =-NH • CH2CH2 • NH2).
deionizedwater usedto preparethis solution shouldbe as free as possiblefrom

theseforms of N.
Controlsmust be included in eachseriesof analysesto allow for the color
producedby the reagentsemployed(including the reagentsusedin extractionof
the soil sample).
Nitrite
Principles. In the proceduredescribed,NO z in a 2 M KCI soil extract is
determinedby a modification of the Griess-Ilosvaymethod. An aliquot of the
extract is treatedwith a diazotizing reagent(sulfanilamidein HCI solution) to
z
convert the NO to a diazonium salt, and then with a coupling reagent[N-(1-
naphthyl)-ethylenediamine]to convert the diazoniumsalt to an azo compound.
z
The concentrationof NO is proportional to the intensity of the reddish purple
color that developsas a resultof thesetreatments.The diazotizationand coupling
reactionsin this methodof analysisare illustratedby Fig. 38-4.
The sensitivity and accuracyof the methodusedfor colorimetric determi-
z
nation of NO are greatest whenabsorbancemeasurements are madeat 540 nm
(the wavelengthof maximal molar absorptivity), and the method obeys Beer's
law up to at least61lg of NOz-N in 50 mL of solution (the volume recommend-
ed for analysis)when measurementsare performedat this wavelengthusing a
light pathof 1 cm. A spectrophotometer is employedin the methoddescribed,but
a colorimetercan be usedif equippedwith a greenfilter having maximal trans-
mittance in the range of 520 to 550 nm. A 5-cm light path provides a fivefold
increasein sensitivity, with a correspondingdecreasein dynamicrange.
1160 MULVANEY
Method5
SPECIAL APPARATUS
1. Spectrophotometer,
equippedto provide a 1-cm light path and capable
of absorbancemeasurements
at 540 nm.
REAGENTS
1. Diazotizing reagent.Dissolve 0.5 g of sulfanilamidein 100 mL of 2.4

M HCI. Storethe solution in a refrigerator.
2. Coupling reagent. Dissolve 0.3 g of N-(l-naphthyl)-ethylenediamine
dihydrochloride in 100 mL of 0.12 M HCI. Store the solution in an
amberbottle in a refrigerator.
3. Standardnitrite solution. Dissolve 0.2463g of sodiumnitrite (NaN02)
in deionizedwater, and dilute the solution to 1 L in a volumetric flask.
If pure, dry NaN02 is used, this solution contains 50 J..lg of NOz-N
mL-1. Storethe solutionin a refrigerator.To preparea working standard
that contains1 J..lg of NOz-N mL-l, dilute 20 mL of the concentrated
solution to 1 L in a volumetric flask with deionizedwater.
PROCEDURE.Pipette an aliquot (usually 2 mL) of the extract into a 50-mL
volumetricflask, and adddeionizedwaterto makethe total volume about45 mL.
Add 1 mL of the diazotizing reagent(Reagent1), and mix the solution. After 5
min, add 1 mL of the coupling reagent(Reagent2). Mix the solution, and allow
it to standfor 20 min. Then make the solution to volume, mix it thoroughly,and
measureits color intensity at 540 nm againsta reagentblank solution.
1,2,3,4,5,and 6 Ilg of NOz-N. Pipette0-, 1-,2-,3-,4-,5-,and 6-mL aliquots
z
of the working standardNO solution into 50-mL volumetricflasks, and measure
the absorbances obtainedby the proceduredescribedfor analysisof the extract.
CALCUlATIONS. Determinethe NOz concentrationof the sampleusing the
the correspondingabsorbancemeasurements. Alternatively, make this determi-
nation by referenceto a calibrationcurve preparedfrom analysesof standards.
COMMENTS. Although otherextractantshavebeenemployedfor analysisof
z
NO by the Griess-Ilosvaymethod(Bremner,1965), no difficulty is encountered
in the analysisof extractsobtainedusing2 M KCI, and this reagenthasthe advan-
tageof extractingexchangeableNHt in addition to NO}" and NO which allows z,
all threedeterminationsto be madeby a single extraction.Extractsobtainedwith
2 M KCl are sometimescolored,but this doesnot normally interferewith analy-
ses by the method describedfor determinationof NOz, becausethe extract is
diluted extensivelywith water before analysis.If necessary,a correctioncan be
made for color interferenceusing the techniqueof Montgomery and Dymock
(1961). With this technique, an additional control analysis is performed that
involves treatmentof the samplewith the diazotizing reagentas in the normal
5 After Barnesand Folkard (1951), Bremner(1965), and Keeneyand Nelson (1982).

procedure,but the treatment with N-(l-naphthyl)-ethylenediamineis omitted.

The optical density of this sampleis then measured against distilled (or deion-
ized) water, and the value obtainedis subtractedfrom the absorbancedetermined
for the sampleby the normal procedure.
z
Little information is available concerningthe stability of NO in extracts
obtainedby the proceduredescribedin "Extraction of ExchangeableAmmonium
and Nitrate and Nitrite" or by other methodsthat have beenproposedfor extrac-
z.
tion of soils for colorimetric determinationof NO Therefore,extractsshouldbe
analyzedimmediatelyafter their preparationwheneverpossible;otherwise,they
should be storedin a refrigeratoror freezer.
The accuracyof the resultsobtainedby modified Griess-Ilosvaymethods
z
of determiningNO dependson proper adjustmentof pH for the diazotization
and coupling reactions(Rider & Mellon, 1946; Barnes& Folkard, 1951). There-
fore, the sampletaken for analysisshould not be strongly buffered, and samples
that are acidic or alkaline should be neutralized(by addition of NaOH or HCI)
beforeanalysis.In the methoddescribed,the pH after addition of the diazotizing
reagentshould be about 1.6, and the pH after addition of the coupling reagent
should be 1.5 (Barnes& Folkard, 1951).
The rate of color developmentin the methoddescribedis very rapid, full
color developmentbeing achievedin 10 min at 25°C. The color formedis stable
for severalhours. Barnesand Folkard (1951) observedno fading of the colors
obtainedby this methodor by other modificationsof the Griess-Ilosvayproce-
dure using lightproof vessels,and concludedthat the fading sometimes observed
using thesemethodsis probably due to exposureof the azo compoundto light.
Therefore, samples prepared for colorimetric measurementby the method
describedshouldbe storedin the dark if they cannotbe analyzedsoonafter devel-
opmentof the color.
z
The Griess-Ilosvaymethodof determiningNO is highly sensitiveand spe-
cific, and it is not subjectto interferenceby high concentrationsof variouscations
and anions(Rider & Mellon, 1946).The substances that havebeenfound to inter-
fere with the method(Boltz & Taras,1978) are unlikely to occur in soil extracts
and will not be discussedhere. However,sincecoppersaltshave beenemployed
z,
to preparesoil extractsfor colorimetricdeterminationof NO.3 and NO and since
copperand mercurysaltsare sometimesusedto preventmicrobial activity in soil
extracts,attentionmay be drawn to the fact that both Cu2+ and Hg2+ have been
z
found to interfere with the determinationof NO by Griess-Ilosvaymethods.
Mercury causeshigh results,and Cu2+ catalyzesthe decompositionof the diazo-
nium salt and causeslow results.Interferenceby high concentrationsof NHt also
hasbeenreported.
The coupling reagentemployedin the methoddescribedhas the disadvan-
tagethat it becomesdiscoloredand tendsto precipitateevenwhen it is kept in the
dark. This increasesthe reagentblank and decreasesthe sensitivity and precision
of the method.However,the reagentis usablefor at least2 mo if it is storedin a
refrigerator.
z
Experiencehas shown that the standardNO solution is stablefor at least
6 mo if storedin a refrigerator,and that there is no needto add CHCl3 or to take
other precautionsgenerallyrecommendedfor preparationand storageof standard
1162 MULVANEY
N02" solutions.The N02" standardcanbe calibratedby titration with KMn04 (Jef-

fery et al., 1989),or by the MgO-Devarda'salloy steam-distillationor microdif-
fusion methodsdescribedin "Methods"under"Steam-DistillationMethods"and
"Methods" under"Microdiffusion Methods."
Controlsmust be includedin eachseriesof analysesto allow for the color
producedby the reagentsemployed(including the reagentsusedfor extractionof
the soil sample).
Ion-SelectiveElectrodes
Introduction
Upon first consideration,the useof electrodesto determineNH4", NO)", and
N02" in soil extractswould appearto offer many advantagesover other methods
for thesedeterminations.The proceduresinvolved are extremelysimpleandcon-
venient,requiring only an electrodeand a suitablepH meter.However,the elec-
trodesare expensive,and the membraneor sensormodule has a limited opera-
tionallifetime (a few daysto severalmonths).Moreover,the electrodesfor mea-
suring NO)" lack sensitivity, requirecontinualrestandardization,and are subject
to numerousinterferences.Information regardingthe theory and designof ion-
selectiveelectrodescan be found in publicationsby Carlsonand Keeney(1971),
Covington(1974), Yu (1985), andTalibudeen(1991).
Ammonium. An NH4 electrodeutilizing a cation-sensitiveliquid mem-
braneis commerciallyavailable(Unicam Ltd., Cambridge,UK), but it is subject
to seriousinterferenceby K+, which precludesanalyseswhen KCI or K 2S04 are
usedto extract exchangeable NH4" from soil. A much more practical alternative
is the NH3 gas-sensingelectrodewith an internal referenceelectrode,which is
availablefrom severalcommercialsources(Table 38-5). With the latter type of
electrode,the solution underanalysisis madealkaline (PH > 11) to convertNH4
to NH3, and the activity of NH3 is determinedby measuringthe pH of an inter-
Table 38-5. Commonlyavailableelectrodesfor measuringNH4, NOj", and NOi.

Concentration Model
Type of electrode range Interferences Manufacturert no.
mgNL-l
Ammonium
Gas-sensing(NH3) 0.Q1-14oo Volatile amines,Hg2+ Coming 476130
Orion 95-12
Unicam 80003
Nitrate
Liquid membrane:/: 0.14-14()()(} C10", 1-, CIOj", CN-, NOi, HS-, Coming 476134
Br, HCOj", CI-, COr, pot Orion 93-07
Unicam 80113
Nitrite
Gas-sensing(NO..) 0.07-140 CO2, HCOj", col-, volatile acids Orion 95-46
t Addresses:Coming,Inc., Coming,NY 14831;Orion Research,Inc., The SchrafftCenter,529 Main
Street,Boston,MA 02129; Unicam Ltd., York Street,Cambridge,UK CBl 2PX.
:/: Requiresexternalreferenceelectrode.
nal filling solution into which NH3 diffusesthrough a semipermeablemembrane.

The pH measurementmay be madeusing any meterwith a resolutionof 0.1 mY.
Several studies have indicated that the NH3 electrodeis satisfactoryfor
determinationof NH.t in soil extracts(Banwartet aI., 1972; Hofman et aI., 1980;
du Preezet aI., 1989). The most comprehensiveevaluationhasbeenthat of Ban-
wart et al. (1972). No difficulty was encounteredin analysisof extractsprepared
using 2 M KCI, and the resultsfor samplescontaining0.05 to 3 mg of NH.t-N per
liter agreed closely with those obtained by the steam-distillation method
describedin "Ammonium-Nitrogen"under "Proceduresin Absenceof Nitrite."
Interferencetestsshowedthat analysesby the NH3 electrodewere unaffectedby
any commonconstituentof soil extracts,including inorganiccationsand anions,
amino acids, amides,hexosamines,purines, and pyrimidines. However, serious
interferenceoccurredwith volatile amines(methylamineand ethylamine),which
the electrodedetectsas NH3, and with Hg2+, which forms a complex with NH3
under alkaline conditions.According to the manufacturer(Orion Research,Inc.,
1989), the latter interferencemay be eliminated by treating the sample with
iodide to complex Hg2+.
A commercially available automatedinorganic N analyzer (Model 380;
Alltech Associates,Inc., Deerfield,IL) utilizes many of the sameprinciplesasthe
NH3 electrode.In this instrument,the sampleis treatedwith alkali to convertNH.t
to NH3, which diffuses through a semipermeablemembraneinto a stream of
water and is determinedfrom the resulting increasein electrical conductivity
(Carlson, 1978). The diffusion-conductivity method has been successfully
employedfor determinationof NH.t in Kjeldahl digests(Carlson,1978) and nat-
ural waters(Scott et aI., 1989),and for determinationof NH.t and N03" in soil and
plant tissueextracts(Carlsonet aI., 1990).
Nitrate_ Electrodeswith selectivity for N03" are available from manufac-
turersin the USA and UK (Table 38-5). Theseelectrodesutilize a NO) -selective
ion exchanger,either a long-chainalkyl NH.t salt (e.g., Coming, Inc.) or a metal
salt of orthophenanthroline (e.g.,Orion Research,Inc.). The ion exchangeris dis-
solved in an organic solvent and impregnatedin a thermo-settingplastic mem-
brane, which separatesthe solution under analysis from an internal reference
solution. Whenthe electrodeis immersedin a solutioncontainingN03" , the N03"
binds to the ion exchangerand is transportedacrossthe membrane,such that an
electricalpotentialdevelopsbetweenthe two sidesof the membrane.The magni-
tude of this potential dependson the differencein N03" contentbetweensample
and referencesolutions,and is measuredrelative to a constantpotentialgenerat-
ed with an externalreferenceelectrode.
The N03" electrodemeasuresthe level of free N03" in solution, or N03" ion
activity (aN03"), which is relatedto molal concentration(mN03") by
aN03" = (yN03")(mN03"), [1]
whereyN03" is the activity coefficient.The measuredpotential,E, is relatedto the

activity by a modification of the Nernstequation,
E = E' - Slog (aN03" + 'LKjaj), [2]

1164 MULVANEY
whereE' is a constant(determinedby the particularNO)" and referenceelectrodes

employed);S is the electrodeslope (at 25°C, 59.16 mV per decadechangein
ionic activity); K j is the selectivity coefficientfor interfering anion, i, with activ-
ity aj; and L denotessummationof Kjaj for all interfering ions. According to
Orion Research,Inc. (1990a), a Nemstianresponseis obtainedwith NO)" con-
centrationsas low as 1.4 mg of N L-l. Measurementsmay be madeat somewhat
lower concentrations(the detectionlimit is specifiedas 0.14 mg of NO)"-N L-l)
(fable 38--6), but calibrationwill be nonlinear.
In the absenceof interference,aNO)" closely approximatesmNO)" at con-
centrationsup to about 150 mg L -1 (~34 mg of NO)" -N L -1) (Table 38--6). At
higher concentrations,the ionic strengthincreasessufficiently that aNO)" under-
estimatesmNO)", and with concentratedsalt solutions(e.g., 2 M KCI), the effect
is so severethat the electrodebecomesunresponsiveto NO)" (Myers & Paul,
1968). Therefore,such solutionscannotbe employedas extractantsfor analyses
with the NO)" electrode,althougha dilute salt solution may sometimesbe desir-
able, especiallywith soils having significant anion-exchangecapacity (Black &
Waring, 1978), or to ensurethat all analysesare carried out at the same ionic
strength.A variety of solutions have been employed for the latter purpose,
including 0.025 M Al z(S04h (Baker & Smith, 1969); 0.01 M CUS04 (0ien &
Selmer-Olsen,1969; Bartuzi et aI., 1976; Hadjidemetriou,1982); a mixture of
Al Z(S04)3(0.01 M), AgzS04 (0.01 M), H3B03 (0.02 M), and sulfamic acid (0.02
M) (Milham et aI., 1970); 0.1 M sodiumcitrate(C6HsNa307)(Raveh,1973); 0.01
M or saturatedCaS04(Bound, 1977; Pedrazziniet aI., 1979; Li & Smith, 1984);
0.04 M KAl(S04)Z (Hofman et aI., 1980); and 0.12 or 0.04 M (NH 4)zS04 (Li &
Smith, 1984; Orion Research,Inc., 1990a).
Severalanionshavesufficiently high selectivity coefficients(Kj in Eq. [2])
that they interfere at very low concentrations.The most serious interference
occurswith CI04", 1-, CIO)", and CN-, but theseions are seldompresentin soil
extractsor otherbiological samples.The major interferingions in soil extractsare
N02", HCO)", Cl-, and C01-, although PO]- (and also HPOi- and HzPO,n can
interfere if the concentrationis exceptionally high (Table 38-7). Organicanions
also can interfere, and such interferencehas been observedin analysisof plant
extracts(Paul & Carlson,1968) and naturalwaters(Csiky et aI., 1985).
A correctioncannotnormally bemadefor anion interference,sinceevenif
anionconcentrationsare known, ionic strengthmay vary amongsamples,and this
Table 38--6. Concentrationand activity of NO) in KN0 3 solutionsat 25°C (Langmuir & Jacobson,
1970).
Concentration Activity Concentration Activity
mgL-l
1 1.00 200 188
5 4.95 300 278
10 9.86 400 367
25 24.4 500 454
50 48.4 750 668
100 95.5 1000 876
150 142
will affect ion activities. Therefore,interfering anions must be removed before

analysis.ExtractantscontainingAg2S04 may be used to reduceCl- interference
(Milham et aI., 1970; Hulanicki et aI., 1974; Orion Research,Inc., 1990a).Paul
and Carlson (1968) suggestedthe use of a silver-treated ion-exchangeresin
(Dowex 50-X8), but this techniquewas found to be unsatisfactoryin subsequent
work by Bakerand Smith (1969). Dilute H2S04 may be usedto removeCOj- and
HCOj" (Orion Research,Inc., 1990a). Nitrite interferencecan be eliminated by
decomposingNOi with sulfamicacid (Mahendrappa,1969; Milham et aI., 1970;
Orion Research, Inc., 1990a), by complexing NOi with sulfanilamide
(C6HsN302S)(Francis& Malone, 1975), or by oxidizing NOi to NOj with acid-
ified KMn04 (Morie et aI., 1972). The latter approachis more complicated,but
permitsdeterminationof both NOj and NOi. Interferenceby organicanionsmay
be reducedthrough acidification to suppressionization (Baker & Smith, 1969;
Orion Research,Inc., 1990a).Csiky et ai. (1985) have describeda simple proce-
dure using disposableadsorptioncolumnsto removehumic substancesfrom nat-
ural watersbefore analysiswith the NOj electrode.
The nitrate electrodeis relatively unaffectedby pH. The operationalrange
is from pH 2 to 12 (Davieset aI., 1972; Hulanicki et aI., 1974); however,a much
narrower range is required for analysesat low NOj concentrations«2 mg of
NOj-N L- 1) (Potterton& Shults, 1970; Davies et aI., 1972), and, in somecases,
measurementshave been carried out with pH buffered at 2.2 or 3.0 (Baker &
Smith, 1969; Milham et aI., 1970; Hulanicki et aI., 1974). This practicedoesnot
appear necessaryfor analysis of soils or soil extracts (Bremner et aI., 1968;
Raveh, 1973), but may be adviseablewith plant tissue extractsto reduceinter-
ferenceby organicacids (Baker & Smith, 1969).
The referenceelectrodemust be chosenwith care, as it can be a major
source of error and variability (Potterton & Shults, 1967; Carlson & Keeney,
1971; Dahnke, 1971). The referenceelectrodeshould give a stable and repro-
ducible output that is unaffectedby stirring, and there should be no contamina-
tion of the solution under analysisby interfering anionsfrom the internal filling
solution. Theserequirementscannotbe met with most of the referenceelectrodes
usedto measurepH.
A variety of referenceelectrodeshave been employedwith nitrate elec-
trodes, including silver/silver chloride electrodeswith a single (Bremneret aI.,
Table 38-7. Concentrationsof anionscommonly presentin soil extractsrequiredto causea +10%

error in measurement
of NO:) with the Orion Model 93-07electrode(Orion Research,Inc., 1990a).
NO:) concentration(mg L-I)t
Anion 10 100
mgL-I
NO z 2 23 230
RCO:) 44 440 4400
Cl- 76 760 7600
COl- 86 860 8600
pot 339 3390 33700
S01- 6857 68570 685700
t One milligram of NO:) L-I =0.22 mg of NO:)-N L-I.
1166 MULVANEY
1968;Keeneyet al., 1970;Sommerfeldtet aI., 1971;Simeonovet al., 1977; Black

& Waring, 1978; du Preezet aI., 1989)or double (Goodman,1976; Pedrazziniet
aI., 1979; Hadjidemetriou,1982;Li & Smith, 1984)sleevejunction, and calomel
electrodeswith a fiber (Baker & Smith, 1969; Mahendrappa,1969; Onken &
Sunderman,1970; Francis& Malone, 1975)or ceramic(Morie et aI., 1972)junc-
tion. Double-junctionelectrodesarenow generallypreferred,as the useof single-
junction electrodescan lead to significantcontaminationby Cl- from the internal
filling solution (Potterton& Shults, 1967; Keeney et aI., 1970; Dahnke, 1971),
whereascalomel electrodesare affected by the rate of stirring and may cause
readingsto drift (Sommerfeldtet aI., 1971).To avoid contaminationby Cl- from
the double-junctionelectrode,the outerchamberis fIlled with a dilutesolutionof
(NH4)zS04 instead of KCl (Hadjidemetriou, 1982; Li & Smith, 1984; Orion
Research,Inc., 1990a).Bound and Fleet (1977) havedescribeda solid-stateref-
erenceelectrodethat usesno filling solution, but the potential is affectedby pH
and ionic strength.
Numerousprocedureshavebeendescribedto determineNO)" in soils with
the NO)" electrode.Most of theseproceduresinvolve an extraction withwater or
a dilute salt solution (Bremneret al., 1968; Mahendrappa,1969; 0ien & Selmer-
Olsen, 1969; Milham et aI., 1970; Onken & Sunderman,1970; Raveh, 1973;
Black & Waring, 1978; Knittell & Fischbeck,1979; Pedrazziniet aI., 1979; Hof-
man et aI., 1980; Li & Smith, 1984), but analyseshave often been performed
without extraction, using a well-mixed soil suspension(Bremner et aI., 1968;
Simeonovet aI., 1977;Hadjidemetriou,1982;du Preezet aI., 1989)or a soil paste
(Bound, 1977). Bremneret a1. (1968) obtainedexcellent agreementwhen NO)"
analyseswere performed on stirred soil suspensionsand on the filtrates from
thesesuspensions,whereasMack and Sanderson(1971) found that the presence
of dispersedcolloidal material in water extractsof soils led to erroneouslyhigh
measurements with a nitrate electrode.The latter finding can likely be attributed
to the fact that a saturatedcalomelelectrodewas employedas the referenceelec-
trode. Bound (1977) reportedsimilar difficulties when a calomel electrodewas
used with soil pastes,but the problem was eliminated by using a silver/silver
chloride referenceelectrodewith a nonporousliquid junction (Bound & Fleet,
1977).
The responsetime for the nitrate electroderangesfrom a few secondsat
high concentrationsof NO)" to severalminutesat low concentrations(Potterton&
Shults, 1967; Davieset aI., 1972). A period of 1 to 2 min is generallyemployed
(Dahnke,1971),althougha longerperiod may be requiredwhen analysesareper-
formed on samplesof widely differing NO)" content(Potterton& Shults,1967).
The nitrate electrodeis subject to drift (Knittel & Fischbeck,1979) and
shouldbe standardizedat regularintervals(normally after every 10-20samples).
Standardizationcan be accomplishedby constructinga plot of the measured
potential(E) againstthe logarithmof NO)" concentrationfor a seriesof standards,
and if the meteremployedis equippedwith a logarithmic scalefor direct readout
of concentration,calibrationmay be carriedout usingonly two standardsolutions
that differ by tenfold in concentrationand encompassthe rangeof concentrations
to be encounteredwith samples(Potterton& Shults, 1967;Dahnke,1971). Static
electricity cancauseseriousinstability in readingswhen relative humidity is low,
in which casethe electrodesshouldbe coveredwith a shieldof aluminumfoil and

groundedto the meter(Onken & Sunderman,1970; Sommerfeldtet aI., 1971).
Severaltechniqueshave beenproposedto increasethe speedand conve·
nienceof analyseswith the nitrate electrode.Milham (1970) hasdescribeda sim·
pIe flow-through cell for manualanalyses.Flow-injection techniquesutilizing the
nitrate electrode have been employed for automatedanalysis of soil extracts
(Hansenet aI., 1977; Ruzicka et aI., 1977; Schalschaet aI., 1981). Goodman
(1976) has describedan apparatusthat automaticallyextractsup to 60 soil sam-
ples, and then carries out an analysisof each extract with the nitrate electrode.
Besidesbeing rapid and convenient,thesetechniquestend to give better accura·
cy and precisionthan can otherwisebe attainedwith the nitrate electrode,since
operationalvariablesare standardized.
A compactmeterfor measuringNO)" hasbeendevelopedby Horiba Instru·
mentsCo., Kyoto, Japan.This unit, known as the Cardy nitrate meter, utilizes a
miniature version of the nitrate electrodein a replaceablesensormodule. The
entire assembly,consistingof the sensormodule and a digital meterwith readout
in partsper million (mg of NO)" -N L- 1), is so compactthat it easily fits in a shirt
pocket. Analysesare readily performedon soil extracts,and the extractionsare
simplified considerablyby using a speciallydesigned plastic filter bag developed
by SpectrumTechnologies,Inc., Plainfield, IL. The Cardy meter has been rec-
ommendedfor on-farm monitoring of soil and crop N status(Hartz et aI., 1993),
but no evaluationof analytical performancehas yet beenpublished.
A simple type of ion-selectiveelectrodecan be madeby immersingcopper
wire in an electroactive reagentcontaininga liquid ion exchanger.The result is
commonly referred to as a coated-wireelectrode(Cattrall & Hamilton, 1984).
Lee et ai. (1986) have describeda coated-wireelectrodefor NO)" that utilizes a
graphite-epoxy paste, and their design has subsequentlybeen evaluated by
Goodroadand Shuman(1990) for analysis of soil extracts. The results of this
evaluationindicate that performanceis adequatefor applicationsnot requiring a
high degreeof accuracy,and that the analysesare subject to the same interfer-
encesas a conventionalnitrate electrode.The low cost of coated-wireelectrodes
makesthem disposable,and this can be a major advantage,particularly when the
membranehasbeencontaminatedby exposureto a high concentrationof an inter-
fering ion (Goodroad& Shuman,1990).
Siegel (1980) has describeda methodfor measuringNO)" in soil extracts
that utilizes the ammoniaelectrode.In this method,NO)" is determinedas the dif-
ferencein the concentrationof NH3-N obtainedfor duplicate aliquots of extract
made alkaline with NaOH, one of which also is treatedwith Devarda'salloy to
convert NO)" to NH 3. Sensitivity and precision are about the same as can be
obtainedwith the nitrate electrode,but most interferencesare eliminated, and
analysescan be performedon KCl extracts.The major limitation appearsto be
the needfor careful control of temperature,which affectsthe rate of reductionof
NO)", the rate of NH3 loss from solution, and the measuredNH3 activity.
Nitrite. Nitrite in aqueoussolution can be determinedwith a gas-sensing
electrodefor nitrogen oxides(NO x ) availablefrom Orion Research,Inc., Boston,
MA (Table38-5). With this electrode,the solution underanalysisis acidified (pH
z
1.1-1.7)to convertNO to NOx species(NO, N02, N20 3, N20 4), which diffuse
1168 MULVANEY
througha gas-permeable membraneinto an internalfilling solutionand are deter-

mined by measuringthe pH of this solution againstan internal referenceelec-
trode. According to the manufacturer(Orion Research,Inc., 1990b), the NOx
electrodeexhibitsa linear responsein the range,5 x 10-6 M to 10-2M, which cor-
respondsto a N02" concentrationof 0.07 to 140 mg of N L-1.
The only studyin which the NOx electrodehasbeenusedto determineN02"
in soil extractshasbeenthat of Tabatabai(1974),who found analysesby the elec-
trode to be in good agreementwith those by the Griess-Ilosvaymethod (see
"Nitrite" under"Colorimetric Methods")when extractionswere performedusing
water, 1 M KCI, 0.1 M LiCI, 0.01 M CaCI2, 0.008M Ca(H2P04)2 • H20, or satu-
ratedCaS04'Seriousoverestimationoccurredwith 2 M KCI, but sucherror can
be eliminated by increasingthe concentrationof the electrodefilling solution
(Orion Research,Inc., 1990b).No interferencewas observedby cationsor anions
commonly presentin soil extracts,or by other substancesthat may be present
under some conditions, such as Cu2+ or Hg2+, both of which interfere in the
Griess-Ilosvaycolorimetric method of analysis for N02" (see "Comments" in
"Nitrite" section under "Colorimetric Methods"). Interferencedid occur when
acidified samplesof extracts were treated with calcareoussoil, which can be
attributedto the formation of CO2 bubbles.Accordingto the manufacturer(Orion
Research,Inc., 1990b), any compoundthat reactswith water to form an acidic
solution will interfere, including CO2, S02' and volatile weak acids [acetic acid
(~H402)' formic acid (H2C02), HF, lactic acid (C3H 60 3), pyruvic acid
(C3H40 3)]·
Methods
Ammonium
Principles. The soil extract is made alkaline by addition of NaOH (pH
11-12) to convertNHt to NH3, and the electromotiveforce (emf, in mY) is mea-
suredwith an NH3 electrode.The concentrationof NHt -N is estimatedby com-
parison to the values obtained in analysis of NHt -N standardsby the same
method.
MethotP
SPECIAL APPARATUS
1. Ammonia electrode.
2. pH-millivolt meterwith resolutionof 0.1 mY.
REAGENTS
1. Sodium hydroxide, 0.25 M. Dissolve 10 g of NaOH in 800 mL of

deionizedwater, and dilute to 1 L in a volumetric flask.
2. Standardammoniumsolution. Dissolve 0.3818 g of ammoniumchlo-
ride (NH4CI) in 1 L of 2 M KCI in a volumetric flask. If pure, dry
NH4CI is used,the solutioncontains100 Ilg of NHt-N mL-l. Storethe
solution in a refrigerator.To preparea seriesof working standardsthat
6 After Banwartet al. (1972),and Keeneyand Nelson (1982).

contain0.1, 0.5,1,5,and 10 Ilg ofNH.t-N mL-l, pipette1-,5-,10-,50-,

and 100-mL aliquots of the concentrated solution into 1-L volumetric
flasks, and dilute to volume with 2 M KCI. If an extractantotherthan 2
M KCI is used,preparethe standardsin this solution.
PROCEDURE.Pipettea 20-mL aliquot of a 2M KCI soil extract into a 30-or
50-mL beakercontaininga Teflon-coatedstirring bar. Placethe beakeron a mag-
netic stirrer, add 2 mL of 0.25 M NaOH, and immersein the solution the ammo-
nia electrodeconnectedto a pH-millivolt meter. Stir the solution for 1 min, and
then recordthe meterreading(in mV). To calibratethe meter,carry out the same
procedureusing 20-mL aliquotsof the working standardNH.t solutions.
CALCULATIONS. If the meteris designedfor direct readoutof concentration,
determinethe NH.t concentrationof the samplefrom the meter reading. Other-
wise, calculatethe concentrationusing the equationobtainedby regressionof the
concentrationsof the standardson the correspondingmillivolt readings,or deter-
mine the value by referenceto a calibrationcurve preparedfrom analysesof stan-
dards.
COMMENTS. The exactprocedurefor measuringelectrodepotentialdepends
on the particularmeteremployed,and referenceshouldbe madeto the operating
instructionsprovidedby the manufacturer.The metershouldbe calibratedimme-
diately beforeeachseriesof analyses.If excessivedrift is observed,or if the elec-
trode slope deviatesmarkedly from 57 to 59 mV per decadechangein NH.t-N
concentration,the electrodemembraneshouldbe replaced.To avoid lossof NH 3,
measurements must be madewithin 1 to 2 min after the addition of NaOH. The
electrodeshouldbe held at a 20° anglewith respectto the vertical, so that air bub-
bles cannot be entrappedunder the electrode.The rate of stirring during the
analysisshould not be so rapid as to form a vortex. The samplesand standards
analyzedin eachseriesof analysesshouldbe at the sametemperature.
Nitrate
Principles. The concentrationof NO)"-N is estimatedby comparisonof the
electromotiveforce (emf, in mV) for the sampleto the valuesobtainedin analy-
sis of NO)"-N standardsby the samemethod.
Method'
SPECIAL APPARATUS
1. Nitrate electrode.
2. Double-junction referenceelectrode(Orion Model 90-02 or equiva-
lent). Fill outer chamberwith 0.02 M (NH4)zS04.
REAGENTS
1. Ammonium sulfate, 2 M (for ionic strengthadjustment).Dissolve 264

g of reagent-grade(NH4)zS04in 1 L of deionizedwater.
7 After Bremneret al. (1968), and Orion Research,Inc. (1990a).

1170 MULVANEY
2. Standard nitrate solution. Dissolve 0.7218 g of potassium nitrate

(KN03) in 1 L of deionizedwater in a volumetric flask. If pure, dry
KN0 3 is used,the solution contains100 Jlg of N03"-N mL-l. Store the
solution in a refrigerator.To prepareworking standardsthat contain 1
and 10 Jlg of N03"-N mL-l, pipette10- and 100-mL aliquotsof the con-
centratedsolution into 1-L volumetric flasks, and dilute to volume with
deionizedwater.
PROCEDURE.Pipettea 20-mL aliquot of an aqueoussoil extractinto a 30- or
netic stirrer, add 2 mL of 2 M (NH4)zS04,and immersein the solution the N03"
and referenceelectrodesconnectedto a pH-millivolt meter. Stir the solution for
1 min, and recordthe meterreading(in mY). To calibratethe meter,carry out the
sameprocedureusing 20-mLaliquots of the working standardN03" solutions.
determinethe N03" concentrationof the samplefrom the meter reading. Other-
wise, calculatethe concentrationusing the equationobtainedby regressionof the
mine the value by referenceto a calibrationcurve preparedfrom analysesof stan-
dards.
COMMENTS. To ensureproper performanceand extend electrodelife, care
should be taken to follow manufacturer'sguidelinesconcerningthe preparation,
storage,and standardizationof the N03" and referenceelectrodes.Calibration
shouldbe carriedout immediatelybeforeeachseriesof analyses,and after every
10 to 20 samples.If excessivedrift is observed,or if the electrodeslopedeviates
markedlyfrom 57 to 59 mY per decadechangein NHt-N concentration,the sen-
sor moduleshouldbe replaced.Chemicaltreatmentof the extractmay be required
to eliminateinterferenceby solublesalts,Cl-, or NOz. The extractmustbe stirred
for 1 min to ensurecompletemixing. Stirring shouldnot be so rapid that the elec-
trode tip is exposedto air bubbles.Use of a thermal-insulatingbarrier between
the beakerand the stirrer is adviseableto avoid temperature fluctuations
(Keeney
et aI., 1970).If desired,analysescan be performedon soil suspensions,using the
proceduresdescribedby Bremneret a1. (1968) or Keeneyand Nelson(1982).
Nitrite
Principles. The soil extract is acidified by addition of a Na2S04-H2S04
buffer (pH 1.2) to convertNOz to gaseousNOx , and the emf (in mY) is measured
with an NOx electrode.The concentrationof NOz-N is estimatedby comparison
to the valuesobtainedin analysisof NOz-N standardsby the samemethod.
Method 8
SPECIAL APPARATUS
1. NOx electrode(Orion Model 95-46).
8 After Tabatabai(1974).
REAGENTS
1. Acid buffer reagent. Dissolve 190 g of anhydroussodium sulfate
(Na2S04)in approximately800 mL of deionizedwater in a 1-L volu-
metricflask, slowly add53 mL of 18 M (concentrated)H2S04, andcool
and dilute the solution to volume with deionizedwater.
2. Standardnitrite solution. Dissolve0.4926g of sodiumnitrite (NaN02)
in 1 L of deionizedwater in a volumetric flask. If pure, dry NaN02 is
used,the solution contains100 Ilg of N02"-N mL-l. Store the solution
in a refrigerator.To prepareworking standardsthat contain0.1, 1, and
10 Ilg of N02"-N mL-l, pipette1-, 10-, and 100-mLaliquotsof the con-
centratedsolutioninto l-L volumetricflasks, anddilute to volumewith
1 M KCl. If an extractantother than 1 M KCI is used,preparethe stan-
dardsin this solution.
PROCEDURE.Pipettea 20-mL aliquot of aIM KCI soil extractinto a 30- or
netic stirrer, add 2 mL of acid buffer, and immersein the solution the NOx elec-
trode connectedto a pH-millivolt meter. Stir the solution for 1 min, and record
the meterreading(in mV). To calibratethe meter,carry out the sameprocedure
using 20-mL aliquotsof the working standardN02" solutions.
determinethe N02" concentrationof the samplefrom the meter reading.Other-
wise, calculatethe concentrationusingthe equationobtainedby regressionof the
mine the valueby referenceto a calibrationcurvepreparedfrom analysesof stan-
dards.
COMMENTS. Referenceshould be made to the instruction manual for the
NOx electrodefor details regardingpreparation,storage,and standardizationof
the electrode.Calibrationshouldbe carriedout immediatelybeforeeachseriesof
analyses.If excessivedrift is observed,or if the electrodeslopedeviatesmarked-
ly from 57 to 59 mV per decadechangein N02"-N concentration,the electrode
membraneshould be replaced.To avoid loss of NOv measurementsmust be
madewithin 1 to 2 min after the additionof the acid buffer. The electrodeshould
be held at a 20° angle with respectto the vertical, so that air bubblescannotbe
entrappedunderthe electrode.The rate of stirring during the analysisshouldnot
be so rapid asto form a vortex. The samplesandstandardsanalyzedin eachseries
of analysesshouldbe at the sametemperature.
DETERMINATION OF NONEXCHANGEABLE AMMONIUM
Introduction
Threetypesof methodshavebeenusedto determine nonexchangeable NH4
in soils. In one,the soil sampleis distilled with NaOH, and a duplicatesampleis
distilled with KOH, nonexchangeable NHS being estimatedas the difference
betweenthe amountsof NH4 releasedby the two distillations. In another,the soil
1172 MULVANEY
sampleis heatedto remove exchangeable NUt and organicN, and non-exchange-

able NHt is determinedfrom analysisof the residueby the Kjeldahl procedure.
In the other, the soil sampleis treatedwith HF to decomposethe clay minerals
containingthe nonexchangeable NHt, and the NHt releasedby this treatmentis
determined.ExchangeableNHt is removedbefore the treatmentwith HF or is
estimatedseparatelyand subtractedfrom the NHt releasedby HE
The duplicate distillation procedurefor estimatingnonexchangeable NH,t
in soils was originally proposedby Barshad(1951). This procedurehas not been
widely used because:(i) distillation with NaOH does not effect quantitative
releaseof nonexchangeable NHt from soils and minerals, (ii) trace amountsof
K+ effectively block the releaseof nonexchangeable NHt during distillation with
NaOH, and (iii) the amountof NHt releasedfrom organic-Ncompoundsis like-
ly to differ for distillations with NaOH and KOH.
Mogilevkina (1964) suggesteda methodof determiningnonexchangeable
NH,t in soils, in which the sampleis heatedat 400°C for 24 to 72 h dependingon
the organic matter content, and nonexchangeable NHt is estimatedby total-N
analysisof the residuefrom this heat treatmentusing a Kjeldahl procedure.This
methodinvolves the assumptionthat the heat treatmentusedeffects quantitative
removalof organicN and exchangeableNH,t but doesnot releasenonexchange-
able NHt, in which caseN in the residuewill occur exclusivelyas nonexchange-
able NHt andwill representall of the nonexchangeable NHt originally presentin
the sample.However,Nelson and Bremner(1966) showedthis assumptionto be
invalid, as the heattreatmentusedled to loss of nonexchangeable NHt, and their
work further showed that the Kjeldahl procedureused failed to quantitatively
recover N in the residue from this treatment.Mogilevkina's method has been
found to give much lower values than other methodsof determining nonex-
changeableNHt in soils (Nelson& Bremner,1966; Bremneret aI., 1967; Mogil-
evkina, 1969; Moyano & Gallardo, 1988) and is unsatisfactory.
A variety of methodshavebeendescribedthat involve the useof HF to esti-
matenonexchangeable NHt in soils. In the first study to provide evidencefor the
presenceof substantialamountsof nonexchangeableNHt in soils, Rodrigues
(1954) showedthat a significant amountof NHt was releasedfrom tropical soils
by treatment withHF and concludedthat this NHt was derived from NHt held
by clay minerals.In his work, nonexchangeable NHt was extractedby treatment
of the soil samplewith a mixture of four volumesof 20 M (40%) HF and onevol-
ume of 9 M (50%) H2S04, NHt in the extract being determinedby distillation
with NaOH after removalof HF with H2S04, This is a drasticprocedure,and sub-
sequentwork by Bremner(1959) showedthat it causesextensivedecomposition
of organic-N compoundsto NHt. Milder HF procedureshave been adoptedin
most investigationsconcerningthe occurrenceand distribution of nonexchange-
able NH,t in soils. The most commonextractantshavebeen5 M HF-0.75M Hel-
0.3 M H2S04 solution (Dhariwal & Stevenson,1958), 1 M HF-1 M HCI solution
(Bremner,1959), and 5 M HF-1 M HCI solution (Silva & Bremner,1966).
Various pretreatmentshavebeenemployedto removeorganicmatter inHF
methodsof estimatingnonexchangeable NHt in soils and therebyreducethe risk
of interferenceby decompositionof organic-Ncompoundsto NHt. In the method
describedby Dhariwal and Stevenson(1958) for estimationof nonexchangeable
NHt, the soil sampleis heatedwith 1 M KOH in an autoclavefor 8 h beforetreat-

ment with HF-HCI-H2S04 solution. Schachtschabel (1960, 1961) suggestedthe
useof H20 2 to oxidize organicmatterbeforeextractionof nonexchangeable NHt
with HF-H2S04' and further recommendedthat the H20 2 treatmentbe carriedout
in the presenceof KCl to preventfixation of the NHt producedby this treatment
and that correctionbe madefor any releaseof NHt from organiccompoundsdur-
ing extractionwith HF-H2S04. SIlva and Bremner(1966) describeda methodfor
estimationof nonexchangeable NHt, in which the soil sampleis treatedwith a
mixture of KOBr and KOH before extraction with HF-HCI. The KOBr-KOH
mixture is more effective for removal of organic matter than hot KOH or H20 2
(Bremneret aI., 1967), and the high concentrationof K+ eliminatesany possibil-
ity of NHt fixation by soil mineralsduring treatmentwith this reagent.Moreover,
the KOBr readily convertsNHt to N2, thereby ensuringrapid removal of ex-
changeableNHt and NHt derivedfrom organic-Ncompounds.
The method of Silva and Bremner(1966) is describedbecauseit has no
apparentdefectsand has beenusedwidely to estimatenonexchangeable NHt in
soils. However, there is no way of verifying the accuracyof this method, and
attentionshould be drawn to the possibility that soil mineralsmay contain high-
ly labile organic-Ncompoundsthat are not removedor decomposedby treatment
with KOBr-KOH, but are releasedby treatmentwith HF-HCI and decompose
extensively to NHt during this treatmentor upon subsequentdistillation with
KOH. Freney(1964) observeda closecorrelationbetweenorganic-Cand total-N
analysesof residuesobtainedby the KOH-HF methodof Dhariwal and Steven-
son (1958) and suggestedthat nonexchangeable NHt estimatedby this methodis
an artifact arising from the decompositionof labile organic-Ncompounds.How-
ever, there is little supportfor this view amongsoil scientists,and all indications
are that the greatmajority of NHt liberatedby KOH-HF or KOBr-HF methodsis
nonexchangeable NHt from silicate minerals. A secondpossible defect of the
procedure described by Silva and Bremner (1966) for estimation of nonex-
changeableNHt is that clay mineralsmay be partially dissolvedby the KOBr-
KOH pretreatmentusedto decomposeorganic-N compoundsbefore addition of
HF-HCl, in which casesomeof the nonexchangeable NHt might be lost. Loss of
NHt also could occur from the presenceof taranakitesor other complex phos-
phatesthat are subjectto quantitativeremovalby treatmentwith KOH or KOBr-
KOH (Bremneret aI., 1967).However,thereis no evidencethat soils containsig-
nificant amountsof NHt in the form of suchcompounds.
Principles
In the methoddescribed,the soil sampleis treatedwith alkaline potassium

hypobromite(KOBr-KOH) solution to removeexchangeable NHt and organic-N
compoundsthat may yield NHt under the conditionsemployedfor releaseand
estimation of nonexchangeableNHt, and the residue from this treatment is
washedwith 0.5 M KCl and shakenwith 5 M HF-1 M HCl solution for 24 h to
decomposemineralscontainingnonexchangeable NHt. The NHt releasedby the
HF-HCI treatmentis determinedfrom the amountof NH3 liberatedby stearndis-
tillation of the soil-acid mixture with KOH.
1174 MULVANEY
The alkaline KOBr solution employedto eliminate interferenceby organ-

ic-N compoundsoxidizessoil organicmatteralmostcompletelyunderconditions
that are unlikely to causesignificant releaseof nonexchangeableNHt and that
involve no risk of fixation of NHt by soil minerals.The NHt formed on oxida-
tion of soil organicmatterby this reagentis immediatelyconvertedto N2 by the
following reaction
2NH3 + 3KOBr ~ N2 + 3KBr + 3H20.
Method9
SpecialApparatus
1. Steam-distillationapparatusdescribedin "Special Apparatus" under
"Steam-DistillationMethods"(Fig. 38-1).
2. Distillation flasks. The flasks employedare 250-mL Pyrex round bot-
tom boiling flasks with standard-taper joint (T24/40) that havebeenfit-
ted with a socketjoint (~ 28/15) for attachmentto the distillation appa-
ratus. Their dimensionsshould be such that when the flasks are con-
nectedto the steam-distillationapparatus,the distancebetweenthe tip
of the steam-inlettube and the bottom of the flask does not exceed4
mm.
3. Microburette(5-mL, graduatedat O.Ol-mL intervals)or automatictitra-
tor.
Reagents
1. Potassiumhypobromitesolution. Dissolve 22 g of KOH in 200 mL of
deionizedwater in a 250-mL Erlenmeyerflask containinga Teflon- or
glass-coatedmagneticstirring bar. Then immerse the flask in a con-
tainer of crushedice, and allow the solution to cool to 5°C. With con-
stant stirring, add 6 mL of bromine (Br2) dropwisefrom a burette at a
rate of approximately 0.5mL min-I. Storethe solution in a refrigerator.
2. Potassiumchloride solution, approximately0.5 M. Dissolve 186 g of
reagent-gradeKCI in 5 L of deionizedwater.
3. Hydrofluoric acid-hydrochloricacid solution (approximately5 MHF-1
M HCI). To 1.5 L of deionized water in a 2.5-L polyethylene or
polypropylenebottle markedto indicate a volume of 2 L, add 167 mL
of 12 M (concentrated)HCI (sp. gr. 1.19) and 325 mL of 31 M (con-
centrated,~52%) HE Then dilute the solution to the 2-L mark, and mix
it thoroughly.
4. Potassiumhydroxidesolution, approximately10 M. Dissolve2.8 kg of
KOH in approximately4 L of deionizedwater, and dilute the cooled
solution to a volume of 5 L. Storethe solutionin a tightly stopperedbot-
tle.
9 After Bremner(1965), and Silva and Bremner(1966).

5. Boric acid-indicator solution. Prepare this reagent as described in

"Reagents"under"Steam-DistillationMethods."
Procedure
Placea 1-g sampleof finely ground «150!lm) soil in a 200-mL tall-form
beaker,and add20 mL of KOBr solution. Swirl the beakerto mix the soil and
KOBr, cover the mouth of the beakerwith a watch glass,and allow the covered
beakerto standat room temperaturefor 2 h. Then add60 mL of deionizedwater,
and after replacingthe watch glass, heat the beakeron a hotplate until the soil-
KOBr mixture hasboiled vigorously for 5 min. Allow the mixture to cool and set-
tle in the covered beaker (preferably overnight); then decant and discard the
supernatantliquid, and transferthe residuewith 0.5 M KCI to a 100-mL polyeth-
ylene or polypropylenecentrifugetube that is markedto indicatea volume of 80
mL. Use a wash bottle containing0.5 M KCI to perform this transfer,and bring
the contentsof the tube to the 80-mL mark with the washingsobtainedby rins-
ing the beakerseveraltimes with 0.5 M KCI. Fit the neck of the tube with a poly-
ethylenecap or rubberstopper,and after shakingthe tube manuallyfor a few sec-
onds,centrifugeit (1100 x g) for 10 min. Decantthe clearsupernatant liquid,add
0.5 M KCI to the 80-mL mark, andshakeandcentrifugethe tube as describedpre-
viously. Decantthe clear supernatantliquid, and add 20 mL of 5 M HF-1 M HCl
solution from a 25-mL polyethylenegraduatedcylinder. Then stopperthe tube,
and shakeit for 24 h on a reciprocatingshaker.
After completionof the treatmentwith HF-HCl solution, transfer15 mL of
10 M KOH to a 250-mL distillation flask, and place a long-stemmedpolyethyl-
ene filter funnel in the neck of the flask so that the stem extendsbelow the sur-
face of the KOH solution. Transferthe contentsof the centrifugetube tothe dis-
tillation flask through this funnel, and completethe transferby rinsing the cen-
trifuge tube and funnel with approximately25 mL of deionized water from a
wash bottle. Then stopperthe flask, swirl it to mix the contents,and allow it to
standfor a few minutes.During this period of standing,add 10 mL of boric acid-
indicator solution to a 250-mL beakerthat is marked toindicatea volume of 100
mL, and place the beakerunderthe condenserof the steam-distillationapparatus
so that the tip of the condenseris in contactwith the side of the beakerand about
1 cm below the top. Attach the distillation flask to the steam-distillationappara-
tus as shownin Fig. 38-1, and immediatelycommencesteamdistillation by clos-
ing the lower stopcockon the steam-bypass assembly.When the distillate reach-
es the 100-mL mark on the receiverflask (~12 min is required),rinse the tip of
the condenser,and stop the distillation by opening the lower stopcock on the
steam-bypassassembly.Titrate the distillate with 0.0025M H2S04, At the end-
point, the color changes fromgreento a permanent,faint pink.
Calculations
Calculatenonexchangeable NHt-N in the samplefrom the expression(S-

C) x T, whereS is the volume of H2S04 usedin titration of the sample,C is the
volume usedin titration of a control (obtainedby steamdistillation of 20 mL of
1176 MULVANEY
5 M HF-l M HCI solution with 15 mL of 10 M KOH solution), and T is the titer

of the titrant (for 0.0025M H2S04, T = 70 J..lg N mL- 1).
Comments
Potassiumhypobromitetendsto decomposeto form KBr and O2 (2KOBr

~ 2KBr + O2), so this reagentshouldbe preparedimmediatelybeforeuse.How-
ever, the rateof decompositionis markedly reducedat low temperaturesin the
absenceof light, and the reagentmay safely be usedfor up to 1 mo if stored ina
refrigerator.
Studiesby Silva and Bremner(1966) have shown that the KOBr pretreat-
ment employedin the proceduredescribed removes 96 to 99% of the organicN
in soils containingup to 150 g of organicmatterkg-1 and that the residuesfrom
this pretreatmentrelease only trace amounts of NHS under the conditions
employedfor distillation of the NHS releasedby the treatmentwith HF-HCI solu-
tion. The 5-min boil after treatmentwith KOBr-KOH solution at room tempera-
ture for 2 h increasesthe amountof soil organicN convertedto Nz, and removes
material that releasesNHS under the conditionsemployedfor distillation of the
NHS releasedby treatmentwith HF-HCI solution. Boiling also causesfloccula-
tion of the soil solids and permits the KOBr-KOH solution to be removedby
decantationwithout preliminary centrifugation.To ensurecompleteremoval of
labile organic-Ncompounds,boiling shouldbe vigorous.
Testsusingseveralsoils haveshownthat the resultsobtainedby the method
describedare not significantly affected if the period of treatmentwith KOBr-
KOH solution is increasedfrom 2 to 12 h, or if the period of shakingwith 5 M
HF-l M HCI solution is increasedfrom 24 to 48 h (Silva & Bremner,1966).
Careis requiredto avoid lossof suspendedmaterialduring the decantations
in the proceduredescribed.The useof a rubberpolicemanis recommendedto aid
removal of adheringsoil particles during washing in transferof the potassium
hypobromite-treated soil to the centrifugetube and of the soil-HF-HCI mixture to
the distillation flask.
Hydrofluoric acid is a volatile reagentand interfereswith the determination
of NHS by distillation with alkali (Bremner & Harada, 1959). In the method
described,this interferenceis eliminatedby the techniqueusedfor addition of the
HF-treatedsampleto the distillation flask, and by swirling the flask and allowing
it to standfor a few minutesafter this addition. No loss of NH3 has beendetect-
ed using this procedure,but the risk of loss is reducedif the 10 M KOH solution
employedfor distillation is kept cold by storing it in a refrigerator.
The apparatusused for separationof NHS by steamdistillation, and the
reagentsusedfor estimationof NHS in the distillate are describedand discussed
in "Methods"under"Steam-DistillationMethods,"and referenceshouldbe made
to this sectionfor additionalinformation concerningthe procedurefor separation
and estimationof NHS.
REFERENCES
Adamsen,FJ., D.S. Bigelow, and G.R. Scott. 1985.Automatedmethodsfor ammonium,nitrate, and
nitrite in 2 M KCI-phenylmercuricacetateextractsof soil. Commun. Soil Sci. Plant Anal.
16:883-898.
Allen, S.E., and H.M. Grimshaw. 1962. Effect of low-temperaturestorageon the extractablenutrient
ions in soil. J. Sci. Food Agric. 13:525-529.
Al- Wehaid,A, and A. Townshend.1986. Spectrophotometric flow-injection determinationof nitrate
basedon reductionwith titanium (III) chloride. Anal. Chim. Acta 186:289-294.
American Public Health Association. 1992. Standardmethods for the examination of water and
Ananth, S., and J.T. Moraghan.1987. The effect of calcium and magnesiumon soil nitrate determi-
nation by automated segmented-flow methods.Soil Sci. Soc. Am. 1. 51:664-667.
Axelrod, H.D., 1.E. Bonelli, and J.P. Lodge, Jr. 1970. Fluorimetric determinationof trace nitrates.
Baker, AS. 1967. Colorimetric determinationof nitrate in soil and plant extractswith brucine. J.
Agric. Food Chern. 15:802-806.
Baker, AS., and R. Smith. 1969. Extracting solution for potentiometricdeterminationof nitrate in
plant tissue.1. Agric. Food Chern. 17:1284-1287.
Balks, R., and I. Reekers.1955. Bestimmungdes Nitrat- und Ammoniastickstoffsim Boden. Land-
wirtsch. Forsch.8:7-13.
Banwart, w.L., M.A Tabatabai,and 1.M. Bremner. 1972. Determination of ammonium in soil
extracts and water samples by an ammonia electrode. Commun. Soil Sci. Plant Anal.
3:449-458.
Barak, P., and Y. Chen. 1987.Three-minuteanalysisof chloride, nitrate, and sulfateby single column
anion chromatography.Soil Sci. Soc. Am. J. 51:257-258.
Barker, AV. 1974. Nitrate determinationsin soil, water and plants. Massachusetts Agric. Exp. Stn.
Res. Bull. 611. Univ. Massachusetts, Amherst.
Barnes,H., and AR. Folkard. 1951. The determinationof nitrites. Analyst (London) 76:599-603.
Barshad,I. 1951. Cation exchangein soils: I. Ammonium fixation and its relation to potassiumfixa-
tion and to determinationof ammoniumexchangecapacity.Soil Sci. 72:361-371.
Bartuzi, J., I. Dechnik, and Z. Stepniewska.1976. Zastosowanieelektrod selektywnychdo pomiaru
aktywnoscichlorkow i azotanoww glebie. Rocz. G1ebozn.27:15-26.
Baveja, AK., and v.K. Gupta. 1982. Extractive spectrophotometricdeterminationof trace amounts
of nitrite and nitrate in irrigation water and soil. Fert. Technol. 19:80-84.
Berthelot,M.P.E. 1859. Violetd'aniline. Rep. Chim. Appl. 1:284.
Best, E.K. 1976. An automatedmethodfor determiningnitrate-nitrogenin soil extracts.Queensland
1. Agric. Anim. Sci. 33:161-166.
Best, E.K., and E.T. Craswell. 1985. Interferenceby magnesiumin the determinationof nitrate in 2
M KCl extractsof soil by steamdistillation. Commun.Soil Sci. Plant Anal. 16:1189-1198.
Biggar, J.W. 1978. Spatialvariability of nitrogen in soils. p. 201-211.In D.R. Nielsen and J.G. Mac-
Donald (ed.) Nitrogen in the environment.Vol. 1. Acad. Press,New York.
Black, AS., and S.A Waring. 1978. Nitrate determinationin an oxisol using K 2S04 extraction and
the nitrate-specificion electrode.Plant Soil 49:207-211.
Boltz, D.F., and M.J. Taras.1978. Nitrogen. p. 197-251.In D.F. Boltz and J.A. Howell (ed.) Colori-
metric determinationof nonmetals.2nd ed. Wiley-Intersci., New York.
Botha,AD.P., and J.C. Johnson.1988.A vacuumfiltration and leachingmethodfor the colorimetric
determination of ammonium and nitrate nitrogen in soils. S. Afr. Tydskr. Plant Grond
5:196-198.
Bound, G.P. 1977. Determinationof nitrate in soil pastesby ion selective electrodes.J. Sci. Food
Agric. 28:501-505.
Bound, G.P., and B. Fleet. 1977. The developmentof a solid statereferenceelectrodefor use in soil
measurements. J. Sci. Food Agric. 28:431-435.
Bradfield, E.G., and D.T. Cooke. 1985. Determinationof inorganicanionsin water extractsof plants
and soils by ion chromatography.Analyst (London) 110:1409-1410.
Breimer, T., and J.H.G. Slangen.1981. Pretreatmentof soil samplesbefore NOrN analysis.Neth. 1.
Agric. Sci. 29:15-22.
Bremner,J.M. 1959. Determinationof fixed ammoniumin soil. 1. Agric. Sci. 52:147-160.
Bremner,1.M. 1965. Inorganic forms of nitrogen. p. 1179-1237.In C.A Black et al. (ed.) Methods
of soil analysis.Part 2. Agron. Monogr. 9. ASA, Madison,WI.
Bremner,J.M., and L.G. Bundy. 1973. Use of ferrous hydroxide for determinationof nitrate in soil
extracts.Commun.Soil Sci. Plant Anal. 4:285-291.
Bremner,1.M., L.G. Bundy, andAS. Agarwal. 1968. Use of a selectiveion electrodefor determina-
tion of nitrate in soils. Anal. Leu. 1:837-844.
1178 MULVANEY
Bremner,J.M., and A.P. Edwards.1965. Determinationand isotope-ratioanalysisof different forms

of nitrogenin soils: I. Apparatusand procedurefor distillation and determinationof ammoni-
um. Soil Sci. Soc. Am. Proc. 29:504-507.
Bremner,J.M., and T. Harada.1959. Releaseof ammoniumand organic matter from soil by hydro-
fluoric acid and effect of hydrofluoric acid treatmenton extractionof soil organic matter by
neutraland alkaline reagents.J. Agric. Sci. 52:137-146.
Bremner,J.M., and D.R. Keeney.1965. Steamdistillation methodsfor determinationof ammonium,
nitrate and nitrite. Anal. Chim. Acta 32:485-495.
Bremner, J.M.,and D.R. Keeney. 1966.Determinationand isotope-ratioanalysisof different forms of
nitrogen in soils: 3. Exchangeableammonium, nitrate, and nitrite by extraction-distillation
methods.Soil Sci. Soc. Am. Proc. 30:577-582.
Bremner,J.M., D.W. Nelson, and J.A Silva. 1967. Comparisonand evaluationof methodsof deter-
mining fixed ammoniumin soils. Soil Sci. Soc. Am. Proc. 31:466-472.
Bremner, J.M., and K Shaw. 1955. Determinationof ammonia and nitrate in soil. J. Agric. Sci.
46:320-328.
Brown, M.W. 1973. A highly sensitive automatedtechniquefor the determinationof ammonium
nitrogen.J. Sci. Food Agric. 24:1119-1123.
Buckett,J., W.D. Duffield, and R.E Milton. 1955.The determinationof nitrate and nitrite in soil. Ana-
lyst (London) 80:141-145.
Burton, D.L., DA Gower, P.M. Rutherford,and W.B. McGill. 1989. Amino acid interferencewith
ammoniumdeterminationin soil extractsusing the automatedindophenolmethod.Commun.
Carlson,R.M. 1978. Automatedseparationand conductimetricdeterminationof ammoniaand dis-
solvedcarbondioxide. Anal. Chern. 50:1528-1531.
Carlson,R.M., R.I. Cabrera,J.L. Paul.,J. Quick, and R.Y. Evans.1990. Rapid direct determinationof
ammonium and nitrate in soil and plant tissue extracts. Commun. Soil Sci. Plant Anal.
21:1519-1529.
Carlson,R.M., and D.R. Keeney.1971.Specificion electrodes: Techniques and usesin soil, plant, and
water analysis.p. 39-65. In L.M. Walsh (ed.) Instrumentalmethodsfor analysisof soils and
plant tissue.SSSA,Madison,WI.
Cataldo,D.A, M. Haroon,L.E. Schrader,and V.L. Youngs. 1975. Rapid colorimetric determination
of nitrate in plant tissueby nitration of salicylic acid. Commun.Soil Sci. Plant Anal. 6:71-80.
Cattrall, R.W., and I.e. Hamilton. 1984. Coated-wire ion-selective electrodes.Ion-Select.Elec. Rev.
6:125-172.
Cawse,P.A 1967. The determinationof nitrate in soil solutions by ultraviolet spectrophotometry.
Cbaube,A, AK Baveja,and V.K Gupta.1984.Determinationof ultra traceconcentrationsof nitrite
in polluted watersand soil. Talanta31:391-393.
Clarke, AL., and A.C. Jennings.1965. Spectrophotometricestimationof nitrate in soil using chro-
motropic acid. J. Agric. Food Chern. 13:174-176.
Clausen,e., B.R. Bock, G.A Peterson,and R.A Olson. 1980. The magnesiumproblem in nitrate
determinationby steamdistillation. Soil Sci. Soc. Am. J. 44:1326-1327.
Clausen,e.R., M.P. Russelle,A.D. F1owerday,and R.A Olson. 1981. Problemsin direct steamdis-
tillation of soil for mineral nitrogen determinationdue to carbonates.Soil Sci. Soc. Am. J.
45:1238-1240.
Conway,E.J. 1947. Microdiffusion analysisandvolumetricerror. 2nd ed. CrosbyLockwood, London.
Covington,AK 1974. Ion-selectiveelectrodes.CRC Cril. Rev. Anal. Chern.4:355-406.
Csiky, I., G. Marko-Varga,and J. A. Jonsson.1985. Use of disposableclean-upcolumnsfor selective
removal of humic substancesprior to measurementswith a nitrate ion-selectiveelectrode.
Dahnke,w.e. 1971. Use of the nitrate specific ion electrodein soil testing. Commun.Soil Sci. Plant
Anal. 2:73-84.
Davies, J.E.w., G.J. Moody, and J.D.R. Thomas. 1972. Nitrate ion selective electrodesbasedon
poly(vinyl chloride) matrix membranes.Analyst (London) 97:87-94.
Dhariwal, AP.S., and EJ. Stevenson.1958. Determinationof fixed ammonium in soils. Soil Sci.
86:343-349.
Dick, W.A., and M.A Tabatabai.1979. Ion chromatographicdeterminationof sulfate and nitrate in
Dorich, R.A, and D.W. Nelson. 1983. Direct colorimetric measurementof ammoniumin potassium
chloride extractsof soils. Soil Sci. Soc. Am. J. 47:833-836.
Dorich, R.A, and D.W. Nelson. 1984. Evaluationof manualcadmium reductionmethodsfor deter-
mination of nitrate in potassiumchloride extractsof soils. Soil Sci. Soc. Am. J. 48:72-75.
du Preez,c.c., R. du T. Burger, andOJ. Laubscher.1987.Evaluationof steamdistillation procedures

for the routine determination of inorganic soil nitrogen.Commun. Soil Sci. Plant Anal.
18:483-493.
du Preez,C.C., OJ. Laubscher,and R. du T. Burger. 1989. Determinationof nitrate in 2,0 M KCI soil
extractsusing ion chromatography.Commun.Soil Sci. Plant Anal. 20:113-120.
du Preez,c.c., OJ. Laubscher,and R. du T. Burger. 1989. Evaluationof specific ion electrodesfor
the determinationof extractableNlij and NO) from soil. Appl. Plant Sci. 3:1--4.
Elliott, c.L., G.H. Snyder,and 1.L. Cisar. 1989. A modified AutoanalyzerII methodfor the determi-
nation of NOrN in water using a hollow-Cd reductioncoil. Commun. Soil Sci. Plant Anal.
20:1873-1879.
Fakhri, N.A, S.A Rahim, and W.A Bashir. 1983. Indole as a chromogenicreagentfor tracesof
nitrate in aqueoussolution. Int. J. Environ. Anal. Chern. 16:131-138.
Fisher,EL., E.R. !bert, and H.E Beckman.1958.Inorganicnitrate,nitrite, or nitrate-nitrite.Rapidcol-
orimetric determination of microgram quantities in aqueous solution. Anal. Chern.
30:1972-1974.
Fox, J.B. 1985. The determination of nitrite: A critical review. CRC Crit. Rev. Anal. Chern.
15:283-313.
Francis,C.W., and C.D. Malone. 1975. Nitrate measurements using a specific ion electrodein pres-
enceof nitrite. Soil Sci. Soc. Am. Proc. 39:15(}-151.
Freney, J.R. 1964. An evaluation of naturally occurring fixed ammonium in soils. J. Agric. Sci.
63:297-303.
Freney,1.R. 1971. Phosphateinterferencein the Devarda'salloy reductionmethodfor nitrate. Com-
mun. Soil Sci. Plant Anal. 2:479--484.
Freney,J.R.,and R. Wetselaar.1967.The determinationof mineral nitrogenin soil with particularref-
erenceto nitrate. Div. Plant Indust. Tech. Pap. no. 23. CSIRO, Melbourne,Australia.
Gasser,J.KR. 1958. Use of deep-freezingin the preservationand preparationof fresh soil samples.
Nature (London) 181:1334-1335.
Gaugush,R.E, and R.T. Heath. 1984. A rapid manualmethodfor nitrate determinationin small vol-
umesby a modification of the cadmiumreductionmethod.Water Res. 18:449--450.
Gentry, C.E., and R.B. Willis. 1988. Improved methodfor automateddeterminationof ammoniumin
soil extracts.Commun.Soil Sci. Plant Anal. 19:721-737.
Gerlach,A 1980. Ein Vergleich von Methodenzur Bestimmungvon Ammonium- und Nitratstick-
stoff in Boden. Acta Ecol.lEcol. Plant. 1: 185-200.
Goodman,D. 1976. Automatic apparatusfor the determinationof pH and nitrate in soils. Analyst
(London) 101:943-948.
Goodroad,L.L., and L.M. Shuman.1990. Nitrate-N determinationin soils using coatedwire elec-
trodes.Commun.Soil Sci. Plant Anal. 21:1559-1567.
Greweling,T., and M. Peech.1960. Chemicalsoil tests.Cornell Exp. Stn. Bull. 960.
Griess, P. 1879. Bemerkungenzu der Abhandlungder HH. Weselsky und Benedikt 'Uber einige
Azoverbindungen.'Chern. Ber. 12:426--428.
Haby, V.A 1989. Soil NOrN analysisin Ca(OH)zextractsby the chromotropicacid method.Soil Sci.
Soc. Am. 1. 53:30&.-310.
Hadjidemetriou,D.G. 1982. Comparativestudy of the determinationof nitratesin calcareoussoils by
the ion-selectiveelectrode,chromotropicacid and phenoldisulphonicacid methods.Analyst
(London) 107:25-29.
Hansen,E.H., AK Ghose,and J. Ruzicka. 1977. Flow injection analysisof environmentalsamples
for nitrate using an ion-selectiveelectrode.Analyst (London) 102:705-713.
Harding, D.E., and DJ. Ross. 1964. Somefactors in low-temperaturestorageinfluencing the miner-
alisable-nitrogenof soils. J. Sci. Food Agric. 15:829--834.
Hartz, T.K, R.E Smith, M. LeStrange,and KE Schulbach.1993. On-farm monitoring of soil and
crop nitrogen status by nitrate-selective electrode. Commun.Soil Sci. Plant Anal.
24:2607-2615.
Hauck, R.D. 1982. Nitrogen-isotope-ratioanalysis.p. 735-779.In AL. Pageet al. (ed.) Methodsof
Heanes,D.L.1975.Determinationof nitrate in soil and waterby an adaptationof an orangeI method.
Heinzmann,EX., M. Miyazawa,and M.A Pavan.1984.Determina~iio de nitrato em extratosde solos
acidospor esprectrofotometria de absor~iio ultravioleta. Rev. Brasil. Ci. Solo 8:159-163.
Henriksen,A, and AR. Selmer-Olsen.1970. Automatic methodsfor determiningnitrate and nitrite
in water and soil extracts.Analyst (London) 95:514-518.
1180 MULVANEY
Henzell, E.F., I. Vallis, and J.E. Lindquist. 1968. Automatic colorimetric methodsfor the determina-
tion of nitrogen in digestsand extractsof soils. p. 513-520.In J.w. Holmeset al. (ed.) Trans.
Int. Congr. Soil Sci., Vol. 3, 9th, 1968. Am. ElsevierPubl. Co., New York.
Hem, J.A, G.K. Rutherford,and G.w. vanLoon. 1983. Determinationof chloride, nitrate, sulphate
and total sulphur in environmentalsamplesby single-columnion chromatography.Talanta
30:677-682.
Hofman, G., C. Ossemerct,and G. Ide. 1980. Interactiesbij de ammonium-en nitraatstikstofbepalin-
gen in grondmonstersmet behulp van ion-specifiekeelectroden.Meded. Fac. Landbouw.,
Rijksuniv. Gent 45:1305-1314.
Horvai, G., and E. Pungor. 1987. Theoreticalbackgroundsof flow analysis. CRC Crit. Rev. Anal.
Chern. 17:231-264.
Huffman, S.A, and K.A. Barbarick. 1981.Soil nitrate analysisby cadmiumreduction.Commun.Soil
Hulanicki, A, R. Lewandowski,and M. Maj. 1974. Determinationof nitrate in water with a new con-
structionof ion-selectiveelectrode.Anal. Chim. Acta 69:409-414.
I1osvay,M.L. 1889. L'acide azoteuxdansla salive et dansI'air exhale.Bull. Soc. Chim. 2:388--391.
Jackson,W.A, C.E. Frost, and D.M. Hildreth. 1975. Versatile muItirange analytical manifold for
automatedanalysisof nitrate-nitrogen.Soil Sci. Soc. Am. Proc. 39:592-593.
Jeffery,G.H., J. Bassett,J. Mendham,and R.e. Denney.1989. Vogel's textbookof quantitativechem-
ical analysis.5th ed. Longman,London.
Jones,M.N. 1984. Nitrate reduction by shaking with cadmium. Alternative to cadmium columns.
Water Res. 18:643-646.
Kamphake,L.J., S.A Hannah,and J.M. Cohen. 1967. Automatedanalysisfor nitrate by hydrazine
reduction.Water Res. 1:205-216.
Keay, J., and P.M.A Menage.1969. Automateddistillation procedurefor the determinationof nitro-
gen. Analyst (London) 94:895-899.
Keeney,D.R., and J.M. Bremner.1966.Determinationand isotope-ratioanalysisof different forms of
nitrogen in soils: 4. Exchangeableammonium,nitrate, and nitrite by direct-distillation meth-
ods. Soil Sci. Soc. Am. Proc. 30:583-587.
Keeney,D.R., B.H. Byrnes,andJ.J.Genson.1970.Determinationof nitrate in waterswith the nitrate-
selectiveion electrode.Analyst (London) 95:383-386.
Keeney,D.R., andD.W. Nelson.1982. Nitrogen-inorganicforms. p. 643-698.In AL. Pageet al. (ed.)
Methodsof soil analysis.Part 2. 2nd ed. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Kempers,AJ. 1974. Determinationof sub-microquantitiesof ammoniumand nitrates in soils with
phenol,sodiumnitroprussideand hypochlorite.Geoderma12:201-206.
Kempers, AJ., and C.J. Kok. 1989. Re-examinationof the determinationof ammonium as the
indophenolblue complex using salicylate.Anal. Chim. Acta 221:147-155.
Kempers,AJ., and A Zweers. 1986. Ammonium determinationin soil extracts by the salicylate
method.Commun.Soil Sci. Plant Anal. 17:715-723.
Knittel, H., and G. Fischbeck.1979. Die potentiometrischeBestimmungvon Nitrat im Boden mit
einer Membranelektrode.Z. Pflanzenernaehr. Dung. Bodenkd.142:669-678.
Kowalenko,e.G.,and L.E. Lowe. 1973. Determinationof nitratesin soil extracts.Soil Sci. Soc. Am.
Proc. 37:660.
Krom, M.D. 1980. Spectrophotometricdeterminationof ammonia:A study of a modified Berthelot
reactionusing salicylateand dichloroisocyanurate.Analyst (London) 105:305-316.
Krug, F.J.,J. Ruzicka,and E.H. Hansen.1979.Determinationof ammoniain low concentrationswith
Nessler'sreagentby flow injection analysis.Analyst (London) 104:47-54.
Kuchnicki, T.C., L.P. Sarna,and G.R.B. Webster. 1985. Determinationof nitrate, nitrite, and phos-
phateat 214 nmby reversephaseHPLe. J. Liq. Chromatog.8:1593-1609.
Kuchnicki, T.C., and G.R.B. Webster.1986. A comparisonof HPLC analysisof nitrate in soils with
the phenoldisulfonicacid and hydrazinesulfate methods.Can. J. Soil Sci. 66:151-157.
Langmuir, D.,and R.L. Jacobson.1970.Specific-ionelectrodedeterminationof nitrate in somefresh-
watersand sewageeffluents.Environ. Sci. Technol.4:834-838.
Lambert,R.S., and R.J. DuBois. 1971. Spectrophotometric determinationof nitrate in the presenceof
chloride. Anal. Chern.43:955-957.
Lee, Y.-K., J.-T. Park, e.-K. Kim, and K.-J. Whang. 1986. Carbonpastecoatedwire selectiveelec-
trode for nitrate ion. Anal. Chern.58:2101-2103.
Lewis, D.G. 1961. Determinationof inorganicnitrogen in soil. J. Sci. Food Agric. 12:735-742.
Li, L.T. 1968. Determination of nitrate by microdiffusion. (In Chinese.) J. Taiwan Agric. Res.
17:49-50.
Li, S., andK.A. Smith. 1984.The rapid determinationof nitrateat low concentrationsin soil extracts:
Comparisonof ion-selectiveelectrodewith continuous-flow analysis. Commun. Soil Sci.
PlantAnal. 15:1437-1451.
Lowe, R.H., and M.C. Gillespie. 1975.An Escherichia coli strainfor use in nitrate analysis.1. Agric.
Food Chern.23:783-785.
Lubochinsky,B., andI.-P. Zalta. 1954.Microdosagecolorimetriquede I'azoteammoniacal.Bull. Soc.
Chim. BioI. 36:1363-1366.
Mack, A.R., and R.B. Sanderson.1971. Sensitivity of the nitrate-ionmembraneelectrodein various
soil extracts.Can.1. Soil Sci. 51:95-104.
Mahendrappa,M.K. 1969.Determinationof nitratenitrogenin soil extractsusinga specificion activ-
ity electrode.Soil Sci. 108:132-136.
Markus, D.K., I.P. McKinnon, and A.F. Buccafuri. 1985. Automatedanalysisof nitrite, nitrate, and
ammoniumnitrogenin soils. SoilSci. Soc.Am. 1. 49:1208-1215.
McNeilly, B.A., and P.I. Howard. 1973. Limitations on the useof chromotropicacid for determining
nitrate in woodlandsoils. SoilBioI. Biochem.5:689-693.
Milham, PJ. 1970.Potentiometric nitrateanalysis:A flow-through electrodeunit. Analyst (London)
95:758-759.
Milham, P.I., A.S. Awad, R.E. Paull, and I.H. Bull. 1970. Analysis of plants, soils and watersfor
nitrate by using an ion-selectiveelectrode.Analyst (London) 95:751-757.
Mogilevkina, lA. 1964. Fixation of ammoniumin the soil and methodof determiningit. Sov. Soil
Sci. 2:185-196.
Mogilevkina, lA. 1969. Comparisonof methodsof determiningfixed ammoniumin the soil. Sov.
Soil Sci. 2:229-238.
Montgomery,H.A.C., and I.F. Dymock. 1961. The determinationof nitrite in water. Analyst (Lon-
don) 86:414-416.
Morie, G.P.,C.I. Ledford, ande.A. Glover. 1972.Determinationof nitrateandnitrite in mixtureswith
a nitrate ion electrode.Anal. Chim. Acta 60:397-403.
Moyano,A., and I.F. Gallardo. 1988. Fixed ammoniumdeterminationin someclay soils. Commun.
Soil Sci. PlantAnal. 19:225-238.
Mulvaney, R.L. 1986.Comparisonof proceduresfor reducingcross-contamination during steamdis-
tillations in nitrogen-15tracerresearch.Soil Sci. Soc.Am. 1. 50:92-96.
Myers, R.I.K., and E.A. Paul. 1968. Nitrate ion electrodemethodfor soil nitratenitrogendetermina-
tion. Can.1. Soil Sci. 48:369-371.
Nakamura,M. 1980. Resorcinolas fluorimetric reagentfor the determinationof nitrate. Anal. Lett.
13:771-779.
Nakamura,M. 1981. Rapidspectrophotometric determinationof nitratewith 4,5-dihydroxycoumarin.
Nawratil, B., M. Marcantonatos,and D. Monnier. 1974. A spectrophotometric methodfor the deter-
minationof tracesof nitrate.Application to wateranalysis.Anal. Chim. Acta 68:217-221.
Nelson,D.W. 1983. Determinationof ammoniumin KCI extractsof soils by the salicylatemethod.
Commuo.Soil Sci. PlantAnal. 14:1051-1062.
Nelson,D.W., and I.M. Bremner.1966.An evaluationof Mogilevkina'smethodof determiningfixed
ammoniumin soils. Soil Sci. Soc.Am. Proc. 30:409-411.
Nelson,D.W., andI.M. Bremner.1969. Factorsaffectingchemicaltransformationsof nitrite in soils.
Soil BioI. Biochem. 1:229-239.
Nelson,D.W., andI.M. Bremner.1972. Preservationof soil samplesfor inorganicnitrogenanalyses.
Agron.1. 64:196-199.
Nieto, K.F., and W.T. Frankenberger,Ir. 1985a.Single column ion chromatography:l Analysis of
inorganicanionsin soil. Soil Sci. Soc.Am. 1. 49:587-592.
Nieto, K.F., and W.T. Frankenberger,Ir. 1985b.Single column ion chromatography:II. Analysis of
ammonium, alkali metals, and alkaline earth cations in soils. Soil Sci. Soc. Am. 1.
49:592-596.
Nommik, H., and K. Vahtras. 1982. Retentionand fixation of ammoniumand ammoniain soils. p.
123-171.In F.I. Stevensonet al. (ed.) Nitrogen in agriculturalsoils. Agron. Monogr. 22. ASA
and SSSA,Madison,Wl
Norman, R.I., I.e. Edberg,and I.W. Stucki. 1985. Determinationof nitrate in soil extractsby dual-
wavelengthultraviolet spectrophotometry. Soil Sci. Soc. Am. 1. 49:1182-1185.
Norman,R.I., andJ.W. Stucki. 1981.The determinationof nitrate and nitrite in soil extractsby ultra-
violet spectrophotometry. Soil Sci. Soc.Am. 1. 45:347-353.
Nydahl, F. 1976.On the optimumconditionsfor the reductionof nitrateto nitrite by cadmium.Talan-
ta 23:349-357.
1182 MULVANEY
Obrink, KJ. 1955. A modified Conway unit for microdiffusion analysis.Biochem.J. 59:134-136.
0ien, A, and AR. Selmer-Olsen.1969. Nitrate determinationin soil extractswith the nitrate elec-
trode. Analyst (London) 94:888--894.
Olson, R.J. 1980. Phosphateinterference in the cadmium reduction analysis of nitrate. Limnol.
Oceanogr.25:758-760.
Onken,AB., and H.D. Sunderman.1970. Use of the nitrate electrodefor determinationof nitratesin
soil. Commun.Soil Sci. Plant Anal. 1:155-161.
Orion Research,Inc. 1989. Instruction manual. Model 95-12 ammoniaelectrode.Orion Res., Inc.,
Boston,MA.
Orion Research,Inc. 1990a. Instruction manual. Model 93-07 nitrate electrode.Orion Res., Inc.,
Boston, MA.
Orion Research,Inc. 1990b. Instruction manual. Model 95-46 nitrogen oxide electrode.Orion Res.,
Inc., Boston, MA.
Paul,1.L., and R.M. Carlson.1968. Nitrate determinationin plant extractsby the nitrate electrode.J.
Agric. Food Chern. 16:766-768.
Pedrazzini,E, A Castelli, and P. Nannipieri. 1979. Determinationof soil nitrate by meansof specif-
ic ion electrode:Comparisonamongdifferent extractingsolutions. Commun.Soil Sci. Plant
Anal. 10:883--893.
Potterton,S.S.,and W.D. Shults. 1967.An evaluationof the performanceof the nitrate-selectiveelec-
trode. Anal. Lett. 1(2):11-22.
Premi, P.R., and AH. Cornfield. 1967. The use of iron(II) sulphatefor the reduction of nitrate to
ammoniain the microdiffusion methodfor determiningnitrate in soil extracts.Analyst (Lon-
don) 92:196-197.
Puttanna,K., and E.V.S. PrakasaRao. 1981. Elimination of chloride interferencein the phenoldisul-
fonic acid method of nitrate determination in soils. Commun. Soil Sci. Plant Anal.
12:711-718.
Puttanna,K., and E.V.S. PrakasaRao. 1986. Modified methodof nitrite determinationin soils by sul-
phanilic acid/N-(1-naphthyl)ethylenediamine.Z. Pflanzenernaehr. Dung. Bodenkd.
149:517-521.
Qasim,M., and T.H. Flowers. 1989. Errors in the measurementof extractablesoil inorganicnitrogen
causedby impurities in filter papers.Commun.Soil Sci. Plant Anal. 20:747-757.
Qiu, x.-c., G.-P. Liu, and Y.-Q. Zhu. 1987. Determinationof water-solubleammoniumion in soil by
spectrophotometry. Analyst (London) 112:909-911.
Raveh,A 1973. The adaptationof the nitrate-specificelectrodefor soil and plant analysis.Soil Sci.
116:388-389.
Reynders,L., and K. Vlassak. 1981. A rapid and sensitivedeterminationmethodfor soil nitrate sta-
tus. Z. Pflanzenernaehr.Dung. Bodenkd.144:628-636.
Rice, C.W., M.S. Smith, and I.M. Crutchfield. 1984. Inorganic N analysisof soil extractsby auto-
matedand distillation procedures.Commun.Soil Sci. Plant Anal. 15:663--672.
Rider, 8.E, andM.G. Mellon. 1946.Colorimetricdeterminationof nitrites. Ind. Eng. Chern.Anal. Ed.
18:96-99.
Robinson,1.8.0. 1967. The preservationunaltered,of mineral nitrogen in tropical soils and soil
extracts.Plant Soil 27:53--80.
Robinson,J.B.D., M. de V. Allen, and P. Gacoka. 1959. The determinationof soil nitrates with a
brucine reagent.Analyst (London) 84:635--640.
Rodrigues,G. 1954. Fixed ammoniain tropical soils. 1. Soil Sci. 5:264-274.
Roskam,R.T., and D. de Langen.1964.A simple colorimetricmethodfor the determinationof ammo-
nia in seawater.Anal. Chim. Acta 30:56-59.
Rowland,AP. 1983.An automatedmethodfor the determinationof ammonium-Nin ecologicalmate-
rials. Commun.Soil Sci. Plant Anal. 14:49--63.
Rowland, A.P., H.M. Grimshaw,and O.M.H. Rigaba. 1984. Control of soil solution interferencesin
an automatednitrate method.Commun.Soil Sci. Plant Anal. 15:337-351.
Rubio, S., A Gomez-Hens,and M. Va1carcel.1984. Spectrofluorimetricdeterminationof nitrite with
pyridoxal-5-phosphate-2-pyridylhydrazone. Anal. Lett. 17:651--663.
Ruzicka,J., and E.H. Hansen.1988. Flow injection analysis.2nd ed. Wiley-Intersci., New York.
Ruzicka,J., E.H. Hansen, andE.A Zagatto.1977. Flow injection analysisPart VII. Use of ion-selec-
tive electrodesfor rapid analysisof soil extractsand blood serum.Determinationof potassi-
um, sodiumand nitrate. Anal. Chim. Acta 88:1-16.
Saghir, N.S., R.L. Mulvaney, and E Azam. 1993. Determinationof nitrogen by microdiffusion in
Mason jars. I. Inorganic nitrogen in soil extracts. Commun. Soil Sci. Plant Anal.
24:1745-1762.
Sahrawat,K.L. 1982. Error causedby carbondioxide in detenninationof ammoniumby direct steam

distillation of tropical wetland rice soils. Plant Soil 69:283-285.
Sahrawat,K.L., and F.N. Ponnamperuma.1978. Measurementof exchangeableNHt in tropical rice
soils. Soil Sci. Soc.Am. J. 42:282-283.
Schachtschabel,P. 1960. Fixierter Ammoniumstickstoffin LOss- und MarschbOden.p. 22-27. In
Trans. Int. Congr. Soil Sci., Vol. 2, 7th, 1960. ElsevierPubl Co., Amsterdam.
Schachtschabel,P. 1961. Bestimmungdes fixierten Ammoniums im Boden. Z. PfIanzenemaehr.
Dung. Bodenkd.93:125-136.
Schalscha,E.B., T. Schirado,and I. Vergara.1981.Flow injection analysisof nitrate in soil extracts-
Evaluationof a nitrate-selectiveflow electrodemethod.Soil Sci. Soc. Am. J. 45:446-448.
Scheiner,D. 1974. A modified version of the sodium salicylatemethodfor analysisof wastewater
nitrates.Water Res.8:835-840.
Scott, T.J., M.J. Mitchell, A Santos, andP. Destaffen.1989. Comparisonof two methodsfor mea-
suring ammoniumin solution samples.Commun.Soil Sci. Plant Anal. 20:1131-1144.
Searle,P.L. 1984.The Berthelotor indophenolreactionandits usein the analyticalchemistryof nitro-
gen. A review. Analyst (London) 109:549-568.
Selmer-Olsen,AR. 1971. Detenninationof ammoniumin soil extractsby an automatedindophenol
method.Analyst (London) 96:565-568.
Selmer-Olsen,AR., A 0ien, R. Baerug,andI. Lyngstad.1971.Pretreatmentand storageof soil sam-
ples prior to mineral nitrogen detennination.Acta Agric. Scand.21:57-63.
Shinn, M.B. 1941. Colorimetric method for detenninationof nitrite. Ind. Eng. Chern. Anal. Ed.
13:33-35.
Shukla, G.c., and M. Singh. 1968. A new photoelectriccolorimetric method for the estimationof
nitrate nitrogenin soils. J. Ind. Soc.Soil Sci. 16:77-81.
Siegel,R.S. 1980.Detenninationof nitrate andexchangeable ammoniumin soil extractsby an ammo-
nia electrode.Soil Sci. Soc. Am. J. 44:943-947.
Silva, J.A, and J.M. Bremner. 1966. Detenninationand isotope-ratioanalysisof different fonns of
nitrogen in soils: 5. Fixed ammonium.Soil Sci. Soc.Am. Proc. 30:587-594.
v.,
Simeonov, I. Asenov,and V. Diadov. 1977. Rapid detenninationof nitrate nitrogenin soils. Talan-
ta 24:199-200.
Sims, 1.R., and G.D. Jackson.1971. Rapid analysisof soil nitrate with chromotropicacid. Soil Sci.
Soc. Am. Proc. 35:603-606.
Singh, 1.P. 1988. A rapid method for detenninationof nitrate in soil and plant extracts.Plant Soil
110:137-139.
Skeggs,L. T. 1957. An automaticmethodfor colorimetric analysis.Am. 1. Clin. Pathol.28:311-322.
Skjemstad,J.~., and R. Reeve.1978.The automaticdetenninationof ppb levels of ammonia,nitrate
plus nitrite, and phosphatein water in the presenceof addedmercury(II) chloride. J. Environ.
Qual. 7:137-141.
Smith, K.A., and A Scott. 1991. Continuous-flowand discreteanalysis.p. 115-169.In K.A. Smith
(ed.) Soil analysis.Modem instrumentaltechniques.2nd ed. Marcel Dekker, New York.
Snell, ED., and C.T. Snell. 1949. Colorimetric methodsof analysis.Vol. 2. 3rd ed. D. Van Nostrand
Co., New York.
Snyder,L.R. 1980. Continuous-flowanalysis:Presentand future. Anal. Chim. Acta 114:3-18.
Soil ScienceSocietyof America. 1987. Glossaryof soil sciencetenns.SSSA,Madison,WI.
Soloway, S.,and A. Santoro.1955. Detectionof unsubstitutedparaposition in phenols.Anal. Chern.
27:798-800.
Sommerfeldt,T.G., R.A. Milne, and G.C. Kozub. 1971. Use of the nitrate-specificion electrodefor
the detenninationof nitrate nitrogen in surfaceand ground water. Commun.Soil Sci. Plant
Anal. 2:415-420.
Sparrow,S.D., and D.T. Masiak. 1987. Errors in analysesfor ammoniumand nitrate causedby cont-
aminationfrom filter papers.Soil Sci. Soc. Am. J. 51:107-110.
Stanford,G., 1.N. Carter,E.C. Simpson,Jr., and D.E. Schwaninger.1973. Nitrate detenninationby a
modified Conway microdiffusion method.J. Assoc. Off. Anal. Chern.56:1365-1368.
Stein,S.N., L. Li, R.L. Mulvaney, and EW. Simmons.1993. Detenninationof nitrogenby microdif-
fusion in Mason jars: III. Nitrogen and nitrogen-15 in Kjeldahl digests.Commun. Soil Sci.
Plant Anal. 24:2765-2776.
Stevenson,F.J., and A.P.S. Dhariwal. 1959. Distribution of fixed ammoniumin soils. Soil Sci. Soc.
Am. Proc. 23:121-125.
Stewart,B.M. 1987.Ion chromatographicdetenninationof nitrate in 2 M KCI soil extracts.J. Soil Sci.
38:415-419.
1184 MULVANEY
Stock, W.O. 1983.An evaluationof somemanualcolorimetric methodsfor the determinationof inor-

ganic nitrogen in soil extracts.Commun.Soil Sci. Plant Anal. 14:925-936.
Storrier, RR 1966.The pre-treatmentand storageof soil samples fornitrogenanalyses.1. Aust. Inst.
Agric. Sci. 32:106-113.
Swann,M.H., and M.L. Adams. 1956. Rapid colorimetric methodfor nitrates.Anal. Chern.28:1630.
Tabatabai,M.A 1974. Determinationof nitrite in soil extractsand water samplesby a nitrogenoxide
electrode.Commun.Soil Sci. Plant Anal. 5:569-578.
Tabatabai,M.A, and W.A Dick. 1979. Ion chromatographicanalysisof sulfateand nitrate in soils. p.
361-370. In J.D. Mulik and E. Sawicki (ed.) Ion chromatographicanalysisof environmental
pollutants.Vol. 2. Ann Arbor Sci. Publ., Ann Arbor, MI.
Talibudeen,0.1991.Ion-selectiveelectrodes.p. 111-182.In K.A. Smith (ed.) Soil analysis.Modern
instrumentaltechniques.2nd ed. Marcel Dekker, New York.
Tanaka,A., N. Nose,and H. Iwasaki. 1982. Spectrophotometric determinationof nitrate in vegetable
productsusing 2-sec-butylphenol.Analyst (London) 107:190-194.
Tel, D.A, and C. Heseltine.1990.The analysesof KCl soil extractsfor nitrate, nitrite and ammoni-
um using a TRAACS 800 analyzer.Commun.Soil Sci. Plant Anal. 21:1681-1688.
Van Meirvenne, M., and G. Hofman. 1989. Spatial variability of soil nitrate nitrogen after potatoes
and its changeduring winter. Plant Soil 120:103-110.
Van Slyke, D.D., and A Hiller. 1933. Determination of ammonia in blood. J. BioI. Chern.
102:499-504.
Vendrell, P.E, and J. Zupancic.1990. Determinationof soil nitrate by transnitrationof salicylic acid.
Vilsmeier, K. 1984. Bestimmung von Dicyandiamid, Nitrit und Nitrat in Bokenextraktenmit
Hochdruckflussigkeitschromatographie. Z. Pflanzenernaehr.
Dung. Bodenkd.147:264-268.
Wang, L., and A 0ien. 1986. Determinationof Kjeldahl nitrogen and exchangeableammoniumby
the indophenolmethod.Acta Agric. Scand.36:60-70.
Weatherburn,M.W. 1967. Phenol-hypochloritereactionfor determinationof ammonia.Anal. Chern.
39:971-974.
West, P.w., and G.L. Lyles. 1960. A new methodfor the determinationof nitates.Anal. Chim. Acta
23:227-232.
Westfall, D.G., M.A Henson,and E.P. Evans. 1978. The effect of soil samplehandling betweencol-
lection and drying on nitrate concentration.Commun.Soil Sci. Plant Anal. 9:169-185.
White, C.S., andJ.R Gosz. 1981. Organicnitrogen interferencewith automatedammoniumanalyses.
Can. J. For. Res. 11:739-741.
Willis, R.B., and C.E. Gentry. 1987. Automatedmethodfor determiningnitrate and nitrite in water
and soil extracts.Commun.Soil Sci. Plant Anal. 18:625--636.
Wolf, B. 1944. Determinationof nitrate, nitrite, and ammoniumnitrogen. Rapid photometricdeter-
mination in soil and plant extracts.Ind. Eng. Chern.Anal. Ed. 16:446-447.
Wood, E.D., EAJ. Armstrong,and EA. Richards.1967.Determinationof nitrate in seawaterby cad-
mium-copper reductionto nitrite. J. Mar. BioI. Assoc. 47:23-31.
Young, J.L. 1962. Inorganicsoil nitrogenand carbon:nitrogenratios of somePacific Northwestsoils.
Soil Sci. 93:397-404.
Yu, T.R. 1985. Application of ion-selective electrodesin soil science. Ion-Select. Electr. Rev.
7:165-202.
Yuen, S.H., and AG. Pollard. 1953. Determinationof nitrogen in soil and plant materials:Use of
boric acid in the micro-Kjeldahl method.J. Sci. Food Agric. 4:490-496.
Published 1996
Chapter39
Nitrogen-Organic Forms
F. J. STEVENSON,University of Illinois, Urbana, Illinois
INTRODUCTION
Chapter32 (Stevenson,1982a)of the secondedition of ASA publicationMeth-

ods ofSoil Analysis (Pageet aI., 1982) hasbeenmodified and updated.The read-
er is referredto this publicationand Chapters85 (Bremner,1965),96(Stevenson,
1965a),and 97 (Stevenson,1965b)of the first edition (Blacket aI., 1965)for de-
tailed background information,including a historicalaccountof the development
of methodsfor determining organic forms of N in soils. Detailed reviews of
organicN compoundsin soil havebeenprovidedby Stevenson(1982b).
Most studieson the forms of organicN in soils are basedon the use of hot
mineral acids(or bases)to liberatenitrogenousconstituentsfrom organiccolloids
and clay minerals.In a typical procedure,the soil is heatedwith 3 or 6 M hydro-
chloric acid (HCI), after which the N is separatedinto severaldiscretefractions
(Table 39-1). Identifiable organic N compoundsare the amino acids and amino
sugars. The N remainingin the soil residueis usually referredto as acid-insolu-
ble N; that recoveredby distillation with magnesiumoxide (MgO) is ammonia-N
(NH3-N). The nitrogen not accountedfor in the aboveforms is referredto as the
HUN fraction (hydrolyzable unknown N).
In addition to amino acidsand amino sugars,soils containtracequantities
of nucleic acids and other known nitrogenousbiochemicals.However, special-
ized techniquesare required for their separationand identification. Only one-
third to one-halfof the organic N in soils can be accountedfor in known com-
pounds.
The methodfor hydrolyzing the soil has not beenstandardized,and many
variations in hydrolytic conditionshave been employed.The variablesinclude:
(i) type and concentrationof acid, (ii) time and temperatureof hydrolysis, (iii)
ratio of acid to soil, and (iv) pretreatment.In general,hydrolysis is done under
reflux with 6 M HCI for 12 to 24 h.
A large amountof soil N, usually about 25 to 35%, is recoveredas acid-
insoluble N (i.e., N recoveredin soil residue).At one time it was thOUght that this
fraction was an artifact resulting from the condensationof amino acids with
reducing sugarsduring hydrolysis, but it is now believed that some of this N
Book Seriesno. 5.
1185
1186 STEVENSON
Table 39-1. Steamdistillation methodsfor determiningthe variousforms of N in a soil hydrolysate.

Form of N Methodt
Total hydrolyzableN Steamdistillation with NaOH after Kjeldahl digestionwith H2S04 and a
K2S04-catalystmixture
Amino acid-N SteamdistiJIationwith phosphate-borate buffer after treatmentwith
NaOH at 100°Cto removeaminosugarsplus NH4 and with ninhy-
drin (PH 2.5, 100°C)to converta-amino-Nto NHi
Amino sugar-N SteamdistiJIationwith phosphate-borate buffer at pH 11.2; correction
for NH3-N
Ammonia-N SteamdistiJIationwith MgO
Acid-insolubleN Obtainedby difference(total soil N-hydrolyzableN)
HydrolyzableunknownN Obtainedfrom the differencebetweentotal hydrolyzableN and the N
(HUN fraction) accountedfor as (NH3 + amin,! acid + amino sugar)-N
t In eachmethod,the NH3 liberatedby steamdistiJIationis collectedin a H3B03-indicatorsolution
and determinedby titration with standard(0.0025M) H2S04•
occursas a structuralcomponentof humic substances.Chenget at. (1975) found

that the percentageof the soil N recoveredin acid-insolubleforms could be re-
ducedby pretreatingthe soil with hydrofiouric acid (HF) beforeacid hydrolysis.
Freney(1968)and Griffith et at. (1976)showedthat part of the insolubleN could
be dissolvedwith dilute baseand subsequentlysolubilized by acid hydrolysis,
therebyreducingthe acid-insolublefraction to about 10% of the total N.
Another uniquefeatureof soil N fractionationschemesis that a large pro-
portion of the soil N, about 20 to 25% for surfacesoils, is recoveredas NH3 by
distillation with MgO. In the older literature, this form of N was referredto as
the "amide-N" of proteins, but it is now known that very little of the NH3 is
derived from the amino acid amides, asparagine(C4HgN20 3) and glutamine
(CSHlON203). Some of the NH3 is derived from indigenousfIxed NHt, part
comesfrom partial destructionof amino sugars.It also is known that NH3 can
arise from the breakdownof certain amino acidsduring hydrolysis.Tryptophan
(Cll H 12N20 2) is lost completely;others,suchas serine(C3H7N03) and threonine
(C4H9N03) are partially destroyed(Stevenson,1982a).
Methods commonly used for determiningamino acid-N in soil hydroly-
satesare thosebasedon the ninhydrin (CgHJI4) reaction(Fig. 39-1). They in-
clude measurements for the CO2 and NH3 formed throughReactionA and for the
blue-coloredproductformed through ReactionB.
The ninhydrin-C02 methodfor amino acid-N is highly specific in that it
requiresthe presenceof both a COOH and an adjacentNH2 or NH-CH2 group.
This procedureis difficult and time-consumingand requiresmanometricequip-
ment that demandsboth skill and experiencefor successfuloperation. On the
otherhand,the ninhydrin-NH3methoddescribedherein(see"Amino Acid-Nitro-
gen") is relatively simple and doesnot require highly specializedequipment.Of
equalimportanceis the fact that the N is recoveredin a form suitablefor isotopic-
ratio analysis.A colorimetric procedurefor amino acid-N also is describedhere-
in (see"Colorimetric Method for Amino Acid-N").
Two methodsare given for amino sugar-N,an alkaline decompositionpro-
cedurein which the N is recoveredand measured as NH3 (see"Amino Sugar-
Nitrogen") and a colorimetric methodbasedon the Elson-Morganreaction(see
"Colorimetric Method for Amino Sugars").
NITROGEN-ORGANICFORMS 1187
A o
((I
II
pH C, /H
R-CH-COOH --";'---l~~ C + R-CHO + cO 2 + NH3
I <2.5 ~ C/ 'OH
NH2 •
blue-colored product
Fig. 39-1. Reactionsof a-aminoacidswith ninhydrin. ReactionA typifies the reactionat pH 2.5, in
which caseNH3 occursas a stablereactionproduct.When the reactionis carriedout at pH 5 (B),
the NH3 combineswith reducedand oxidizedforms of ninhydrin to form a blue-coloredproduct.
Quantitativedeterminationof individual amino acidsand amino sugarsin

soil hydrolysateshasbeenaccomplishedby applicationof chromatographicpro-
cedure(see review of Stevenson,1982b). This work can be greatly facilitated
through the use of suchtechniquesas high pressureliquid chromatographyand
gaschromatography.
RECOVERY OF VARIOUS FORMS OF NITROGEN AS AMMONIA

BY STEAM-DISTILLATION PROCEDURES
Principles
In this procedure,the different forms of N in a soil hydrolysateare con-

vertedto, andestimatedas,NH3. The only specialapparatusrequiredis the steam
distillation unit describedlater in "SpecialApparatus."Since all forms of N are
recoveredas NH3, the procedurescan be usedin tracerinvestigationsusing 15N_
enrichedcompounds.
The hydrolysis procedureutilized herein is basedon the observationof
Bremner(1965) that maximal releaseof amino acid-N from surfacesoils, and
nearly maximal releaseof total N, was obtainedby hydrolysis under reflux for
about12 h using 3mL of 6 M HCl per gram of soil. Hydrolysis for a longer time
would lead to excessivelyhigh losses of amino sugars,which are partially
destroyedby acid hydrolysis.
Methodsfor determiningthe different forms of N are describedin Table
39-1. All forms are convertedto NH3 (trappedas NHt), which is recoveredby
distillation under alkaline conditions,collected in a boric acid (H3B03)-mixed
indicatorsolution, and determinedby titration with standard(0.0025M) H2S04,
Amino acid-N is determinedby distillation of the NH3 formed by reaction
with ninhydrin at pH 2.5 (Fig. 39-1, ReactionA). Interferencefrom NHt and
1188 STEVENSON
DI water
3' 28115 O-ring

ball Joint
d f)
~flaSk(100mL)
KJeldahl
Sleam-
inlet lube
To waste
o
Electric heating
4 8
em L.LJ
mantle
/
To va riable
transformer
Fig. 39-2. Steam-distillationapparatus.Adapted from Fig. 38-1 (Mulvaney, 1996) and provided
through the courtesyof R.L. Mulvaney.
alkali-labile organic N compounds(e.g., amino sugars)is eliminated by treat-

ment with sodium hydroxide (NaOH, lOO°C) before the ninhydrin reaction is
carriedout.
The method for NH3-N is based on the observation that alkali-labile
nitrogenousconstituents(e.g., amino sugars)are not deaminatedby short-term
distillation with MgO. A separatealiquot of the hydrolysateis distilled with a
phosphate-borate buffer at pH 11.2, yielding (NH3 + amino sugar)-N. Amino
sugar-Nis takenas the differencebetweenthe amountsof NH3 recoveredby the
two distillation procedures.If desired,a separatefraction of the hydrolysatecan
be analyzedfor (serine+ threonine)-N(Bremner,1965) but this is seldomdone.
Method
SpecialApparatus
1. Micro-Kjeldahl digestionunit.
2. Steamdistillation apparatus(Fig. 39-2). Steamfor distillation is gen-
eratedby boiling distilled, deionizedwater in a 5-L flask containinga
few Teflon boiling chips and equippedwith an electric heatingmantle
controlledby a variabletransformer.Before use,the apparatusshould
be steamedout for about10 min, during which time the transformeris
adjustedso that distillate is collectedat the rate of 7 to 8 mL min-I, at
a temperatureno higher than 22°C.
3. Distillation flasks, 50- and lOO-mL Pyrex Kjeldahl flasks with stan-
dard-taper(124/40)ground-glassjoints and fitted with glasshooksso
that they can be connectedto the steamdistillation apparatusby spiral
steel springsas shown in Fig. 39-2.
4. Automatic titrator or microburette(5 mL, graduatedat O.OI-mL inter-
vals).
NITROGEN·ORGANICFORMS 1189
Reagents
1. Hydrochloric acid, approximately6 M: Add 513 mL of concentrated
HCI (sp. gr. 1.19) to about500 mL of water, cool, and dilute to a vol-
ume of 1 L in a volumetric flask.
2. Sodium hydroxide, approximately10 M: Dissolve 2 kg of NaOH in
approximately4 L of deionizedwater, and dilute the cooled solution
to a volume of 5 L. Storein a tightly stopperedbottle.
3. Sodiumhydroxide,approximately5 M: Dilute 500 mL of reagent2 to
1 L, and store in a stopperedbottle.
4. Sodiumhydroxide,approximately0.5 M: Dilute 50 mL of reagent2 to
1 L, and storein a stopperedbottle.
5. Potassiumsulfate (K2S04)-catalystmixture: Mix 100 g of powdered
K 2S04with 1 g of coppersulfate(CUS04• 5H20) and 1 g of Se. Grind
the mixture in a mortar to powderthe cakethat forms during mixing.
7. Boric acid-indicator solution: Prepare as described in Chapter 38
(Mulvaney, 1996).
9. Citric acid: Grind 100 g of reagent-gradecitric acid (C6H80 7 • H20)
in a mortar, and store in a small widemouthbottle.
10. Ninhydrin (triketohydrindenehydrate; indane-trionehydrate):Grind
10 g of reagent-gradeninhydrin in a mortar. Store in a small wide-
mouth bottle.
11. Phosphate-boratebuffer mixture: Mix 100 g of sodium phosphate
(Na3P04• 12H20) with 25 g of borax (Na2B407• lOH20) and grind
to a fine powder. When dissolvedin H20 (1 g per 100 mL), the pH
shouldbe 11.2.
12. Magnesiumoxide, carbonatefree: Prepareas describedin Chapter38
(Mulvaney, 1996).
13. Standard(NHt + amino sugar + amino acid)-N solution: Dissolve
0.189g of (NH4)2S04,0.308g of glucosamine(C6H13NOS)• HCI, and
0.254 of alanine(C3H7N02) in water. Dilute the solution to a volume
of 2 L in a volumetric flask, and mix the solution thoroughly. If pre-
paredfrom pure, dry reagents,this solution contains20 Ilg of NHt -N,
10 Ilg of amino sugar-N,and 20 Ilg of a-aminoacid-N per milliliter.
Storethe solution in a refrigeratorat 4°C.
14. n-Octyl alcohol.
Preparationand Samplingof Soil Hydrolysate

Placea sampleof finely ground«150Ilm) soil containingabout 10 mg of
N in a round-bottomflask fitted with a standard-taper(124/40) ground-glass
joint. Add two dropsof octyl alcohol and 20 mL of 6 M HCI, and swirl the flask
until the acid is thoroughlymixed with the soil. Placethe flask in an electricheat-
ing mantle,connectthe flask to a Liebig condenserfitted with a 124/40ground-
glass joint,and heat the soil-acid mixture so that it boils gently under reflux for
12 h.
1190 STEVENSON
After completion of hydrolysis, wash the reflux condenserwith a small

quantityof distilled water,allow the flask to cool, andremovethe flask from the
condenser.Filter the hydrolysis mixture through a Buchnerfunnel fitted with
Whatmanno. 50 filter paper,usinga suctionfiltration apparatusthat permitscol-
lection of the filtrate in a 200-mL tall-form beakermarkedto indicatea volume
of 60 mL. Washthe residuewith 5- to lO-mL portionsof distilled water until the
filtrate reachesthe 6O-mL mark on the beaker.Immersethe lower half of the
beakerin crushedice (or cool in a refrigeratorto lO°C) and partially neutralize
the hydrolysateby cautiousaddition of 5 M NaOH, using a pH meterto follow
the courseof neutralization.In eachstep,addthe alkalislowly with constantstir-
ring to ensurethat the hydrolysatedoesnot becomealkaline at any stageof the
neutralizationprocess.Transferthe neutralizedhydrolysateby meansof a small
funnel to a l00-mL volumetric flask, and dilute to volume with washings
obtainedby rinsing the beaker,electrodesand stirrer severaltimes with small
quantitiesof distilled water. Stopperthe flask and invert severaltimes to mix the
contents.
In the methodsemployedto determinedifferent forms of N, a 5- to lO-mL
sampleof the neutralizedhydrolysate(room temperature)is pipettedinto a 50-
or lOO-mL distillation flaSK, and after appropriate treatment of the sample,the
flask is connectedto the steamdistillation apparatus(seeFig. 39-2). The form
of N under analysisis determinedfrom the NH3 liberatedby steamdistillation
for 2 to 4 min. To obtain a representativesampleof the hydrolysate,it is essen-
tial to mix the contentsof the volumetricflask thoroughlyby inverting the flask
severaltimes immediately before sampling.It also is necessaryto use pipettes
with wide tips that permit rapid delivery of the sampleto the distillation flask.
Pipetteswith fine tips can causesampling errors, becausethey tend to retain
someof the suspendedmaterialin the sample,and they are easily clogged.
Total HydrolyzableNitrogen
Place5 mL of the neutralizedhydrolysate(see"Preparationand Sampling
of Soil Hydrolysate")in a 50-mL distillation flask, add 0.5 g of K2S04-catalyst
mixture and 2 mL of concentratedH2S04, and heat the flask cautiously in a
micro-Kjeldahl digestion unit until the water is removed and frothing ceases.
Then increasethe heat until the mixture clears,and completethe digestion by
boiling gently for 1 h.
After digestion, allow the flask to cool, and add about 10 mL of water
(slowly and with shaking).Cool the flask undera cold-watertap, and placeit in
a beakercontainingcrushedice. Add 5 mL of boric acid-indicatorsolution to a
100-mLgraduatedbeaker,andplacethe beakerunderthe condenserof the steam
distillation apparatusso that the tip of the condenser is about4 cm abovethe sur-
face of the H3B03• Connectthe cooleddistillation flask to the distillation appa-
ratus,place 10 mL of 10 M NaOH in the entry funnel, and run the alkali slowly
into the distillation flask by raising the funnel plug. When about0.5 mL of alka-
li remainsin the funnel, rinse the funnel rapidly with about5 mL of water, and
allow about 2mL to run into the distillation flask beforesealingthe funnel.
Commence steam distillation by closing the stopcockon the steambypass
tube of the distillation apparatus,and stop the distillation by opening thisstop-
NITROGEN-ORGANIC FORMS 1191
cock when the distillate reachesthe 35-mL mark on the receiverflask (period of
distillation, approximately 4 min). Then rinse the end of the condenser,and
determineNHt -N in the distillate by titration with 0.0025 M H2S04 from an
automatictitrator or microburette(1 mL of 0.0025M H 2S04 =70 fJ,g NHt -N).
The color changeat the end-pointis from greento a faint, permanentpink.
Acid-Insoluble Nitrogen
This form of N is the differencebetweentotal soil N and total hydrolyz-
able N (see "Total Hydrolyzable Nitrogen"). Methods for the determinationof
total soil N are given in Chapter37 (Bremner,1996).
Amino Acid-Nitrogen
Place 5 mL of the hydrolysate(see "Preparationand Sampling of Soil
Hydrolysate")in a 50-mL distillation flask, add 1 mL of 0.5 M NaOH, and heat
the flask in boiling water until the volume of the sampleis reducedto 2 to 3 mL
(approximately20 min). Allow the flask to cool, add 500 mg of citric acid and
100 mg of ninhydrin, and place the flask in a vigorously boiling water bath, so
that its bulb is completely immersedin boiling water. Mter about 1 min, swirl
the flask for a few secondswithout removing it from the bath, and allow it to
remainin the bath for an additional9 min. Then cool the flask, add 1.25 g of the
phosphate-borate buffer mixture and 10 mL of deionizedwater. Finally, add 1
mL of 5 M NaOH and immediately connectthe flask to the steamdistillation
apparatus.Determine the amount of NH3-N liberated by steam distillation as
describedin "Total HydrolyzableNitrogen" (period of distillation, approximate-
ly 4 min).
Ammonia-Nitrogen
Place 10 mL of the hydrolysate(see "Preparationand Sampling of Soil
Hydrolysate") in a 50- or 100-mL distillation flask, add 0.07 ± 0.01 g of MgO,
and connectthe flask to the steamdistillation apparatus.Determinethe amount
of NH3 liberatedby steamdistillation as describedin "Total HydrolyzableNitro-
gen," but discontinuedistillation when the volume of distillate reachesthe 20-
mL mark of the receiverflask (period of distillation, approximately2 min).
(Ammonia + Amino Sugar)-Nitrogen

Place 10 mL of the hydrolysate(see"Total Hydrolyzable Nitrogen") in a
100-mL distillation flask, add 1.25 g of phosphate-borate buffer mixture, and
connectthe flask to the distillation apparatus.Determinethe amountof NHrN
liberated by steam distillation as describedin "Total Hydrolyzable Nitrogen"
(period of distillation, approximately4 min).
Amino Sugar-Nitrogen
This form of N is takenas the differencebetweenthe amountsof N recov-
ered in the precedingtwo sections.
1192 STEVENSON
HydrolyzableUnknown Nitrogen Fraction

This form of N is takenas the differencebetweentotal acid-solubleN and
the N accountedfor as (NH3 + amino acid + amino sugar)-N.
Comments
A completediscussionof the methodsoutlined in this section has been

given by Bremner(1965), and only the major points are coveredherein. Refer-
enceshouldbe madeto Chapters37 (Bremner,1996) and 38 (Mulvaney, 1996)
for additionalinformationconcerningtechniquesfor the determinationof total N
and for the estimationof NH3 by steamdistillation.
The recommendedhydrolysis procedure (see"Preparationand Samplingof
Soil Hydrolysate")can be modified at the discretionof the individual investiga-
tor. However,methodsfor determiningthe variousforms of N were designedfor
analysisof neutralizedhydrolysates,andpoor recoveriesof someforms of N may
result if the quantity of acid usedfor hydrolysis, or the amountof N in the soil
sample,is eitherconsiderablylargeror smallerthan the amountspecified.Accu-
racy also may be affected if recommendationsconcerningsamplevolume for
analysisare not observed(Bremner,1965).
Addition of octyl alcohol to the hydrolysis mixture preventstroublesome
frothing that is somethingencounteredwith calcareous soilsand increasesthe
precisionof resultsfor soils that do not wet readily with acid. Calcareoussoils
shouldbe acidified by cautiousaddition of HCL beforeaddition of the 20 mL of
6MHCl.
It is often expedientto make the hydrolysateto volume before neutraliza-
tion, such as when the hydrolysateis not to be analyzedwithin a day or so.
Unneutralizedhydrolysatecan be storedfor long periodswithout changesdue to
microbial activity. This techniquealso permits replication of the entire analysis
procedure.In this case,neutralize20- or 25-mL aliquotsof the hydrolysate,and
adjustthe volume of eachneutralizedsampleto 50 mL before analysis.
The hydrolysate need not be neutralized for the determinationof total
hydrolyzableN, but bumpingwill be less likely to occur during Kjeldahl diges-
tion. A 30- to 60-min digestionperiod usually is adequate.
The hydrolysismethodusedhereincausesgreaterdecompositionof amino
sugarsthan conventionalhydrolysis proceduresfor amino sugar-N(see"Colori-
metric Method for Amino Sugars'').Not only is hydrolysis carried out for a
longer time, but thesoil-acid mixture is heatedunderreflux ratherthan at 100°C.
The correctionfactor to allow for hydrolysislossesis about1.4 (Bremner,1965).
The variousmethodscan be checkedby analyzingaliquotsof Reagent13.
Suitablealiquots are 5 mL for NH3-N and amino acid-N and 10 mL for amino
sugar-NoThe reagentis stablefor severalmonthsif storedin a refrigerator.
Controls should be included in eachseriesof analysesto allow for any N
derived from the reagents (including those used for preparation of the
hydrolysate).It also is recommendedthat the techniquesfor analysisbe checked
using aliquots of the neutralizedhydrolysatebefore and after addition of known
amountsof NH.t-N [as (NI4)zS04 or NH4Cl), amino sugar-N(as glucosamine-
HCl), and amino acid-N (as alanine).Resultsreportedby Bremner(1965) show
NITROGEN·ORGANIC FORMS 1193
that the methodsdescribedare not subject to interferenceby various inorganic

and organicsubstanceslikely to be presentin neutralizedsoil hydrolysates.
Bremner's(1965) original proceduresfor amino acid-N and NHTN have
been slightly modified in that, for distillation, the phosphate-boratebuffer is
added as a solid rather than as a solution. I (unpublishedobservations)have
observedthat extra ninhydrin is sometimesrequired for complete recovery of
amino acid-N as NHTN. For suspectedproblem soils, a duplicatesampleof the
hydrolysateshouldbe run using 200 mg ratherthan the recommended100 mg of
ninhydrin. The color of the hydrolysis mixture should be observedduring distil-
lation, and any samplegiving evidencefor the formation of a blue-coloredprod-
uct (Fig. 39-1, ReactionB) should be discardedand the analysisrepeated.
Recoveriesof the different forms of soil N are affectedby time of hydrol-
ysis of the soil with 6 M HCI (Yonebayashi& Hattori, 1980). Hydrolysis in an
autoclavehasbeensuggestedas an alternativeto the reflux method(Lowe, 1973).
Duplication of results during amino acid separationswas improved, and auto-
clave hydrolysiswas the more convenient.
COLORIMETRIC METHOD FOR AMINO ACID·NITROGEN
The ninhydrin-NH3 method(see"Amino Acid-Nitrogen") is bestsuitedfor

the analysisof soils and sedimentscontainingrelatively large amountsof amino
acids.Methodsbasedon the colorimetricdeterminationof the blue-coloredprod-
uct obtainedby carrying out the ninhydrin reaction at pH 5.0 (Fig. 39-1, Reac-
tion B) havegreatersensitivity, and they can be usedfor analysisof samplescon-
taining low amountsof amino acids, such as the Band C horizonsof terrestrial
soils and marine sediments(Stevenson& Cheng,1969, 1970).
Principles
The method outlined herein is basedon the observationthat interference

from metal cations(Fe3+, Al3+, Ca2+, etc.) in an acid hydrolysateof soils orsedi-
mentscan be eliminatedby carrying out the reactionin the presenceof a chelat-
ing agent.Ammonium and alkali-labile amino compounds(i.e., amino sugars)are
removedfrom the hydrolysateby an alkali treatmentbeforethe reactionof amino
acids with ninhydrin. Conventionalhydrolysis with 6 M HCI has beenobserved
to give low recoveriesof amino acids from argillaceoussedimentscontaining
small quantitiesof amino acids,and for this reason,an HF pretreatmenthasbeen
incorporatedinto the hydrolysis procedure(Cheng et aI., 1975; Stevenson&
Cheng,1970).
Method
SpecialApparatus
1. Constanttemperatureoil bath (110°C).
2. Freezedryer.
3. Spectrometer.
1194 STEVENSON
Reagents
1. Hydrofluoric acid-hydrochloricacid solution (5 M HF/0.1 M HC1):
Placeabout300 mL of water in a 500-mL polyethylenegraduatedcyl-
inder. Cautiously add86 mL of 49.2% (wt/vol.) HF (approximately29
M) and 4.3 mL of concentratedHC1, and dilute to the 500-mL mark
with water. This reagentshouldbe preparedin a well-ventilatedfume
hood and storedin a polyethylenecontainer.
2. Hydrochloric acid, 6 M: Add 513 mL of concentratedHCl (sp. gr.
1.19) to about500 mL of water, cool, and dilute the solution to 1 Lin
a volumetricflask. For samplescontainingvery low concentrationsof
amino acid-N «30 Ilg of amino acid-N/g) constantlyboiling, redis-
tilled HCl (1:1 HCl • HzO mixture) shouldbe used.
3. Phenolphthaleinindicator, 0.1% ethanolicsolution: Dissolve 100 mg
of phenolphthaleinin 100 mL of 95% ethyl alcohol.
4. Sodiumhydroxide,SM: Prepareas directedin'''Reagents''for "Recov-
ery of Various Forms of Nitrogen or Ammonia by Steam-Distillation
Procedures."
5. Sodium citrate dihydrate (C6HsNa307• 2H zO) solution, 0.4 M: Dis-
solve 177.6 g of sodium citrate dihydrate in about 300 mL distilled
water and dilute to 1 L in a volumetric flask. Add 1 mL of toluene
(C7HS) to protect frommicrobial activity.
6. Sodium acetatebuffer, pH 5.0: Dissolve 500 g of crystalline sodium
acetatetrihydrate(CH3COONa·3H zO) in about400 mL of water, add
100 mL glacial acetic acid (CH3COOH), and dilute to 1 L in a volu-
metric flask.
7. Ninhydrin reagent:Convert 100 g of Dowex-50cation exchangeresin
(Dow ChemicalCo., Midland, MI) (38-75 11m) to the W-form, wash
first with water and then with methyl cellosolve(Kodak) and add to 1
L of a 4% (wt/vol) solution of ninhydrin in methyl cellosolve.Storein
a dark bottle underN z. The reagentis stablefor at least 1 mo.
8. Stannouschloride dihydrate(SnCl z • 2HzO), reagentgrade.
9. Ethyl alcohol (CzHsOH), 50% aqueoussolution (vol/vol): Dilute 1 L
of absoluteethyl alcohol with an equalvolume of distilled water.
10. Standardleucine (C6H 13NO z) solution: Dissolve 0.262 g of leucine in
100 mL of 0.1 M HC1, and dilute the solution to a volume of 1 L in a
volumetric flask, using NHt -free water. If preparedfrom pure, dry
reagent,this solution will contain 141lg of amino acid-N/0.5mL.
Procedure
Placea 50- to 125-mgsampleof finely ground soil or sedimentinto a 50-
mL polypropylenecentrifugetube. Add 2.5 mL of 5 M HF/0.1 M HCI solution,
and shakethe sampleon a reciprocal shakerfor 24 h. For calcareoussamples
acidify with 6 M HCI before addition of HFIHCI. Add 5 mL of distilled water,
and freeze-drythe sampleto removeHE Suspendthe residuein 10 mL of 6 M
HCI, and place the tube in an oil bath at a temperatureof 110°C for 16 h with a
cold-fingercondenserinsertedinto the tube to preventlossof liquid througheva-
NITROGEN·ORGANICFORMS 1195
poration. Removethe tube from the bath, filter the suspensionthrough Whatman
no. 42 filter paper,and washthe residuewith 10 to 15 mL of distilled water. Col·
lect the filtrate and washingsin a second50-mL polypropylenecentrifugetube,
and freeze-dry to remove HCI. Dissolve the residue,in 5 mL of distilled water,
add two to three drops of phenolphthaleinindicator, and titrate with 5 M NaOH
to the pink color of the indicator (about pH 11.0). Placethe tube in an oil bath at
100°C, and after 10 min, passa slow streamof air over the surfaceof the solu-
tion until the volume has beenreducedto about 2 mL. This procedureremoves
NHt and destroysamino sugars.Add 6 M HCl dropwise until metallic hydrox-
ides are dissolved(solution assumesa pale yellow color), and adjust the volume
to 10 mL with NHt -free water.
For colorimetric analysis,place a O.5-mL aliquot of the solution into a 18-
by 160-mmtest tube, add 0.5 mL of sodiumcitrate solution, mix thoroughly,and
add 2 mL of a solution preparedby combiningthe following: (i) 25 mL of sodi-
um acetatebuffer, (ii) 50 mL of ninhydrin reagent,(iii) 25 mL of water, and (iv)
80 mg of SnCl2 • 2H20. Placea capover the tube to preventlossof liquid through
evaporation,and place the tube in a boiling water bath for 30 min. Removethe
samplefrom the bath,cool the solution by immersingthe tube in cold running tap
water, and add5 mL of 50% ethyl alcohol. Finally, obtain optical density at 570
nm in a suitablespectrometer.For samplesthat readabove1.00,dilute with addi-
tional 50% ethyl alcohol. Computethe amountof amino acid-N in the unknown
from a calibrationcurve obtainedby running a seriesof leucine standards.
Comments
Accuratequantitativevaluesare obtainedfor soils containingas little as 20

mg of amino acid-N per gram. The procedureis applicablefor analysisof surface
and subsurfacesoils, marine and lake sediments,peat, and isolatedsoil organic
matterfractions. With slight modification, the colorimetricprocedurecan be used
for the determinationof amino acid N in aqueoussamples,as well as for biomass
N by analysis of ninhydrin-reactiveN compoundsin chloroform-treatedsoil
(Joergensen& Brookes,1990).
The colorimetric method,like the ninhydrin-NH3 method,fails to take into
account lossesof amino acids by acid hydrolysis (see "Introduction"). Losses
during the HF treatmentto destroysilicatemineralswould be expectedto be neg-
ligible under the conditions specified. Some amino acids, such as proline
(CSH9N02) and hydroxyproline (C3H9N03), give low colorimetric equivalents
with ninhydrin (Moore & Stein, 1948). The colorimetric methodusedherein is a
modification of the method by Jacobs(1956) but other colorimetric procedures
basedon the ninhydrin reaction (Fig.39-1, reactionB) shouldwork equally well.
Becauseof the rathersmall samplesize (125 mg), considerablecareshould
be exercisedduring sample preparation.Bulk samplesshould be crushedand
ground to passthrough a 150-l1m (lOO-mesh)screen,and precautionsshould be
taken to prevent samplecontamination.Using the fingers to assistsieving can
result in markedincreasesin the amino acid contentof the experimentalsample.
The freeze-dryingtechniquefor removingHF providesa convenientmeans
of eliminating this acid from the samplewhile at the sametime minimizing chem-
1196 STEVENSON
icallossesof amino acids.The freeze-dryerfor removingexcessacid shouldbe

thoroughly cleaned after use and washed with dilute sodium bicarbonate
(NaHC03) solution.
Errors due to spuriousbackgroundreadingsfrom mineral matter solubi-
lized during extractionappearnegligiblefor samplescontaining>70 Ilg of amino
acid-N per gram. Correctionscan be made,if necessary,by running a duplicate
samplethrough the entire extraction procedurebut with inclusion of a nitrous
acid (HN03) treatmentto destroyamino acids(Stevenson& Cheng,1970).
COLORIMETRIC METHOD FORAMINO SUGARS

Methodsfor the determinationof amino sugarsin soil include alkaline de-
composition(see"Amino Sugar-Nitrogen")and the classicalcolorimetricproce-
dure of Elson and Morgan (1933).A limitation of the alkaline decomposition
method is that amino sugar-N is calculatedfrom the difference betweenthe
amount of NH3 recovered by two distillation procedures.Accordingly, the
method is unsuitablefor analysisof samplescontaining low concentrationsof
amino sugarsin the presenceof relatively high concentrationsof total hydrolyz-
able NH3 (e.g., many subsurfacesoils).
Principles
With the Elson-Morganmethod and its modifications,the sampleis first
heatedwith an alkalinesolution of acetylacetone(CSHS0 2), followed by addition
of an acid, alcoholicsolutionofp-dimethylaminobenzaldehyde (4HllNO) (Ehr-
lich's reagent).A chromogenis formed by the first reaction,following which
addition of the aldehydeproducesa red solution.
Many substances,including Fe and mixtures of amino acids and sugars,
producecolorsthat interferewith the determinationof aminosugarsby the above
method.In soil hydrolysates,dark-coloredhumic materialsalso interfere.
The substancesin soil hydrolysatesthat interferewith color formation by
this method are removedby treating the hydrolysatewith an anion exchange
resin, followed by a cation exchangeresin. The anion exchangeresin retainsthe
dark-coloredhumic materialsaswell as Fe andAI. Thesecationsare retainedon
the resin bed, becausethey are precipitatedwhen the acid becomesneutralized.
The amino sugarsare then recoveredby elution with a 0.02 M solution of
NaHC03, which also servesto preventpeptizationof Fe andAI.
An aliquot of the effluent from the anionexchangeresinis thenpassedonto
a cation exchangeresin. Amino sugarsand amino acidsare retained,but neutral
sugarspassthrough.The amino sugarsare then recoveredby elution with 2 M
HCI, and their amountsare determined(Stevenson,1957).
Method
SpecialApparatus
1. Anion resin column: Wash the anion exchangeresin [Amberlite IRA-
400 (Rohm and HassCorp., Philadelphia,PA) or equivalent,150--300
mm or 50- to 100-mesh]sequentiallywith 2 M NaOH, 2 M HCI, water,

0.5 M Na2C03,and water. Preparea slurry of the resin in water, and
pour it into a 2.5- by 20-cmglasstube with the lower end taperedto an
openingof 1 to 2 mm and providedwith a filter madefrom a plug of
Pyrex glass wool. Adjust the height of the resin bed to 10 cm by
removing excessresin from the top of the bed.
2. Cation resin column: Wash the cation exchangeresin (Dowex-50 or
equivalent,38-75 J..Lm or 200-400mesh)sequentiallywith 2 M NaOH,
2 M HCI, and water. Repeatthe washingcycle severaltimes. Prepare
a slurry of the resin in distilled water, and pour it into a 1.0 by 15 cm
chromatographiccolumn fitted with a fritted glassdisc of coarsepor-
osity. Adjust the height of the resin bed to 2.5 cm by removing excess
resin from the top of the bed.
3. Flashevaporator.
Reagents
1. Anion exchangeresin (Amberlite IRA-400 or equivalent,150-300J..Lm
or 50-100mesh).
2. Cation exchangeresin (Dowex-50 or equivalent,38-75 I1ID or 200-
400 mesh).
4. Sodiumhydroxide,approximately2 M: Dissolve 80 g of NaOH in 1 L
of distilled water.
5. Hydrochloric acid, approximately2 M: Add 171 mL of concentrated
HCI (sp. gr. 1.19) to about 300 mL of distilled water, cool, and dilute
to 1 L in a volumetric flask.
6. Sodiumcarbonate(Na2C03),0.5 M: Dissolve53.0 g of Na2C03in 1 L
of distilled water.
7. Sodiumbicarbonate(NaHC03), 0.5 M: Dissolve53.0 g of NaHC03 in
1 L of distilled water.
8. Acetic acid, approximately1.0 M. Dilute 56.9 mL of glacial aceticacid
(approximately17.6 N molarity) to 1 L with distilled water.
9. Hydrochloric acid, 0.03 M. Dilute 15 mL of reagent5 to 1 L with dis-
tilled water.
10. Acetylacetonereagent:Dilute 1 mL of acetylacetoneto 50 mL with 0.5
MNa2C03'
11. Ethyl alcohol, absolute.
12. Ehrlich's reagent:Dissolve2.67 g of p-dimethylaminobenzaldehyde
in
a mixture of 50 mL of absolute ethylalcohol and 50 mL of concen-
trated HCI.
Procedure
Placea 5- to lO-g sampleof soil in a 200-mL round-bottomflask, add 50
mL of concentratedHCI, stopperthe flask, and allow it to standfor 60 h at room
temperature,with occasionalshaking. Add 50 mL of distilled water, attach the
flask to a reflux condenser,and heat in a constanttemperaturebath at 100°C for
1198 STEVENSON
6 h. Cool the solution, filter it througha Buchnerfunnel fitted with Whatmanno.

50 filter paper,washthe residuethoroughlywith distilled water, and combinethe
filtrate andwashings.RemoveHCI by evaporatingthe solution to neardrynessin
a flash evaporatorat 40°C, Add 10 mL of distilled water, and evaporate thesolu-
tion to drynessagain.Dissolvethe residuein distilled water, and dilute the solu-
tion to 25 mL.
Add an aliquot of the hydrolysate(estimatedto contain5 mg of amino sug-
ars) to the anion exchangecolumn. Agitate the upper 3 to 5 cm of the resin bed
vigorously with a glassstirring rod until the HCI in the sampleis neutralized,as
evidencedby cessationof CO2 evolution. Allow the hydrolysateto flow slowly
throughthe resin, and then rinse the sidesof the column several timeswith small
portions of 0.02 M NaHC03 solution. Do not allow the surfaceof the resin to
becomedry, becausebubblesof air may becometrappedin the resin bed. Fill the
tube with 0.2 M NaHC03 solution., and attachthe column to an overheadsupply
of 0.02M NaHC03 solution. Collect 180 mL of effluent in a 200-mL volumetric
flask containing5 mL of 1.0 M aceticacid, and adjustthe volume to 200 mL with
distilled water.
Add an aliquot of the effluent from the anion resin column (estimatedto
containfrom 0.1-0.2mg of amino sugars)to the cation resin column. Allow the
solution to drain completelyinto the resin, and then passthrough 10 mL of dis-
tilled water. Discard the effluent and washings.Elute the column with 2 M HCI,
and collect 10 mL of effluent in a volumetric flask.
Pipettea 1.0-mL aliquot of the effluent from the cation resin column into a
10-mL, glass-stoppered, volumetricflask. Placethe flask in a vacuum desiccator
over KOH, evaporatethe solution to dryness,and then dissolvethe residuein 1
mL of 0.03 M HCl. This solution containsthe amountof HCI calculatedto give
a pH ranging from 9.6 to 9.8 during acetylation. Add 1 mL of acetylacetone
reagent,suspendthe flask in a water bath maintainedat 89 to 92°C, and after
about2 min stopperthe flask. After heatingfor 45 min, removethe flask, add 2.5
mL of absoluteethyl alcohol (C2H50H), and then add 1 mL of Ehrlich's reagent.
Adjust the volume to the mark with ethyl alcohol,allow the flask to standat 30°C
for 1 h, and transferthe solutionto a spectrometertube. Readabsorbanceor trans-
missionat a wavelengthof 530 nm using a suitablespectrometer.
Calculations and Interpretation

Computethe amount of amino sugarsin the unknown from a calibration
curve obtainedby running a seriesof glucosaminestandardsin the sameway and
at the sametime as the unknown. Multiply the result by the factor 1.14 to adjust
for lossesof amino sugarsduring hydrolysis (Stevenson,1957). A different fac-
tor will be requiredif hydrolysis is performedunder conditionsother than those
specified (see "Comments"for "Recovery of Various Forms of Nitrogen and
Ammonia by SteamDistillation Procedures").The extentto which amino sugars
in soils are degradedduring hydrolysis is unknown, and thus the use of the cor-
rection factor is of uncertainvalidity.
Comments
With this method,as little as 0.1 mg of aminosugarper gram of soil can be
detected.By reducingthe size of the anion exchangecolumn, or by using a larg-
er volumeof effluent from the cationexchangeresin for the color test,the amount
of amino sugarsrequired can be reducedconsiderably.If many determinations
are to be carried out simultaneously,a rack provided with slots from which a
seriesof lO-mL volumetric flasks can be suspendedis recommended.The reac-
tion also can be carriedout in closedtubesor in tubesprovidedwith cold-finger
condensers.
The cationexchangeresin columncanbe usedindefinitely. Regenerationis
accomplishedby eluting the resin with 15 mL of 2 M NaOH, followed by 15 mL
of 2 M HCl. Before the column is usedagain,acid is permittedto run throughto
removethe small amountof resin that ordinarily goesinto solution on standing.
The resin is then washedwith distilled water to removeHCl. A new anion ex-
changeresin column must be preparedfor eachdetermination.
The colorimetric analysisof amino sugarsin the effluentfrom the cation
resin column can be performedwithout prior removal of HCl. Boas (1953) and
Stevenson(1957) describeproceduresfor neutralizing the acid prior to colori-
metric assayby the Elson-Morganmethod.
Several factors are of importance in the colorimetric determinationof
amino sugarsby the aboveprocedure.Variations in the conditionsduring acety-
lation appearto be of greaterimportancein color formation than incubationwith
Ehrlich's reagent.The temperatureduring acetylationmust be constant,and the
pH shouldbe between9.6 and 9.8 (Schloss,1951).The temperatureduring acety-
lation can be higher than 89 to 92°C, but the sensitivity will be decreased(Boas,
1953). Colorless acetylacetone shouldbe used,and redistillation in vacuoof the
commercialproductis recommended.
Stevenson(1957) pointed out that interferencefrom mixtures of amino
acidsandsugarsin the effluent from the anion resin column is negligible and that
for many soils, treatmentwith a cation exchangeresin is unnecessary.In caseof
direct analysis of the effluent from the anion resin column, a 0.5- to 2.0-mL
aliquot of the effluent is used.
Recoveryof amino sugarsfrom the anion exchangeresin is 100 ± 1.8%,
recoveryfrom the cation resin column is 97 ± 3.0%. The coefficient of variation
for measurementwithin the rangeof 1.0 to 4.0 mg of amino sugarsper gram of
soil is about5%.
Specific instructionscannotbe given for the hydrolysis of soils for amino
sugaranalysis,becausesoils differ in the easewith which amino sugarscan be
separatedfrom the mineral matter(Stevenson,1957).Also aminopolysaccharides
differ in their resistanceto acid hydrolysis. For the most accuratework, hydroly-
sis curvesshouldbe preparedfor eachtype of soil examined,using different con-
centrationsof HCl. The conditions best suited for releaseof amino sugarsare
determinedby plotting the amountof aminosugarsliberatedagainsttime. Amino
sugars are partly destroyedby acid hydrolysis; hence, the amount recovered
underany given set of conditionsrepresentsa balancebetweenamino sugarlib-
1200 STEVENSON
eration and destruction.For clay-rich samplescontaininglow amountsof amino

sugars,an HF pretreatmentmay be required.
My unpublishedwork indicatesthat amino sugarsin desaltedhydrolysates
of soil can also be determinedby the indole-Hel colorimetric procedureof Dis-
che and Borenfreund(1950).
REFERENCES
Black, C.A, D.O. Evans,J.L. White, L.E. Ensminger,and EE. Clark. 1965. Methodsof soil analysis.
Part 2. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Boas, N.E 1953. Method for the determination of hexosaminesin tissues. J. BioI. Chern.
204:553-563.
Bremner,J.M. 1965. Organic forms of soil nitrogen. p. 1238-1255.In C.A Black et al. (ed.) Meth-
ods of soil analysis.Part 2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Bremner, J.M.1996. Nitrogen-total.p. 1085-1121.In D.L. Sparkset al. (ed.) Methodsof soil analy-
sis. Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Cheng,C.-N., R.C. Shufeldt, and FJ. Stevenson.1975. Amino acid analysisof soils and sedimnets:
Extraction and desalting.Soil BioI. Biochem.7:143-151.
Dische, Z., and E. Borenfreund.1950. A spectrophotometricmethod for the microdeterminationof
hexosamines.J. BioI. Chern. 184:517-528.
Elson, L.A, and W.TJ. Morgan. 1933. A colorimetric methodfor the determinationof glucosamine
and chondrosamine.Biochem.J. 27:1824-1828.
Freney,J.R. 1968. The extraction and partial characterizationof non-hydrolyzablenitrogen in soil.
Trans. Int. Congr. Soil Sci. 9th. (Adelaide)3:531-539.
Griffith, S.M., F.J. Sowden,and M. Schnitzer.1976.The alkaline hydrolysisof acid-resistantsoil and
humic acid residues.Soil BioI. Biochem.8:529-531.
Jacobs,S. 1956. An improved methodfor the quantitativedeterminationof amino acid by meansof
indanetrionehydrate.Analyst (London) 81:502-503.
Joergensen,R.G., and P.C. Brookes. 1990. Ninhydrin-reactivenitrogen measurements of microbial
biomassin 0.5M K 2S04 soil extracts.Soil BioI. Biochem. 22:1023-1027.
Lowe, L.E. 1973. Amino acid distribution in forest humuslayers in British Columbia.Soil Sci. Soc.
Am. Proc. 37:569-572.
Moore, S., and W.H. Stein. 1948. Photometricninhydrin method for use in the chromatographyof
amino acids.J. BioI. Chern. 176:367-388.
Mulvaney, R.L. 1996. Nitrogen-inorganicforms. p. 1123-1184.In D.L. Sparkset al. (ed.) Methods
of soil analysis.Part 3. Chemicalmethods.SSSABook Ser.5. SSSAand ASA, Madison,WI.
Page,AL., R.H.Miller, and D.R. Keeney.(ed.). 1982.Methodsof soil analysis.Part 2. 2nd ed. Agron.
Monogr. 9, ASA and SSSA,Madison,WI.
Schloss,B. 1951. Colorimetric determinationof glucosamine.Anal. Chern. 23:1321-1325.
Stevenson,EJ. 1957. Investigationof aminopolysaccharides in soils: I. Colorimetricdeterminationof
hexosaminesin soil hydrolysates.Soil Sci. 83:113-122.
Stevenson,FJ. 1965a.Amino sugars.p. 1429-1436.In CA Black et al. (ed.) Methodsof soil analy-
sis. Part 2. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Stevenson,FJ. 1965b.Amino acids. p. 1437-1451.In C.A Black et al. (ed.) Methodsof soil analy-
sis. Part 2. Agron. Monogr. 9. ASA and SSSA,Madison,WI.
Stevenson,FJ. 1982a.Nitrogen-organicforms. p. 625-{)41.ln AL. Pageet al. (ed.) Methodsof soil
Stevenson,F.J. 1982b.Organicforms of soil nitrogen. p. 67-122.In EJ. Stevenson(ed.) Nitrogen in
agricultural soils. Agron. Monogr. 22. ASA, CSSA, and SSSA, Madison,WI.
Stevenson,EJ., and C.-N. Cheng.1969. Amino acid levels in the Argentine Basin sediments:Corre-
lation with Quaternaryclimatic changes.J. Sediment.Petrol. 39:345-349.
Stevenson,EJ., and C.-N. Cheng.1970. Amino acids in sediments:Recoveryby acid hydrolysisand
quantitativeestimationby a colorimetric procedure.Geochim.Cosmochim.Acta 34:77-88.
Yonebayashi,K., and T. Hattori. 1980. Improvementsin the method for fractional determinationof
soil organic nitrogen.Soil Sci. Plant Nutr. 26:469-481.
Published 1996
Chapter40
Cation Exchange Capacity and Exchange

Coefficients
M. E. SUMNER AND W. P. MILLER, University of Georgia,
Athens, Georgia
INTRODUCTION
Origin of Chargeson Soil Particles

The origin of cation exchangecapacity (CEC)lies in the negativechargescarried
by soil particles,usually clay, organicmatterand sesquioxides.A full discussion
of the origin and natureof thesechargesis presentedin Chapter41 (Zelazny et
aI., 1996). Basically thesechargesfall into two distinct categories,being either
permanentor variable (i.e.,pH dependent)dependingon whetheror not ambient
conditions(pH or salts) in the soil solution affect their magnitude.Much confu-
sion in the literature concerningthe measurementand interpretationof CEC has
stemmedfrom the lack of recognitionthat thesechargesfall into two distinct cat-
egoriesexhibiting different behavior. These problemswill be addressedin the
discussionof the methodsfor the determinationof CEC.
OperationalDefinition of Cation ExchangeCapacity

Until relatively recently,CEC was defined in rather simple terms in ele-
mentarytexts as"the sum total of the exchangeable cationsthat a soil can absorb"
(sic) (Brady, 1984)or "the capacityof a soil to hold cations"(Jones,1982),which
doesnot recognizethe importanceof the procedureusedin determiningthe out-
come of the CEC measurement.The negativechargeson soil particles are neu-
tralized by an excessof cations and a deficit of anions (negativeadsorptionor
anion repulsion) over that which would be presentin the absenceof the solid
phase.This essentiallymeansthat the major part of the negativechargeattracts
cationsfor neutralizationand the remainderis involved in repelling anions,which
is equivalentto attractingcations.Thus CEC should be defined as
CEC =Mt~cess + Ab-;;ficit [1]
Book Seriesno. 5.
1201
1202 SUMNER & MILLER
where MX+ and AX- are the cations and anions in the system, expressedon a
chargebasis(Le., cmolekg-1 soil for cationsretained,cmola kg-1 for anions).For-
tunately the magnitudeof the anion deficit is quite small in many cases,varying
from 5 to 20% of the CEC at an equilibrium electrolyteconcentrationof 0.1 M to
0.5 to 2% at 0.001 M, and thus can be and often is ignored. In addition, all soils
contain salts in the soil solution in equilibrium with the exchangesites. During
the determinationof CEC, thesesaltsare extractedwith the exchangeable cations
and can result in errors. Thus M~teess in the systemwould be equal to the total
MX+ extractedless the product of the moisturecontentand equilibrium concen-
tration of the soil solution with respectto MX+. In systemslow in salt, thiscor-
rection is very small and often can be neglected.To be unambiguousin the defi-
nition of CEC, one should statethat it is the total chargeexcessof cationsover
anions in the system.Although this appearssimple, many complicationsarise
during measurement.
The discussionso far has assumedthat the chargeson the soil particlesare
not affectedby conditionsin the ambientsolution. From Chapter41 (Zelazny et
aI., 1996) on charging,the variable chargecomponentis very definitely altered
by both the concentrationand valenceof the ions in the equilibrium solution. In
addition, specific adsorptionof both cationsand anionscan have markedeffects
on CEC. It thereforebecomesclear that the CEC of a soil dependsrather criti-
cally on the mannerand conditionsin which it is determined.Consequently,CEC
always must be operationallydefined in terms of variablessuch as pH, concen-
tration, nature and valenceof the ions and buffer capacityof the extractingsalt
solution, the nature of the washing liquid to removeentrainedsalt solution and
the temperature.In addition, the purposefor which the CEC is being determined
must be taken into account. For example, the method selected dependson
whetherone seeksa value at the "field" pH and electrolyteconcentrationof the
soil, or a value reflecting the conditionsafter the soil hasbeenlimed to pH 7.
Historical Review of Cation ExchangeCapacityMethods
Following Thompson's(1850) discoveryof the cation exchangephenome-

non, Way (1850, 1852) developedmethodologywhich demonstratedboth the
permanentand variable chargecharacterof soils. But it was not until 1887 that
the first quantitativemethodfor its measurementwas developed(Kelley, 1948)
involving extractionwith saturatedNH4CI by repeateddecantationand estima-
tion of the CEC as the loss of NUt from the solution; this methodproved to be
subjectto substantialerror. In nonsalinesoils, the sum of cationsin the leachate
would give the CEC. This value, including exchangeableAl3+ determinedby an
unbufferedsalt solution, later becameknown as the Effective Cation Exchange
Capacity(ECEC). Subsequentimprovementsfeaturedleachingwith 1 M NH4CI
and washing,initially with waterand later with methyl (CHzOH) or ethyl alcohol
(CzHsOH) to remove the excessNH4CI entrainedin the soil prior to estimation
of NHt adsorbed(Kelley & Brown, 1924, p. 1-34). Although not realizedat the
time, the use of an unbufferedsalt in this methodologywas appropriatefor the
estimationof CEC at "field" pH, which is particularly importantin highly weath-
ered acid soils as we shall seelater. At this stage,the problemspresentedby the
CATION EXCHANGE CAPACITY & EXCHANGE COEFFICIENTS 1203
presenceof salts, both soluble and insoluble, in the stoichiometry of cation

exchangewere known.
Schollenberger& Dreibelbis(1930) drew attentionto the so-calledadvan-
tages of using a buffered salt such as CH3COONH4 (ammonium acetate,
NH40Ac) at pH 7 and washingwith neutralethanolratherthan water to prevent
"hydrolysis" of part of the NHt adsorbed(Kelley, 1948; Schollenberger&
Simons, 1945). In hindsight, this was a particularly unfortunate selection of
extracting solution for acid, highly weatheredsoils becausethe buffered salt
automaticallyraisesthe pH of the soil to 7, greatly inflating the CEC likely to
exist in the soil underfield conditions.The reasonfor the selectionof pH 7 and
a buffered salt was probably an accidentof location, as at this time most of the
work on CEC was being conductedin westernEurope,Russiaand the western
and midwesternUSA, which were the most important areasof agricultural pro-
duction and where the soils are generallyneutral to alkaline in reaction.In addi-
tion, the natureof soil acidity was not clearly understoodevenas late as 1948,as
illustrated by the statementsof Kelley (1948): "the exchangecapacity of a soil
dependson the pH of the solution usedin the determination,"and "is now wide-
ly usedin America as a measureof the CEC of acid, neutral and alkaline soils,"
and "is especiallywell adaptedto this determination."Unfortunately, many of
thesesentimentspersistevento this day, and the 1 M NH40Ac (pH 7) methodis
still widely used even on highly weathered,variable charge,acid soils, under
which conditions, it gives highly inflated values for CEC (Grove et aI., 1982;
Gillman & Hallman, 1988). Despite the fact that it was selectedas a standard
methodfor the classificationof soils (Soil Survey Staff, 1975), the continuing
useof this method onacid soilsis inappropriatefor a clearunderstandingof their
exchangechemistry.An excellentparallel discussionof how the views on soil
acidity developedduring this period is presentedby Thomas (1977), which
should be consultedfor an appreciationof their impact on CEC estimation.It is
interestingto speculatewhat course CECmethoddevelopmentmight have taken
had the work been conductedin the humic tropical areas of Latin America,
Mrica and Asia where the soils are generallyhighly acid.
At this stagein the developmentof CEC methods,there seemedto be a
greaterrealization of the potential for underestimationof the CEC using 1 M
NH40Ac (pH 7) on alkaline soils than for its grossoverestimationin the caseof
acid soils (Kelley, 1948). Again, this seemsto be a quirk of location in that Kel-
ley conductedmost of his work in the westernUSA where many soils are alka-
line. During this period, many workersdevelopedvariationsof the methodology
for useon calcareoussoils to reducethe effect of CaC03 solubility in the extract-
ing solutionon the estimationof exchangeable Ca2+ with the mostsuccessfuland
widely adoptedtechniquebeingdevelopedby Mehlich (1942). This involved the
useof 0.1 M BaCl2 bufferedto pH 8.2 with triethanolamine(C6H1SN03) (TEA),
in which CaC03 is practically insoluble. Becausethe Ba2+ adsorbedby the soil
is a measureof its CEC, the presenceof SOa-in the soil resultsin inflated val-
ues. Bascomb(1964) modified this approachto incorporate the "compulsive
exchange"principle to replacethe adsorbedBa2+ efficiently with MgS04. Unfor-
tunatelyMehlich (1945) recommendedthe useof the BaCITTEA method onacid
soils using a back titration of the leachateto estimateexchangeableH+, and this
haslead to much confusionamongusersnot entirely familiar with the principles

involved. Most of the W replacedby BaCI2-TEA do not exhibit acid character
at the "field" soil pH, and are merely a reflection of the magnitudeof the poten-
tial variable chargeon the soil. During this period, basesaturation([ exchange-
able cations/CEqx 100) becamea much-usedparameterfor assessingthe acid
characterof soils. With our current knowledgeof the variable chargecharacter
of many acid soils, it becomesclear why the useof CEC valuesobtainedby the
1 M N~OAc (PH 7) and BaCI2-TEA methodsresultedin unrealisticallylow val-
uesfor basesaturation.
In the caseof soils containingsolublesalts,no specialproblemswere pre-
sentedin terms of measuringCEC by a replacingcation provided that the salts
were removed in the process.However, it was usually impossible to arrive
unequivocallyat the CEC of such soils by summationof exchangeablecations
(Kelley, 1948).
The next step forward was taken by Schofield(1939), who revisited vari-
able chargeson soil particlesand developeda method for the measurementof
positive and negativechargesat different pH valuesusing a saturatingsolution
of 0.2 M NH4CI, weighing the entrainedexcessfor which correctionwas made
and replacingwith 0.2 M KN03 (Schofield,1949). Subsequently,Sumner(1963)
comparedthe effectsof correctingfor the entrainedsolution and its removal by
washingwith ethyl alcohol (CH30H)/watermixtureson the CEC obtained.The
acid soils studiedlost adsorbedcation and anion in equivalentamountsas wash-
ing proceeded,indicating that as the ionic strengthdecreased,positive and neg-
ative chargeswere mutually neutralizingeachother. Becausea low electrolyte
concentrationis likely to be the normal situation in soils underfield conditions,
this chargeneutralizationwill lead to a more realistic measureof the actualCEC
of the soil. Unfortunately, this washingprocedureresults in a final electrolyte
concentrationwhich is unknownand may vary considerably.In addition, thereis
always the possibility that some of the exchangeablecations might be
hydrolyzedfrom the exchangesites as the electrolyteconcentrationapproaches
zero. Nevertheless,this washingstepwith water was shown to measurethe net
chargeon the soil which, for most soils, correspondsto the ECEC as originally
proposedby Colemanet al. (1959) as the sum of exchangeableCa2+, Mg2+, K+,
Na+, andAl3+ (Groveet aI., 1982).The ECECapproachis not valid for soils con-
taining appreciablelevels of salts. In such casesit is impossibleto distinguish
unambiguouslybetweenthe cations of the salt and thosewhich are exchange-
able.
The above work set the stage for the developmentof cation exchange
methodswhich, in addition to consideringthe effectsof buffering and pH on the
results, also recognizedthe profound effect of the eqUilibrium electrolytecon-
centration.The first methodwas developed byGillman (1979) and considerably
improved by Gillman & Sumpter (1986a). It is based on the "compulsive
exchange"principle originally enunciatedby Bascomb(1964). Basically,the soil
is saturatedwith a 0.2 M BaCl2 solution followed by reductionof the electrolyte
concentrationto 0.002 M BaCl2 (I = 0.006 M), estimation of the amount of
entrainedsolution and then compulsiveexchangewith 0.005 M MgS04, which
resultsin Ba2+ being quantitativelyreplacedby Mg2+ on the exchangesites.The
CEC is calculatedby differenceafter measuringMg2+ remainingin solution after

the compulsiveexchangeand correctingfor the Ba2+ in the entrainedsolution.
The value obtainedis called the CECCE and is equalto the capacityof the soil to
retain basiccations(CECB) measuredby Ca2+ in soils not containingAl3+ (Gill-
man & Hallman, 1988). Under acid conditions when Al3+ is present,the total
CEC (CEC T) is calculatedas CECB + Al3+. It also is possibleto measurecorre-
spondingvaluesfor anion exchangecapacities(AEC B andAECcE) by measuring
the adsorption of Cl·. In principle, this compulsive exchangemethod should
prove to be universally applicableto all soils provided an appropriateequilibri-
um ionic strengthis selected.
Criteria for Selectionof Cation ExchangeCapacityMethodology
Today it is clearly understoodthat CEC is operationallydefined and tai-

lored to the purposeto which the resultsare to be applied and to the peculiarities
of various catagoriesof soils (Bache,1976; Rhoades,1982; Uehara& Gillman,
1981).To facilitate the following discussion,the criteria for methodologyselec-
tion will be divided on a final use basis.
Characterizationof Soils in "Field" State

From the point of view of describingprocessesin field soils that may be
affected by CEC, there is little doubt that the best approachis to use methods
involving unbuffered extracting solutions so that the CEC is measuredat the
"field" pH value of the soil. Methodsinvolving water/alcoholwashesto remove
entrainedelectrolyteare acceptable,but thosewhich control and correctfor the
ionic strengthof the final equilibrium solution are likely to give more meaning-
ful results.Dependingon the soil,the ionic strengthof the final equilibrium solu-
tion should be adjustedto approximatethat of the naturalsoil solution. For soils
which are at or above pH 7 or contain little variable chargematerial, appropri-
ately buffered extracting solutionscan be used with water/alcoholwashing. In
soils which do not containappreciablelevelsof soluble saltsor free CaC03, CEC
can be estimatedas the sum of exchangeableCa2+, Mg2+, K+, Na+, and Al3+,
expressedin moles of cation chargeper soil mass(e.g., cmoi.: kg-1 soil).
Soil Classification
If the objective is to compareCEC valuesof soils as an aid to soil series
classification,a strong casecan be madefor conductingthe measurements at a
standardpH value. If this is not done, anthropogeniceffectssuch as liming and
fertilization could causethe samesoil to havewidely different CEC values,par-
ticularly if it containsappreciableamountsof variablechargesurfaces.It should
be realizedthat the measurement of the CEC valuesof acid variablechargesoils
at a standardneutralto alkaline pH value will result in valuesvery different from
thoseextantin the field. In suchcircumstances,a strongcasecould bemadefor
measuringthe CEC at both the standardand field pH values.
Having optedto measureCEC at a standardpH, its value must be decided.
Good argumentscan be madein favor of both the commonly usedpH 7 and 8.2
standardswhich wereselectedbasedon considerationsof the equilibria between

CO2 and free CaC03. The pH of a fully basesaturatedsoil in equilibrium with
atmosphericCO2 and excessCaC03 is 8.2, whereasthat of a biologically active
topsoil in equilibrium with a higher CO2 content(1%) and free CaC03 is pH 7.3
(Bache,1976).
Cation ExchangeSelectivity Coefficients
The study of cation exchangereactionshasheld a specialplaceof interest

within soil chemistrydating from the early part of the twentiethcentury.Someof
the first applicationsof chemicalthermodynamicsto soil systemswere madeto
cationexchangereactions,in an effort to understandthe equilibrium relationships
betweenionic compositionof the solution and the surface(adsorbed,exchange-
able)cations(Sposito,1981a).Interesthascontinuedto this day, asevidencedby
the continuedpublicationof paperson this topic in the scientific journals.
In the contextof this discussion,the term "cation exchangereaction"is to
be understoodas the relatively rapid, reversiblereplacementof one cation held
by the negativechargeof a soil colloid by anothercation."Selectivity" results
from somecombinationof propertiesof the particularcation and/or the colloid
exchangerthat may enhanceadsorptionof one ion in preferenceto another.The
studyof exchangeable selectivityis centralto the descriptionof the cationiccom-
positionof the soil exchangephase,which itself is a key propertyin the manage-
mentof soils. The studyof soil K reactions,including the quantity/intensity(Q/l)
studiesof Beckett (1964) which were basedin cation exchangetheory, have
attemptedto understandthe K fertility statusof soils. The continuedinterestin
Na exchangestemsfrom the importanceof Na in determiningthe physicalprop-
ertiesof semiaridandirrigatedsoils with respectto dispersionand hydrauliccon-
ductivity (Shainberg& Letey, 1984). And more recently, studiesof trace metal
adsorptionandsolutetransporthaveusedcationexchangereactionsto predictthe
solubility and transportof contaminantsin an environmentalcontext(Gaston&
Selim,1990).
The continuedinterestin cation exchangeselectivity, both in practicaluse
and in theoreticalstudies,hasbeenhamperedto somedegreeby varying proce-
dural approachesto datacollection and manipulationusedin the literature.This
variety of methodsis basedin historical differencesin conceptualizationof the
exchangeprocess,andwhile noneis necessarilysuperiora priori to another,clear
delineationsneedto be madebetweenthem. This sectionwill attemptto define
thesedifferent approaches,and suggesta critical methodologyfor datacollection
applicableto each.
Formulationof ExchangeReactions
Binary cation exchangereactionsmay be formulatedin a numberof ways;
for the caseof monovalent/divalentexchange,the replacementof onecationheld
on an exchangesurface(symbolizedasX- a monovalentexchangesite) by anoth-
er cationfrom the solution phasemay be formulatedas
2NaX + Ca 2+ ? caX2 + 2Na+ [2]
whereX- represents1 mole of surfacenegativecharge.If we assumethis is a

reversiblereactionthat canbe treatedthermodynamically,an equilibrium expres-
sion for the constantof exchangecan be written as
[3]
If we specify in this equationthat ( ) are to indicateactivities of the varioussolu-

tion and surfacespecies,then this expressionrepresentsthe true thermodynamic
equilibrium constantfor the reaction.Activities of solubleions are readily calcu-
lated using the Daviesapproximationof the Debye-Huckeltheory as
[4]
where
log y, = -1).51 z,[ 1~'I - 0.31 1 [5]
and
1= 0.5 Li(ZrCi) [6]
whereai =activity of speciesi (molesL- 1), Ci =concentrationof i (molesL- 1), Yi

= activity coefficient of speciesi, Zi = valenceof cation i, and I = ionic strength
of the solution.
Activities for adsorbedspeciesmay be expressedanalagouslyas as.i = ~/;,
where surface quantities are representedas mole fractions (Mi> moles/total
adsorbedmoles)and/isthe activity coefficientofthe adsorbedphase.The useof
molar quantities and mole fractions for adsorbedspecieshas been termed the
Vanselow convention by Sposito (1977), after the early pioneer in exchange
chemistry, while if equivalents of charge and equivalent fractions (Ei) are
employed,suchformulationsmay be referredto as using the Gapon convention.
Valuesof/can be computedfrom exchange data (Sposito,1981a),but in practice
surfaceactivity coefficientsareoften neglectedby factoringthemout of the equi-
librium expression.Thus, Eq. [3] may be written
K=(~lK
eq E
J(NaX)
2 c [7]
where
[8]
1.0
0.9
____ -::;:---; f:},7
/," ........ "-/ / / ,/,,/ .. .-"'. /
0.8 /" // .,./ ./
'§' /' ,/ ./ /
0.7
/,/,/ /' /
~ / / /
"c::
//
U)
0.6 / /
/
/
//
/
/
~ I / / ./
u
0.5 / I / /
""'5,.
<1)
I // /
/ ...... /
.' /
0-
0.4 I / /1 ...... / . Total Normality
~ 1/ / .... / 0.02SM.Na·Ca-
0.3 / I 1 .' - / D.OSOM,Na·Ca - -
UJ/j
/1 / ... / 0.100M. Na.Ca - - ·
0.2 I I ... / 0.250M. Na·Ca - - _.
(I I . / 0.500 M, Na·Ca
0.1
1/ )' All cone., Mg-Ca - -
I/./
(/
0.0
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0
ECa (equiv. fraction, soln)
Fig. 40-1. "Ideal" (nonpreferenceexchangeisothermsfor Na+-Ca2+ and Mg2+_Ca2+exchange(after

Sposito,1981b).
Kc is a conditional equilibrium constant,in that it may vary with the composi-

tion of the exchangephase(Le., as M j variesfrom 0 to 1); this variation is due to
the fact that the surfaceactivity coefficients(h) may changewith different surface
coverages(Mj ) of the cationsinvolved.
The expressionfor Kc in Eq. [8] was first proposedby Vanselow (1932),
and is referred to as the Vanselow selectivity coefficient, Kv' The term "coeffi-
cient" is usedto reflect the conditional,or variablenatureof this value with com-
position of the exchangephase.A plot of Kv vs. E j (equivalentfraction of com-
ponenti) or M j may show variation in Kv with exchangercomposition,indicating
that the ratio of activity coefficients(fNa}l!caX) is not constantas the composition
of the exchangephasevaries. This "non-ideal" behavior,defined as/; i; t i; 1
(Sposito& Mattigod, 1979), is commonly observedfor exchangeon both refer-
encemineralsand soil clays; Sposito (1981b),in fact, points out that mixturesof
minerals presentin soil clays will never behaveideally due to the differential
selectivity of the componentsover a rangeof cationcompositions.A value of Keq
may neverthelessbe evaluatedas the integral of In Kv over the rangeof E j or M j
from 0 to 1 (that is, the areaunderthe curve of a plot of Kv vs. E j ). Freeenergies
of exchange(~G) and other thermodynamicparametersmay then be computed
using this value of Keq.
Preferentialadsorptionof one cation relative to anotheris indicatedwhen
the value of Keq 1= 1. A plot of Ej (equivalentfraction of cation i in solution) vs.
E j (equivalentfraction of i on surface)also may be usedto evaluatecation pref-
erence.For mono-monovalentor di-divalent exchange,the result should be a
straight line of slope = 1 when Keq = 1 (Fig. 4~1); for the mono-divalentcase,
the equilibrium expressionand cation massbalanceequationscan be rearranged
to give
E
Ca
=1- ( 1 +2- - [ 1,
A TN (1 - ECa)2 - (1 -Eca)
1 ]]-112 [9]
where
2
A = 'YNa [10]
'YCa
and TN = total salt normality (Le., total anion chargein molarity of the solution).
Thus, for exchangereactionsinvolving unequalvalencecations, the nonprefer-
ence isotherm is actually exponentialin shape,and varies with the total ionic
strengthof the equilibrium solution, approachinga slopeof one only at high salt
concentration(Fig. 40-1; Sposito,1981b).Exchangedatamust thereforebe com-
paredto the nonpreferenceisothermcomputedat a similar salt level in order to
evaluatethe preferenceof the exchangerfor one cation over another.
Other ExchangeCoefficients
Other coefficients have been proposedto representthe cation exchange
process.Gapon(cited in Sposito,1981a)wrote an expressionfor the equation
Cao.sX+ Na+ ~ NaX + 0.5 Ca+2 [11]
using equivalentfractions,Ei = equivalents/totaladsorbedcharge,as
[12]
This Gapon coefficient, KG, is commonly used in the literature for expressing
cation selectivity, and has beenusedby the Salinity Laboratory in work on irri-
gationwaterquality (Salinity Lab. Staff, 1954).Defining sodium adsorption ratio
(SAR) = (Na)/(Ca)l!2 and exchangeable sodium ratio (ESR) = (ENaX/ECaX)' Eq.
[12] becomes
ESR
KG = SAR' ESR =KG SAR [13]
If exchangeable sodium percentage, ESP = 100 X E Na , is used,then
[ ESP )
[14]
KG = SAR (100 - ESP)
While SAR was originally formulated as an activity ratio by Gapon,field

applicationof this equationhas often revertedto the use of total cation concen-
trationsdeterminedin saturatedpastes,often including Mg2+ in the denominator
(Le., [Na+]/([Ca2+] + [M g2+])1!2, where bracketsindicate total concentrationsin
mmol L-l). The averagevalue of KG = 0.0145determinedfor a rangeof Califor-
nia soils by the Salinity LaboratoryStaff (1954)usingthis approachhasbeencrit-

icized as potentially in error dueto the effectsof ionic strengthandcomplexation
on activities of the ions (Sposito& Mattigod, 1977). Completeanion and cation
analysesand computerspeciationof solutionsis neededto accuratelycalculate
activity-basedSAR values,which may not alwaysbe justified in practice,given
that SAR values basedon concentrationare often within 10% of the activity-
basedvalues,and consideringthe ratherwide rangeof KG valuesreportedin the
literaturefor varioussoil materials(Levy et aI., 1988).
The useof equivalent,ratherthanmole fractionsin the Gaponequationalso
hasbeencriticized as nonthermodynamic,sincean activity coefficient(f;) cannot
be meaningfullyassignedto a "formal" quantity suchas an equivalent(Sposito,
1977; Ogwada& Sparks,1986).Sposito(1977),usingthe reversereactionof Eq.
[11], derived a new expressionfor the Gapon equationthat is mathematically
consistentwith the Vanselowequation, and thus thermodynamicallycorrect,as
[15]
At ENaX <0.2, the expressionin parentheses on the right handside of eq. [15] is
nearly equalto one,andENaX alsois approximatelyequal to ESR [ENaX = ESR/(1
+ ESR)]. Thus, this equationreducesto KG = SAR/(2 ESR) at low exchangeable
Na, which is a linear relation differing only by a factor of two from the "tradi-
tional" Gaponequationof Eq. [13].
The questionof the "correct" units for expressingexchangeable cationshas
introduced some confusion in the literature, as other exchangeexpressions
(notablythat of GainesandThomas,describedby Sposito, 1981a)also haveused
equivalentfractions,and derivedthermodynamicparameters(e.g.,IlG valuesfor
exchange)basedon them. While any correctly formulatedexchangecoefficient
may be usedto describea given cationexchangereaction,it might be arguedthat
the useof molesand mole fractionshasgreaterthermodynamicsignificance,and
any standardizationwould allow much more ready comparisonof coefficients
publishedin the literature.On the otherhand,the utility of the equh'alent,or mole
of charge,conceptin CEC hasbeenprovenpracticallyand historically, and is not
likely to soondisappear.
PROBLEMS IN MEASUREMENT OF
CATION EXCHANGE CAPACITY
Presence of Soluble Salts and Carbonates
The first stepin CEC determinationstypically involves the replacementof

exchangeablecationsby a saturatingsolution. Cationsother than that of the sat-
urating solution may be present,arising from the solubility of the salts and
CaC03 in the extractingsolution.Soluble salts are readily removedduring the
extraction,but sparinglysolublesaltssuchas CaS04and CaC03 will continueto
dissolvefor prolongedperiods.Polemio& Rhoades(1977) haveshownthat such
dissolution can be appreciablein commonly used saturatingsolutions.In addi-

tion, such salts can be soluble in the washing solution designedto remove the
entrainedsolution. As a result, not all the exchangesiteswill be occupiedby the
saturatingcation, and this will lead to an underestimationof the CEC. The accu-
rate determinationof exchangeable cationsin salineand calcareoussoils is clear-
ly compromisedbecauseof this problem of quantitatively separatingsoluble or
sparingly soluble from exchangeablecationsduring the extractionprocedureby
any method. As a result in such cases,CEC cannot be estimatedby summing
exchangeablecations,and often not by simple saturationtechniques.
Solublecationsarising from dissolutionreactionsalsocauseerrorsin selec-
tivity coefficient determinations.Moderatelysolublecompoundssuchas gypsum
and calcite must either be removedprior to initial cation saturation,or suchdis-
solution accountedfor during the measurement(Amrhein & Suarez,1990). Dis-
solution of the clay itself also may be a concernif very low ionic strengthsare
usedduring washingor equilibrationsteps,releasingMg2+, K+ and other cations
to the solution phase(Frenkel & Suarez,1977).
Effect of Cation and Anion Type
In soils containingmicaceousminerals,problemsmay arise when cations

suchas K+ and NHt are usedin the saturationprocess dueto their fixation in the
interlayer positions of the minerals. This can result in underestimationof CEC
(Bower, 1950). In addition, somemonovalentions usedfor extractionare some-
what less efficient than divalent ions in removing Al3+. Many polyvalent ions
suchas Al3+ and trace metalsform hydroxy ions, which may result in an overes-
timation of ECEC if they are assumedto be presentas the simple ions. However,
for Ba2+ at pH valuesbelow nine, only very small amountsof hydroxy ions are
formed, and thus errors in CEC estimationwould be minimized (Bache, 1976).
When monovalent ions such as NHt and Na+ are used to saturatethe CEC,
hydrolysis may occur to a considerable extent as the electrolyteconcentrationis
reduced,resulting in a loss of cation and underestimationof CEC.
In selectivity coefficient determinations,choiceof cationsis dictatedby the
study objectives;it shouldbe noted, however,that the theory of cation exchange
assumesexchangereactionsare reversible,and that there are a fixed numberof
exchangesitesaccessibleto both cations (assuming fixed ionic strengthand pH).
Trace metals (Cu, Zn, Mn, etc.) may violate these assumptionson clays with
oxidic or organic components,where metals may exchangewith protons on
unchargedsurfacefunctional groups(Sposito& Fletcher,1985; Sposito,1981a).
As notedabove,K+ and NHt also are strongly and irreversibly adsorbedby some
clay minerals.Such"specific adsorption,"by involving proton exchangeand lim-
ited reversibility, is bettermodeledusing site-bindingmodelswhich accountfor
the stoichiometryand competingions (i.e., H+) involved in such reactions.
Anions suchas SO~- and pog- are specifically adsorbedby variablecharge
surfaces,which can result in an increasein CEC; hencethe preferencefor Cl- or
Cl04 salts in CEC measurements(Matsue & Wada, 1985; Hendershot& Du-
quette,1986).In addition,formation of solublecomplexeswith anionsmay result
in errors in both CEC and selectivity coefficient determinations.Sposito et al.
(1983) havesuggestedthat their observedincreasesin CEC at higher Ca cover-

ageson bentonitemay be dueto CaCI+ complexformation, alsowhich influenced
the selectivity of the surfacefor Ca vs. Na. Suarez& Zahov (1989), however,
found no anioneffect on CEC usingCl-, SO~-, andCI04" on montmorillonite,and
little differencein Ca-Mg selectivity.
pH EtTectsand the Use of ButteredSolutions
Almost all topsoils and many subsoils contain variable charge surfaces
(organic matter, sesquioxidesand clay mineral edges)which can associateand
dissociateH+ dependingon the ambient pH value (Uehara& Gillman, 1981).
Thuswhen a solutionbufferedat a particularpH valuesuchas 1 M NH40AC (pH
7) or BaClz-TEA (PH 8.2) is usedduring the extractionprocess,the pH of the soil
is broughtto the pH valueof the solution.The magnitudeof the error incurredin
the CEC measurementdependson the differencein pH valuesbetweenthe soil
and extracting solution. Such buffered solutions cannot be used to estimate
exchangeable Al3+. Thusif estimatesof CEC underfield conditionsare required,
methodsinvolving unbufferedsalt solutionsshouldbe used.
Variation in pH also may affect cation selectivity of soil clays, as function-
al groupsmay show differencesin selectivity as they dissociatewith increasing
pH (Miller et aI., 1990). Somestudies,however,show little effect of pH on Ky
(Sposito& Fletcher,1985).It is probablydesirableto measureselectivity coeffi-
cientsat nearfield pH, and to avoid pH levels below 4.5 to 5, where clay disso-
lution releasesAl3+ to solution to greatly complicatethe situation.Buffers con-
taining phosphateor acetatemust be avoided due to complex formation with
divalent cations.
Ionic StrengthEtTectsand Removalof EntrainedElectrolyte
The magnitudeof the chargeon variablechargesurfacesalso is a function

of the concentrationof the equilibrium solution (Uehara & Gillman, 1981).
Thereforeit is importantto fix or measurethe ionic strengthof the solutionat the
end of the extraction process during CEC measurement.Usually the ionic
strengthof the solution is selectedto approximatethat of the soil solution under
field conditions. In methodswhich involve a water/alcohol wash to remove
excesssaturatingsolution,errorsin the CEC measuredcanariseif all the electro-
lyte is not removed.Suchwashingalso resultsin an unknownvaluefor EC (elec-
trical conductivity) of the final equilibrating solution. Furthermore,prolonged
washingcan result in the hydrolysis of adsorbedsaturatingions, giving rise to
low valuesfor CEC.
Variation in ionic strength has an effect on the distribution of cations
betweensolution and surfacefor mono-divalentexchange,as predictedby theo-
ry (Sposito,1981b);however,thereis no effect on the value of Ky obtained,pro-
vided that activity correctionof solubleions is performed.Many reportedKG val-
ues prior to 1980 did not employ suchcorrectionsin mono-divalentexchangeat
appreciableionic strengths,andthus may be in error(Sposito& Mattigod, 1977).
Most measurements of exchangecoefficientsare made at 0.01 to 0.05 M ionic
strength, largely in order to provide sufficient solution cations to effect the

desiredexchangewith the solid phase.
CATION EXCHANGE CAPACITY OF SOILS CONTAINING SALTS,

CARBONATES OR ZEOLITES
Introduction
Arid region soils often containcarbonatesand othersolublesalts,resulting

in complicationswith respectto the quantitiesof exchangeable cationsextracted.
A numberof methodshave beenproposedto overcomethesedifficulties involv-
ing the use of double extractionsand LiEDTA (Begheyn, 1987), BaClz-TEA
(Mehlich, 1939), CaCl2 (Papanicolaou,1976), and NaCI-NaOAc (Gupta et aI.,
1985).
The method of Amrhein & Suarez(1990) which has been selectedhere,
was developedto facilitate the measurementof CEC and the exchangeable
cationsin calcareousand gypsiferoussoils by taking into accountthe dissolution
of calcite and gypsumduring the saturationand extractionsteps.In addition, it
accountsfor anion exclusion,but doesnot correctfor primary weatheringwhich
is assumedto be negligible in comparisonto that of calcite and gypsum.The soil
is first saturatedwith 0.2 M CaCl2 solution adjustedto pH 8.2 and then extracted
with 0.5 M Mg(N03)2, correctingfor the entrainedCaCl2 solution. Corrections
for gypsumand calcite dissolution are madefrom the SO~- and HCO) contents
of the soil solution prior to extraction,and the saturatingand extractingsolutions.
Method
Apparatus
1. Atomic absorptionspectrometer.
2. Centrifuge.
3. Centrifugetubes(30 mL).
4. Reciprocatingor end-over-endshaker.
5. Vortex stirrer.
Reagents
1. Saturatingsolution, 0.2 M CaCI2/O.0125 M CaS04' Dissolve 29.41 g
CaCl2 • 2H20 and 2.15 g CaS04 • 2H20 in approximately 900 mL
deionizedwater and adjustto pH 8.2 using saturatedCa(OHhsolution.
Make up to 1 L with deionizedwater.
2. Dilute saturating solution, 0.025 M CaCI2• Dissolve 3.68 g CaCl2 •
2H20 and make up to 1 L with deionizedwater.
3. Extracting solution, 0.5 M Mg(N03h- Dissolve 91.19 g Mg(N03h •
2H20 and make up to 1 L with deionizedwater.
4. 0.01 M KH(I03h solution. Dissolve3.8994g KH(I03h and make up to
Procedure
Weigh 5 g soil into a preweighed30-mL centrifugetube and add20 mL of
the saturatingsolution. Shakefor 5 min, centrifuge,decantandsavesupernatant.
The Vortex stirrer is usedto resuspendthe soil. Repeatthis processfour times,
combining supernatantsin a 100-mL volumetric flask for determinaionof ex-
changeableMg, K and Na. Add 20 mL of the dilute saturating solution,shakefor
5 min, centrifugeand decantsupernatant.Repeatthis processtwice. Decantand
savethe last supernatantfor determinationof Ca, S04,CI and alkalinity (HC03),
and then reweigh the tube plus contentsto obtain the weight of entrainedsolu-
tion. Add 20 mL of the extractingsolution, shakefor 5 min, centrifugeand save
supernatant.Repeatthis processa further two times.Combineall the supernatants
in a 100-mL volumetricflask and determineCa, S04,CI and alkalinity (HC03).
The cationsCa, Mg, K and Na are determinedby atomic absorptionspec-
trophotometry[Chapters19 (Helmke& Sparks,1996),21(Suarez,1996)], CI by
Chloridometer(LABCONCO, KansasCity, MO) [Chapter31 (Frankenbergeret
aI., 1996)], S04by turbidimetry [Chapter33 (Tabatabai,1996)] and alkalinity by
titration to pH 4.40 using the 0.01 M KH(I0 3)2'
Calculations
CEC =10 x {Tea - THcOJ - Ts04 + V([HC03] + [S04] - [Ca])
- (To - V[Cl]) - [S04] (To - V[Cl]/[CI] - [HC0 3](To - V[Cl])/[Cl]}
whereT denotesions in the Mg(N03h extractin millimoles of cationchargeper

kilogram, [ ] denotesion concentrationin the fmal rinse with dilute saturating
solutionin millimoles of cationchargeperliter, V is the volumeof entrainedsolu-
tion/weightof soil in liters per kilogram, CEC is the cationexchangecapacityin
centiniolesof cation chargeper kilogram.
Comments
The primary advantageof this methodis that it permits the estimationof
exchangeable cationsand CEC simultaneouslywhile correctingfor the presence
of solublesaltsand the dissolutionof calcite and gypsum.This methodalso en-
ablesselectivitycoefficientsto be calculated.The correctionfor calcite and gyp-
sum is basedon the assumptionthat any HC03" and SOI- found in the extracting
solution in excessof that in the entrainedsolution is due to dissolution.The sat-
urating solution containssaturatedgypsumto reducegypsumdissolutionduring
the saturationprocess,so that whentracesarepresentthey are not removedprior
to extraction.
The methodavoids the useof barium saltswhich increasecalcite dissolu-
tion resultingfrom the precipitationof BaC03•
A much simplermethod,requiringfewer analyses,but which measuresthe
CEC only of soils containingcarbonates,gypsumand zeoliteswas presentedin
the previousedition of this volume (Rhoades,1982).
CATION EXCHANGE CAPACITY OF ALL ornERSOILS
CompulsiveExchangeMethod
Introduction
This methodwas originallyproposedby Gillman (1979) and subsequently
modified by Gillman & Sumpter (1986a) to measure CECcE (compulsive
exchangeCEC). It hasbeenshownto be a measureof the basiccation exchange
capacity (CECB), which is defined as the capacity of the soil to retain basic
cationsunderfield conditions(simulatedby a standardionic strengthof 0.006M)
(Gillman & Hallman, 1988; Gillman & Sumpter,1986b). For over 200 soils of
widely varying origin and for 22 Andisols, the relationshipbetweenCECB and
CECCE was very similar for the two cases
CECB =-0.14 + 1.09 CECCE ,.z =0.82 (200 soils) [14]
CECB =-0.09 + 1.05 CECCE ,.z =0.88 (22 Andisols) [15]
The total cation exchangecapacity(CEC r) is obtainedby adding exchangeable

AI obtainedby extractionwith 1 M KCl (Bertsch & Bloom, 1996; see Chapter
18) to the value for CECB (Gillman & Sumpter,1986b). Inaddition,the anion ex-
changecapacity(AEC) also can be estimated.This methodhasbeensuccessful-
ly applied to all types of soil including saline and calcareousversionsalthough
originally developedfor use on highly weathered,variablechargesoils.
The soil is initially saturatedwith Ba2+ and then broughtto an equilibrium
solutionionic strengthsimilar to that of the original soil solution.The Ba2+ is then
exchangedby Mg2+ by addition of MgS04, which precipitatesBaS04(S).After
readjustmentof the ionic strengthto a value comparableto that of the soil solu-
tion, the quantity of Mg2+ adsorbed(= CEC) is estimatedasthe lossof Mg2+ from
the MgS04 solution added.
Apparatus
1. Bench-top centrifuge equipped for 30-mL polypropylene centrifuge
tubes withcaps.
2. pH meterwith combinationelectrode.
3. Conductivity meter, preferably with the facility to operate in "ratio
mode" using two electrodes.
4. Vortex stirrer.
5. End-over-endshakeror reciprocatingshaker.
6. Top loading balancereadingto 0.01 g.
7. Dispensersand micropipettes.
Reagents
1. Saturatingsolution,0.2 M BaClzlO.2M NH4Cl. Dissolve48.9 g BaCl2 •
2H20 and 10.7 g NH4CI and make up to 1 L with deionizedwater.
2. 0.05 M BaCl2 solution. Dissolve 12.2 g BaCl2 • 2H20 and make up to 1

3. Equilibrating solution,0.002M BaCI2. Dissolve0.4889g BaCl2 • 2H20
and make up to 1 L with deionizedwater.
4. Reactantsolution, 0.005 M MgS04. Dissolve 1.2324 gMgS04 • 7H20
and make up to 1 L with deionizedwater.
5. 0.05 M MgS04 solution. Dissolve 12.3240g MgS04 • 7H20 and make
up to 1 L with deionizedwater.
6. Ionic strengthreferencesolution, 0.0015M MgS04. Dissolve 0.3697g
MgS04 • 7H20 and make up to 1 L with deionizedwater.
7. Sulfuric acid (H2S04), 0.1 M.
Procedure
Weigh a 30-mL centrifugetube (Column A, Table 40--1), add approximate-
ly 2 g soil, and reweighto determinethe exactsoil mass(Column B). Add 10 mL
deionizedwater and shakefor 1 h. Measurethe suspensionelectricalconductiv-
ity (EC) and pH. If the soil containssoluble salts as indicatedby EC, wash with
20 mL 70% ethanol in water and then 20 mL 10% ethyleneglycol (C2H60 2) in
water and discard solutions. Add 10 mL 0.2 M BaCIz/0.2 M NH4Cl solution,
shakefor a further 2 h, centrifuge and retain supernatantfor estimationof ex-
changeablecations [Chapter 19 (Helmke & Sparks, 1996) and 20 (Suarez,
1996)]. Add 20 mL 0.05 M BaCl2 to the tube, mix thoroughlywith Vortex stirrer,
centrifuge and discard supernatant.Care should be taken to avoid loss of soil
material, which can be effected by removal of the supernatantby suction.To
bring the soil to the standard0.006 M ionic strength,wash three times with 20-
mL portions of 0.002M BaClz solution. During the last washingafter thorough
mixing, measurethe suspensionpH (pHBacI2) prior to centrifugation.If AEC is to
be determined,retain the supernatantfor Cl- determination(Variable C2). Weigh
the tube andcontents(Column C) to estimatethe volume of entrainedBaClz solu-
tion. Add 10 mL 0.005 M MgS04 solution to begin the compulsiveexchangeof
Table 40-1. Worksheetfor computationof CEC by the compulsiveexchangemethod.

Number
Weighings ofO.5-mL Volume
increments MgS04
Tube + of expressed
Tube + soil + Entrained 0.05M as Final
Sample pHBaclz Tube soilt BaCI2 Final volume MgS04 0.005M volume CECCE +
A B C E VI = D V2 =10 + V3 =
C-B 5xD E -B
1589 5.37 13.00 15.01 16.56 40.82 1.55 5 35 25.81 13.56
1625 4.82 13.18 15.23 16.86 33.18 1.63 0 10 17.95 2.24
1696 5.14 13.19 15.26 17.54 28.37 2.28 2 20 13.11 7.76
t If contentshave been transferredto a beaker,substitute"wI. beaker+ soil" for "wI. tube + soil."
AEC (cmola kg-I) =50(CI V3 - C2V I); where C I =concentrationof CI- in final solution, and C2 =
concentrationof CI- in entrainedsolution millimoles per milliliter.
+ CECCE (cmole kg-I) = 100(Mg added- Mg remaining)/weightsoil =(l00/wl. soil) • (0.01 V2 -
0.003 V3)
Mg for Ba. Mix thoroughly and allow to standfor 1 h. Comparethe EC of the

suspensionwith that of the reference0.0015M MgS04 solution. If the conduc-
tivity ratio (CR =ECsusr!ECref) < 1.0, add O.5-mL incrementsof 0.05 M MgS04
and record the number of such additions. If initial CR > 1.0, take no further
action. MeasurepH of the suspension.If pHsusp> pHBaCl2 by more than 0.2 to 0.3
units, add0.1 M H2S04 dropwiseuntil pH =pHBaC12andallow to standfor at least
1 h. ReduceCR to 1.0 by adding deionizedwater and allow to standovernight.
RecheckpHsuspand CR. If necessary,readjustto pHBaC12with 0.1 M H2S04, and
CR to 1.0 ± 0.05 with deionizedwater. When satisfiedthat the appropriatecon-
ditions of pH and ionic strengthhavebeenestablished,reweighthe tube(Column
E). If AEC is to be determined,centrifugeand determineCl- concentrationin the
supernatant(Variable Ct ) and in the solution retainedabove(Variable C2 ) [Chap-
ter 31 (Frankenbergeret aI., 1996)].
Calculation
The worksheetshown in Table 40--1 gives examplecalculationsfor this
method,and its useconsiderablysimplifies the record-keepingnecessarywith the
procedure.Such a table might readily be constructedin a computerspreadsheet
application,greatly facilitating computations.
Comments
Matsue& Wada(1985) criticized this method onthe groundsthat it could
not be appliedto soils which specifically adsorbSOa-.They proposedusing 0.01
M SrCl2 insteadof BaCl2 and extractingwith 0.5 M HCI or 1 M NH40Ac. Hen-
dershot& Duquette(1986) found that the CEC of predominantlyvariablecharge
soils measuredby compulsiveexchangewith MgS04 was higher than when the
Ba was replacedby MgCI2, suggestingthat specific adsorptionof SO~- may be
responsible. Subsequently,Wada & Matsue (1987) questioned Gillman &
Sumpter's(1986a)use of H2S04 to reducethe pH of the MgS04 soil suspension
to its value in BaCI2, andcalculatedthat the CEC may be overestimatedby 10 to
15% usingthis method.Gillman & Hallman (1988) addressedthesecriticisms by
using CaCl2 insteadof BaCl2 and extracting with 1 M N~OAc; their results
showedthat on a rangeof Andisols, specific adsorptionof SOi- was not a prob-
lem. As far as the use of H2S04 is concerned,they indicatedthat the magnitude
of the error was small and would only be significant when a soil had >5 cmol+
kg-1 of both CEC and AEC. None of the soils they studiedshowedany of these
characteristicsevenremotely.
If this methodis to be usedto estimateexchangeablecations,care should
be taken to preparestandardsin the same matrixas the unknownsolutions;Ca2+
in particular suffers some interference from sOi- with air-acetyleneatomic
absorptiondetermination.When soils contain soluble salts, pretreatmentshould
be usedto removethem, but this can causeerrorsin the estimationof exchange-
able cations.Howeverthe presenceof soluble saltswill havelittle or no effect on
the measurementof CECCE•
The reasonfor the use of an equilibrating solution of divalent cationsof
ionic strengthequalto 0.006M is becauseit approximatesthat of the "soil solu-
tion" of many highlyweatheredsoils (Gillman & Bell, 1978). In soils where the
ionic strengthof the soil solution differs substantiallyfrom 0.006M, an appro-
priate value shouldbe used.
Although BaCl2 is not usually used as an electrolyte to measuresoil pH,
pHBaCI2 shouldbe a reliable estimateexceptfor salinesoils wherethe removalof
solublesaltsgenerallyresultsin an increasein pH.
For greaterconvenience,a conductivity meter which has the capability of
being operatedin a ratio mode is preferred,althougha meterwith a single elec-
trode will suffice.
With experience,it becomespossiblewith highly bufferedsoils to overad-
just pHBaCI2 with 0.1 M H2S04, knowing that an upward drift will occur. Simi-
larly, adjustmentof CR to 1.0 by the addition of deionizedwater is soil-depen-
dent. Shouldthe capacityof the centrifugetube be exceeded,simply transferthe
contentsto a weighed beaker and continue. A modification to the procedure
which makesit lesstime-consuminghasbeenproposedby Sumneret aI. (1994),
in which insteadof dilution with water to bring the CR to 1.0 in the final step,the
sampleis centrifugedprior to water addition and the EC of the supernatantis
measured.From a calibrationcurve relatingEC of the supernatantto wateradded,
the amountof water that would have beenaddedcan be estimated.
Cation exchangecapacity values obtained by the compulsive exchange
methodare similar to thoseobtainedby the silver thioureamethod(Searle,1986;
Gillman et aI., 1983),sumof exchangeable cations(ECEC) (Gillman et aI., 1983;
Grove et aI., 1982), and 0.2 M NH4CI (Grove et aI., 1982) methods.
UnbufferedSalt ExtractionMethod
Introduction
This method is basedon the original proposalof Schofield (1949) which
enabledthe measurementof the CEC of a soil at its "field pH" value. An un-
bufferedsalt solution is usedin placeof the bufferedsolutionssuch as NH40Ac
and BaCl2-TEA which were in vogue at that time for saturatingthe exchange
complex.The methodpresentedhereis a modificationof the proceduredescribed
by Grove et aI. (1982). It involves the saturationof the exchangesiteswith NHt
using an unbufferedNH4CI solution, reducingthe ionic strengthto an appropri-
ate value (or removingthe entrainedsalt with water),assessingthe volume of the
solution which is entrainedand then displacingNH4+ with a solution of KN0 3.
The quantitiesof NHt and CI- in the final extract are correctedfor the amounts
in the entrainedsolution. If the volume of entrainedsolution is measured,this
methodalso permitsthe estimationof the anion exchangecapacityfrom the quan-
tity of Cl- adsorbed.
Apparatus
1. Bench-topcentrifuge.
2. 50-mL centrifugetubeswith caps.
3. Vortex stirrer.
4. End-over-endor reciprocatingshaker.
5. Top-loadingbalancereadingto 0.01 g.
6. Dispensers.
Reagents
1. Saturatingsolution, 0.2 M NH4CI. Dissolve 10.7 g NH4Cl and make up
to 1L with deionizedwater.
2. Equilibrating solution, 0.04 M NH 4CI. Dissolve 2.1 g NH4Cl and make
up to 1 L with deionizedwater.
3. Extracting solution, 0.2 M KN0 3. Dissolve 20.2 g KN0 3 and make up
Procedure
Weigh 5 g of soil into a preweighed50-mL centrifugetube. Add 30 mL of
0.2 M NH4CI and shakefor 5 min, centrifugeand decantsupernatantinto a 250-
mL volumetric flask, being careful to avoid loss of soil. Add 30 mL of 0.2 M
NH 4CI, resuspendthe soil using the Vortex stirrer (Scientific Industries,Bohe-
mia, NY), shake for 5 min, centrifuge and decant supernatantinto volumetric
flask. Repeatthis processthreemore times, combiningsupernatants prior to mak-
ing up to volume with 0.2 M NH4CI. Savethis solution for the determinationof
exchangeableNa, K, Ca, Mg and AI [Chapter 18 (Bertsch & Bloom, 1996), 19
(Helmke & Sparks,1996), and 20 (Suarez,1996)]. Two options are possibleat
this point: EITHER washthreetimes with deionizedwater and discardthe super-
natant,OR add 3 x 30-mL portionsof 0.04M NH4CI, resuspend,shakefor 5 min,
centrifuge and discard the supernatanteach time, and then weigh tube to deter-
mine volume of entrainedsolution. Add 30 mL 0.2 M KN0 3, resuspend,shake
for 5 min, centrifuge and collect supernatantin a 250-mL volumetric flask.
Repeatthis processa further four times, combiningthe supernatants. Analyze this
solution for NHt [Chapter38 (Mulvaney, 1996)] and, if entrainedsolution was
measuredand AEC is desired,for Cl- [Chapter31 (Frankenbergeret aI., 1996)].
Calculations
With water wash,
CEC = (NHt x 5)/18
where NHt = NHt in KN0 3 extract in milligrams per liter.

CEC = cation exchangecapacityin centimolesof cation chargeper kilo-
gram.
With correctionfor entrainedsolution,
CEC = 0.2775 x NHt - 0.80 X VEn
where NHt = NHt in KN03 extractin milligrams per liter.

VEn = volume of entrainedsolution in milliliters
AEC =0.14 x Cl- - 0.8 X YEn
where CI- =CI- in KN03 extractin milligrams per liter

AEC = anionexchangecapacityin centimoles ofanionchargeperkilogram
Comments
Whenwaterwashesare usedinsteadof estimatingthe volume and concen-
tration of the entrainedsolution, the soil may begin-to disperse,and a higher
speedcentrifugemay be necessaryto separatethe phases.The concentrationof
0.04 M NH4CI wasselectedas that which would preventthe deflocculationof the
clay in mostsoils. It is in the middle of the rangeusedby Matsue& Wada(1985).
The valuesof CEC obtainedat this concentrationare similar to thoseobtainedin
the compulsiveexchangemethodat a concentrationof 0.001M BaCI2• Grove et
al. (1982) suggestedthat the net chargeon the soil was given by the value for
CEC obtainedusingthe water wash,which wasessentiallyequalto the valueob-
tainedwhenthe AEC wassubtractedfrom the CEC. This value alsowas equalto
the sum of exchangeable cations(ECEC).
AmmoniumAcetate(pH 7) Method
Introduction
Although this methodhas beenusedfor many years,it overestimatesthe
"field" CEC of soils with a pH <7. Nevertheless,it is a standardmethodusedin
the classificationof soils (Soil Surv. Lab. Staff, 1992)andconsequentlywarrants
citation here. Becausethe NH40Ac usedduring the procedureis bufferedat pH
7, the methodcausesvariablechargesitesin acid soils not active at the field pH
to becomeionizedandconsequentlymeasured.Therearea numberof variantsof
this methodusingboth batchandcontinuousleachingtechniques.The methodol-
ogy selectedhereis a leachingtube methodproposedby the Soil SurveyLabora-
tory Staff (1992).
Apparatus
1. Mechanicalvacuum extractor(Centurion International,Inc., Lincoln,
NE Model 24).
2. loo-mL volumetric flasks.
3. Top-loadingbalanceweighing to 0.01 g.
Reagents
1. Saturatingsolution, 1 M NH40Ac pH 7.0. Mix 68 mL N~OH (sp gr.
0.90) and 57 mL 99.5% CH3COOH per liter of solution desired.Cool,
adjust to pH 7.0 with CH3COOH or NH40H and dilute to 1 L with
deionizedwater.
2. Ethanol,95%.
3. Replacingsolution, 1 M KCI. Dissolve 74.5 g KCI, dilute to 1 L of
deionizedwater.
Procedure
Preparethe leachingtubesby placing either filter paperor filter paperpulp
into the syringebarrelsand compressingit with the plunger.Weigh 5.0 g soil into
the tube and place on mechanicalvacuum extractor.Add 25 mL 1 M NH40Ac,
stir and leach. Add an additional 25 mL 1 M NH40Ac and allow to stand
overnight by using a pinch clamp or by stopperingthe leachingtube. Leach and
make up to volume and save for determination of exchangeablecations if
required [Chapter 19 (Helmke & Sparks, 1996) and 20 (Suarez, 1996)]. Add
about 10 mL ethanolto the soil pad, stir and leach. Leach with 100 mL ethanol
and check for NHi in leachate.If NHi is present,leach with an additional 100
mL ethanol.Discard leachate.Now leachwith a total of 60 mL M KCl and make
up to 100 mL. DetermineNH4 in the leachateby an appropriatemethod [Chap-
ter 38 (Mulvaney, 1996)].
Calculations
where NH4 is the concentrationin the leachatein milligrams per liter
Summationof Cations(Effective Cation ExchangeCapacity)
Introduction
The conceptof effective cation exchangecapacity (ECEC) was first for-
malizedby Colemanet al. (1959) as the sum of the exchangeable Ca, Mg and Al
displacedfrom the soil using 1 M KCl but hasevolvedto include Na and K. There
is considerableevidence to show that the quantities of exchangeablecations
extractedfrom nonsalinenoncalcareoussoils by any of the common extracting
solutionsare very similar (Grove et aI., 1982). Thus it is possibleto use this tech-
nique toestimat>!cation exchangecapacity in soils which do not containsaltsand
carbonates.
Apparatus
As listed under"Preparations"for "Cation ExchangeCapacityof All Other
Soils," or "Apparatus"for "UnbufferedSalt Extraction Method."
Reagents
As listed for "Reagents"for "Cation Exchange Capacity of All Other
Soils," or "Reagents"for "Unbuffered Salt Extraction Method."
Procedure
DetermineCa, Mg, K, Na andAl by atomic absorptionspectrometryasout-
lined in Chapters 19 (Helmke & Sparks, 1996), 20 (Suarez, 1996), and 18
(Bertsch& Bloom, 1996).
Calculations
Calculateexchangeableions (W+) in centimolesof cation chargeper kilo-
gram as
M"+ =(M"+ x Vx n)/(WxA)

whereM"+ = concentrationof cation in extract in milligrams per liter
V =volume of extract(mL)
n = valenceof cation
W =weight of soil (g)
A = atomic weight of cation
ECEC = Ca + Mg + K + Na + Al
where Ca =exchangeableCa in centimolesof cation chargeper kilogram

Mg =exchangeableMg in centimolesof cation chargeper kilogram
K =exchangeableK in centimolesof cation chargeper kilogram
Na = exchangeableNa in centimolesof cation chargeper kilogram
AI = exchangeableAI in centimolesof cation chargeper kilogram
Comments
For soils containingsalts and carbonates,this procedureresults in highly
inflated valuesfor cation exchangecapacitybecauseof the appreciablesolubili-
ty of thesematerials in the extracting solutions. For all other soils, the values
obtainedare very similar to those measuredby the methodsdesignedto deter-
mine the CEC at "field pH" as describedin "UnbufferedSalt ExtractionMethod"
and "CompulsiveExchangeMethod" above.This agreementis to be expectedon
theoreticalgrounds.
Over a wide variety of soils, valuesfor the sum of basic cationsextracted
by a varietyof extractantssuchas 1 M NH 40Ac, 0.2 M NH4CI, 0.2 M BaCI2> 0.2
M CaClz, 1 M BaClz-TEA and 0.01 M SrClz (Bache, 1976; Grove et aI., 1982;
Gillman & Hallman, 1988; Hendershot & Duquette, 1986; Matsue & Wada,
1985) plus AI extractedwith 1 M KCI or 0.2 M NH 4CI [Skeen & Sumner,
1967a,b;Chapter18 (Bertsch & Bloom, 1996)] were essentiallythe same,indi-
cating that almost any extractantis suitablefor estimationof ECEC.
MEASUREMENT OF SELECTIVITY COEFFICIENTS

Introduction
A wide rangeof methodsand computationalapproacheshavebeenusedin
measurementsof selectivity coefficients,despite a common objective: to bring
solutions containing varying ratios of two competing cations into equilibrium
with an exchangerphaseat fixed pH and ionic strength,and to measureboth solu-
tion and exchangercompositions.In arriving at a method to suit a particular
objective,severalchoicesmust be made:
ExchangerPhase
In using referenceclays or standardexchangermaterials,size-fractionated
suspensionsare typically preparedfor use in the exchangeexperiments.With
soils, whole soil materialmay be used,or the colloid fraction separatedandhan-
dled as a suspension.
SaturatingTechnique
Homoionicclaysor soils may be useddirectly in the exchangeexperiment
by addingmixed salt solutionsand allowing exchangeto occur. Or, the exchang-
er may be preadjustedto a fractionalcoverageof onecationby washingwith con-
centratedsolutionscontainingactivity ratios (AR) equivalentto desiredcation
equivalentratios on the clay. Using a homoionicexchangeris more convenient,
but requirescarefulselectionof solution ionic strengthand solution/soilratio, in
orderto providesufficient solublecationsto exchangewith the homoionicclay.
Rangeof SurfaceCoverage
Most studieswill attemptto spanthe entire rangeof 0 to 100% coverageof
the exchangerwith eachcation; however,choiceof a more limited rangeof inter-
est is acceptable,althoughwill not allow computationof Keq (see"Formulation
of ExchangeReaction").
Determinationof ExchangeableCations
After equilibrium is attained(2-12 h), cationsheld on the exchangermay
be measuredby displacementwith an addedsalt, with or without a washingstep
to removeentrainedsolution.Or, total dissolutionof the clay may be performed,
after accountingfor entrainedsolution and the presenceof cations in the clay
matrix (Sposito et at, 1981). Alternately, at low surfacecoverages,the disap-
pearanceof one cation from solution may be usedas a measureof adsorptionif
a homoionicexchangersaturatedwith the competingion is used(Milberg et aI.,
1978).
ComputationalMethod
Any or all of the exchangecoefficientsmentionedearlier(see"Formulation
of ExchangeReactions")may be computedwith the soluble and exchangeable
cationdata.Activity correctionsshouldbe madeif the competingions are of dis-
similar valance,or appreciableS04 or other complexinganions are presentin
solution.
The following methodis an exampleof a combinationof the abovechoices
used in published coefficient determinations.Possible modifications are dis-
cussedat the end of the methoddescription.
Use of Homoionic Clay Fractionsto Determinethe Vanselow

Selectivity Coefficient
This method, modeledafter Spositoet ai. (1981) and Sposito& Fletcher

(1985), uses<2 Ilm clay (eitherseparatedfrom soil or referencegeologicmater-
ial) to computeKv. This clay fraction shouldbe separatedby standardtechniques

(Jackson,1974), and storedas a concentrated(>20 g L- 1) homoionic suspension
containingat least0.1 M salt to preventhydrolysisand clay dissolution.
Procedure
Ten or moremixed solutionsof the cationsspanningthe rangeof 1 to 100%
exchangercoverageare suggested,with at least three replicatesof each. In for-
mulating solution concentrations,assumenonpreferentadsorptionand use Fig.
4~1 to choosesolution equivalentratios (E) that yield roughly evenly spaced
exchangerequivalentratios(E). The formulas in Table 4~2 may then be usedto
calculateconcentrationsfor the individual cations at eacht and a given ionic
strength(1).
Sufficient solution cationsmust be addedto the homoionicclays to ensure
that exchangedoesnot overly depletethe solution of one cation. Choiceof mass
of addedclay and concentrationand volume of salt addedare the importantvari-
ables. For example,in equilibrating a calcium-saturatedclay with a 100% Na
solution, the total equivalentsof Na addedshould be 6 to 10 times the equiva-
lents of exchangeableCa. Thus, 0.25 g of clay with a CEC of 50 cmole kg-1 con-
tains 0.125 cmole Ca2+, 50 mL of 0.025 M NaCI contains1.25 cmole, providing
a 10:1 ratio of addedsolution cationsover thoseinitially exchangeable.
Before use, the clay suspensionshould be washedto a lower solution I
«0.01 M), and the clay concentrationcarefully measuredby drying an aliquot at
105°C; the salt contentalso should be measuredin the solution phase.Pipetting
from a vigorously stirred suspensionmay be used to dispensethe clay into
weighedcentrifugetubes(50 or 100 mL) after computingthe volume neededto
deliver the neededmassof clay. Water and concentrated(i.e., 0.5 M) salt solution
of the individual cations are then addedto achievethe required concentrations
computedfrom Table 4~1. Rememberto accountfor the cation transferredwith
the clay suspensionin the calculations.
After equilibration (typically with shaking,8-24 h), centrifuge the tubes
and fully decantthe solution phaseand savefor cation analysis.Weigh the tubes
to determinethe volume of entrainedsolution, then displacethe exchangeable
cationsby addinga volume of concentrated(0.2-1.0M) salt containinga differ-
ent cation than the two of interest (see "Problems in Measurementof Cation
Exchange Capacity"). Corrections should be made for the contribution of
entrainedsolution to the cationlevelsin the displacementsolution(see"Problems
in Measurementof Cation ExchangeCapacity").
Table 40-2. Solutionequivalenceconditions,ionic strength,and concentrationformulas forexchange
reactionsof Cationsi at a given solution equivalentratio (E) and ionic strength(l).:f:
Type of exchange
Equation mono)-monoZ
Equiv. condition E =c;I(c) + cz)t
j Ej =2cd(2c) + 2cz) =Cj /(c) + cz) Em =cm/(cm + 2cd)
Ionic strength I =c) + Cz 1= 3c) + 3cz 1= cm + 3Cd
Solve for Cj, Cm cj=EI Cj =E;I/3 cm = zEml/(3 - Em)
t Cj is expressedin moleslliter.
:f: c =concentration,d =divalent cation, m =monovalentcation; anionsare assumedmonovalent.
Determinecationsin the equilibrium and displacementsolutionsby atom-

ic absorption spectroscopy or othersuitablemethod[seeChapters19 (Helmke &
Sparks,1996) and20 (Suarez,1996)]. Extremecareshouldbe exercisedin these
analyses,including the use of standardsmade in matricesto match samples,in
order to obtain the most accurateresultspossible.
Calculations
Solution concentrationsmay be expressedin moles per liter or millimoles
per liter (mmol L- 1); activity ratios and coefficientsfor mono-divalentexchange
will differ by a factor of 31.6 (i.e., -V1000) when computedusing thesetwo sets
of units. Exchangeableions may be computedas mole fractions,using the sum of
the moles of exchangeableions as denominatorsfor M;, or as equivalentfrac-
tions. For mono-monovalentor di-divalent exchange,Ky may be calculated
directly from
[17]
using free concentrations(i.e., correctedfor complexationusing the approachof

Sposito& Mattigod, 1977) of cationsi andj ratherthan activities,sincethe activ-
ity coefficients of the two cations are equal. For mono-divalentexchange(i.e.,
Na-Ca),
[18]
individual'Y valuescan be calculateddirectly using eqs.4-6, or solutionsmay be

speciatedusing geochemicalspeciationprogramssuch as GEOCHEM (Sposito
& Mattigod, 1977) or MINTEQ (Allison et aI., 1991).
The Ky valuesobtainedover a range of M; may be plotted as In Ky vs. M;
to examine the dependenceof Ky on exchangercomposition. Integrals for the
curve will ~ield Keq. Plots similar to Fig. 40-1 also may be made after comput-
ing E; and E; and comparedwith the nonpreferenceisothermsshown.
Comments
Rieu et al. (1991) have pointedout the large analyticalerrors in determina-
tion of selectivity coefficients,stemminglargely from uncertaintyin exchange-
able cation measurementsat low M; or E;. They recommend an empirical
approachto relating E; and E; ratherthan a thermodynamicone due to their esti-
matesof 15 to 25% error in computedK values;adequatereplication (3-5), con-
scientious measurements,and careful choice of initial solution concentrations
will minimize theseproblems.Overall error should be computedfor eachset of
replicatesas a standarddeviation in K, and reportedor usedin statisticaltesting.
Many variations onthe proposedprocedureare possible,dependingon per-
sonal preference,experimental objectives, etc. Whole soils may readily be
employed,with soil masschosenbasedon the expectedCEC of the clay fraction.
Samplingfrom soil containersshould be done carefully to avoid samplingerror,

and salt releaseshould be checkedby analysesof selected equilibriumsolutions
for releaseof cations(K, Mg, AI) from the solid phase.
It is possibleto preadjustM j to a value close to that expectedat equilibri-
um for eachsolution compositionby presaturatingthe solid phasewith concen-
trated salt solutions. This eliminatessubstantialchangein composition of the
solution phaseafter equilibration. The concentratedsolutions(0.5 M), chosento
span therangeof M j of interest,are subsequentlyreplacedwith more dilute solu-
tions of the sameactivity ratio [i.e., (Na)/(Ca)l/2 for Na-Ca exchangeusing the
Gaponequation] until the final desiredionic strength(typically 0.05-0.01M) is
reached.This approach has advantagesin that a wider and morepredictablerange
of E j is obtained,comparedto beginningthe exchangeprocesswith a homoionic
clay. However, the processis considerablymore time-consumingin that each
tube must go through numerous washingstepswith the progressivelymore dilute
solutions.Details of this method are describedby Levy et al. (1988) and Miller
et al. (1990).
Any of the otherexchangecoefficientsdiscussed(Gapon,Gaines-Thomas)
may be computedusing the dataobtained,in order to compareresultswith other
suchvaluesin the literature. However, it should be noted that much early work
did 'not correctsolution concentrationsto activities, and may in somecasesthere-
by be in error.
Anion exclusionduring selectivity coefficient measurements hasthe poten-
tial to reduceCEC, particularly with high-chargeclays (>50 cmole kg-I) at equi-
librium ionic strengths<0.05 to 0.10 M (Amrhein & Suarez,1990). Anion con-
centrations in equilibrium solutions will increase above initial levels if this
occurs, but this was not observedin one study on montmorillonite (Sposito &
Fletcher,1985).
Binary exchange coefficients have been used to model tertiary-level
exchangesystems,where three or more cationsare presentin the system;while
there is some questionas to the theoreticalbasis of this practice, the resulting
model predictionsappearto be consistent(Chu & Sposito,1981).
ACKNOWLEDGMENTS
Contribution fromthe Departmentof Crop and Soil Sciences,University of

Georgia,Athens, GA 30602.
REFERENCES
Allison, J.D., D.S. Brown, andK. Novo-Gradac.1991. MINTEQA2-PRODEF,a geochemicalassess-
ment model for environmentalsystems:Ver. 3 usersmanual.USEPA 600/3-91/021.USEPA,
Athens,GA.
Amrhein, c., and D.L. Suarez.1990. Procedurefor determiningsodium-calciumselectivity in cal-
careousand gypsiferoussoils. Soil Sci. Soc. Am. J. 54:999-1007.
Bache, B.W. 1976. The measurementof cation exchangecapacity of soils. J. Sci. Food Agric.
27:273-280.
Bascomb,c.L. 1964. Rapid methodfor the determinationof cation exchangecapacityof calcareous
and non-calcareous soils. J. Sci. Food Agric. 15:821-823.
Beckett, P.H.T. 1964. Studieson soil potassium:II. The immediateQ/I relationsof labile potassium
in the soil. 1. Soil Sci. 15:9-23.
Begheyn,L.Th. 1987.A rapid methodto determinecation exchangecapacityand exchangeablebases
in calcareous,gypsiferous,saline and sodic soils. Comm. Soil Sci. Plant Anal. 19:911-932.
Bertsch,P.M., and P.R. Bloom. 1996. Aluminum. p. 517-549.In D.L. Sparket al. (ed.) Methodsof
soil analysis.Part 3. Chemicalmethods.SSSABook Ser. 5. SSSAand ASA, Madison,WI.
Bower, c.A. 1950. Fixation of ammoniumin difficultly exchangeableforms under moist conditions
by somesemi-aridregion soils. Soil Sci. 70:375-383.
Brady, N.C. 1984. Nature and Propertiesof Soils. MacMillan Co., New York.
Chu, S.-Y. and G. Sposito. 1981. The thermodynamicsof ternary cation exchangesystemsand the
subregularmodel. Soil Sci. Soc. Am. I. 45:1084-1089.
Coleman,N.T., S.B. Weed,and R.I. McCracken.1959. Cation-exchangecapacityand exchangeable
cationsin Piedmontsoils of North Carolina. Soil Sci. Soc. Am. Proc. 23:146-149.
Frankenberger,Ir., W.T., M.A. Tabatabai,D.C. Adriano, and H.E. Doner. 1996. Bromide, chlorine,
and fluorine. p. 833-867.In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3. Chemi-
cal methods.SSSABook Ser.5. SSSAand ASA, Madison,WI.
Frenkel,H., and D.L. Suarez.1977. Hydrolysis and decompositionof calcium montmorillonite. Soil
Sci. Soc. Am. 1. 41:887-89l.
Gaston,L.A., and H.M. Selim. 1990. Predictingcation mobility in montmorillonitic media basedon
exchangeselectivitiesof montmorillonite.Soil Sci. Soc. Am. 1. 54:1525-1530.
Gillman, G.P. 1979.A proposedmethodfor the measurement of exchangepropertiesof highly weath-
ered soils. Aust. I. Soil Res. 17:129-139.
Gillman, G.P., and L.c. Bell. 1978. Soil solution studies on weatheredsoils from tropical North
Queensland.Aust. 1. Soil Res. 16:67-77.
Gillman, G.P., R.C. Bruce, B.C. Davey,I.M. Kimble, P.L. Searle,and 1.0. Skjemstad.1983. A com-
parison of methodsusedfor determinationof cation exchangecapacity.Commun. Soil Sci.
Plant Anal. 14:1005-1014.
Gillman, G.P.,and M.I. Hallman. 1988. Measurementof exchangepropertiesof Andisolsby the com-
pulsive exchangemethod.Soil Sci. Soc. Am. 1. 52:1196-1198.
Gillman, G.P., and E.A. Sumpter.1986a.Modification to the compulsiveexchangemethodfor mea-
suring exchangecharacteristicsof soils. Aust. 1. Soil Res. 24:61-66.
Gillman, G.P.,and E.A. Sumpter.1986b.Surfacechargecharacteristicsand lime requirementsof soils
derived from basaltic,granitic and metamorphicrocks in high-rainfall tropical Queensland.
Aust. 1. Soil Res. 24:173-192.
Grove,1.H., C.S. Fowler, and M.E. Sumner.1982. Determinationof the chargecharacterof selected
acid soils. Soil Sci. Soc. Am. J. 46:32-38.
Gupta, R.K., C.P. Singh, and I.P. Abro!. 1985. Determiningcation exchangecapacityand exchange-
able sodium in alkali soils. Soil Sci. 139:326-332.
Helmke, P.A., and D.L. Sparks. 1996. Lithium, sodium, potassium, rubidium, and cesium. p.
551-574. In D.L. Sparkset a!. (ed.) Methodsof soil analysis.Part3. Chemicalmethods.SSSA
Hendershot,W.H., and M. Duquette.1986. A simple barium chloride methodfor determiningcation
exchangecapacityand exchangeablecations.Soil Sci. Soc. Am. I. 50:605-608.
Jackson,M.L. 1974. Soil chemical analysis--advanced course. 2nd ed. Dep. Soil Sci., Univ. Wis-
consin, Madison,WI.
lones,U.S. 1982. Soil fertility and fertilizers. RestonPub!. Co., Reston,VA.
Kelley, w.P. 1948. Cation exchangein soils, Reinhold Pub!. Corp., New York.
Kelley, w.P., and S.M. Brown. 1924. Replaceablebasesin soils. California Agric. Exp. Stn. Tech.
Pap. 15:1-39.
Levy, GJ., H.v.H. van der Watt, I. Shainberg,and H.M. du Plessis.1988. Potassium-calcium andsodi-
um-calciumexchangeon kaolinite and kaolinitic soils. Soil Sci. Soc. Am. I. 52:1259-1264.
Matsue, N., and K. Wada. 1985. A new equilibrium method for cation exchangecapacity measure-
ment. Soil Sci. Soc. Am. I. 49:574-578.
Mehlich, A. 1939. Use of triethanolamineacetate-bariumhydroxide buffer for the determinationof
some base-exchangeproperties and lime requirementsof soil. Soil Sci. Soc. Am. Proc.
3:162-166.
Mehlich, A. 1942. Rapid estimationof base-exchange propertiesof soils. Soil Sci. 53:1-14.
Mehlich, A. 1945. Effect of type of colloid on calcium adsorptioncapacity and on exchangeable
hydrogenand calcium as measuredby different methods.Soil Sci. 60:289-304.
Milberg, R.P.,D.L. Brower, andJ.V. Lagerwerf.1978.Exchangeadsorptionof tracequantitiesof cad-

mium in soils treated with calcium and sodium:A reappraisal. Soil Sci. Soc. Am. J.
42:892-894.
Miller, W.P., H. Frenkel, and KD. Newman. 1990. Flocculationconcentrationand sodium/calcium
exchangeof kaolinitic soil clays. Soil Sci. Soc. Am. 1. 54:346-351.
Mulvaney, R.L. 1996. Nitrogen-inorganicforms. p. 1123-1184.In D.L. Sparkset al. (ed.) Methods
of soil analysis.Part 3. Chemicalmethods.SSSABook Ser.5. SSSAand ASA, Madison,WI.
Ogwada,R.A, and D.L. Sparks.1986. Use of mole or equivalentfractions in determiningthermody-
namic parametersfor potassiumexchangein soils. Soil Sci. 141:268-273.
Oster,J.D., and G. Sposito. 1980. The Gaponcoefficient and the exchangeablesodium percentage-
sodiumadsorptionratio relation. Soil Sci. Soc. Am. J. 44:258-260.
Papanicolaou,E.P. 1976. Determinationof cation exchangecapacityof calcareoussoils and their per-
cent basesaturation.Soil Sci. 121:65-71.
Polemio, M., and 1.0. Rhoades.1977.Determiningcation exchangecapacity:A new procedurefor
calcareousand gypsiferoussoils. Soil Sci. Soc. Am. 1. 41:524-528.
Rhoades,1.0. 1982. Cation exchangecapacity.p. 149-158.In AL. Pageet al. (ed.) Methodsof soil
analysis.Part 2. Agron. Monogr. 9. ASA and SSSA, Madison,WI.
Rhoades,1.0. 1996. Salinity: Electrical conductivity and total dissolvedsolids. p. 417-435.In D.L.
Rieu, M., J. Touma, and H.R. Gheyi. 1991. Sodium-calciumexchangeon Brazilian soils: Modeling
the variation of selectivity coefficients.Soil Sci. Soc. Am. J. 55:1294-1300.
Salinity LaboratoryStaff. 1954. Diagnosisand improvementof saline and alkali soils. USDA Agric.
Handb.60. U.S. Gov. Print. Office, Washington,DC.
Schofield,R.K 1939. The electricalcharges onclay particles.Soils Fert. 2:1-5.
Schofield, R.K 1949. Effect of pH on electric chargescarried by clay particles.J. Soil Sci. 1:1-8.
Schollenberger,C.J., and F.R. Dreibelbis. 1930. Analytical methodsin base-exchange investigations
in soils. Soil Sci. 30:160-173.
Schollenberger,C.J., and R.H. Simons. 1945. Determinationof exchangecapacityand exchangeable
basesin soils. Soil Sci. 50:13-24.
Searle,P.L. 1986.The measurementof soil cation exchangepropertiesusing the single extractionsil-
ver thiourea method:An evaluationusing a range of New Zealandsoils. Aus!. J. Soil Res.
24:193-200.
Shainberg,I., and J. Letey. 1984. Responseof soils to sodic and saline conditions.Hilgardia 52:2.
Skeen,J.B., and M.E. Sumner. 1967a. Exchangeablealuminum: I. The efficiency of various elec-
trolytes for extractingaluminum from acid soils. S. Afr. J. Agric. Sci. 10:3-10.
Skeen,J.B., and M.E. Sumner.1967b. Exchangeablealuminum: II. The effect of concentrationand
pH value of the extractanton the extractionof aluminium from acid soils. S. Afr. J. Agric. Sci.
10:303-310.
Soil Survey Staff. 1975. Soil taxonomy:A basic systemof soil classificationfor making and inter-
pretingsoil surveys.USDA-SCSAgric. Handb.436. U.S. Gov. Print. Office, Washington,DC.
Soil Survey LaboratoryStaff. 1992. Soil survey laboratorymethodsmanual.Soil Surv. Invest. Reps.
42. USDA-SCS,Washington,DC.
Sposito, G. 1977. The Gapon and the Vanselow selectivity coefficients. Soil Sci. Soc. Am. J.
41:1205-1206.
Sposito,G. 1981a.Cationexchangein soils: An historical and theoreticalperspective.p. 13-30.In M.
Stelly (ed.) Chemistryin the soil environment.ASA, Madison,WI.
Sposito,G. 1981b.The thermodynamicsof soil solutions.Oxford Univ. Press,Oxford, England.
Sposito,G., and P. Fletcher. 1985. Sodium-calcium-magnesium exchangereactionson a montmoril-
lonitic soil: III. Calcium-magnesium selectivity. Soil Sci. Soc. Am. J. 49:1160-1163.
Sposito,G., KM. Holtzclaw, L. Charlet,C. Jourany,and AL. Page.1983. Sodium-calciumand sodi-
um-magnesiumexchangeon Wyoming bentonitein perchlorateand chloride backgroundionic
media. Soil Sc. Soc. Am. J. 47:51-56.
Sposito,G., K.H. Holtzclaw, C.T. Johnston,and C.S. LeVesque-Madore.1981. Thermodynamicsof
sodium-copper exchange on Wyoming bentonite at 298 K Soil Sci. Soc. Am. J.
45:1079-1084.
Sposito,G., and S.V. Mattigod. 1977. On the chemicalfoundationof the sodiumadsorptionratio. Soil
Sci. Soc. Am. J. 41:324-329.
Sposito,G., and S.V. Mattigod. 1979. Ideal behaviorin Na+-tracemetal cation exchangeon Camp
Berteaumontmorillonite. Clays Clay Miner. 27:125-127.
Suarez,D.L., and M.E Zahow. 1989. Calcium-magnesiumexchangeselectivity of Wyoming mont-

morillonite in chloride, sulfate,and perchloratesolutions.Soil Sci. Soc. Am. 1. 53:52-57.
Suarez,D.L. 1996. Beryllium, magnesium,calcium, strontium, and barium. p. 575-601. In D.L.
Sumner,M.E. 1963. Effect of alcohol washingand pH value of leachingsolution on positive and neg-
ative chargesin ferruginoussoils. Nature(London) 198:1018-1019.
Sumner,M.E., L. de Ramos,and U. Kukier. 1994. Modification to compulsiveexchangemethodfor
determiningcation exchangecapacityof soils. Comm. Soil Sci. Plant Anal. 25:567-572.
Tabatabai,MA 1996. Sulfur. p. 921-960.In D.L. Sparkset al. (ed.) Methodsof soil analysis.Part 3.
Chemicalmethods.SSSABook Ser. 5.SSSAand ASA, Madison,WI.
Thomas,G.w. 1977. Historical developmentsin soil chemistry:Ion exchange.Soil Sci. Soc. Am. J.
41:230-238.
Thompson,H.S. 1850. On the absorbentpower of soils. J. R. Agric. Soc. England11:68-74.
Uehara,G., and G. Gillman. 1981.The mineralogy,chemistry,and physicsof tropical soils with vari-
able charge clays.Westview PressInc., Boulder, CO.
Vanselow, A.P. 1932. Equilibria of the base-exchange reactionsof bentonites,permutites,soil col-
loids, and zeolites.Soil Sci. 33:95-113.
Wada,K., and N. Matsue.1987. Commentson "Modification of the compulsiveexchangemethodfor
cation exchangecapacitydetermination".Soil Sci. Soc. Am. J. 51:841.
Way, J.T. 1850. On the power of soils to absorbmanure.J. R. Agric. Soc. England 11:313-379.
Way, J.T. 1852. On the power of soils to absorb manure. J. R. Agric. Soc. England13:123-143.
Zelazny, L.w., L. He, and A. Vanwomhaudt.1996. Chargeanalysisof soils and anion exchange.p.
1231-1253. In D.L. Sparkset al. (ed.) Methods of soil analysis.Part e. Chemical methods.
Published 1996
Chapter41
ChargeAnalysis of Soils and Anion Exchange

LUCIAN W. ZELAZNY, LIMING HE,
ANDAN VANWORMHOUDT, Virginia Polytechnic Institute
and State University, Blacksburg, Virginia
Most soil surfacesand colloidal systemsare chargedto some degree.Soil and

surface scientists have put forth extensiveefforts to measureand model the
chargesfrom thesesurfacesin order to betterunderstandadsorptioncharacteris-
tics of the solid and colloidal stability. However, eachscientistmust establisha
consistentterminology base,choosean appropriateexperimentalmethod,devel-
op a relevantmodel, and subsequentlyinterpretand reconcilethis datawith all of
the above.Often the soil scientistis further confrontedwith a mixture of miner-
als in which individual particlescan contain both constantand variable surface
chargesof varying strengthand densityresultingfrom severalprocesses,and fur-
thermorethe effectsof lateral heterogeneitiesin the form of intraplanar,interpla-
nar and interparticleinteractionsare unknown.The complexity of thesefactors is
a major sourceof the variancein the methodsof chargemeasurementand in the
modelsdevelopedto examinethe chargemagnitudeand distribution from solid
surfaces.
In order to satisfy the requirementof system electroneutrality,surface
charge is satisfied by positive adsorptionof counterions(oppositecharge)and
negativeadsorption(exclusion)of coions (samecharge)as shown in Fig. 41-1.
The chargedsurfaceincluding the counterbalanced and excludedions is referred
to as the electricdoublelayer (Grahame,1947).It also is termedthe Gouy-Chap-
man model since its analysiswas conductedby Gouy (1910) and independently
by Chapman(1913). The electric double layer obtainsits namefrom the charged
surfacelayer and its associatedlayer of counterions andcoions,while the electric
modifying term resultsfrom the electroneutralityreqliirement.Sincethe solution
chargesare to someextentfree to diffuse, the systemalso hasbeenreferredto as
the diffuse doublelayer. The diffuse modifying term resultsform the mobile solu-
tion chargesattainingequilibrium betweenthermal and electrostaticforces. This
is in contrastto the surfacechargesthat are assumedto be immobile. Specific
adsorptionof counterchargesalso are possibleand will be discussedlater. The
region of solution chargeimbalanceperpendicularto the chargedsurfacecan be
quite significant (i.e., 200 nm) relative to the size of the particle itself and is
Book Seriesno. 5.
1231
1232 ZElAZNY ET AL.
+ a
+ +
+ +
'"av
-"
'-
C/)
+
+
+
+
+
+
Bulk
solution
+
+ +
-0.15
~
N
I
E -0.10
U
~
b
'"
Ol
'-
-0.05
a
.s::
U
0.00
> -200
E
,g
c -100
c
'0'"
CL
0
10'
~
::::; 10°
10-'
c
0
10-2
~C
10-3
'"
V
C
10-4 d
0 10-5
U
0 10 20 30 40
Distance from surface (nm)
Fig. 41-1. A negativeconstantchargesurfacewith a diffuse double layer (ion concentration=0.001
M, ion valence=1, surfacecharge=-1.11 mole kg-I): a) distribution of positive (counterion)and
negative(coion) ions at the particle-solutioninterface, b)variation of chargewith distancefrom the
surface,c) variation of electric potentialwith distancefrom the surface,d) variation of cation and
anion concentrationwith distancefrom the surface.
called the double layerthickness.Double layer thicknessis an indication of ion

distribution from the surfaceand is important in determiningcolloidal stability.
Thus the concentrationof counterionsdecreases withincreasingdistancefrom
the surfacewhile the concentrationof coions increases.The diffuse layer charge
density (<1d) can be given by
<1D = 102 m l:Zd [Ci (x) - Ci (00)] dv [1]

i v
where
<1D =chargedensityof the diffuse layer (cmolc kg-I),
Zi =valencyof diffuse-layerIon i (dimensionlessbut carriessign),
SOILS & ANION EXCHANGE CHARGE ANALYSIS 1233
Constant charge surface Variable charge surface

10'
: ,>«_ C~tlons
10'
10 2 h
:~: . f i c ot;ons
.>-.~.:.~.. ::""'..........
c
o
2
: .:;:.........................
c
_::== = .. ----
Q)
u
C
o Anions
u
Anions
c
o
10- 7
10-' L-- - ' _---'-_-'-_ --'---' L----'_--'-_-'-_-'---'
1.0 ~----------, .-----------,
:: \,/\q/
0.8 I [ 1~(.lro l yte eoncenlr olion ( M)
I
c ···· · 0. 1
o Canons . - - 0.01
- 0.001
c
Q)
Ca tio ns
Z = 1
u
C
o
u
Anions
10 20 30 40 500 10 20 30 40 50
Distance from surface (nm)
Fig. 41-2. Ion distribution in the diffuse double layer of a negativeconstantchargesurfaceas con·
trastedto a negativevariablechargesurface(surfacecharge= -1.11 mole kg-I). Note that the dis-
tribution curvesfor cationsare symmetricalto thosefor anionsif a log scaleis used. But they are
asymmetricalif a linear scaleis used.
m =massof solid adsorbent(kg),

V = volume of aqueoussolution contactingadsorbent(L),
C,(x) = concentrationof Ion i at solution distancex (M), and
C;(00) = concentrationof Ion i in bulk solution (M).
On a log concentrationscalethis observeddistribution of counterionsand
coins is symmetrical,while on a linear concentrationscalecounterionchargefar
exceedscoion exclusion(Fig. 41-2). An exponential decrease of counterioncon-
centration,potential,and counterchargeoccurswith increasingdistancefrom the
surfacewith the magnitudeof the decreasedependingon assumptionsthat are
used (Fig. 41-1). In bulk solution,counterionconcentrationequalscoion con-
centration.Water moleculesas well as unchargedmoleculesalso may be present
in the diffuse layer, however, they are normally disregardedsince they do not
directly contribute to the local charge density except for possibly altering the
electric field by influencing the dielectric constant.Several models have been
developedto describeparticle surfacechargeand its associatedcountercharge
distribution.The fundamentaldifferencesfor all modelsare the processof charge
development,the location of particle surface,and the choice of countercharge
layersand their associatedinteractionwith particle charge.
1234 ZElAZNY ET AL.
SOURCEOF SURFACECHARGE
ConstantCharge
There are two major processesof developingsurfacechargethat result in
quite different behavior particularly at high ionic strengths(Gast, 1977). One
processis the developmentof permanentcharge,better knownas constantcharge
(a ee), which occurswith minerals such as montmorillonite and vermiculite and
arisesalmostentirely in phyllosilicateand zeolite minerals.The constantsurface
charge for these minerals results from the substitution of ions with differing
valence within the crystal units or from crystal imperfections,both of which
almostuniversallycreatesnegativelychargedsurfaces
[2]
where
aee =surfacechargedensityof constantchargecomponents(Cmole kg-I)
X = layer chargeof mineral (mole mol-I), and
MFW =formula weight of mineral (kg mol-I).
The magnitudeand locationsof this substitutionvary with phyllosilicatespecies
with further discussionof theseeffects on surfacechargegiven by White and
Zelazny (1986) and Bolt and Van Riemsdijk (1991). Often effective surface
chargeor the quantity of ions neutralizingsurfacecharge,which can be readily
exchangedwith anotherion [Le., cation exchangecapacity(CEq], is more im-
portantthan total structuralcharge.The absolutequantity of the mineral compo-
nentswith this processof chargedevelopmentis most significant for many soil
systemssince this is a structuralor bulk charge.Less importanceis given to sur-
face area since constantchargeis not derived from surfacereactions.Constant
surface charge is often expressedon a weight basisbut also can be given on a sur-
face areabasisfor normalizationpurposes.The ions balancingchargeon constant
charge surfacesare usually consideredas a homogeneouslayer or as being
smearedout on the surfacesincecounterchargeoccursat somedistancefrom the
point of substitutionin the crystal. Therefore,site numbersare not importantand
adsorptionis "mobile" being parallel to the surface.
Variable Charge
The other type of surfacechargeis pH-dependentor variablecharge(aye),
which occurson componentssuch as organicmatter,oxides,hydroxides,oxyhy-
droxides, and edgesof phyllosilicates.Hence,phyllosilicatescontain both con-
stantand variablesourcesof surfacechargewith the significanceof eachsource
being phyllosilicategroup dependent.Andersonand Sposito(1991) haveindicat-
ed that 14% of the total surfacechargeof Wyoming montmorillonite was of the
variablechargetype. Variable chargecan result from eitherion adsorption,or the
dissociationof surfaceionizablegroups.
The classicalprocessof variablechargedevelopmentresultsfrom sparsely
solubleionic solids adsorbingaqueousions that are constituentsof the solid. The
adsorptionof solution Ag+ and I- by solid AgI is the foremost example.These

adsorbedions are consideredpotential determiningions (PDIs) since they pro-
duce a measurablesurfacepotential that follows the Nemst equation
RT CPDI
"'0 = ZF In Co [3]
where
"'0 =surfacepotential(volts),
R =molar gas constant(8.314 J mol-l K-I),
T = absolutetemperature(K),
Z = ion valence(dimensionlessbut carriessign),
CPD! = concentrationof PDI (M), and
Co =concentrationof PDI (M), at which =O. "'0
Systemsthat obey the Nemst equation include silver halides, silver sulphide
(Ag2S), calcium sulfate(CaS04),calciumcarbonate(CaC03), and calcium phos-
phateCa(H2P04)2' Thesesystemsprovide a familiar O.059/z volts surfacepoten-
tial increasefor every decadeconcentrationincreaseof PDI adsorbedat 25°C
(298 K). Sincechargeon thesemineralsis only dependenton adsorbedPDIs that
createa surfacepotential, thesemineralsalso have beenlabeledconstantpoten-
tial surfaces.
The adsorptionof (solid-unlike) ions that are not constituentsof the solid
or the ionization of surfacegroupssuchas H+ or OH- also will result in variable
charge.This chargeusually exhibits a surfacepotentiallessthan Nemstianbehav-
ior, and in order to avoid confusion these adsorbedions should be labeled as
chargedeterminingions (CDIs). Upon adsorption,the CDIs may becomeindis-
tinguishablefrom the solid matrix, however,like PDIs variablesurfacechargeis
developed.Variable chargeresultsfrom the sum of proton transferreactions(OH)
and other surfacecoordinationreactions(OeDI) and is given by
[4]
where
ove = surfacecharge densityof variable chargecomponents(cmole kg-I),
oH =
surfacechargedensity of proton transferreactions(cmole kg-I), and
0CD! =
surfacechargedensity from CDI adsorptionreactionsother than H+ or
OW (cmolc kg-I).
The 0H is defined as
[5]
where
r H =adsorptiondensity of protons(cmole kg-I), and
r OH = adsorptiondensityof hydroxyls (cmolc kg-I).
The OCD! is defined as
1236 ZELAZNY ET AL.
(JCD! =L (ZCD! r CD!) [6]
where
ZCD! =valenceof adsorbedCDI's other than H+ or OH- (dirnensionlessbut
carriessign), and
r CDl = adsorptiondensityof CDI's otherthan W or OH- (crnolc kg-I).
Systernsnot obeyingthe Nemstequationinclude rnost naturalsurfacespar-
ticularly the rneal oxides, hydroxides,and oxyhydroxideswith the possibleex-
ception of Ti02• The non-Nemstianbehaviorgenerally results frorn an activity
changeof the adsorbedion as the surfacebecornescharged.It is obviousthat the
sourceof variable chargesfrorn both PDIs and CDIs is due to surfacereactions
with functional groupsand for that reasonsurfaceareaof the cornponentrniner-
al becornesan irnportantpararneterand of lesserirnportanceis the absolutequan-
tity of the rnineral. Variable chargesare biilancedby both cationsand anionsand
can be expressedon a weight basis(as given in Eqs. [5] and [6]), but are often
expressedon a surfaceareabasisfor cornparativepurposes.The chargeon vari-
able chargesurfacesis usually consideredto be "discrete" or to occur at a spe-
cific site. Therefore,site nurnberson the particle surfaceare quite irnportantwith
adsorptionlocalized at recognizablesites.
Charge/Potential Relationships
We can relate the surfacechargedevelopingfrorn thesetwo very different

processesto surfacepotential accordingto the Gouy-Chaprnantheory for syrn-
rnetrical electrolytesas follows
(J =(8.6 X 10-3 RTEf:OC)I/2 sinh ze'l'o

-- [7]
2RT
where
(J =surfacechargedensity (crnol kg-I),
C =electrolyteconcentration(M),
e =electroniccharge(1.6021 x 10-19 C),
10 = dielectric constantof rnediurn (for water at 298 K = 78.5), and
Eo =permittivity of free space(8.854 x 10-12 C y-I rnol-I).
By definition the surfacechargefor constantchargernineralsis not influencedby

pH, electrolyteconcentration,solventcharacteristics,ionic rnakeupof the ernbed-
ding solution, or ternperature,howeverthe surfacepotentialfor thesernineralsis
affected by all of the above pararneters.Alrnost the reverseoccurs for variable
chargesurfaces.Again by definition surfacepotential for variablechargerniner-
als doesnot vary with electrolyteconcentration,solventcharacteristics,ternpera-
ture or ionic rnakeupof the ernbathingsolution providedthe ions are not PDIs or
CDls, howeversurfacechargedoes.It is irnportantto note that the surfacepoten-
tial on variablechargernineralsdoeschangewith adsorptionof PDls or CDls, or
a pH change.Although the surfacechargefor constantcharge rninerals is not
influenced by electrolyte concentration,the ratio of counterionsadsorbedto
coionsexcludedis greaterat low concentrationsthan at high concentrations(van

Olphen,1963).For variablechargeminerals,the ratio of counterionadsorptionto
coion exclusion remains unaltered with increasing electrolyte concentration,
while the ratio of surfacechargevaries as a squareroot function of electrolyte
concentration(van Olphen, 1963). One also should note the larger double layer
thicknessfor constantcharge surfacesthan for variable charge surfaceswith
increasingelectrolyteconcentration(Fig. 41-2). Thus the processof developing
surface charge results in very different behavior and is a major factor in the
methodof chargemeasurement and model development.
COUNTERION ADSORPTION
Specific VersusIndifferent
The adsorbedcounterionscan either havea specific affinity for the particle

surfaceor be indifferent. Specifically adsorbedions form surfacecomplexesof
varying strengththat for certain ions (i.e., organic) may be enhancedby hydro-
phobic interactions,hydrogen-bondformation, or London dispersion.Specifical-
ly adsorbedions can be either located at some distancefrom the surface, or
assumedto coincidewith the surfaceplane(Hunter, 1993). For most naturalsys-
temsthat are very complex,this separationis not alwayseasily madeand is ulti-
mately determinedby defmition. Indifferent ions are not specifically adsorbed
but exhibit strictly coulombic interactionswith the surfaceand are often placed
in the diffuse layer. It is frequentlyassumedthat simple 1:1 electrolytesare indif-
ferent particularly at low electrolyte concentrations.Exceptions,however, do
occur such as surfacecomplexesof Na+ and Cl- with kaolinite and oxide soil
minerals(Ferris & Jepson,1975; Davis et aI., 1978).
Inner-spbereVersusOuter-spbere
Two broadcategoriesof specific adsorptioncan be distinguishedon struc-

tural grounds.If no moleculeof the bathingsolventis interposedbetweenthe sur-
face functional group and the molecule unit it bounds,the complex formed is
called an inner-spherecomplex. If at least one solvent molecule is interposed
betweenthe functional group and the bound molecule, the complex formed is
called an outer-spherecomplex. As a general rule, outer-spherecomplexes
involve electrostaticbondingprocessesand thereforeare less stablethan inner-
spherecomplexes,which necessarilyinvolve either ionic or covalentbondingor
somecombinationof theseprocesses.Spectroscopicor nuclearresonancetech-
niquesthat can measurealterationsin the solvation statusof an ion can be used
to distinguish between these two complexes. Assessingthe effects of ionic
strengthon surfacecomplexationequilibria is the usual indirect methodfor dis-
tinguishingbetweenthesetwo complexes.A strongdependence on ionic strength
is typical for outer-spherecomplexesbut ionic strengthhas minimal effects on
inner-spherecomplexes.Likewise, orbital overlap in outer-spherecomplexesis
much smaller than in inner-spherecomplexes.Generally specifically adsorbed
1238 ZELAZNY ET AL
-1600 . - - - - - - - - - - - - - - - - - - - - ,
____ Stern thickness
~~~
-1400
~'Surface potential
-1200
E -1000
""§ -800
~ ""~~ __ ~~_ Stern layer
-0
Cl.
-600
-400
I~ /,/ Stern potential

-200 V
DIstance from surface (nm)
Fig. 41-3. Potentialas a function of distancefrom the surfaceaccordingto the Stemmodel (ion con-
centration=0.001M, ion valence=1, surfacecharge= -1.11 mole kg-I).
ions also have a longer time scale of residenceon the surface as comparedto
indifferent ions which are presentin the diffuse layer. However,this separationis
not that distinct for comparingouter-spherecomplex ions to diffuse layer ions
since both speciesare solvated.
SternModel
Stem(1924) recognizeda specifically adsorbedlayer (Stemlayer) that had

moleculardimensionsfrom the surface(Stem thickness)and a specific adsorp-
tion potential that linearly decreasedthe surfacepotential to much lower values
(Stem potential (Fig. 41-3). This treatmentprovidesgreaterreliability for high
surfacepotentialmineralsand high electrolyteconcentrationsystems,and is fur-
ther able to separateadsorptiondifferencesobservedbetweensimilar valency
systems.The Gouy-Chapmanmodel is unable to differentiate betweensimilar
analogsystemsandpredictsexcessivelyhigh counterionconcentrationat the sur-
face even at moderatesurfacepotentials(e.g., 250 mV). Theselimitations result
from the assumptionthat counterionsbehaveas point chargesin solution and can
thereforeapproachthe surfacewithout limit.
Solid ParticleVersusEmbathingSolution
Intuitively it would appearthat identifying the terminationof the solid par-

ticle andthe beginningof its embathingsolutionwould be simple. Howeveragain
the experimentalistand modeler must make choices.All inorganic solids have
broken bonds on at least some of their crystal faces. A few common minerals
(Le., the entire phyllosilicate group) have no broken bonds on one face, while
other commonminerals(Le., quartz and feldspars)have broken bondson every
face. Surfacescontainingno broken bonds(Le., the siloxanesurfaceof 2:1 layer
phyllosilicates)are the simplestfor establishinga definitive boundarybetween

solid and embathingsolution. Brokenbond surfacesthat adsorbor releaseW or
OH- are dependentupon bond characteristicsand solution composition. The
adsorbedH+ or OH- is generallyconsideredas part of the surfacewith the embat-
hing solutioncontainingthe counterions.However,the chemisorptionof Cu2+ or
H2P04" on oxide surfacesor the edgesof kaolinite is moredifficult to resolvepar-
ticularly at the molecularscale.Often it is simplestto considerall inner-sphere
complexesincluding W or OH- as well asothereDIs as part of the surface,and
outer-spherecomplexesand diffuse ions as part of the embathingsolution.It
should be noted that W or OH- also can exist in the outer-sphereor diffuse
region.
POINTS OF ZERO CHARGE
A vast array of terms have beenusedto define the points of zero charge
(pzq that havecreatedmuch confusion.Part of this confusionis relatedto the
differencesin the two processesof developingsurfacecharges,many experimen-
tal methodsof determiningpze,and the assumptionsneededfor datainterpreta-
tion. Although nomenclatureconfusionexists,the importanceof PZC measure-
mentsin understandingadsorptioncharacteristicsof solidsand colloidal stability
is well recognized.
The surfacechargeon a particle has been conceptuallycharacterizedby
Sposito (1981, 1992) by genericdefinitions of various componentsof surface
chargeand the law of surfacechargeconservation.This simple but unique tech-
nique characterizesthe chargeon a particle with no molecularstructureimplied
or specific planesindicated,and providesan opportunity to define various PZC
parameters.For conveniencesurfacechargedensitiesaregroupedinto the intrin-
sic surfacechargedensity(Oint) to emphasizethe importanceof structuralcharg-
ing processes
[8]
and the Stem layer surfacechargedensity (ss) to emphasizethe importanceof

specifically adsorbedspecies
[9]
where
0is =inner-spherecomplexchargeotherthan H+ or OH- (cmolc kg-I),
Oos = outer-spherecomplexchargeotherthanW or OH- (cmolc kg-I).
The net total particlecharge(Ope) can now be separatedinto the following types
of surfacecharge
[10]
where
ape = total net particle surfacecharge(cmolc kg-I).
1240 ZELAZNY ET AL.
The O'pe as well as O'H, O'is, and 0'08 can be either positive, zero, or negative
depending on solution composition.The O'ce is almost always negative and
rangedfrom -0.02 mole kg-1 for hydrousoxidesand kaolinite to -2.5 mole kg-1
for vermiculite. The SH rangesfrom -9 to +1 mole kg-1 for soil organic matter,
and from -0.7 to +0.4 mole kg-1 for soil mineralscontainingsurfaceOH groups
(Sposito,1984, 1989).
For an individual particle in suspensionor an entire suspension,the gener-
al conservationlaw requiresthat the total netparticlesurfacechargebe equaland
oppositeto the diffuse charge(O'D) so that
O'pe + O'D = o. [11]
Although soil particlesare often charged,soils or aqueoussuspension themselves

must alwaysbe electrically neutral.Thus,
[12]
The PZC definitions establishedby Sposito(1982,1992)are consistentand

extremely useful. His defining conditions include pH values or properties at
which one or more of the surfacechargecomponentsvanishat fixed temperature,
pressureor aqueoussolution composition(Table 41-1). The PZC (Everett, 1992)
is the pH value at which net particle chargevanishes(O'pe =0). Thus there is no
diffuse layer (O'd) and all adsorbedions are surfacecomplexedand considered
immobile. The point of zero net proton charge (PZNPC) (Sposito, 1981) is
definedas the pH value at which the net protonsurfacechargedensityis zero (O'H
= 0). It alwaysdecreasesas pH increasesregardlessof soil solution composition
or ionic strength.The point of zero net charge(PZNC) (Parkeret aI., 1979) is the
pH value at which the intrinsic surfacechargedensityis zero. Thus it also equals
the condition wherenet adsorbedion chargeother than O'H vanish(O"is + O'os + O'D
=0). For the adsorptionof an indifferent electrolyteit is the pH valuewhereCEC
is equal to AEC. The point of zero salt effect (PZSE) (Parkeret aI., 1979) is the
pH value where net proton surface chargedensity is unaffectedby changesin
ionic stiength(I). Often it is definedas the crossoverpH value of potentiometric
acid-basetitration analysesperformedat variousionic strengths.Relating PZSE
to the vanishingof a surfacechargecomponentis indirect and difficult to inter-
pret. The pH value where electrophoreticmobility (~) vanisheshas beentermed
the isoelectricpoint (IEP) (Everett, 1992).
Table 41-1. Pointsof zero charge(Sposito1981).

Symbol Name Defining condition
PZC Point of zero charge ape =0

PZNPC Point of zero net proton charge aH =0
PZNC Point of zero net charge aint =0
IEP Isoelectricpoint Electrophoreticmobility (11) =0
PZSE Point of zero salt effect aaH/aI =0
MEASUREMENT OF SURFACECHARGE
Severalexperimentalmethodsare availablefor measuringsurfacecharge.

Particle surfacechargecan be analyzedby itself as well as the accompanying
counterchargethat is equalbut oppositein sign. Surfacechargeson 2:1 type phyl-
losilicateshavebeendeterminedby extensivechemicalanalysisaccompaniedby
chargebalancecalculations.This method ignores chargesdevelopedfrom edge
effectsand is only applicablefor monomineraliccompositions.Particlescontain-
ing variable chargeresulting from proton transferreactions«jH) have beensuc-
cessfully examinedby measuringthe proton excessor deficiency at the surface.
Often potentiometricacid-basetitration analyseshave been successfulin mea-
suring surfacechargeon oxides, hydroxides,and organic matter. This analysis
assumesthat H+ or OH- are the only CDIs that createcharge.This method also
is most applicablefor monomineraliccompositions.
Accompanyingcounterchargesare often determinedby cation and anion
adsorptionanalysis.The adsorptionanalysiscan be determinedby measuringthe
changein solution concentrationafter addition of the solid phaseas is typical for
"pH edge" determinations,or by measuringthe quantity of solute that is associ-
ated with the solid as is typical for CEC and AEC analysis. Often these latter
methodsrequire some formof ion saturation,removal of excesssalts, and solid-
liquid separation(i.e., centrifugationor filtration), which may result in problems
and artifacts. Shifts of the pH edgeat high sorbate/sorbentratios can result from
saturationof availablesitesor multiplicity of site types.Also the resultingcalcu-
lated surfacechargefrom theseadsorptionmeasurements often neglectsnegative
adsorptionand possibleadjoining site neutralizationof oppositecharge.
It is apparent thatsurfacechargeis an experimentallyaccessibleparameter
but surfacepotential is not. Thus, knowledgeof the chargedistribution from the
surface,which is essentialin exploring colloidal stability, cannot be obtained
from adsorptionmeasurements alone.It requirespotentialmeasurements for con-
firmation or appropriatemodeling with assumptionsregardingcharge-potential
relationships.Potentialmeasurements at specifieddistancesfrom the surfacecan
be obtainedfrom electrokineticstudies,however, severalassumptionsmust be
involked (Hunter, 1981) in regardsto the shearor slipping plane, the viscosity
and permittivity of the solution near the surface,the particle geometry,and the
relaxation effect at high potentials.The potential at the slipping plane has been
calculated asthe zetapotential (~) and it is often assumedto occur at the bound-
ary betweenthe Stem layer and the beginningof the diffuse layer. In pure sys-
temszetapotentialscan provide somemodel constraintsbut their usefulnessdra-
matically decreaseseven in a mixture of two mineral species (Anderson &
Bertsch,1993).
Although no universally acceptabletechniqueexistsfor measuringthe sur-
face chargeof soils, the measurements of PZNC, PZNPC, PZSE, and IEP, for
monomineralicsystemshave most often beenreported(Table 41-2). The PZSE
measurementsare generally easiestto experimentallyperform but are theoreti-
cally ambiguous.The PZNC measurementsare experimentally attainablewith
the firmest theoretical background.The PZNPC and IEP measurementshave
been successfullydeterminedon pure mineral componentsbut their interpreta-
1242 ZELAZNY ET AL
tions are ambiguousfor heterogeneous naturalparticles.Sposito(1992) indicates

that his PZC theorems..."suggestthat the point of zero net charge(PZNC) is the
most accessibleexperimentallyand the most versatileconceptually."Presented
herearethe methodsfor determiningPZNC, PZSE,PZNPCandconstantcharge-
densityfor soils.
POINT OF ZERO NET CHARGE

Introduction
The principle of determiningPZNC is identical to that of CEC measure-

ments. However, PZNC determinationsincludes analysisfor ABC as well as
CEC. The methodfor measuringPZNC wasfirst developedby Schofield(1949).
Soils weresaturatedwith 0.2 M NH4CI, and subsequentlythe adsorbedNH.t and
CI- were replacedby K+ and NOj" in 0.2 M KN03 solution. This methodwas
improvedandmodified by manylaterresearchers (van Raij & Peech,1972;Wada
& Okamura, 1977; Marsh et aI., 1987). The key to determiningPZNC is the
selectionof both a cationand anion that are usedto saturatesoils at a concentra-
tion and pH that are realistic to natural conditions. Theseions should not be
specificallyadsorbedby soils and ideally shouldbe easyto analyze.It seemsthat
K+ and Cl- are the bestchoicesfor saturationsinceNa+ can sometimesdisperse
soils and causedifficulties in solid-liquid separation(van Raij & Peech,1972).
The methoddescribedhere is modified from Ueharaand Gillman (1982), with
KCI as a saturatingsolution and NaN03 insteadof NH4N03 as a replacingsolu-
tion, sinceNILJ+ can displacefixed K+, overestimatingCEC if the soil contains
soil mica and vermiculite.
Table 41-2. Point of zero chargefor selectedoxides,hydroxides,oxyhydroxides,and phyllosilicates

~~er Sposito (1984),and Stumm (1992)]t.
Material PZNPC PZNC PZSE IEP
Corundum(a-Al20 3) 9.1 9.1 ± 0.2

Alon (y-Al 20 3) 8.2 ± 0.5 8.5 8.7
Gibbsite(a-Al(OH)J) 8.5 ± 0.5
Boehmite(y-AlOOH) 8.2
Hematite(a-Fe203) 8.4 ± 0.1 8.5
Magnetite(a-Fe304) 6.5
Goethite(a-FeOOH) 7.7 ± 0.2 7.3 ± 0.2 6.1 ±0.6
Amorphousiron (Fe(OH)J)* 8.0 ± 0.1 8.0 ± 0.1 8.1 ± 0.1
Quartz(a-Si02) 2.9 ± 0.9 2.0 ± 0.3
Pyrolusite(p-MnO:z) 7.2
Birnessite(8-MnO:z) 2.2 ± 0.7 1.9 ± 0.5 2.3:!: 1.1 1.7 :!: 0.4
Calcite (CaC03) 9.5 10:!: 1
Feldspars 2.2 ± 0.2
Kaolinite 4.6 4.8 4.7
Montmorillonite 2.5
t The valuesgiven herevary with investigatorsand materialsused.
* Dzombakand Morel, 1990.
Apparatus
1. Centrifuge.
4. pH meter.
Reagents
1. 1.0 and 0.01 M KCl.

2. 1.0 M KOH and HCl.
3. 0.5 M NaN03•
Procedure
Place5 g (383K oven dry-weight basis)of air-dried soil sampleinto each
of 10 100-mLprelabeledand preweighedcentrifugetubesto 0.001 g. Add 50 mL
of 1M KCI and shakefor 1 h using a reciprocalshakerwith a speedof 100 oscila-
tions min-I. Centrifugeand discardthe supernatant.Washfive times with 50 mL
of 0.01 M KCl. At the final washing,adjust the pH valuesof the suspensionsto
spanthe expectedPZNC by adding 1 M KOH or HCl. Shakefor 4 h, centrifuge,
determineand recordsupernatantpH. Centrifugeand decantthe final supernatant
but retain for K+ and Cl- determination(CI). Note that the supernatantfrom each
centrifugationshould be clear. Otherwise, the removal of suspensionparticles
will produceexperimentalerrors. Weigh the tubes to obtain the volume (VI) of
entrainedsolution in the sedimentin order to determineentrainedK+ and Cl-.
When weighing, a foam holder shouldbe usedto hold tubesin a vertical position
to reduceweighing errors resultingfrom variableorientationof tubeson the bal-
ance pan. It is assumedthat the concentrationsof the entrainedK+ and Cl- are
equal to thoseof the final washingsolution. Washfour times with 20 mL of 0.5
M NaN03 to displace the adsorbed K+ and Cl-. Centrifuge and decant the
exchangingsolution into a 100-mL volumetric flask (V2) that is brought to vol-
ume and subsequentlyanalyzedfor K+ and Cl- (C2). Calculatethe CEC (K+) and
the AEC (Cn as centimolesof chargeper kilogram. A plot of CEC and AEC vs.
pH will reveal the crossoverpoint where AEC = CEC, which correspondsto
PZNC.
Calculations
[13]
I 0_._1~(C..::.2.-:V2::..--_C.....:I,--V=I)
AEC (cmole kg- ) = 35.5 W [14]
where
C] =concentrations(mg L-l) of K+ andCI- in final washingsolutionof 0.01
MKCI,
1244 ZELAZNY ET AL
C2 = concentrations(mg L-1) of K+ andCl- in the displacingsolutionof 0.5

MNaN03,
V1 =volume (mL) of the solution entrainedin sedimentsafter the final
washingof 0.01 M KCl,
V2 =total volume (mL) of the displacingsolutionof 0.5 M NaN03,
39 =atomic weight of K+,
35.5 =atomicweight of Cl-, and
W =oven-driedsoil sampleweight (g).
Comments
Although the sourcesof errors in conventionalCEC methodsare applica-

ble to PZNC determinations(seeSumner& Miller, 1996,Chapter40), otherdou-
ble layer detailsalso are importantto consider.It is recognizedthat both valence
and concentrationof the saturatingions influencesurfacechargedensity,which
in tum affectsPZNC. Positive,negative,and net chargedensities,determinedby
divalent ions, are generallyhigher than thoseobtainedwith NaCI (van Raij &
Peech,1972), and this effect is evengreaterwhen both the cation and the anion
are divalent (i.e., MgS04). Upon decreasingthe electrolyte concentration,the
magnitudeof both positive and negativechargesdecreases(van Raij & Peech,
1972; Espinozaet aI., 1975). Suchdecreasesin the surfacepositive and negative
chargeswith decreasingelectrolyteconcentration,at a constantpH, are in agree-
ment with double layer theory for variable chargesurfaces,and may be in part
due to the mutual cancellationof adjacentoppositechargewhen the distanceof
separationof the chargesbecomeslessthandoublelayerthickness.Therefore,the
observationof Marsh et a1. (1987) that the amountsof surfacecharge(positive
and negative)measuredin 0.1 M NaCI were similar to thosemeasuredin 0.025
M CaCl2 is reasonable.ComparisonamongPZNC determinationfrom different
researchers shouldbe madecautiously.If <Jis is significant, ions like K+ and Cl-
acting as probeelectrolytesare unableto displacetheseinner-spherecomplexes.
As a result, the valuesof CEC and ABC determinedwill be lower than those
expected(Sposito,1992).
Errors in CEC andABC determinationoften result from disregardingneg-
ative adsorptionand its influenceon the ion concentrationin the entrained solu-
tion. As a result,ABC valueslessthan zero havebeenobservedby van Raij and
Peech(1972),Laverdiereand Weaver(1977), Parkeret a1. (1979),andAnderson
and Sposito (1992). A correction for negativeadsorptionwould increaseboth
positive and negativecharges,but the net chargewould be lessaffectedbecause
of their mutual cancellation.
POINT OF ZERO SALT EFFECT
Introduction
Potentiometrictitration analysesoften have been performed at selected

ionic strengthsfor measuringa PZSE. From thesemeasurements a PZNPCalso
can be obtainedfor monomineralicsolids such as oxides, hydroxides,and oxy-

hydroxidesthat consistentirely of variablechargeresultingfrom proton transfer
reactions.Theseanalyseshave beenperformedon soils, howeverexperimental
and interpretationaldifficulties must be overcome.A soil sampleis equilibrated
in a known backgroundelectrolyte solution, containingsequentialadditions of
acid or base.Temperature,pressureand soiVsolution ratio are maintainedcon-
stant while severalionic strengthsare selectedto bracket the area of interest.
After equilibration, the soil suspensionpH is determinedwith a glassand refer-
ence electrode.The protons adsorbedor releasedare often calculated as the
amountof acid or baserequiredto bring the pH of a blank solution of the same
backgroundelectrolyteconcentrationto that of the soil equilibratedin this same
solution. From theoreticalgrounds,it requiresa calibration in terms of H+ con-
centrationinsteadof H+ activity, the knowledgeof surfaceproton concentration
at the beginningof the titration, and the minimization of side reactionssuch as
dissolution,precipitation,and junction potential errors (Sposito, 1992). Often it
is assumedthat surfaceprotonexcessis zeroat the beginningof a titration, or that
the crossoverpoint of a potentrometrictitration performedat severalsalt contents
representszero surfaceproton excess.Theseassumptionsmay exist but mustbe
experimentallyverified. Charletand Sposito(1987) have establishedthe surface
adsorbed proton scalethroughindependentmeasurement of proton adsorptionat
the PZNC and the assumptionof knowledge that ape is zero. The ape is or
approacheszero for many Oxisols, Histisols, and Spodic horizons or must be
measured(Anderson& Sposito,1991).
EMF errors due to a junction potential can be minimized by inserting the
referenceelectrodeinto the clear supernatant.A supernatantbacktitrationtech-
nique has been developed to correct for pH-dependentsolubility effects
(Schulthess& Sparks,1986). Other dissolutionor precipitationreactionscan be
monitored during titration analysis through chemical analysis of the solution
(Chorover& Sposito,1995)or minimized byshort sampleequilibrationtimes or
rapid automatedpotentiometricanalytical techniques.The automatedpotentio-
metric techniquehas the advantageof providing a continuousanalysisthrough
the pH rangeexamined,but it requiresan automatictitrator and has largerjunc-
tion potential and variable solid/solutionratio errors. Thus, severalPZSE tech-
niquesare presented.
Batch Method (Discontinuous)
The batch methodconsistsof equilibratingseveralaliquotsof soil over an

extendedperiod of time with a fixed volume of an indifferent electrolyteat vari-
ous concentrationsthrough a rangeof pH values. Most equilibration times used
for soil systemsrangebetween2 and 4 d, but Choroverand Sposito(1995) indi-
catedpH stability of Oxisols after 0.3 h and Andersonand Sposito(1992) useda
1-h equilibrationtime to measurethe proton surface-charge densityof soils. Pre-
liminary trials can be usedto establishthe point in time where no changein pH
is observed.Sakuraiet al. (1989) examinedseveralsoiVsolutionratios and found
1:10 most suitable.Other authorshave successfullyusedthe ratios of 1:5 (Van
Raij & Peech,1972; Parkeret aI., 1979; Schulthess& Sparks,1986). Adjusted
1246 ZELAZNY ET AL
suspensionpH often rangesfrom threeto nine, howeverdissolutionand precipi-

tation reactionsalso aregreatestat the extremelimits of this rangeand shouldbe
avoidedif possible.Backgroundelectrolyteconsistsof indifferent cations(Li+,
Na+, K+, and NHt) and anions(CI-, NO)", or HCIO.$} at a concentrationranging
from 1 to 0.001M. Often NaCI is employedasthe backgroundelectrolyte.The 1
M ionic strength is generally not representativeof natural conditions, while
experimentallyit is difficult at an ionic strengthof 0.001M to vary pH and keep
ionic strengthconstant.
Another importantquestionto considerfor potentiometrictitration analy-
sesis whetherone is interestedin the surfacecharacteristicsof the naturalsoil or
if onewantsto comparedifferent soils. In the former case,an untreatedsoil sam-
ple is preferred.For the lattercase,the soil samplescanbe saturatedwith an indif-
ferent cation suchas Na in order to bring all samplesto an equalstateof satura-
tion.
Apparatus
1. pH meter with a combination electrodeand appropriatecommercial
standardbuffer solutions(in the acid range,use pH 4 and 7 buffers; in
the basicrange,usepH 7 and 10 buffers).
4. Centrifuge.
5. Automatic titrator for back titration procedureand automatedtitration
procedure.
Reagents
1. Standardbaseof 0.02 and 0.005 M NaOH [standardizedagainst0.02
and 0.005M potassiumacid phthalate(CgHsK04), respectively].
2. Standardacid of 0.02 and 0.005M HCI (standardizedagainststandard
baseof 0.02 and 0.005M NaOH, respectively).
3. StandardNaCI solutionsof 0.5, 0.1, 0.05, 0.02,0.01, and 0.005M.
Soil SamplePreparation
1. Untreatedsoil. Passthe air dried soils througha 2-mm sieve,thorough-
ly mix and usefor all determinations.
2. Sodium-saturated soil. wash5 g of sievedsoil with five 50-mL aliquots
of 0.5 M NaCI. This is followed by three50-mL washeswith either0.05,
0.01, or 0.005M NaCI to bring the occludedsolution to the desiredsalt
content.
Procedure
Place 5 g (383K oven dry-weight basis) of air-dried soil in each of 39
cappedplastic tubes,arrangedin threerows of 13. Designatethe middle tube in
eachrow aszero-additionof acid or base.Eachtube in a row will containa NaCI
solution of the sameionic strength(Le., 0.005, 0.01, and 0.05 M, respectively).
To establisha pH range(i.e., 3-9) add increasingamountsof respectively0.005

M HCI and0.005M NaOH (standardized)to the tubeson the left and on the right
of the zero tube (e.g., from 0.5-25 mL). For the 0.005M ionic strengthrow, add
0.005 M NaCI to bring the total volume in each tube to 50 mL. The remaining
ionic strengthsolutions(i.e., 0.05 and 0.01 M) havea similar sequentialaddition
of acid and basewith further additionsof ionic strengthsalt (0.02 and 0.1 M) and
distilled waterto provide a 50-mL solutionaliquot at a fixed ionic strength(Table
41-3). Dependingon samplecharacteristics, experimental design,and interpreta-
tional needs,othercombinationsmay be necessary to increaseor decreasethe pH
or ionic strengthrange.
Allow to equilibratefor 4 h while shakingsampleson a reciprocalshaker
with a speedof 100 oscillationsmin-to Centrifugeand determinethe supernatant
pH. Electrode/solutionequilibrium is consideredcompletewhen responsevaries
less than 0.01 pH units. If a chemicalanalysisof the supernatantis desired,it is
recommendedthat a subaliquotbe retainedbefore pH determination.It also is
recommendedthat conductivity measurementsbe performed to determine the
precisefinal ionic strengthof the equilibrium solution.
Performa blank titration at eachionic strengthstudiedaboveby using elec-
trolyte solutionswithout soils added.Similarly to the procedureof the soil sus-
pensiontitration, known amountsof electrolyte,standardizedacid or base,and
deionizedwater are mixed to producea solution of desiredpH and ionic strength.
The blank solutions have the sametotal volume as used in the soil suspension
titrations. Equilibration and pH measurements are all performedsimilar to those
describedabove.Calculatea dH-dOH (cmole kg-I) for eachsampleat eachionic
strengthexaminedand plot vs. equilibrium pH.
Table 41-3. Amount of acid, base,ionic strengthsalt awl distilled water necessaryto establisha 50-
mL aliquot consistingof a sequentialpH changewhile maintainingan ionic strengthof 0.005,0.01
and 0.05Mt.
0.005M O.OlM 0.05M
0.005M 0.005M 0.005M 0.02M Distilled O.lM Distilled
Tube no. HCI NaOH NaCI NaCI H2O NaCI H2O
mL
1 25 0 25 18.8 6.2 23.2 1.8
2 15 0 35 21.2 13.8 24.2 10.8
3 8 0 42 23.0 19.0 24.6 17.4
4 4 0 46 24.0 22.0 24.8 21.2
5 2 0 48 24.5 23.5 24.9 23.1
6 0.5 0 49.5 24.2 25.3 25.0 24.5
7 0 0 50 25 25 25 25
8 0 0.5 49.5 24.2 25.3 25.0 24.5
9 0 2 48 24.5 23.5 24.9 23.1
10 0 4 46 24.0 22.0 24.8 21.2
11 0 8 42 23.0 19.0 24.6 17.4
12 0 15 35 21.2 13.8 24.2 10.8
13 0 25 25 18.8 6.2 23.2 1.8
t Valuesgiven to 0.1 mL significanceonly.

1248 ZELAZNY ET AL.
Calculations
[lO-PHB - lO-PHS] - [lO-<I4-pHB) - lO-<I4- pHS)] 0.1

MI-!lOH = x- [15]
'Y W
where
MI-!lOH = apparentprotonsurfacechargedensitycalculatedby titration at
given ionic strengthand pH in cmo1.Jcg-I,
MI = the differencebetweenthe final H+ concentrationof a suspension
and that of the blank,
!lOH =the differencebetweenthe fmal OH- concentrationof a suspen-
sion and that of the blank,
pHB =pH of the blank solution,
pHs =pH of the solutionequilibratedwith the sample,
14 =conditionaldissociationproductof water,
r = single ion activity coefficientcalculatedwith the Daviesequa-
tion (Davies,1962),and
W = oven-driedsoil sampleweight (g).
BacktitrationMethod
After the soil sampleshave beenequilibrated,centrifugedand the super-

natantpH measured,asin the batchmethod,the alkalinity or acidity canbe deter-
minedby backtitratingto pH 7 asan end-pointwith an autotitrationsystemusing
0.02 M HCI asa titrant for the alkalinesupernatantsand0.02M NaOH asa titrant
for the acid supernatants(Schulthess& Sparks,1986).An aliquot (i.e., 40 mL) of
the supernatantmust be taken since solution lossesoccur due to entrainingby
soils. The supernatantmust be free<of all solids and can be filtered if necessary.
Calculatea !lH-AOH (cmolc kg-I) for eachsampleat eachionic strengthexam-
ined and plot vs. eqUilibrium pH.
Calculations
where
subscriptA =acid,
subscriptB =base,
V =volume of acid or baseaddedto the soil (mL),
C =concentrationof acid or baseaddedto the soil (M),
VBr =volume of acid or baseaddedin backtitration(mL),
CBr =concentrationof backtitrant(0.02M),
SV =volume of supernatantbacktitrated(mL), and
W =oven-driedsoil sampleweight (g).
Automatic PotentiometricTitration (Continuous)
The automaticpotentiometrictitration proceedurehas the distinct advan-

tageof a continuouspH analysiswith the disadvantageof greaterjunction poten-
tial errors,kinetic effectsandvarying soil/solutionratios. Five grams(383K oven
dry-weight basis)of air-dried soil sampleis placed in the titrating vesselalong
with 50 mL of the appropriateionic strengthcontrolling salt solution (Le., 0.05,
0.01, 0.005 M). The sampleis stirred for 2 min for pre-equilibration(Sakuraiet
aI., 1989). Subsequentlywith continuousstirring, titrate at a certaininterval with
0.02 M Hel or NaOH made up in the ionic strengthcontrolling solution. There
are many choices concerningthe interval of acid/baseadditions. Overall, the
interval increaseswith increasingelectrolyteconcentrationand increasingcharge
density of the variable chargecomponents(Sakurai et aI., 1989). Sakurai et al.
(1989)suggests the following intervalsof titrant additions:For volcanic ashsoils,
use 12 s for 0.01 M systemsand 120 s for 0.1 M systems;for strongly weathered
soils, use 24 s for 0.01 M systemsand 240 s for 0.1 M systems.
As in the batch method, a titration curve of the blank electrolyte is per-
formed in both the acid and basedirection and the differencebetweenthe sample
and the blank is assumedto be the W or OH- adsorbedby the sample.
Calculations
In this technique,an equationrepresentingthe blank titration curve needs

to be obtainedby fitting the blank titration datato an equation.The AH-AOH val-
ues at a given pH can be calculatedas follows.
Mf - AOH = 100[(NA,sample-NB,sample)- (NA, Blank -NB, Blank)]/W [17]
where
NA , Sample = cmol of acid addedto the soil suspension,
N B, Sample = cmol of baseaddedto the soil suspension,
NA, Blank = cmol of acid addedto the blank solution to bring it to the samepH
value as the suspension.
This value may only be obtainedfrom the
fitted equationrepresentingthe blank titration curve,
N B, Blank = cmol of baseaddedto the blank solution to bring it to the samepH
value asthe suspension.This value may only be obtained fromthe
fitted equationrepresentingthe blank titration curve, and
W = oven-driedsoil sampleweight (g).
Comments
Differences in results obtained by the various proceduresfor measuring

PZSE are due to observeddifferencesin reaction rates (Hendershot,1978) and
variation in side reactions(Parkeret aI., 1979; Sposito, 1992). The presenceof
inner-sphereand outer-spherecomplexesgreatly influencesthe measuredPZSE
(Rymanet aI., 1979). Generallythe presenceof theseions on the exchange com-
1250 ZElAZNY ET AL
plex increasesthe measuredPZSEfor cationswhile lowering them for anions.

The oppositechangein measuredPZSEoccursif theseions constituteelectrolyte
solutions.
The AH and AOH quantity is the amountof H+ or OH- that is lost from
solution. It can createcharge or be absorbedon the soil colloid. Parker et al.
(1979) observedthat approximately30 to 90% of the acid or baselost from solu-
tion did not contributeto a surfacecharge.Other reactions,suchas ion exchange
betweenH+ and other cationsbalancingnegativechargesites, the dissolutionof
solids,aswell as the mechanismof partial neutralizationof adsorbed hydroxy AI,
could consumeH+ without affecting surfacecharge.It is thereforeimportant to
determinethe mechanismsresulting in the measuredAH-AOH.
Breeuwsma(1973) found it essentialto maintain a small nitrogen over-
pressurein the titration vesselto prevent CO2 absorptionfrom the atmosphere.
The absorptionof CO2 can indeedbe a sourceof error, especiallywith long equi-
libration times. This error can be reducedby cappingthe vessels orpurging with
N2·
POINT OF ZERO NET PROTONCHARGE
The mostcommonprocedureto estimatethe PZNPCis to equateit with the

pH value at the crossoverpoint of potentiometrictitration curvesobtainedat dif-
ferent ionic strengths,i.e., PZSEas describedabove.However,many researchers
indicate that the PZSEis unequalto PZNPC, for solid phasemixtures such as
soils. Direct measurementof the PZNPC for soils was proposedby Charlet and
Sposito(1987).
In the PZSEdetermination,we obtainedAH-AOH, the apparentprotonsur-
face chargedensity. To obtain the PZNPC,the net proton surfacechargedensity
(OH) for a given ionic strength andpH, must be determinedand can be calculat-
ed as follows.
0H =(Mf - AOH) - (Mf - AOH)PZNPC [18]
where (Mf - AOH)PZNPC is the apparentproton surface chargedensity at the

PZNPC(Plot 0H againstpH and determinethe pH value at 0H =0, which is the
PZNPC).The (AH-AOH)PZNPC may be calculatedby the following equation
(Mf - AOH)PZNPC =(Mf - AOH) + ace+ (CEC -AEC) [19]
where ace = the constantchargedensity of a soil and can be measuredby the

cesiumadsorptionmethod(cmol kg-I) of Andersonand Sposito(1991).
This procedureis necessaryto accountfor the surfaceproton excessthat
occurswhen potentiometrictitration analysisbegins.Thus the AH-AOH scaleis
adjustedby the initial surfaceproton concentrationto become0H. This centi-
molesper kilogram scalealso can be convertedto centimolesper meterssquared
scalethroughthe knowledgeof surfacearea(m2 g-I) that is importantwhencom-
paring variousvariablechargeminerals.
The valuesof ABC and CEC may be measuredby the methoddescribedin

"Point of Zero Net Charge."The direct determinationfor permanentcharge,O"ce,
is given below.
CONSTANT CHARGE DENSITY
To measurethe constant charge density of soils (Anderson & Sposito,

1991), a soil sampleis saturatedwith CsCl. The cesium-saturated sampleis then
dried to promote the formation of inner-spheresurfacecomplexesand washed
oncewith a dilute solution of LiCl. The lithium washpreferentiallydisplacesCs
from variablechargesitesbut not from constantchargesiteson the basalplanes
of 2:1 clay mineralsthat havea very low affinity for Li. Finally, Cs adsorbedon
constantchargesitesis extractedand quantifiedby reactingthe soil severaltimes
with NH40Ac solution.
Apparatus
1. Centrifuge.
4. pH meter.
Reagents
1. 0.05, 0.1, and 0.5 M CsCl.

2. 95% (VN) ethanol(~H50H)/water.
3. 0.01 M LiCl.
4. 1.0 M NH40Ac.
Procedure
Place1 g of air-driedsoil sampleinto a 1oo-mL prelabeledand preweight-

ed centrifugetube. Add 50 mL of 0.5 M CsCI and shakefor 30 min. Centrifuge
and discardthe supernatant.Washtwice with 50 mL of 0.1 M esCIandoncewith
0.05 M CsCl. This solution of 0.05 M CsCI shouldbe adjustedto a pH value of
near 7. The esCI saturatedsample is then washedonce with 50 mL of 95%
ethanolby shakingfor 1 min. Centrifugethe ethanolicsuspensionand retain the
supernatantfor cesiummeasurement. Weigh the tube to determinethe volume of
entrainedethanolsolution in orderto obtain entrainedCs. Dry the soil in the tube
at 338K for 48 h to promotespecific adsorptionof Cs+ on basalsurfacesites.
After drying, add50 mL of 0.01 M LiCI solution andshakefor 30 min with a re-
ciprocal shakerat a speedof 100 oscilationsmin-1 to displaceCs+ from variable
charge sites. Centrifuge and save the supernatantfor Cs+ measurement.The
entrainedsolution is weighed as describedabove.Extract the LiCI-washedsoil
four timeswith 30 mL of 1 M NH40Ac. Shakefor 30 min andcentrifugefor each
extraction.The supernatantfrom eachof the extractionsis combinedand saved
for cesiumanalysis.
1252 ZELAZNY ET AL.
Calculations
The constantchargedensity,O'ee> cmol kg-I, is given by the equation
O'zc = (Ncs.tot - MCs(LiCl) Ventr(LiCl» X HP [20]
where
Ncs.tot =total molesof NH40Ac-extractableCs,
MCs(LiCl) = concentration(M) of Cs in the LiCI solution, and
Ventr(LiCl) = volume of entrainedLiCI solution.
Comments
This procedurehasnot beenextensivelytestedon many soils but alternate

proceeduresare unknown.
REFERENCES
Anderson,M.A., and P.M. Bertsch. 1993. Electrophoreticmobility and particle size of clays using
laser Doppler velocimetry-photon correlation spectroscopy. Soil Sci. Soc. Am. J.
57:1641-1643.
Anderson,S.1., and G. Sposito.1991. Cesium-adsorptionmethodfor measuringaccessiblestructural
surfacecharge.Soil Sci. Soc. Am. 1. 55:1569-1576.
Anderson,S.J.,and G. Sposito. 1992. Proton surface-chargedensity in soils with structuraland pH-
dependentcharge.Soil Sci. Soc. Am. J. 56:1437-1443.
Bolt, G.H., and WH. Van Riemsdijk. 1991. The electrified interface of the soil solid phase.A. The
electrochemicalcontrol system.p. 37-80.In G.H. Bolt et al. (ed.) Interactionsat the soil col-
loid-soil solution interface.Kluwer Acad. Publ., Dordrecht,The Netherlands.
Breeuwsma,A. 1973.Adsorptionof ions on hematite(<x-Fez03):A colloid-chemicalstudy. Commun.
Agric. Univ., Wageningen,The Netherlands.
Chapman,D.L. 1913. A contribution to the theory of electrocapillarity.Philos. Mag. and J. Sci. 25
(Ser.6):475-481.
Charlet, L., and G. Sposito. 1987. Monovalent ion adsorptionby Oxisols. Soil Sci. Soc. Am. 1.
51:1155-1160.
Chorover,J., and G. Sposito. 1995. Surfacechargecharacteristicsof kaolinitic soils. Geochim.Cos-
mochim. Acta 59:875-884.
Davis, J.A., RD. James, and J.~. Leckie. 1978. Surface ionization and complexation at the
oxide/water interface. I. Computationof electrical double layer propertiesin simple elec-
trolytes. J. Colloid InterfaceSci. 63:480--499.
Dzombak,D.A., and EM.M. Morel. 1990. Surfacecomplexationmodeling: Hydrous ferric oxide. A
Wiley-IntersciencePublication.John Wiley & Sons,New York.
Espinoza,W, R.G. Gast, and RS. Adams, Jr. 1975. Chargecharacteristicsand nitrate retention by
two andeptsfrom South-centralChile. Soil Sci. Soc. Am. Proc. 39:842-846.
Everett, D.H. 1992. Manual of symbols and terminology for physiochemicalquantities and units.
Appendix II. Definitions, terminology and symbolsin colloid and surfacechemistry. Part I.
Butterworths,London.
Ferris, A.P., and W.B. Jepson.1975. The exchangecapacitiesof kaolinite and the preparationof
homoionicclays. J. Colloid InterfaceSci. 51:245-259.
Gast, RG. 1977. Surfaceand colloid chemistry. p. 27-73. In J.B. Dixon et al. (ed.) Minerals in soil
environments. SSSA, Madison,WI.
Gouy, G. 1910. Sur la constitutionde la chargeelectriqueala surfaced'un electrolyte.J. Phys. Radi-
um (serie4) 9:457-468.
Grahame,D.C. 1947. The electrical double layer and the theory of electrocapillarity. Chern. Rev.
47:441-501.
Hendershot,W.H. 1978. Measurementtechniqueeffects of the value of zero point of chargeand its

displacementfrom zero point of titration. Can. J. Soil Sci 58:439-442.
Hendershot,W.H., and L.M. Lavkulich. 1983. Effect of sesquioxidecoatingson surfacechargeof
standardmineral and soil samples.Soil Sci. Soc. Am. J. 47:1252-1260.
Hunter, R.I. 1981. Zeta potential in colloid science.Acad. Press,New York.
Hunter, R.I. 1993. Introduction to moderncolloid science.Oxford Univ. Press,New York.
Laverdiere,M.R., and R.M. Weaver. 1977. Chargecharacteristicsof Spodic horizons. Soil Sci. Soc.
Am. J. 41:505-510.
Marsh, K.B., R.W. Tillman, and J.K. Syers. 1987. Chargerelationshipsof sulfate sorption by soils.
Soil Sci. Soc. Am. J. 51:318--323.
Parker,J.e.,L.w. Zelazny,S. Sampath,and w.G. Harris. 1979. A critical evaluationof the extension
of zero point of charge(ZPC) theory to soil systems.Soil Sci. Soc. Am. J. 43:668--674.
Ryman,M.A.F., J.W. Bowden,and A.M. Posner.1979.The movementof titration curvesin the pres-
enceof specific adsorption.Aust. J. Soil Res. 17:191-195.
Sakurai,K., Y. Ohdate,and K. Kyuma. 1989.Potentiometricautomatictitration (PAJ) methodto eval-
uate zero point of charge(ZPC) of variable chargesoils. Soil Sci. Plant Nutr. 35:89-100.
Schofield, R.K. 1949. Effect of pH on electric chargescarriedby clay particles.J. Soil Sci. 1:1--8.
Schulthess,c.P., and D.L. Sparks. 1986. Backtitration techniquefor proton isotherm modeling of
oxide surfaces.Soil Sci. Soc. Am. J. 50:1406--1411.
Sposito,G. 1981. The operationaldefinition of the zero point of chargein soils. Soil Sci. Soc. Am. J.
45:292-297.
Sposito,G. 1984. The surfacechemistryof soils. Oxford Univ. Press,New York.
Sposito,G. 1989.The chemistryof soils. Oxford Univ. Press,New York.
Sposito,G. 1992. Characterizationof particle surfacecharge.p. 291-314.In J. Buffle and H.P. van
Leeuwen(ed.) Environmentalparticles.Vol. 1. Lewis Publ., Boca Raton,FL.
Stern,O. 1924. Zur theorie der elecktroyltischendoppelschict.Z. Electrochem.30:508--516.
Stumm,W. 1992. Chemistryof the solid-waterinterface.John Wiley & Sons,Inc., New York.
Sumner, M.E., and W.P. Miller. 1996. Cation-exchangecapacity and exchangecoefficients. p.
Uhara,G., and G. Gillman. 1982. The mineralogy,chemistry,and physicsof tropical soils with vari-
able chargeclays. Westview Press,Boulder, CO.
Van Olphen, H. 1963. An introduction to clay colloid chemistry. 2nd ed. JohnWiley & Sons, New
York.
Van Raij, B., and M. Peech.1972. Electrochemicalpropertiesof some Oxisols and Alfisols of the
Tropics. Soil Sci. Soc. Am. Proc. 36:587-593.
Wada,K., and Y. Okamura.1977. Measurements of exchangecapacitiesand hydrolysisas a meansof
characterizingcation and anion retentionsby soils. p. 811--815. Proc. Int. Seminaron Soil
Environmentand Fertility Managementin intensiveagriculture.Tokyo. 10--14 October.The
Societyof the Scienceof Soil and Manure,Tokyo, Japan.
White, G.N., and L.w. Zelazny. 1986. Chargepropertiesof soil colloids. p. 39--81. In D.L. Sparks
(ed.) Soil physicalchemsitry.CRC Press,Boca Raton,FL.
Published 1996
Chapter42
RedoxMeasurementsof Soils
w. H. PATRICK, R. P. GAMBRELL,
AND S. P. FAULKNER, Wetland Biogeochemistry Institute,
Louisiana State University, Baton Rouge, Louisiana
Soil reduction-oxidation or redox potential measurementshave been used

throughoutmost of the history of soil scienceto characterizethe intensity of
reductionor oxidation and relate this to biological processesoccurring in flood-
ed soils. Along with the disappearance of soil O2, the decreasein redox potential
is the most striking changecausedby flooding a soil. Aeratedsoils havecharac-
teristic redox potentialsin the range+400 to +700 mY. Seasonallyflooded soils
have a much broaderrangeof redox potentialssince they function as both oxi-
dized (+400 to +700 mY) and reduced(as low as -250- to -300 mY) systems.
The narrow range and poor reproducibility of redox potentialsin well-drained
soils limits their value as a tool for characterizingaeration. The poor repro-
ducibility is causedprimarily by a lack of poisingof the reduction-oxidationsys-
tems dominatedby O2 (Ponnamperuma,1955). Redox potential measurements
are most useful if limited to flooded soils and sediments.Generally,the reduc-
tion-oxidationstatusof surfacewatersand oxidized uplandsoils is bettercharac-
terizedby dissolvedO2 and soil atmosphericO2 measurements, respectively.An
exceptionwould be beneaththe thermoclineof a deep-waterbody wherestrong-
ly reducedconditions exist and one may want to know whether the systemis
moderatelyreducing(i.e., ferrous iron is stable)or strongly reducing(sulfide is
stable).Another exceptioninvolves wetlanddelineationstudieswhere a transect
from wetland to upland is being characterizedand redox readingsfrom wetland
and transitionzonescan be comparedto uplandreadings.Also, seasonalchanges
in the oxidation statusof wetlandand transition-zonesoils can be followed.
The redoxpotentialof an aqueouschemicalsystemis a measureof the elec-
tron availability or potential. For redox potential measurementsin biological-
chemicalsystemssuch as soils, a platinum electrodeis usually employed.The
platinum electrodereadily transferselectronseither to or from the medium, but
presumablydoesnot reactchemicallywith the medium.When the platinum elec-
trode is coupledwith a suitable half cell of known potential, reducing systems
tend to transferelectronsto the electrodewhile oxidizing systemstend to take
electronsfrom the electrode.For actual redox potential measurements, electron
flow is prevented,and the potentialbetweenthe half cell composedof the plat-
Book Seriesno. 5.
uss
1256 PATRICK ET AL.
inurn in contactwith the substrateand the known potentialof the referenceelec-

trode half cell is measuredwith a suitable meter that respondsto electromotive
force or potential.
THEORY
Chemical reactionsin soils and sedimentsinvolve some combinationof
proton and electron transfer (Baas Becking et aI., 1960). Electron transfers
involving the lossof electronsare known asoxidation reactionssince,in the early
days of chemistry, the known oxidation reactions(i.e., rust formation) always
involved O2, The gain of electronsis called reductionsincethe addition of nega-
tively chargedelectronsreducesthe overall chargeof cationicspecieswhich have
beenhistorically the most studied.The quantitative measure of the electronavail-
ability in thesereactionsis defined as the electrodepotential and denotedas Eh.
The electrodepotential can be defined as either oxidation or reductionpotential
dependingon how the chemicalreactionis written.
As Bohn (1971) aptly statedover 20 yr ago, the "ambiguity in oxidation-
reductionnomenclatureis long-standingand unresolved."Thereis still confusion
over the many termsusedto describethesereactions:electrodepotential,oxida-
tion-reduction,redox potential, EO, Eh, etc. While oxidation potentialsare more
commonly usedin the chemistryliterature,reductionpotentialsare more applic-
able to soils and sediments.Therefore,we hopeto clarify the terminology in soil
systemsby defining the term redox potential (Eh) as the reduction potential.
Redox reactionsof soil oxidantsare represented by the following half-cell reduc-
tion equation
Ox + mH+ + ne- ~ Red [1]
where Ox is the oxidized componentor electron acceptor,Red is the reduced

componentor electrondonor, m is the numberof hydrogenions involved in the
reaction,and n is the numberof electronsinvolved in the reaction.The electrons
usedin Eq. [1] mustbe suppliedby an accompanyinghalf-cell oxidation reaction.
In soils, organic matter(CH20) is the primary sourceof electrons.Therefore,a
completeredox reactionbalancesthe reductionreactionwith an appropriateoxi-
dation reaction.Equations[2], [3], and [4] illustrate this approach withthe reduc-
tion of Fe(OHh
4Fe(OHh+ 12H+ + 4e- ~ 4Fe2+ + 12H20 (reduction) [2]
CH20 + H20 ~ CO2 + 4W + 4e- (oxidation) [3]
4Fe(OH)3+ CH20 + 8W ~ 4Fe2+ CO2 + 11H20 [4]
The reductionreactioncan be definedquantitativelythroughthe changein

Gibbs free energy(AG)
Red
AG = AGO + RT In - - - - [5]
(Ox)(w)m
REDOX MEASUREMENT OF SOILS 1257
where !!"Go is the standardfree energy change,R is the gas constant,and T is

absolutetemperature.The Nernst equation expressesthe reduction reaction in
termsof electrochemicalenergy(millivolts) using the relationship!!,.G= -nEF
RT (Red) mRT
Eh=Eo--ln- - [6]
nF (Ox) nF
whereEh is the electrodepotential,EO is the standardhalf cell potential,F is the

Faradayconstant,n is the numberof electronsexchangedin the half-cell reaction,
m is the numberof protonsexchanged,and the activities of the various oxidized
and reducedcomponentsare shown in parentheses. Substitutingvaluesof 8.31 J
K- 1 mol-1 for R, 9.65 x 104 C mol-1 for F, and 298 K for T and using the rela-
tionship In(x) = 2.303 log(x), Eq. [6] simplifies to
59 (Red) m
Eh(mV)=Eo--log - - +59-pH [7]
n (Ox) n
Inspection of Eqs. [6] and [7] reveals that redox potential (Eh) increaseswith
increasingactivity of the oxidized component,decreaseswith increasingactivity
of the reducedcomponent,and increaseswith an increasein H+ activity (or a
decreasein pH).
The importantrole of pH in redox reactionsis evidentfrom Eq. [7]. Where
the ratio of protons toelectrons(min) is unity, there is a predicted59 mV change
in Eh per pH unit. This value is often usedto adjustmeasuredredoxpotentialsfor
comparisonat a given pH. However, the Eh/pH slope varies dependingon the
oxidants and reductantsinvolved. The relationshipoften may be linear, but not
always so, and there is little basisfor expectingredox potentiaVpHslopesto be
the samein different naturalsystems.For instance,a changeof +177 m V per pH
unit is the predictedslopefor the reductionof Fe(OH)3to Fe2+ (seeEq. [2]) based
on the min value of three (24/8). This was verified experimentallyby Patrick
(1964).
The Eh/pH slopepredictedfrom the Nemstequationassumesthat the redox
couplecontrolsthe pH of the system.While this may be true for controlledlabo-
ratory solutions, the pH of natural soils and sedimentsis buffered by silicates,
carbonates,and insoluble oxyhydroxideswhich are not involved in redox reac-
tions (Bohn, 1969). Therefore,it is not surprisingthat measuredslopesin natur-
al soils deviatefrom the predictedvalues,and applying the 59 mV correctionfac-
tor may be inappropriateunder those conditions (Bohn, 1969; Ponnamperuma,
1972).
The reductionreactionin Eq. [1] also can be definedby an equilibrium con-
stant
[8]
which can be rewritten in log form

1258 PATRICK ET AL.
log k =10g(Red)- log (Ox) - nlog (e-) - mlog (H+) [9]
The -log (e-) term in Eq. [9] can be definedaspe in the sameway pH is defined
as -log (W). RearrangingEq. [7] yields
pe + pH =log K - 10g(Red)+ log (Ox) [10]
Equation[10] is a convenientform for plotting equilibrium solubility diagrams

becausethe pe + pH sumcan be usedas one axis and speciesactivity as the other
axis (Lindsay 1979, 1988).The Eh is the more commonexpressionof soil redox
potential since it is readily determinedwith a platinum electrode.Soil pe can be
calculatedfrom Eh using Eq. [11].
Eh (mV)
pe = [11]
59
APPLICATION OF REDOX MEASUREMENTS

In saturatedsoils, four factors combine to make the oxidation-reduction
potential the best available measureof the oxidation or reduction statusof the
soil. First, as mentionedabove,the rangeof redox potential in alternatelyflood-
ed and drained soils is much wider, approximately1000 m V as comparedto a
rangeof approximately300 m V in unsaturatedsoils. Second,the higher concen-
trations of reducedcomponentsthat contribute to the mixed potential result in
better poising and better reproducibility of the potential reading, although poor
reproducibility is still one of the main limitations of redox potential measure-
mentsin saturatedsoils. Third, O2 is easily reducedand, therefore,usually absent
from saturatedsoils. Methodsusedto measureO2 contentand O2 diffusion rate
in well-drainedsoils cannotbe usedin saturatedsoils. And fourth, althoughone
could identify reducedsoil conditionsby analysisof selectedchemicalforms of
someelements(Le., the presenceof ferrous iron, manganousmanganese, or sul-
fide sulfur) or the use of certainmicrobial characterizationanalyses,thesemeth-
ods are not alwaysapplicableto everysoil, and many of thesemeasurements may
not indicatethe intensity of reduction.Thus, redox potential remainsa generally
applicable,reasonablyconvenientway to identify the presenceand intensity of
reductionin wetlandsoils and sediments.
Along with pH, redoxpotential(Eh) definesconditionsunderwhich impor-
tant biogeochemicalprocessestakeplace.Pourbaix(1966)and GarrelsandChrist
(1965) demonstratedthe value of using Eh-pH diagramsto characterizeredox
reactionsof importancein natural geochemicalenvironments.This approachis
usually representedby a parallelogramdelineatedby pH valuesof four and nine
and the 021H20 equilibrium line at the oxidizing limit and the WIH 2 line at the
reducinglimit. (Fig. 42-1). With a few exceptions(acid mine drainage,acid sul-
fate soils, and black alkali soils), most of the earth'snatural environmentshave
Eh-pH valueswhich lie within this parallelogram.In a classicpaper,BaasBeck-
ing et al. (1960) used Eh-pH diagramsto delineatethe Eh-pH field at which a
1000 €:i .. /
. -'>"" -So
800 :1",#
800
""'\
400
200
ZERO
Eh
-200
-400
-eoo
0 2 4 e 8 10 12
pH
Fig. 42-1. GenericEh-pH diagramoutlining the limits of most natural environmentswith measured
Eh (mY) and pH valuesof naturalsoils andsediments(Adaptedfrom Garrels& Christ, 1965; Baas
Beekinget aI., 1960).
numberof important water and sedimentredox processestake place(Fig. 42-1).

Their delineationswere basedon publishedvaluesof experimentallydetermined
redox potential and pH values.Using this approach,they were able to differenti-
ate amongnormal (oxic) soils, wet (seasonallysaturated)soils, and waterlogged
(semipermanentlysaturated)soils. Their plot also showedthat the pH rangewas
narrowerin reducedsoils (more negativeredox potentials)than in oxidized soils.
The pH of both acidic and alkaline soils convergeon neutrality as they become
reducing subsequentto flooding (Patrick & Mikkelsen, 1971; Ponnamperuma,
1972).
The quantitativecapability of the Nernst equationto predict the activity of
chemicalspeciesis valid only underequilibrium conditions.Sincesoils and other
biological systemsare rarely at redox equilibrium, redox potentialmeasurements
cannotbe usedto accuratelypredict the activity of specificoxidized andreduced
componentsof the system. The complex natureof soil systemswith many inor-
ganic and organicsystemscontributingto the resultantmixed potentialmeasured
on the platinum electrodeis the primary causeof this disequilibrium (Laitinen,
1960; Bohn, 1968). Even though the redox potential is of limited value in pre-
dicting theactual activities undertheseconditions,the measurementis still valu-
able for characterizingthe generalintensity of oxidation or reductionof biologi-
cal systems.Field measurements are at best semiquantitative,while under prop-
er conditions, laboratorymeasurements of redox potentialcan provide near-quan-
titative predictionsof important redox processes(Fig. 42-2). For both field and
laboratoryapplications,redoxpotentialis extremelyusefulfor signalingthe onset
of reducingconditionscausedby the depletionof oxidantssuch as O2 and N03,
as well as the onset of oxidizing conditions when O2 reentersthe soil. Experi-
mentsby Patrick and Wyatt (1964) clearly showedthe responseof redox poten-
tial to alternateflooding and drying of the soil (Fig. 42-3). Their resultsalso doc-
umentedthe importanceof a microbial energysourcefor the reduction reaction
1260 PATRICK ET AL. .
Z&LZl?C
P,&;&;i)J}.'
S_ _
a&B.ii
HIGHLY REDUCED MODERATELY OXIDIZED
REDUCED REDUCED
-
- 300 - 200 - 100 0 + 100
I
+ 200
~~
+ 300 + 400
Redox Potential (mV) at pH 7
Fig. 42-2. Measuredredox potentialsrequiredto reduceoxidized forms of the variousredox couples

in soils and sediments(from Masscheleyn,1990).
since depletionof organic matter with repeatedcycles of flooding and draining

resultedin less reduction following reflooding of the soil (Fig. 42-3). Organic
matteris the primary electrondonor in natural systems.
Despite the semiquantitativenature of mixed potentialsin soils and sedi-
ments,field redox measurements can be a useful tool for interpretingecosystem
processes.In conjunction with controlled laboratory studies that define the
boundaryconditionsfor specificelements,redox potentialsof contaminatedsoils
can identify conditionsconduciveto increasedbioavailability of potentially dan-
gerous heavy metals (Gambrell et aI., 1977; Reddy & Patrick, 1977). Redox
potentialmeasurements also havebeenusedto identify changesin plant metabo-
900 r-------:-::------::=zr-------.
0- No rice straw
0 - Rice straw added
40 60 80 100 120
Time-Days
Fig. 42-3. Redox potential through four submerged-dryingcycles (from Patrick & Wyatt, 1964).
lism (Mendelssohnet aI., 1981) and to explain observedplant speciesdistribu-

tion (Josselynet aI., 1990).
Redox potential is particularly diagnostic for determining if an area is
functioning as a wetland with distinct patternsobservedin nonwetland,transi-
tional, and wetland habitats(Faulkneret aI., 1989, Megonigal et aI., 1993). The
assessment is evenmore reliable when the redox potentialdataare usedin con-
junction with additionalquantitativedatasuchas depthto watertable and soil O2
content. In a 3-yr study of bottomlandhardwoodforest soils (Faulkner et aI.,
1991), the Eh and O2 contentof nonwetlandsites varied little during the study
(Fig. 42-4). This is a characteristicpatternfor a soil systemdominatedby O2•
Transitional sites alternated between oxidized and reduced conditions in
responseto the rising and falling water table. Redox potentials remainedlow
until the water drainedfrom the profile and O2 diffused into the soil. The redox
patternsat these sites are sometimesinconclusive by themselves,but provide
essentialdata to support other wetland indicators (hydric soil morphology and
hydrophytic vegetation).The low redox potentialsat wetland sites reflect the
extendedperiodsof flooding and soil saturation(Fig. 42-4).
Long-term redox data encompassingseasonaltrends also can facilitate
interpretationof soil morphology and hydrologic regime. Diagnostic soil mor-
phological featuressuch as color and mottling are commonly usedto infer soil
moistureregime (Soil Survey Staff, 1975; Bouma, 1983; Pickering & Veneman,
1984; Buol & Rebertus,1988). Soilcolor and mottling are primarily a function
of iron redox chemistry. When the soil is saturatedfor long periods and iron
oxides are reduced under low redox potentials, gray colors predominate(~2
chroma)and the soil is consideredhydric. More balancedperiodsof alternating
oxidation and reduction result in soils that are mottled. Recentchangesto Soil
Taxonomy have resultedin the term "redoximorphicfeatures"which includesFe
and Mn accumulationsand depletionsthat form from the reduction, transloca-
tion, and oxidation of iron/manganeseoxides (Soil Survey Staff, 1992;
Vepraskas,1992). Conclusionsbasedon the qualitative soil profile characteris-
tics (redoximorphicfeatures)havegenerallybeensupportedby quantitativedata
(soil redox potential, water table depth) in several recent studies(Faulkner &
Patrick, 1992; Griffin et aI., 1992; Hudnall & Wilding, 1992; Megonigal et aI.,
1993). However, many discrepanciesexist including relict morphology,satura-
tion without reduction, and reduction without redoximorphicfeatures(Bouma,
1983; Couto et aI., 1985; Franzmeieret aI., 1983; Wilding & Rehage,1985).
Field redox measurements are the most effectiveway to identify active reduction
associatedwith the current moisture regime and contemporaryredoximorphic
features(Faulkner& Patrick, 1992; Soil SurveyStaff, 1992; Vepraskas,1992).
CONSTRUCTIONOF PLATINUM ELECTRODES
Platinum wire with a diameterof 1.024 mm is cut into 1.3-cm segments

with wire-cutting pliers that are only usedfor cutting platinum. Smaller gauge
platinum wire is not recommendedbecauseit is more likely to bend during
installation in the soil and producespuriouspotentials.The platinum segments
TRANSITIONAL WETLAND ....
NONWETLAND
800 I 1 250 ~
BOOr CD :Dj
9
600 I-o-Eh I 200;g
-'-°2 :0
.§.
:; ::~~Mj~:::
~ 400 I- • ~. - " -1150
400 150 ~
o
a
200 100~
200
1J -o-Eh P
8l. 0 -.-02 50 o 50 r:
~~t -200 II! I "
~
II! 1'11111111111 IIII 1111111' "II
r 0
-200 "'" " " I ,,'TV' " " "ET' " , , _ _ '... ' « 0
SNJMAMJSNJMAMJAOOFAMJ -200 Lu'" 1111'11 I"" 11'"11111111111'1111' II SNJMAMJAOOFMAMJSNJMAJA
~1983+ 1984 1985.

1983 1984 1985 •
1+ + -1. '"~:~ t::~o+~.: -1"
12'
120
80
80
40
~ 40
.r:: soil
! _I r.unace o
-40
~ -40
~ -ao
~ -80
-12
-120!. , '!lIN' III ,,' " '+ MA, ," " ' '" ' ,,' ,,I S NJ MA MJ A 00 FA MJ A 0 0 F MMJ -120 I . , I I! 1111"'111"'11 I I I I I ! ! I I I " I I ' ! ' '" I!
5 NJMAMJ SNJ MAMJAOOFAMJ SNJMAMJAOOFMAMJSNJMAJA ~
~ 1983 +-1984+19~ ~
~ 1983 1984 _~1985. ~ 1983 + 1 9 8 4 - 1 . 1 9 8 5 .
~
+ Date
Date
Date ~
Fig. 42-4. Characteristicredoxpotential,O2 content,and watertable depthpatternsfor nonwetland,transitional,and wetlandsitesin bottomlandhardwoodforests(adapt- F
ed from Faulkner& Patrick, 1992).
(a) (b)
Welded Mercury Junction
_12-gauge _ _
Copper Wire
~ Silicone Sealant Covered

With Heatshrink Tubing
Glass Tubing
Triple Distilled Mercury

4
~ Waterproof Epoxy
- PI~~~gua~~ir"----'
Fig. 42-5. Diagram of the welded (a) and mercury-junction(b) platinum redox electrodes(from
Faulkneret a!., 1989).
are placedin a 1: 1 mixtureof concentratednitric (HN03) and hydrochloricacids

(HCI) for at least4 h to removesurfacecontaminationof metalsother than plat-
inum that could occur during cutting or handling. Then, the segmentsare
"soaked"in distilled, deionizedwater overnight.
Two methodsof constructioncan be used(Fig. 42-5). In the first, the plat-
inum is weldedor fused directly to a 2.053-or 1.628-mmdiam. copperwire that
has had its insulation pushed back several centimeters.During the welding
process,it is desirableto hold the platinumwire with pliers usedonly for this pur-
poseto minimize surfacecontaminationof the platinum with other metals. The
weld is then coatedwith a waterproofepoxy, the wire's insulation is moved back
down to the weldedjunction, and additionalepoxy applied to the end of the insu-
lation. It is importantthat the weld (and the Cu aboveit) are completely isolated
from any contact with the surroundingmedium after the electrodeis installed.
This weldeddesignis most appropriatefor studiesof 1 yr or lessas manyepoxy
cementsare not stablefor extendedperiodsunder continuousexposureto water.
Longer measurementperiods are better served by a glass body electrodede-
scribedbelow.
In the secondconstructionmethod, the electrode is mounted in a glass
body. Pyrex glass tubing (o.d. - 1.0 cm) forms the body of the electrode.The
glasstubing is cut into desiredlengthsand oneend is fire polishedwith a propane
torch just enoughto smooththe break.The lengthof the glasstube is not critical,
though the depth of installation is a factor to consider. Depending on the
researcheror field conditions,the glasstube length can be selectedsuch that the
glass tube extendsabove the soil surface,or only the copper lead wire extends
1264 PATRICK ET AL.
above the surface.The other end of each cut tube is heatedin a propanetorch
flame with continuousrotation until the glassbecomessoft and the openingcon-
stricts to an insidediameterjust greaterthan the platinumwire. After cooling, the
platinum wire is placedin the constrictedend of the glassso that about 0,65 cm
of platinum extendsout of the glass(the length of exposedplatinum is not criti-
cal). This end is heatedagainwith rotation in the flame until the end of the glass
tubing collapsesfor about 0.3 cm along the centerlength of the platinum wire.
The result is about0.65 cm of platinum wire exposedoutsidethe glassbody for
soil contactand about0.3 cm of exposedplatinum wire within the glassbody to
serveas an electricalconnectionto the electrodelead. Theselengthsare not crit-
ical, but should be consideredthe minimum desirableamount. Longer lengths
will be temperedby the cost of platinum wire.
The glass-platinumjunction should be water tight, but can be tested for
leaksby insertingthe end of the electrodeinto a beakerof water and connecting
the top to a low pressureair sourcewhile observingfor bubblesof air. At this
stage,the platinum wire segmentis held firmly in place mechanically.However,
this doesnot insurea permanent,water-tight,electrically insulatingsealbetween
the two ends of the platinum wire. Heating and cooling may causedifferential
expansionand contractionof the glass and platinum causinga leak to develop
betweenthe two andsubsequentlyan electricalcontactbetweenthe soil and inter-
nal metal electricalleadsto the electrode.To permanentlysealthe glass-platinum
junction, a small amountof powderedor shavedparaffin wax, or red sealingwax
(FisherScientific Co., Pittsburgh,PA, Cat. no. 15-530)is droppeddown the open
end of the glasstube and the tube tappedgently until the wax is down aroundthe
internally exposedend of the platinum wire. Then, this end of the electrodeis
heatedgently until the wax just melts. Gravity and capillary action drawsthe liq-
uid wax into the crevicebetweenthe glassand platinum. Caremust be exercised
here as toomuch heat will causethe wax to foam or boil which coatsall of the
platinum wire with an insulating layer of wax. When the wax solidifies, a per-
manentseal is achievedat the glass-platinumjunction that can withstand some
shrinking and expansionof the glassdue to temperaturechangesin the field.
Next, triple-distilled liquid mercury(HgO) is pouredinto the openendof the
electrodeto serveas an electrical contactbetweenthe bare end of the platinum
wire and the strippedend of insulated,2.053-mmdiam., copperwire electrode
lead which will be lowered into the mercury. The lengthof this wire lead will
vary dependingon the installation depth of the electrodeand the flooding depth
of the site. A balancemust be struck as too much abovegroundlead will cause
problemsfrom flooding debris, animals, vandals,etc. Usually 15 to 30 cm of
abovegroundlead is sufficient for most applications.The insulation is stripped
only from the first 5 to 7.5 cm of the copperwire in the glasstube so the lead is
insulatedwhere it comesout of the top end of the electrode.Silicone rubber is
usedto seal this juncture of the electrodeso soil water cannotleak into the top
short-circuiting the electrode and producing erroneousreadings. The silicone
overlapsthe endof the glassand coversabout2.5 cm of the insulatedwire above
the glasstube.
To further insure a permanent,leak-free seal at the top of the electrode,
heat-shrinktubing is placedover this part of the electrodeand heatappliedto col-
lapsethis insulatingmaterialaroundthe silicone rubbercompressingthe silicone

rubber againstthe glass and lead wire's insulation.Two overlappingpiecesof
heat-shrinktubing with different diametersare used: the larger size collapses
aroundthe glasstubing and the smallerdiameterpiececollapsesaroundthe insu-
latedcopperwire and over the top of the collapsedfirst pieceof heatshrink tub-
ing. About 2.5 cm of insulation is strippedfrom the top of the electrodelead to
connectthe electrodewith an alligator clip attachedto the portablepotentiometer
lead.Therearevariouswaysof doing this. Onemustfirst identify the type of con-
nectoron the meter,i.e., "US Standard"or "BNC." Thesecanbe purchasedsep-
arately through sciencesupply distributorsor electronicsupply stores,then an
appropriatelength cableis joined to the connector.It is often more convenientto
locatean old, nonfunctioningpH electrodewith the desiredconnector,cut off the
electrode,and use the cable.An alligator clip must be fastenedto the electrode-
end of the cable.The U.S. Standardconnectorshavesingle conductorsand sim-
plify fabricationof a redox electrodecable.A BNC connectorand the associated
cablehavedual connectors.The alligator clip that connectsto the redoxelectrode
shouldbe connectedto the centerconductorand a separateconnectorattachedto
the braidedwire shield of the cablefor the referenceelectrode.
A variation of the glassbody constructiontechniqueis to fuse or weld the
platinum wire onto a pieceof 2.053-mmdiam. copperwire and slide the insula-
tion backdown to the platinum as previouslydescribedfor the weldedplatinum-
copperwire electrode.Then this is insertedin a glasstube also preparedas pre-
viously describedwith at least 0.65 cm of platinum wire extendingout of the
glasstube. Continueconstructionaccordingto the proceduregiven for the glass
body electrodeexceptthis variation eliminatesthe needfor a mercuryjunction.
Also, a much larger quantity of sealingwax (either powderedor fine wax shav-
ings) is addedand heatedas describedfor the glassbody electrodeto give a seal
at both the glass-platinumjunction, as describedbefore,and to provide an addi-
tional water-tight seal betweenthe top of the electrodeand the exposedcopper
wire nearthe copper-platinumweld.
CLEANING AND TESTING ELECTRODES
Prior to testing new or previously used electrodes,the electrodesneed

cleaning.A numberof satisfactoryprocedureshave beenused.One of our pre-
ferred methodsis to makea pasteof a commercialscouringpowder(suchasAjax
or Comet), then use a toothbrushto scrub the exposedsurfaceof the platinum
wire with the paste.The pasteresidueis then rinsedoff the platinumwith a high
pressurestreamof tap water, while rubbing the platinum surfacewith your fin-
gers.Particularattentionshouldbe given to flushing the scouringpowderresidue
out of the small crevicethat may exist at the platinum wire-glassbody junction
with the high pressurestreamof wateras thesecleaningagentscontainoxidizing
agentsthat could result in poor readingsif the electrodesare not thoroughly
rinsed. The electrodesare then placedin distilled or deionizedwater overnight
prior to testing.
Other methodsof cleaningelectrodesthat have been used with success,
dependingon the constructionof the electrode,include: (i) "burning" the plat-
1266 PATRICK ET AL.
inurn surfacein an ethanolflame, (ii) scrapingthe surfacewith a stainlesssteel

razor bladeor knife blade in the field, (iii) repeatedpushinginto cleansand,(iv)
electrolysis,and (v) soakingin varioussolutionsincluding acidsor hydrogenper-
oxide (H20 2).
With someof thesecleaningmethods,rinsing the electrodewell with water
and allowing the electrodeto standovernightin distilled or deionizedwaterprior
to testingis necessary.
Prior to installation, electrodesshould be testedin a pH-buffered,quinhy-
drone solution (Table 42-1). The concentrationof quinhydrone(C12H lO0 4) in
pH-buffer solution is not critical, but a pinch or about0.05 g in 30 mL of buffer
solution containedin a 50-mL beakershouldgive good results.Stir the mixture,
allow at least 15 min to equilibrate,then stir again. Not all of the quinhydrone
will dissolve.The testsolutionshouldbe goodfor at leasthalf a day. If more than
about5 min is requiredfor the electrodereadingto stabilize,the electrodesshould
be cleanedand checkedagain.It shouldbe noted the readingsin Table 42-1 are
given relative to either a calomel referenceelectrodeof a silver/silver-chloride
referenceelectrode.Dependingon the referenceelectrodeused,readingsshould
be compareddirectly to this table without correction.Any electrodediffering
more than 5 mV from the propervalue should be cleanedand testedagain, but
discardedif cleaningfails to correctthe problem.
Somepeoplehave useda much more strongly poisedtestingsolution con-
sisting of a fairly concentratedmixture of ferrous iron, ferric iron, and sulfuric
acid (H2S04) (Light, 1972). The theoreticalbasisfor this solution is easyto see
as the test potentialcan be calculatedfrom the Nernstequation.However,faulty
or dirty electrodesare more likely to give a properreadingin this highly poised
testing solution than they are in the more weakly poised pH-buffered quinhy-
drone solutions. Therefore,we recommendless well-poisedtesting solutionsto
increasethe chancesof identifying an electrodethat will perform poorly in the
field or lab.
FIELD INSTALLATION
The installation procedurefor the welded design is straightforward. A

sharpenedmetal rod (a O.3-cm steel welding rod works well) is usedto make a
hole in the soil slightly larger than the diameterof the wire and 2.5 cm short of
the desired depth. The electrodeis fed into the hole and the platinum end is
pushedsolidly into undisturbedsoil to the desireddepth.This providesgoodcon-
tact betweenthe platinum and the soil. The top of the hole is closedaroundthe
copperlead.
Table 42-1. EMF of platinum/referenceelectrodepairs in quinhydronedissolvedin pH buffer (adapt-
ed from Bohn, 1971).
pH4 pH7
Referenceelectrode 293K 298K 293K 298K
--------------mV -----------------
Ag-AgCl 268 263 92 86
Saturatedcalomel 223 218 47 41
For the glass body electrode,a sharpenedmetal rod with a diameternear

that of the electrodeis usedto makea hole in the soil to a depthof about 2.5 cm
less than the desireddepthof the exposedplatinum wire. The electrodeis placed
in the hole, then a hollow polyvinyl chloride (PVC) tube of appropriatelengthfor
eachelectrodeis placedover the electrode'slead until the end of the PVC tube
fits snugly againstthe bulge formed by the heat shrink tubing, silicone rubber,
and the top end of the glasstube. This hollow PVC tube facilitates pushingthe
electrode2.5 cm deeperinto the soil (beyondthe depth openedby the sharpened
metal rod) so that the exposedplatinumend is placedsolidly into undisturbedsoil
to the desireddepth insuring a good contactbetweenthe soil and platinum wire.
Onceinstalledto the properdepth,a dry clay materialfree of any organicmatter
is pouredinto the hole and packedaroundthe electrodelead using the PVC tub-
ing to pack this material every few centimeters.Commercially available ben-
tonite works well, however,any predominantlyclay soil low in reactiveiron can
be heatedin a muffle furnace to oxidize organic matter renderingit suitable for
this purpose.Sealingthe hole with a soil containingorganicmatteraddsan ener-
gy sourcefor microbesfairly nearthe desiredmeasuringdepthand it is important
to preventthis below the A horizon. The rest of the hole is filled with mortar mix
and also packedlightly with the PVC tube. The mortar mix drawsmoisturefrom
the surrounding soil and hardensto minimize channelizedflow of water or
enhanceddiffusion of surfaceair down the installation hole to the electrodethat
might contributeto erroneousreadings.
For generalcharacterizationof a site, installation depthsof 15-, 30-, 60-,
and 90-cm are recommended.Studieson soil morphology will likely focus on
individual soil horizons rather than specified depths. Becauseof the impact of
saturatedsoil conditions on plant metabolismand community composition,the
rooting zone(upper30 cm) is an areaof particularimportance.
FIELD MEASUREMENT OF REDOX POTENTIAL
Field measurements of redox potential are increasinglybeing usedto esti-

mate the intensity of oxidation or reduction of soils and sediments.Although
semiquantitative,thesemeasurements permit predictionof a numberof important
biogeochemicalprocessesin the soil, rangingfrom activities of aerobicmicroor-
ganismsat the more oxidizing end of the redox scale to predictionsof sulfate
reductionand methane(CH4) formation on the reducingend of the redox scale.
The statusof the N, Mn, and Fe systemsalso can be inferred from properly mea-
suredredox potentialsand pH values. Alternating aerobicand anaerobiccondi-
tions can be detectedby field measurements of redox potentialmore rapidly than
by any other measurement.
Redox potential measurementsare made in the field using a portable
pH/millivolt meter and a saturatedcalomel or silver/silver-chloride reference
electrode.The reference electrode is pusheda shortdistanceinto wet or moist soil
at the surfaceto insure a good electrical contact.If the soil is relatively dry, a
knife is usedto breakup a small volume of soil, and water addedto form a paste,
then the referenceelectrodeis installed in this to provide good contactwith the
1268 PATRICK ET AL
soil solution. If the soil is dry or highly weathered,a dilute salt solution (i.e., 5 g
KCI in 100 mL H20 canbe usedto moistenthe referenceelectrodehole and pre-
vent a junction potentialfrom being establishedbetweenthe referenceelectrode
and the soil. The position of the referenceelectrodein relation to the redox elec-
trode may not be critical, but it is recommendedthat the referenceelectrode
should be placed within 2 m of the surface position of the redox electrode.
Thoughsomereferenceelectrodesmay be submergedwithout problems,it is rec-
ommendedthe entire referenceelectrodeshould not be submergedwhere water
is deep.Wherewater is deeperthan the length of the referenceelectrode,simply
submergingthe working end of the referenceelectrodein the floodwaterwill be
satisfactory.
In locations where flooding may periodically immerse the aboveground
electrodelead, a thin film of salt or clay may coat the entire electrodelead after
the water has receded.This coating may act as an additional electrical contact
betweenthe alligator clip attached tothe top of the electrodeand the soil surface,
bypassingthe platinumwire. To preventthis problem,the aboveground insulation
on the electrodelead may be cleanedwith a moist tissueand then dried. Also, it
is importantto makea good electrical contactbetweenthe alligator clip and the
electrodelead. It may be necessaryto gently scrapethe exposedcopperleadwith
a knife to insurepropercontact.
Sometimesa drift in the meterreadingis notedwhen the meteris first con-
nectedto the electrodesin the field. The rate of drift decreasesrelatively rapidly
if the electrodesarefunctioningproperly,and,usuallywithin a coupleof minutes,
the drift rate is low enoughto record the Ec readingindicated(Ec is the direct
meterreadingusinga calomelreferenceelectrode).Well-poised soilsystemscon-
taining appreciablesoluble levels of redox couplestend to give minimal drift.
This is typical of many reducedsoils and sediments.Oxidized soils and surface
waterstend to be poorly poisedand may more often exhibit considerabledrift for
extendedperiods.Sometimesit is necessaryto wait for this drift to slow to a low
rate beforerecordinga valuefrom the meter,andsometimesit is not necessaryto
wait. For example,if a meter readingis drifting upward after the Eh (corrected
Ec reading--seebelow for correction)is above450 mV, the systemis clearly oxi-
dizedregardingall redoxcomponentsof interest,andlikely containsdissolvedO2
in the soil solution as well. Thus, for interpretationpurposes,it is not necessary
to wait longer for the drift to stop. If, on the other hand, the Eh readingwere in
the rangeof +100 m V and drifting upward,or -100 m V and drifting downward,
one should wait for the drift rate to approacha low level before recording the
reading. This is becauseseveral redox-activecomponents(Fe, Mn, N03) are
transformedin the redox range of +100 mV to +350 mV (Fig. 42-2) and it is
desirableto know more precisely the soil redox potential.Similarly, should a
downwarddrift be notedfrom around-100 mV, it is useful to know if a soil or
sedimentis sufficiently reducing(about-150 mV) for sulfate(S04) reductionto
sulfide to occur (Connell & Patrick, 1968). For temporarily installed electrodes,
it is desirableto leavethem overnightbeforetaking a reading.
By convention,electrochemistsconsiderall electrodepotential measure-
ments to be made using a standardhydrogenelectrode(SHE) as the reference
electrode. In an electrochemicalcell, the contribution of the SHE has been
definedaszeromillivolts. The SHE is not a convenientelectrodefor field or rou-

tine laboratorywork, and other referenceelectrodessuch as the calomel or sil-
ver/silver-chloridereferenceelectrodesare commonlyused. To converta redox
potentialmeterreadingusing a calomelreferenceelectrodefilled with a saturat-
ed KCl solution(Ec) to a value that would be obtainedhad the SHE beenusedas
a referenceelectrode(Eh), the meterreadingsare adjustedby adding245 mY to
the Ec value.If a silver/silver-chloridereferenceelectrodewere used,the correc-
tion addedto obtain an Eh value would be +199 mY. Thoughthe correctionfac-
tor is temperaturedependent,correctionsfor field temperaturesare generallynot
madeas the error involved from this sourceis relatively small comparedto other
inherenterrorsin the system.
RELIABILITY OF FIELD REDOX MEASUREMENTS
Occasionally,readingsfrom someelectrodesbecomesuspectand the elec-

trodeshaveto be replaced.It is not possibleto testthe permanentlyinstalledelec-
trodesin the field, so a judgmentmust be madeconcerninga suspectelectrode
basedon comparisonswith readingsfrom replicate electrodesand from other
depthsandplots. Triplicate electrodesat eachdepthare recommendedif time and
moneypermit,a minimum of duplicateelectrodesarerequired.It shouldbe noted
that in field applicationswhereapparentbad readingsare obtained,the problem
may be the microzonearoundthe electrode.If the endof the electrodeis installed
in or closeto a living treeroot, a marshplant root, or in an old root channelwhere
decayis occurring(situationsnot discerniblefrom the surface),readingsnot rep-
resentativeof the bulk soil will be obtained.Evenwithin shortdistances,consid-
erablevariability in field measurements can be expectedbecauseof inherentsoil
heterogeneity.The authorsare currently working on a statistical approachand
other proceduresfor identifying the optimum numberof electrodesrequiredto
give an acceptablelevel of accuracyof redox readings,and for identifying "sus-
pect" electrodes.
RETRIEVAL OF ELECTRODESAT END OF STUDY
At the endof a study,electrodesthat areconstructedof platinumwire fused

directly to an insulatedcopperwire leadcanoftenbe recoveredby simply pulling
themout of the ground.Glass-bodyelectrodesconstructedandinstalledsuchthat
a portion of the glassis abovegroundalso may be pulled straightout of the soil
by pulling on the glasstube. Glass-bodyelectrodesconstructedand installedsuch
that only the copperwire leadis abovegroundusuallymustbe dug out of the soil.
CONTROLLED REDOX POTENTIAL STUDIES
Redoxpotentialmeasurements madein the field involve simply measuring

the potentialand not controlling it. It is sometimesbeneficialto study a reaction
1270 PATRICK ET AL
6 8
3 15 1. Pressure regulator
2. Acid or alkali supply
3, Combination pH electrode
4. Acid or alkali inlet
Magnetic stirrer 5. Oxygen inlet
6. Platinum electrodes
7. Serum cap
8. Salt bridge
g. Stirring bar
1Q Air trap
Fig. 42-6. Diagramof the apparatususedfor controlling redox potentialand pH in soil suspensions
(from Patricket aI., 1973).
or processundercontrolled redox potential conditionswhere selectedpotentials

canbe imposed.This approachhasbeenusedextensivelyin this Institute tostudy
processesaffecting nutrient transformationsand availability in soils (Jakobsenet
aI., 1981; Patrick & Mahapatra,1968; Tusneem& Patrick, 1971; Williams &
Patrick, 1971; Van Cleemput & Patrick, 1974), trace metal chemistry in soils
(Patrick & Henderson,1981; Patrick & Jugsujinda,1992), toxic metal immobi-
lization in sediments(Gambrellet al., 1980;Masscheleynet aI., 1990,1992),and
the redox conditionsfavoring degradationof pesticides,petroleumhydrocarbons,
and synthetic industrial organics (Gambrell et aI., 1984; Pardueet aI., 1988;
Delaune et aI., 1990). This section describesa simple system for controlling
redox potential and pH in soil suspensionsover the entire range encountered
undernormal conditions.The majorcomponentsof this control systemare a lab-
oratory pH/millivolt meterand an appropriatemeterrelay. A modification of the
systemalso can be usedto control pH of a soil suspension.
The apparatusfor controlling redox potentialsand pH is illustrated in Fig.
42-6. For controlling redox potential,any commonlyavailablepH meteroperat-
ed in the millivolt mode with a recorderoutput can be used to monitor redox
potential in the systemand provide a signal to a meter relay which activatesan
O2 supply system.The meterrelay shouldbe selectedto haveapproximatelythe
samerangeas the recorderoutput rangeof the pH meter.In recentyears,several
commercially available models of pH/millivolt meters plus controllers have
becomeavailablewith a wide rangeof prices and featuresthat make laboratory
studiesof controlledEh systemsmore convenient toset up. Theseunits combine
the function of a metercoupledto a switching relay that can activatean electri-
cally operatedvalve or pump.
The control system uses O2 or air to prevent the redox potential from
decreasingbelow the set value. In the absenceof air, reductionprocesses,as a
function of microbial activity in the flooded soil or suspension,causethe poten-
tial to decrease.When the set potential is reached,a solenoidvalve is activated

by the relay allowing O2 to enterthe soil andpreventingthe potentialfrom falling
below the set value. Air is usually adequatefor controlling redox potential. But,
for obtainingand controlling low redox potentialsin soils that supportrelatively
low microbial activity, a mixture of N containinga low amountof O2 (0.S-2%)
will provide sufficient O2 to control the potentialwithout excessO2 causingtoo
greata temporarysurgein redox potential.For most applications,adequateaera-
tion for controlling the potentialcanbe maintainedby havingthe meterrelay acti-
vate an aquariumpump to provide air to the suspensionthrough a flow-control
valve. At low redox potentials,the air flow shouldbe low, no more than approx-
imately one bubble per second,to insure that too much O2 is not supplied.The
redox potentialcan be controlledwithin ±S to 8 mV for long incubationperiods.
Two platinum electrodesare usually usedso that electrodeperformancecan be
checkedperiodically by manually connectingfirst one electrodeand then the
otherto the meterto look for variation in the readings.Unlike field studieswhere
the soil can be quite heterogeneouswithin short distances,electrode repro-
ducibility is normally excellentin laboratorystirredsuspensions dueto the homo-
geneousnatureof the soil suspension.
In the redox potentialcontrol systemdescribedhere,O2 is usedto prevent
the potentialfrom decreasingin a systemwherebiological processes tendto drive
the potential down. A low flow of Ar or N is continuouslypurgedthrough the
suspension.With minimum rewiring of the relay, redox potentialcontrol can be
obtainedusing a gasmixture consistingof a few percentageH in a nitrogencar-
rier wherethe potentialtendsto increase.
The systemcan be used for two methodsof maintainingredox potential
control: (i) the soil can be flooded and allowed to becomereducedbeforebeing
placedin the electrodeflask, and the control systemthen usedto raisethe redox
potentialto any selectedvalue that is higher than the potentialof the preincubat-
ed soil, or, (ii) the soil can be placedin a flask, flooded, and the control system
usedto preventthe potentialfrom falling below the set value. Severaldaysmay
be requiredfor the redox potentialof the systemto reachthe set value.
A similar systemmay be usedfor controlling pH in the flooded soil sus-
pensionexceptthat a glasselectrodeis usedinsteadof a platinumelectrode.Acid
or alkali (usually 0.5 M HCI or o.s M NaOH) is addedautomaticallyusing the
meter and relay systemto control a valve or pump to maintain the pH at the
requiredvalue.Situationswill seldomdevelopwhereboth acid and alkali will be
neededduring incubationat a given pH. The systemcan be designedso that the
amountof acid or alkali addedis measured.Control of pH within ±O.OS pH unit
or lesscan be easily maintainedfor long incubationperiods.
Redox potential and pH can be controlled simultaneouslyby using both
control systemswith the samesoil suspension.Both a glasselectrodeand two
platinum electrodeswith appropriatereferencehalf cells are used. Oxygen and
either acid or alkali are provided as requiredto maintain the soil suspensionat
variouscombinationsof redox potentialandpH. After a few daysof adjustment,
the pH doesnot requiremuch additionaladjustment,and it is feasibleto control
the pH manuallyby daily additionsof acid or alkali until the desiredpH is main-
tainedwith little further adjustment.
1272 PATRICK ET AL
REFERENCES
BaasBecking,L.G.M., I.R. Kaplan,and D. Moore. 1960.Limits of the naturalenvironmentin terms
of pH and oxidation-reductionpotentials.J. Geol. 68:243-284.
Bohn, H.L. 1968.Electromotiveforce of inert electrodesin soil suspensions. Soil Sci. Soc.Am. Proc.
32:211-215.
Bohn, H.L. 1969.The EMF of platinum electrodesin dilute solutionsand its relation to soil pH. Soil
Sci. Soc.Am. Proc. 33:639-640.
Bohn, H.L. 1971. Redoxpotentials.Soil Sci. 112:39-44.
Bouma,J. 1983.Hydrology andsoil genesisof soils with aquicmoistureregimes.p. 253-281.In L.P.
Wilding et al. (ed.) Pedogenesis andsoil taxonomy.I. Conceptsand interactions.ElsevierSci.
Publ. B. v., Amsterdam,The Netherlands.
Buol, S.W., and R.A. Rebertus.1988.Soil formation underhydromorphicconditions.p. 253-262.In
D.O. Hook et al. (ed.) The ecologyand managementof wetlands.Vol. 1. Timber Press,Port-
land, OR.
Connell,W.E., and W.H. Patrick,Jr. 1968.Sulfatereductionin soil: Effect of redox potentialand pH.
Science (Washington, DC) 159:86-87.
Couto, w., C. Sanzonowicz,and A. de O. Barcellos. 1985. Factorsaffecting oxidation-reduction
processesin an Oxisol with a seasonalwatertable. Soil Sci. Soc.Am. J. 49:1245-1248.
DeLaune,R.D., R.P. Gambrell,J.H. Pardue,and W.H. Patrick,Jr. 1990.Fateof petroleumhydrocar-
bonsand toxic organicsin Louisianacoastalenvironments.Estuaries13:72-80.
Faulkner,S.P.,and W.H. Patrick,Jr. 1992. Redoxprocessesand diagnosticwetlandsoil indicatorsin
bottomlandhardwoodforests.Soil Sci. Soc.Am. J. 56:856-865.
Faulkner,S.P.,W.H. Patrick,Jr., andR.P. Gambrell.1989.Field techniquesfor measuringwetland soil
parameters.Soil Sci. Soc.Am. J. 53:883-890.
Faulkner,S.P.,W.H. Patrick,Jr., w.B. Parker,R.P. Gambrell,and BJ. Good. 1991. Characterization
of soil processesin bottomlandhardwoodwetland-nonwetlandtransitionzonesin the Lower
Mississippi River Valley. Contract Rep. WRP-91-1. U.S. Army Eng. WaterwaysExp. Stn.,
Vicksburg, MS.
Franzmeier,D.P., J.E. Yahner,G.C. Steinhardt,and H.R. Sinclair, Jr. 1983. Color patternsand water
table levels in someIndianasoils. Soil Sci. Soc. Am. J. 47:1196-1201.
Gambrell,R.P., R.A. Khalid, M.G. Verloo, and W.H. Patrick,Ir. 1977.Transformationof heavymet-
als and plant nutrientsin dredgedsedimentsas affectedby oxidation-reductionpotentialand
pH. Part II. Materials and methods,results and discussion.Contract Rep.DACW-39-74-C-
0076. U.S. Army Eng. WaterwaysExp. Stn., Vicksburg, MS.
Gambrell,R.P., R.A. Khalid, and W.H. Patrick,Jr. 1980. Chemicalavailability of mercury,lead, and
zinc in Mobile Bay sedimentsuspensionsas affectedby pH and oxidation-reductioncondi-
tions. Environ. Sci. Technol.14:431-436.
Gambrell,R.P., C.N. Reddy,V. Collard, G. Green,and W.H. Patrick,Jr. 1984.The recoveryof DDT,
kepone,and permethrinaddedto soil and sedimentsuspensionsincubatedundercontrolled
redox potentialand pH conditions.J. WaterPollut. Contr. Fed.56:174-182.
Garrels,R.M., and C.M. Christ. 1965.Solutions,minerals,andequilibria. FreemanandCooper,New
York,NY.
Griffin, R.W., L.P. Wilding, andL.R. Drees.1992.Relatingmorphologicalpropertiesto wetnesscon-
ditions in the Gulf Coastprairie of Texas.p. 126-134.In J.M. Kimble (ed.) Proc. 8th Int. Soil
Correlation Meet. Characterization,classification,and utilization of wet soils. USDA-SCS,
Natl. Soil Surv. Center,Lincoln, NE.
Hudnall, W.H., and L.P. Wilding. 1992. Monitoring soil wetnessconditionsin Louisianaand Texas.
p. 135-147.In J.M. Kimble (ed.) Proc. 8th Int. Soil CorrelationMeet. Characterization,clas-
sification, and utilization of wet soils. USDA-SCS,Natl. Soil Surv. Center,Lincoln, NE.
Jakobsen,P., W.H. Patrick,Jr., and B.G. Williams. 1981.Sulfide and methaneformation in soils and
sediments.Soil Sci. 132:279-287.
Josselyn,M.N., S.P. Faulkner,and W.H. Patrick,Jr. 1990.Relationshipsbetweenseasonallywet soils
and occurrenceof wetlandplantsin California. Wetlands10:7-26.
Laitinen, HA 1960. Chemicalanalysis.McGraw-Hill Book Co., New York.
Light, T.S. 1972.Standardsolution for redox potentialmeasurements. Anal. Chern.44:1038-1039.
Masscheleyn,P.H., R.D. DeLaune,and W.H. Patrick,Jr. 1990.Transformationsof seleniumasaffect-
ed by sedimentoxidation-reductionpotentialand pH. Environ. Sci. Technol.24:91-97.
Masscheleyn,P.H., J.H. Pardue,R.D. DeLaune,and W.H. Patrick,Jr. 1992. Chromiumredox chem-
istry in a lower Mississippi Valley bottomland hardwood wetland. Environ. Sci. Technol.
26:1217-1226.
Megonigal,J.P., W.H. Patrick, Jr., and S.P. Faulkner. 1993. Wetland hydrology, redox potential, and
soil O2 contentin relation to hydric soil morphologyand wetland delineation.Soil Sci. Soc.
Am. J. 57:140-149.
Mendelssohn,LA., K.L. McKee, and W.H. Patrick, Jr. 1981. Oxygen deficiency in Spartinaalterni-
flora roots: Metabolic adaptationto anoxis. Science(Washington,DC) 214:439-441.
Pardue,J.H., R.D. Delaune,and W.H. Patrick,Jr. 1988. Effect of sedimentpH and oxidation-reduc-
tion potential on PCB mineralization.Water Air and Soil Pollut. 37:439-447.
Patrick, W.H., Jr. 1964. Extractableiron and phosphorusin a submergedsoil at controlled redox
potentials.p. 605-609.In Trans. 8th Int. Congr. Soil Sci., Bucharest,Romania.Acad. Social-
ist Rep. Romania.
Patrick, W.H., Jr., and R.E. Henderson.1981. Reductionand reoxidation cycles of manganeseand
iron in flooded soil and in water solution. Soil Sci. Soc. Am. J. 45:855-859.
Patrick, W.H., Jr., and J. Jugsujinda.1992. Sequentialreductionand oxidation of inorganic nitrogen,
manganese,and iron in flooded soil. Soil Sci. Soc. Am. 1. 56:1071-1073.
Patrick, W.H., Jr., and I.e. Mahapatra.1968. Transformationand availability to rice of nitrogen and
phosphorusin waterloggedsoils. Adv. Agron. 20:323--358.
Patrick, W.H., Jr., and D.S. Mikkelsen. 1971. Plant nutrient behaviorin flooded soil. p. 187-215.In
R.A Olson (ed.) Fertilizer technologyand use.2nd ed. SSSA,Madison, WI.
Patrick, W.H., Jr., and R. Wyatt. 1964. Soil nitrogenloss as a result of alternatesubmergenceand dry-
ing. Soil Sci. Soc. Am. Proc. 28:647-653.
Patrick, W.H., Jr., B.G. Williams, and 1.T. Moraghan. 1973. A simple systemfor controlling redox
potentialand pH in soil suspensions.Soil Sci. Soc. Am. Proc. 37:331-332.
Pickering,E.W., and P.L.M. Veneman.1984. Moisture regimesand mornpologicalcharacteristicsof
a hydrosequence in central Massachusetts.Soil Sci. Soc. Am. J. 48:113--118.
Ponnamperuma, EN. 1955. The chemistry of submergedsoils in relation to the yield of rice. Ph.D.
Diss. Cornell Univ.
Ponnamperuma, EN. 1972. The chemistryof submergedsoils. Adv. Agron. 24:29-96.
Pourbaix,M. 1966. Atlas of electrochemicaleqUilibria. (In French.)Pergamon Press, New York.
Reddy,C.N., and W.H. Patrick, Jr. 1977. Effect of redox potential and pH on the uptakeof Cd and Pb
by rice plants.J. Environ. Qual. 6:259-262.
Soil Survey Staff. 1975. Soil taxonomy:A basic systemof soil classificationfor making and inter-
pretingsoil surveys.USDA-SCSAgric. Handb.436. U.S. Gov. Print. Office, Washington,De.
Soil Survey Staff. 1992. Keys to soil taxonomy.SMSSTech. Monogr. 19. 5th ed. PocahontasPress,
Inc., Blacksburg,VA.
Tusneem,M.E., andW.H. Patrick,Jr. 1971. Nitrogen transformationsin waterloggedsoil. Agric. Exp.
Stn. Bull. 657. LouisianaStateUniv., Baton Rouge,LA.
Van Cleemput,0., and W.H. Patrick, Jr. 1974. Nitrate and nitrite reduction in flooded soils at con-
trolled redox potential and pH. p. 152-159.In A.Y. Sokolov (ed.) Trans. 10th Int. Congr. Soil
Sci. NaukaPubl. House,Moscow.
Vepraskas,M.J. 1992.Redoximorphicfeaturesfor identifying aquic conditions.North CarolinaAgric.
Res. Servo Tech. Bull. 301.
Wilding, L.P., and J.A. Rehage.1985. Pedogenesis of soils with aquic moistureregimes.p. 139-157.
In Wetlandsoils: Characterization,classification,and utilization. Int. Rice Res. Inst., Manila,
Philippines.
Williams, B.G., andW.H. Patrick,Jr. 1971. Effect of Eh and pH on the dissolutionof strengite.Nature
(London) 234:16-17.
Published 1996
Chapter43
Kinetic Methods and Measurements
DONALD L. SPARKS,SCOTT E. FENDORF,

CHARLES V. TONER, IV,
AND THEODORE H. CARSKI, University of Delaware, Newark, Delaware
Soil chemicalprocessescan be studied fromboth equilibrium and kinetic view-

points. While equilibrium measurements are useful in providing information on
the final stateof a system,no dataare furnishedon the ratesof the reactions.Such
kinetic information is extremely important in accuratelypredicting the fate and
transportof plant nutrients, metals, radionuclides,pesticides,and other organic
chemicalsin soil with time.
The study of kinetics of soil chemicalprocessesis a challengingand ardu-
ous exercise.This is in large part due to the heterogeneityof soils and even,soil
componentssuchas clay minerals,metal oxides,and humic substances.One can
either measurethe chemical kinetics or the kinetics of a reaction.Chemical kinet-
ics refers to the molecularaspectsof chemical reaction rates where transportis
not limiting. It is very difficult to eliminate transportphenomenain heteroge-
neous systemssuch as soils and soil components(Aharoni & Sparks, 1991).
Thus, one is usually measuringthe kinetics of reactions,or simply time-depen-
dent phenomena.In fact, with almost all of the batch and flow methodsthat are
being usedto measureratesof soil chemicalprocesses,a combinationof chemi-
cal kinetics and transportprocessesis usually being measured(Aharoni et aI.,
1991). However,chemicalrelaxationmethodsoffer a meansto study the chemi-
cal kinetics of soil chemicalreactions.Thus, the objective of one'sstudy, i.e., to
measurethe chemical kinetics or the kinetics of reactionswill certainly influence
what methodis chosen.
An array of kinetic methodshave been used to measurethe rates of soil
chemicalprocesses.Theseinclude: batch, flow, stirred-flow, stopped-flow(SF),
and otherchemicalrelaxationtechniques.None of thesemethodsis a panaceafor
kinetic analyses.
In this chapter, the virtues and disadvantages,as well as experimental
details of eachmethod,and methodof separationof liquid and solid phasesare
all importantconsiderationsin any kinetic study. Additionally, experimentalpara-
meters such as effects of reactant concentration,temperature,pH, and ionic
strengthalso are importantfactors in kinetic studies(Amacher,1991).
Book Seriesno. 5.
1275
1276 SPARKSET AL.
Ion Association
.Multivalent Ion Hydrolysis.
Gas-Water
41 ...
Ion Exchange
Sorption
Mineral-Solution
III .,
Mineral Crystallization <4----li

Fig. 43-1. Time rangesrequired to attain equi-
librium by different types of reactionsin soil ~s h d ma rr,i\
environments(from Amacher,1991). Time Scale
There are many types of chemical reactionsthat occur in soils. The time
scalesvary considerably,asshownin Fig. 43-1, and the reactionscan occurcon-
secutivelyand concurrently(Amacher,1991). Clearly, the time scaleof a partic-
ular reaction of interestwill be an important, if not overriding factor in deter-
mining which kinetic methodone chooses.For example,reactionsthat occur on
time scales<-15 s, e.g., someion association,ion exchange,and sorption reac-
tions, cannotbe easily monitoredusing batch, continuousflow, and stirred-flow
methods.
Measurementof rapid reactions requires the use of relaxation, e.g.,
stopped-flow techniques.Another advantageof using relaxation techniquesis
that the effects of transportphenomenaare greatly diminishedsince initial rates
can be accurately measuredand mixing effects are minimal. Thus, chemical
kinetics are being measuredand reaction mechanismscan be more definitively
determined.Moreover, experimentallydeterminedrate parameters,such as rate
coefficients, are mechanistic,rather than apparentparameters,independentof
degreeof mixing or reactantconcentration.Reactionsthat are slower, i.e., >-15
s can be studiedusing batch,continuousflow, and stirred-flow techniqueswhich
will be discussedlater.
BATCH TECHNIQUES
Batch methodshave been widely used to measurereaction rates in soils.

Many batch reactorsare closedsystemsin that following the mixing of all reac-
tants, no further reactantsare addedand productsare allowed to accumulate.At
various times an aliquot of solution is removedfrom the reactorfor analysis.A
flow diagramshowing thestepsinvolved in a typical kinetic study using a batch
reactor is shown in Fig. 43-2. Of course,there are numerousvariations of this
generalizeddiagram.For example,mixing may be effectedby stirring or shaking,
aliquots may be sampledby filtration or centrifugation,and reactantconcentra-
tions/activitiesmonitoredin situ, using ion specific electrodes
or spectrophotom-
etry.
Tube Technique
The simplestbatchmethodinvolves placing an adsorbentand adsorptivein

a reactorsuch as a centrifuge tube, stirring or agitating the suspensionusing a
KINETIC METHODS & MEASUREMENTS 1277
Suspend known amount of solid phase in known

volume of background electrolyte solution or
other solution of known composition.
Begin mixing and gas flow and adjust pH to

desired level.
At t=D, add known amount of solute to achieve

desired initial reactant concentrations, ionic
strength, and total volume.
At periodic intervals, sample suspension using

syringe sampler and filter through membrane
filters.
Analyze filtrates and separated solid phase.
Fig. 43-2. Flow diagramoutlining a typical kinetic experimentusing a batchreactor(from Amacher,

1991).
shaker,and then filtering or centrifuging the suspensionto obtain a solution for

analyses.This methodcan be employedin kinetic studiesif: (i) the reactionsare
slow suchthat the reactantconcentration isnot significantly changedduring cen-
trifugation and sampling; (ii) if multiple aliquots are removedfrom the react9r
over time, the aliquots should be small so that the soiVsolution ratio is minutely
altered(microanalyticalor radiotracermethodswould satisfy this caveat);(iii) it
is not necessaryto monitor and adjust pH repeatedly;and, (iv) continuouspurg-
ing with inert gas is not necessary(Amacher,1991).
Stirred-BatchReactors
ExperimentalApparatus
A typical stirred batchreactoris shownin Fig. 43-3. The typical reactoror
vesselcontainingthe sorptive and sorbent,usually an Erlenmeyerflask, can be
S)'iinge Sampler"
Combination
pH Electrode"
Acid or Base
Add it ion ....
Suspens ion
Fig. 43-3. Typical batch reactorconfiguration(from Amacher,1991).

1278 SPARKSET AL.
constructedof any type of material.The vesselmaterialthat is chosenshouldbe

nonreactivewith the sorptiveandsorbent.Thus,glassis often preferablefor stud-
ies involving organics,while plastic may be better suited for inorganic species,
particularly trace metals(Amacher,1991).
The batch reactorin Fig. 43-3 is equippedwith a gasdispersiontube that
enablesone to passN2 or AI through the colloidal suspension,eliminating CO2
and O2 from the suspension.
With this technique,a pH electrodecan be connectedto an autotitration
apparatusto maintaina constantpH (pH-stat)during the experiment.If onewish-
es to monitor changesin pH during the experiment,the pH meter can be con-
nectedto a recorder.
An addition port allows one to add soluble reactantsto the reactionvia a
funnel, if the suspensionis open IO the atmosphere.A serum stopperwith a
syringe needleis usedwhen the atmosphereis controlled(Amacher,1991). The
reactorcontentscan be mixed using eitheran overheadstirrer or a magneticstir-
rer placedunderthe reactor.
While a magneticstirrer providesa convenientway to mix the suspension,
one must be careful that the heatgeneratedfrom the stir plate doesnot increase
the temperatureof the reactorand its contents.This can be avoidedby placing a
heatshield betweenthe stir plate and reactorand/orputting the reactorin a con-
stant-temperature jacket.
A sorptionstudy employingthe apparatusof Fig. 43-3 involves placement
of sorbentin the reactor,addition of sorptivesolution,pH andsuspensionvolume
adjustment,andagitationof the suspension.At varioustimes,suspensionaliquots
are withdrawn using a syringe containingN2 gas to prevent CO2 and O2 from
entering the reaction vessel.The aliquot is then quickly filtered using a mem-
brane filter, and the filtrate weighed and analyzed (Zasoski & Burau, 1978;
Sparks,1989). The reactor should either be placed in a temperature-controlled
chamber,a water bath, or water-jacketedto maintaina constanttemperature.
Usesand Advantages
The batch technique previously describedwas developedand used by
Patrick et al. (1973). Zasoski and Burau (1978) introduceda batch reactor to
study the kinetics of sorption reactions. Variations of the Zasoski and Burau
(1978) methodhavebeenusedby van Riemsdijk and de Haan(1981), Harterand
Lehmann(1983), and Phelanand Mattigod (1987). Van Riemsdijk and de Haan
(1981)studiedP04 sorptionkinetics from soil solutionsat constantsuper-satura-
tion with respectto metal phosphatesutilizing a phosphate-state (P-stat)method.
Phelanand Mattigod(1987) studiedcalcium phosphate[Ca(H2P04hlprecipita-
tion from stablesupersaturated solutionsusing pHlCa-statand pH-stat. The H+
and Ca2+ activities of the reaction mixturewere maintainedat constantvalues
using ion-specificelectrodesconnectedto automatictitrators.
Thereare severaladvantagesto the stirred-batchtechnique.Centrifugation
is avoided and an aliquot can be sampledand filtered about every 15 s. Thus,
reactionsas fast as 15 s can usually be measured.If the reactorcontentsare well
mixed, a constantsolid-to-solutionratio can be maintainedthroughoutan exper-
iment. Zasoski andBurau (1978) have demonstratedthat, even with repeated
sampling,the solid-to-suspension
ratio is not significantly alteredsince the sam-
ple is well mixed.
Effects of Mixing
Effective mixing is critical in batch kinetic studies. Transportprocesses,

particularly mass-transferphenomenainvolving diffusion through the hydrody-
namic film aroundsorbentparticles(film diffusion) and/ordiffusion onto or into
the sorbent particles (particle diffusion), are affected by the rate of mixing.
Increasesin rate coefficientsand energiesof activation as mixing rates increase
are an indication that masstransferphenomenaare diminishedat higher mixing
rates.
While mixing has a beneficial effect on mass transfer, excessivemixing
ratescan result in particle abrasionand alterationof the sorbentsurface.Ogwada
andSparks(1986b)found that excessivemixing rates(>478 rpm with stirring and
>2318 rpm with vortexing) resultedin increasedspecific surfaceareasfor kaoli-
nite, vermiculite, and a Chesterloam soil (fine-loamy, mixed, mesicTypic Hap-
ludults), indicating that particle abrasionoccurred.Thus" with a particularkinet-
ic method,it is importantthat mixing effectson specific surfaceand reactionrate
be avoided.Mixing shouldbe sufficient to diminish transportphenomenabut not
excessiveenoughto causeparticle abrasion.
In addition to particle abrasion,magnetic stir bars and mineral particles
may act to abradeglassreactorwalls, contaminatingthe sorbentwith quartz. Use
of an overheadstirrer can preventthis problem(Amacher,1991).
Disadvantages
There are severalproblemswith using batchmethodsto study the kinetics

of soil chemicalprocesses.If centrifugationis requiredto separatethe solid from
liquid phases,it could causeelectrokineticeffects close to soil constituentsur-
faces that would changethe ion distribution (van Olphen, 1977). Moreover, the
time required for centrifugation is often longer than the equilibration time of
many commonsoil reactions.
To properly measurethe kinetics of a reaction,the reactantconcentration
shouldnot changethroughoutthe courseof the reaction(Zasoski& Burau, 1978),
viz., the sample and suspensionshould have a similar solid to solution ratio
throughoutan experiment.Unfortunately, in most batch studies,there is a wide
solid/solutionratio (Sparks,1985, 1986, 1989).
As mentionedpreviously, most batchtechniques,exceptthosethat employ
ion exchangeresins,are closedsystems.Consequently,desorbedspeciesare not
removed and reversereactionsmust be taken into accountwhen data are ana-
lyzed. Thus, readsorptionof desorbedspeciesmay occur, and desorbedspeciesin
the solution phasemay inhibit further releaseof adsorbate.Secondaryprecipita-
tion reactionsmay occur in solution as well (Sparks,1989; Amacher,1991).
With many batchtechniques,difficulty may be encounteredin maintaining
suspensionuniformity, particularly with soils of large sandand/ororganicmatter
contents.Sandparticleshavea tendencyto sink, whereas,organicmattertendsto
1280 SPARKSET AL.
rise to the top of the reactor, artificially partitioning the various sorbentsin the
mixture.
Advantages
In summary, while batch methods have inherent disadvantages,as with
other kinetic methodsthere are severalsalient attractivefeatures.Batch reactors
are usually inexpensiveto purchaseor constructand they are easy to use. With
somebatchreactors,like that shownin Fig. 43-3, it is possibleto maintaina con-
stantsolid/solutionratio and effectively excludeCO2 and O2 (Amacher,1991).
FLOW METHODS
Flow methods,while less widely usedthan batchmethods,are increasing-

ly gaining popularity in the study of kinetics of soil chemicalprocesses.Table
43-1 provides examplesof studies that have employedeither continuousflow
(miscible displacement)or stirred-flow techniques.
Flow methodsdiffer from batchtechniquesin severalrespects,most impor-
tantly, they are opensystems.Soluteis continuouslyaddedand reactionproducts
are removed.Thus, sorbentswith solution flowing past them may be exposedto
a greatermassof solute(concentrationx flow rate x time) than in a batchsystem
(concentrationx solution volume) by the time equilibrium is reached(Sparks,
1989). Moreover,the removalofreactionproductsinsuresthat readsorptionphe-
nomena,inhibition of desorption,and secondaryprecipitationare minimized.
Another attributeof many flow methodsis that narrow solid/solutionratios
are employedthat more realistically mimic field conditions.In batchtechniques,
solution/soil ratios are typically two orders of magnitudehigher than in flow
methods.
ContinuousFlow Methods
Equipment
A schematicof a typical continuousflow apparatusis shownin Fig. 43-4.
Solid samples(usually 0.5-2g) are depositedon a membranefilter eitheras a dry
sampleor injected as a suspensionwith a syringe, resulting in a thin soil disk.
Miller et a1. (1989) useda disk thicknessof 80 flm for a goethitesystem.The fil-
ter holder is cappedtightly, and connected.toa fraction collectorand a peristaltic
pump to maintain a constantflow rate. Solution is then pumpedthrough the fil-
ter holder and the effluent is collectedat various time intervals.
For sorption/desorptionstudies,the sorption reactionis followed by moni-
toring the increaseor decreasein concentrationof the reactantof interestin the
effluent over time. When an apparentequilibrium is reached,the effluent reactant
concentrationequalsthe eluatereactantconcentration.Any processthat effectsa
changein reactantconcentrationwill be interpretedas sorptionor desorption.
Disadvantages
The continuousflow method is not without problems.Carski and Sparks
(1985) found that the dilution of incoming sorptivesolution by the liquid usedto
Table 43-1. Summary of studies using flow methodsto obtain kinetic and sorption isotherm data
(adaptedfrom Amacher,1991).
Type of flow method Reactionstudied Reference
Miscible displacement AUNH4 exchangeon soils Sivasubramaniam & Talibudeen

(thin disk) (1972)
Miscible displacement K exchangeon soils Sparkset al. (1980)
(thin disk)
Fluidized bed Albite weatheringas a function of pH Chou & Wollast (1984)
Miscible displacement K exchangeon soils Sparks& Jardine(1981)
(thin disk)
Miscible displacement K exchangeon soils Sparks& Rechcigl (1982)
(thin disk)
Miscible displacement K exchangeon soils and clays Jardine& Sparks(1984)
(thin disk)
Miscible displacement K exchangeon soils and clays Sparks& Jardine(1984)
(thin disk)
Soil column P sorption by soils underP-stat van Riemsdijk & van der Linden
conditions (1984)
Stirred-flow reactor K exchangeon kaolinite Carski & Sparks(1985)
Miscible displacement Al sorption by clay mineralsand peat Jardineet al. (1985)
(thin disk)
Miscible displacement K exchangeon soils Ogwada& Sparks(1986a,b,c)
(thin disk)
Miscible displacement Data analysistheory and methods Skopp & McAllister (1986)
(thin disk)
Stirred-flow reactor N~-N releasefrom soils Carski & Sparks(1987)
Miscible displacement S04 sorption/desorptionin soils Hodges& Johnson(1987)
(thin disk)
Stirred-flow reactor Zn, Cd, and Hg sorption by humic Randle& Hartmann(1987)
acids
Stirred-flow reactor Data analysistheory and methods Schnabel& Fitting (1988)
Stirred-flow reactor Al sorption/desorptionon clay Walker et al. (1988)
minerals
Miscible displacement P and Si sorption on goethite Miller et al. (1989)
(thin disk)
Stirred-flow reactor Ca/Mg exchangeon soils Seyfried et al. (1989)
Stirred-flow reactor N03 sorption/desorptionon soils Toner et al. (1989)
Stirred-flow reactor K/Ca exchangeon montmorillonite Eick et al. (1990)
and vermiculite
Stirred-flow reactor K sorption on soils Sadusky& Sparks(1991)
Stirred-flow reactor Cr(lll) oxidation on O-MnOz Fendorf& Zasoski(1992)
Stirred-flow reactor Phenoland aniline sorption/desorption Zhang & Sparks(1993)
on an organo-c1ay
Fig. 43-4. Thin-disk flow apparatus(from Amach-

Reservoir
er,1991).
1282 SPARKSET AL.
I~ •. ~.~ IB
Reservoir
Fig. 43-5. Stirred-flowreactorapparatus(from Amacher,1991).
load the sorbentonto the filter (if the sorbentis addedto the filter as a suspen-
sion), or the washing out of leftover sorptive solution during desorptionwill
result in concentrationchangesnot due to sorption or desorption.When unac-
countedfor, theseeffects could lead one to erroneouslyconcludethat sorption
and desorptionhave occurred.
Incompletedispersionof the samplein a flow reactorcan result in varia-
tions in sorptivetravel times to individual sorptionsitesin the sample.Variations
in particle size and pore size will yield similar results.
With all continuous flow methods,pronouncedmass transfer occurs in
combination with chemical reaction kinetics (Ogwada & Sparks, 1986a).
Thus,onecannotseparatechemicaleffectsfrom physicaleffectsand the resulting
apparentrate coefficientsare dependenton flow rate.
Stirred-FlowReactor
Stirred-flow reactorshave beenusedby chemical engineersand chemists
for many years,but only recentlyhavethey beenemployedto study reactionrates
on soils andsoil components(Carski & Sparks,1985; Randle& Hartmann,1987;
Miller et aI., 1989; Seyfried et aI., 1989; Bar-Tal et aI., 1990; Eick et aI., 1990;
Sadusky& Sparks,1991).
Apparatus
A diagramof a typical stirred-flow kinetic apparatusis shownin Fig. 43-5.

An illustration of the reactionchamberdevelopedby Carski and Sparks(1985),
consistingof a chamberwith influent and effluent ports, a magneticstirring star,
and an adjustableplungerbaseis shownin Fig. 43-6. A small quantity of sorbent
is placedin the chamberand retainedthereinusing a prefilterand a 0.8-)lm mem-
branefilter fitted just below the effluent port. As in the continuousflow system
discussedearlier, a peristaltic pump is used to maintain a constantflow rate of
solution and a fraction collector is usedto sampleeffluent.
Bar-Tal et ai. (1990) and Seyfriedet al. (1989) haveshown that solution to
solid ratios in the reactor ranging from 5:1 to 10:1 are most effective in main-
®
~Outlet
_ ..... Filter
Plunger
Fig. 4~. Stirred-flow reactor chamber (from Carski &

Sparks,1985).
taining suspensionuniformity and constantflow rate. Carski and Sparks(1985)

demonstratedthat solution/soil ratios in excessof 10:1 can lead to difficulty in
separatingdilution from sorptioneffects,particularly for soils with small sorption
capacities.Concentrationchangesduring the dilution of the liquid contentsof the
chamberwithout soil are measuredto constructa dilution curve to distinguishthe
dilution effects from adsorptioneffects on effluent concentrationsfor each run
(Carski & Sparks,1985).
Miller et al. (1989) useda stirred-flow reactorin which stirring was effect-
ed with a propeller connectedto a high-torquemotor. Leakagefrom the reactor
aroundthe shaft of the propelleris preventedby a specialbearing.
Advantages
Stirred-flow reactorscombinesomeof the attributesof both batchand flow
techniques. Because of mixing, mass transfer phenomenaare significantly
reduced.Seyfriedet al. (1989) found that the stirred-flow methodeffectsperfect
mixing, viz., chamberand effluent concentrationsare equal, over a flow rate
rangeof 0.28 to 2.20 mL min-I. The stirred-flowmethodalso allows one to mea-
sure relatively rapid reactions.Seyfried et al. (1989) found that the fastestreac-
tions onecould measurewith the stirred-flow techniquehad half-lives (t1/2) of 0.3
min and reactionswith t1/2 >60 min could not be measured,at a flow rate of 0.83
mL min-I and a chambervolume of 8.3 mL-l.
Disadvantages
Despitethe advantagesof the stirred-flow methodover the continuousflow
and most batch techniquesin measuringratesof soil chemical phenomena, one
of the biggestshortcomingsof this methodis the clogging of the filter, particu-
larly with high clay and organic matter contentsoils and with smectite.Several
attemptshavebeenmadeto alleviatethis problem,including useof doublefilters,
and continuouslyscouringof the filter during the courseof the experiment.No
completeresolutionof this problemhas yet beenachieved.
1284 SPARKSET AL.
DistinguishingInstantaneous
and Kinetic Reactions
In using the stirred-flow method, it is important to establish that time-
dependentand not instantaneousequilibrium reactionsare being measured.To
differentiatebetweenthesereactions,Bar-Tal et al. (1990)suggesteda simple test
wherebyflow throughthe stirred-flow chamberwasstoppedfor a periodof time.
The assumptionof this test is that, if nonequilibrium conditions exist (time-
dependentreactions),stoppingflow for a sufficient time will allow the systemto
attain equilibrium and, the sorptive concentrationin the chambershould drop.
When flow is initiated again, a drop in concentrationwill be measuredwhen a
kinetic model is employed but not for an instantaneousequilibrium model
(Seyfried et aI., 1989). Bar-Tal et al. (1990) recommendedthat flow be stopped
for at leastthe time requiredfor 50% of the reactionto be complete.In their stud-
ies on potassium-calciumexchangeon soils, the stoppingtime was 15 to 60 min.
A massbalanceequationfor a chamberwith sorbentcan be written as,
lCi =lCout + Vc!1C/dt + MdS/dt [1]

where
Cj =influent concentration(mol m-3)
COUI =effluent concentration(mol m-3)
C =chambersolution concentration(mol m-3)
1 =flow rate (m3 min-I)
t =time (min)
Vc =solution volume in the chamber(L)
S = amountsorbed(mol kg-I)
M =sorbentmass(kg)
An additional mass balance equation can be developed when flow is
stopped,
[2]
whereSc is the total quantity (mol) of sorbateion S in the chamber.
Thesemassbalanceequationswere usedin combinationwith the following
models
Instantaneous Equilibrium Model

S/E =(ClCT ) [3]
where
E = sorptionmaximum(mol kg-I)
CT = sum of competingions
Kinetic Model Dependent on Solution Concentration

S/E =(ClCT ) [1- exp (-kt)] [4]
wherek is an apparentforward rate coefficient in minutes-I.
1.0
0.9
0.8
0.7
E(luation
0.6
ci-
0.5
C) Ga
3b
~
Ga Stop
3b Slop
o
o 10 20 30 10
Effluent volume, mL
Fig. 43--7. Relative effluent concentration (C/C j ) as 3 function of effluent volume calculatedfor:
instantaneousequilibrium (Eq. [3]) with calculated line showing results for continuous and
stopped-flowruns (i.e., the resultswere identical for both runs); and kinetic models(Eqs. [4] and
[5]), for continuousand stopped(60 min of stoppingflow after a 5-min reactiontime) flow runs.
Parametervaluesare: flow rate (1) =1.5 mL min-I, solution volume in chamber(Vc) =5 mL, influ-
ent concentration(Cj) =1 mol m-3, adsorbentmass(M) =1 g, adsorptionmaximum(E) =5 mmol
kg-I, apparentforward rate coefficient(k) =0.023 min-I, forward rate coefficient (kl) =0.2 min-I,
and reverserate coefficient (k_l) =0.01 min-I.
Kinetic Model Independent of Solution Concentration

dS/dt = -k-l S + kl (E - S) [5]
where
k-l = reverserate constant
k] = forward rate constant
Equation [1] in combinationwith Eqs. [3], [4], or [5] for the continuous
flow tests and Eq. [2] in combinationwith Eqs. [3], [4], or [5] for the stopped-
flow runs can be solved analyticallyor numerically (D02BBF of Numerical
Algorithms Group) to calculatethe adsorbedfraction and the solution concentra-
tion as a function of stopping time (Numerical Algorithms Group, 1984).The
solution concentrationand adsorbedfraction values obtainedfrom Eq. [2] are
then introducedas initial conditionsfor solving Eq. [1] in combinationwith the
respectivemodel describing the remaining continuous run after the flow was
restarted.
Using the aboveapproachesonecancalculatecurvesrelating relative efflu-
ent concentration(C/Ci ) to effluent volume in milliliters (Fig. 43-7). Thereis dis-
continuity in the curvesfor the two kinetics modelswhen flow is stopped,unlike
the smoothlines observedwith the instantaneous-equilibrium model (Fig. 43-7).
This discontinuity allows one to distinguish betweeninstantaneous-equilibrium
1286 SPARKSET AL.
models(Eq. [3]) and kinetics models(Eqs. [4] and [5]) and providesone with a
simple way to differentiatebetweenequilibrium and kinetic phenomena.
DataAnalysis
With the stirred-flow method, one measuresthe kinetics of the reactions
with and without sorbentin the chamberto separatedilution effects from sorp-
tion. Thus, one can calculatethe amountsorbedusing the relation below(Schna-
bel & Fitting, 1988)
[6]
where,
q(ti) = retentionwhen both effluent fraction and chamberconcentrationsof
sorptiveare considered
c = concentrationof sorptive in fraction collected
C = sorptiveconcentrationin chamber
ti = time at end of samplecollection Period i
At = length of collection period
J = flow rate
V = volume of solution in chamber
m = massof solid in chamber
and subscriptn signifies no sorbentin the chamber,and subscripts denotessor-
bent in the chamber.
Fluidized Bed Reactor

Apparatus
The fluidized bed reactorhas been employedby engineers,chemists,and
geochemiststo study a variety of rateprocesses.With this method,the flow ofthe
fluid is adjustedso that its velocity is equivalentto the settling rate of the solid
particles in the particular suspension.The settling rates of different-sizedparti-
cles are equalizedby collisions with other particles if the suspensiondensity is
high enough.To achievethis, it is preferableto use well-defined size fractions.
With this technique,one can effect a homogeneous,well-mixed system(Sparks,
1989; Amacher,1991).
A diagramof a fluidized bed reactor, that was utilized by Chou and Wol-
last (1984) to study albite weathering,is shown in Fig. 43-8. The flow velocity
requiredto maintainparticlesin suspensionis controlledby the pumping rate PI>
while P2 is the rate of addition of new solution. Rate P2 also is the output veloc-
ity of the reactedsolution. Changingthe renewal rate, P2, will control the resi-
dencetime of the fluid in the reactor.To maintain a small differencein concen-
tration betweenthe input at the bottom of the fluidized bed and the output at the
top of the bed, P2 shouldbe small comparedto the mixing rate Pt. Chou and Wol-
last (1984) maintainedPz between3 and 6% of Pt. The reactionratewas obtained
from the product of the renewalrate, P2 and the solute concentrationdifference
betweeninput and output solutionsnormalizedvis-a-vis the total surfaceareaof
the solid (Amacher,1991).
FIg. 43-8. Schematicrepresentationof the fluidized bed

reactor.PI is the rate requiredto keep the particlesin
Input suspension.P2 is the rate of addition of fresh input
Solution
solution (from Chou & Wollast, 1984).
Uses
The fluidized bed reactor has been employed by several researchersto
investigate rates of chemical weathering(Chou & Wollast, 1984; Holdren &
Speyer,1987). There are severalattributesof this method:(i) mixing is vigorous
enoughto eliminatestrongconcentrationgradientsin the aqueousand solid phas-
es; (ii) soluteconcentrationscan be maintainedat low enoughlevels so that pre-
cipitatesdo not form; and, (iii) onecan study the effect of changein reactioncon-
ditions, such as pH, by altering the input solution compositionwithout changing
the solid phase.
Stopped-FlowTechnique
Background
Stopped-flowkinetic investigationshave been used predominantlyin the
study of solution reactions.The rapid initial mixing of the flow cell and coupling
with various analytical instrumentsmakesthis techniquewell suited for investi-
gationsof a variety of solution reactions.Furthermore,the versatility of stopped-
flow kinetics for studying "dramatically" different reaction time scales,and its
easeof employment,has addedto its popularity. With the use of rapid injectors,
measurementof millisecond reactiontimes is possible(Stachet ai., 1985), while
conventionalsyringe injectorsallow reactiontimes >25 ms to be easily measured
(Klimes et ai., 1980). The maximum time length of measurements is only deter-
mined by the physical constraintsof a given system. However, stopped-flow
kinetics has only found limited application in the study of colloidal reactions
(Ikeda et ai., 1984) due to analytical difficulties with suspensions.
One possiblesystemperturbationis a small changein the reactantconcen-
tration. Thus, stopped-flowkinetics may fall into the broadcategoryof chemical
relaxationtechniques,describedin more detail later in this chapter.By inducing
a small changein either the solid or, more commonly, the solution reactantcon-
centration,a concentration-jumpoccurs.The reactionmust then be monitoredby
a methodwhich is sensitiveto such small perturbations,thus electrical conduc-
1288 SPARKSET AL.
.tivity (EC) has been employedto measurethe chemical relaxation induced by

small concentrationjumps (Ikeda et al., 1984). However, EC only monitors the
reactionindirectly and often equilibrium parametersare necessaryto obtain rate
information. Additionally, the reactionmust be fully reversiblein order to use a
chemicalrelaxationapproach.
A more direct approachis to induce a large systemperturbationwith sub-
sequentmonitoring of the reaction (i.e., addition of a reactant).When coupled
with a methodwhich directly monitors the reactantspecies,this offers a power-
ful kinetic techniquewhich may greatly simplify evaluatingchemical kinetics
and allows for the measurement of a wide rangeof reactiontime scales.In addi-
tion, the reaction need not be reversible, as direct measurementof a reactant
speciesduring a reactionremovesthe restrictionsof having to indirectly measure
reactant concentrationsthat exist with other chemical relaxation techniques.
However, these advantagesare offset somewhatin that the simplifying assets
which result from chemical-relaxationapproaches,e.g., linearizationof reactions
regardlessof molecularityor order, cannotbe utilized (Bernasconi,1976).
Until recently,direct in situ measurement of a reactantpresentin a colloidal
systemhad not been implementedin a kinetic investigation. McBride and co-
workers (Bleam & McBride, 1986; McBride, 1987) have utilized electronpara-
magneticresonance(EPR) spectroscopyfor investigatingreactionsinvolving an
EPR active constituent[e.g., Mn(II) and Cu(II)]. Coupling EPR monitoring with
a stopped-flowkinetic technique allowsone to directly measurean EPR active
constituenton a millisecondtime scalewithout any complicationsinducedby the
presenceof colloidal material,as demonstratedby Fendorfet al. (1993). In addi-
tion, other direct monitoring techniquesare available, and with the quickly
advancingfield of surfacespectroscopies, there will undoubtedlybe many other
analytical techniquescapableof direct measurementof reactantspeciesin situ.
Coupling such monitoring deviceswith a stopped-flowreaction cell will allow
kinetic data to be obtainedfor non-EPRactive materials.Currently, efforts are
being madeto couple x-ray absorptionspectroscopy(XAS) with a stopped-flow
reactioncell.
In the following discussionwe will describean EPR-SFmethod.However,
this approachis applicableto any stopped-flowtechniquewhich directly moni-
tors a reactantin situ, and may thus be adaptedaccordingly.
Apparatus
A stopped-flowapparatusgenerally consistsof two syringe drive pumps
which rapidly inject reactantsinto a mixing cell. However, additional in-flow
ports canbe addedand the pumpingsystemcan be of any type as long as a rapid
flow necessaryfor instantaneousmixing is induced.The mixing cell is designed
to provide efficient mixing of the reactantsvia the injection pressure.Mixing is
usually completedwithin 25 ms, although more rapid mixing can be accom-
plishedwith high-speedinjectors. In many stopped-flowsystemsit is the mixing
time which limits the ability to make rapid measurements.Fortunately,mixing
times are more rapid than most soil chemicalreactiontimes.
The length of the reaction monitoring time is governedby the reactants'
stabilities.Most soil chemicalreactionsinvolve colloidal material;the suspension
ACljy;,ltion
switch
React;ml II
Reactant I
Fig. 43-9. Schematicdiagramof the EPR monitoredstopped-flowkinetic apparatus(from Fendorfet

aI., 1993).
stability (i.e., the settling rate) is the primary factor in determiningthe upperlimit
for a reaction time scale-i.e.,how long one can monitor a reaction. Thus,
stopped-flowkinetic techniqueswhich directly monitor a reactantoffer a large
degreeof versatility in measuringa rangeof reactiontime scales.Such methods
are capableof monitoring rapidsorption/desorptionreactionsin addition to slow-
er redox or precipitation/dissolutionreactions.However,one must be awarethat
mixing in a stopped-flowcell is only accomplishedat the onsetof the reaction;
mixing doesnot take place during the actual reaction. Hence,transportphenom-
enamay be rate-limiting if measurements occur over a prolongedtime period.
ElectronParamagneticResonance-Stopped
Flow Apparatus
A schematicdiagramof an EPR-SFapparatusis shownin Fig. 43-9. In this
instrumentalset-up,the mixing cell is locatedin the EPR spectrometer , allowing
EPR detectionof the cell contents.A single outflow-port is fitted with a 2-mL
effluent collection syringe equipped with a triggering switch. The triggering
switch activates the data acquisition system. Each run consists of filling the
inflow-port syringeswith the desiredreactants,flushing the systemwith the reac-
tants severaltimes, and finally initiating and monitoring the reaction. Consecu-
tive runs can be conductedwith the samereactantssimply by repeatingthis pro-
cedure. In Fig. 43-9, a Hi-Tech Scientif ic Limited syringe (Hi-Tech Scientific
Limited, Salisbury,Wiltshire, United Kingdom) pumping systemwas fitted to a
Wilmad (Model wg-385-b, variable temperatureaqueousmixing cell; Wilmad
GlassCo., Buena,New Jersey)mixing cell which was located in an EPR spec-
trometer. Data were obtained with a Microstar Electronics data acquisition
processorboardmountedin an IBM-AT computer. Furtherdetailsof this appara-
tus along with a demonstrationof its utility can be found in Fendorfet al. (1993).
The signal output from the detectionsystemmust be quantified to the con-
centrationof a reactantspecies.Here we provide an illustration of an EPR-SF
1290 SPARKSET AL.
300-r---------
- -- - - - - - 7 I
J; f
tl
•
~ 200
~
......
,.-."
......
'--"
C
~ 100
O+----~----~----~---~
-5000 o 5000 10000 15000
EPR Signal
Fig_ 43-10. The standardcurve relating the EPR signal intensity of the down field hyperfine peakto
solution Mn(II) concentration.Inset, the sextethyperfine structureof Mn(II)aqueous(from Fendorf
et a\., 1993).
techniquein which the EPR spectrometerwas usedto analytically measurethe

solution concentrationof Mn(II). To obtain quantitativeinformation on Mn(II),
the EPR cell was filled with a known concentrationof Mn(II) (in the absenceof
colloidal material). The magneticfield was then swept, resulting in the Mn(II)
hyperfine sextetstructure(inset in Fig. 43-10). The magneticfield was centered
at the downfield peakand known solution concentrationsof Mn(II) were usedto
relatethe EPR signal to the Mn(lI) concentration.Figure 43-10showsthe linear
relationshipbetweenthe EPRsignaland [Mn(II)] within the rangeof 5 to 500 /-lM
Mn(lI) (bracketsdenoteconcentrationunits).
ANALYSES OF RATE DATA COLLECTED FROM BATCH

AND FLOW TECHNIQUES
In designinga kinetic experimentone must keep in mind the objectivesof
the researchand perform the properexperimentsto obtain the data that are nec-
essaryto make a properkinetic analysis.There are severalapproachesthat one
can use to analyzerate experiments.Theseinclude using integratedrate expres-
sions and graphing the data, employmentof initial rates, and using nonlinear
least-squaretechniques(Sparks,1989).A discussionof thesemethodsand exam-
plesof their applicationto soil chemicalreactionsis providedbelow. Many excel-
lent referencesprovide further details on these approachesand the reader is
referredto these(Gardiner,1969; Lasaga,1981; Bernasconi,1986; Skopp, 1986;
Sparks,1989).
IntegratedRateEquationsand GraphicalAssessment
One way to analyzekinetic datais to use integratedrate equationsdirectly

and then, graphthe ratedata.For example,given the following elementaryreac-
tion (Sparks,1989)
[7]
The time-dependence
of the concentrationof ReactantA is given by,
d[A ]Idt = -kl [A ][Bf + k-l [Y] [8]
If one only considersthe reactionfar from equilibrium, by measuringini-

tial rates,then the forward reactiondominatesand Eq. [8] reducesto,
d [A ]ldt = -kl [A ][B]2 [9]
This is written as a third-order reaction,but can be reducedto a pseudo

first-order irreversiblereactionby maintaininga large excessof ReactantB (e.g.,
a solid), over ReactantA (e.g., a solute), i.e., ReactantB is a constant.Accord-
ingly, dataanalysescan be greatly simplified.
With initial reactionconditionsof A = Ao at t = 0 and integratingEq. [9],
one obtains
AIAo =exp (kct) [10]
Then, by plotting log (AIAo) vs. t, if a straight line results, one could
hypothesizethat the reactionis first-order. However,basedon this result alone,it
is dangerousto concludethat the reactionis first-order. One shoulddetermineif
the reactionis first-order at severalinitial reactantconcentrationsand if it is, and
the calculatedrate constantsdo not change,then one can more definitively con-
clude that the reaction is first-order. Additionally, one should apply the experi-
mentaldatato other kinetic modelssuchas zero-order,second-order,etc.
Another pitfall, in using integratedrate equationsdirectly to analyzerate
data is that secondaryor reversereactionsmay be operational.If one has not
properly accountedfor thesethen Eq. [9] is invalid and plotting data using inte-
gratedrate equationsis inappropriate.
Initial RateMethod
The generalprinciple for the initial rate methodis simply that measuring
the initial reactionrate, so that no extensivechemicalchangeoccursduring the
measurement,allows one to vary a reactant,while keeping the rest constant.
Therefore,the rate of reactionwill be proportionalto that particularreactantcon-
centration,allowing easygraphicalanalysisof the reactionorderas shownbelow,
Rate = d [A] I dt = " [A r [11]
where [A] is the concentrationof ReactantA, n is the reactionorderwith respect

to [A], and" is a constant.ExpressingEq. [11] in a log-log form,
log rate = log" + n log [A] [12]

1292 SPARKSET AL.
The reactionorder with respectto A can be obtainedfrom the slope of Eq. [12].
In order to confirm that the ratelaw obtainedby the initial rate method is com-
pleteandaccurate,the proposedrate expression shouldbe usedto predictthe con-
centrationas a function of time. If diffusion processesare influencing the reac-
tion, a deviationfrom the chemicalreactionpredictionswill occur.
LeastSquaresTechniques
Kinetic dataalso can be analyzedusing leastsquarestechniques.With this

method,the best straight line is fit to a set of data points. The best straight line,
as determinedfrom the least-squaresanalyses,is the one which minimizes the
sum of the squaresof the deviationof the y variablefrom the line.
Exampleof DataAnalysesUsing Initial Rate

and IntegratedRateExperiments
In the following discussionwe presentan exampleof how rate data for

Mn(II) sorption/desorptionon o-Mn02 employinga EPR-SFmethodcan be ana-
lyzed using initial ratesand integratedequations(Fendorfet aI., 1993).
At pH = 5, the following complexationreactionof Mn(II) on o-Mn02 was
suggestedfor low Mn(II) surfaceloadings (the basis for this hypothesisis dis-
cussedbelow)
[13]
where, Mn 2+ is the solution concentrationof Mn(II) ([Mn2+]), S- is the surface

site, and S-Mn is the complexed(sorbed)Mn(II). If a pH of 5 is maintained,the
forward reaction rate will be independentof pH and the H+ term in the reverse
rate expressioncan be incorporatedinto the reactioncoefficient.
Thus, if Eq. [13] is an elementaryreaction,the forward reactionshouldbe
dependenton [Mn2+] and [S-]; as it is first-order in [Mn2+] and [S-], andsecond-
order overall. The reversereactionwill be dependenton pH and the amountof
Mn(II) complexed,[S-Mn].
Reaction[13] is of the type, A + B <==> C + D, and leadsto the follow-
ing second-orderreversiblerate expression,
[14]
By ensuringa large excessof sorbentover sorptive ([S-] » [Mn2+]), and

maintaining a constant pH, the overall reaction should be pseudo first-order
dependingonly on [Mn2+]. Thus, underthe reactionparametersemployedin this
study, the rate expressionwould be simplified to a pseudofirst-order reaction.
d [Mn 2+]/dt = -k{ [Mn2+] + k{ [S-Mn] [15]
where k{= kl [S-] and k_ 1' = k-l [H+]. Integration of the above simplified
reversiblefirst-order rate expression(Eq. [15]) leadsto Eq. [16].
Unfortunately,k l ' and k_ l ' cannotbe independentlydeterminedsolely from rate

information on the sorption of Mn(II) (one equation,two unknowns)without a
further simplifying assumption.If one assumesthat at the onsetof the reaction
the initial reactionrate will be dominatedby the forward reaction,and the initial
reaction rate can be accuratelymeasured,then the forward rate constantcan be
ascertainedusing a first-order nonreversibleanalysis[in which k l ' can be deter-
mined from the time dependentMn(II) sorption data]. The reversereactionrate
constant,k_ l ', can then be calculatedusing k{ and the integratedexpressionfor
a reversiblefirst-order reaction(Eq. [16]).
With a knowledgeof k{ andk_ l ', a rearrangement of of Eq. [16] allows one
to calculateMn(II) sorption as a function of time. Thus, once the rate constants
are determined,the validity of the aboveapproachcan be verified by predicting
the time dependenceof Mn(II) sorptionand comparingthe predictedtrendswith
thosethat are measured.
Becausethe forward rate is much greaterat the onsetof the reaction,mea-
suring initial reactionratesshould allow one to measureonly the forward (sorp-
tion) rate constant,kl . For the systemparametersdefined previously and a site
density of 12 sites/nmz (Morgan & Stumm, 1964), the numberof sorption sites
would be at least IOZ times greaterthan the numberof Mn(II) ions present,even
at the highestMn(II) concentration.Thus pseudofirst-order assumptionshold.
The following reactionis hypothesizedto representthe contributingspecies
of Reaction[13] for the sorptionof Mn(II) by MnOz (i.e., for the initial reaction
rate and conditions employed in the study, i.e., having [S-]»[Mn2+] and main-
taining a constantpH)
Mnz+ ---. S-Mn [17]
Reaction[17] is valid far from equilibrium whereRsorption » Rdesorption(whereR is

the reactionrate, d [Mnz+]/[dt]); thus,kl[Mnz+] - k-l [Mn-S] '" kt[Mn z+]. The rate
dependencecan be describedas
[18]
The first-order dependenceof [Mnz+] can be evaluatedusing the integratedform

of Eq. [18] (Sparks,1989)
[19]
Under pseudofirst-order conditionsfor [Mnz+], the time dependence of [Mnz+] is

thus given by Eq. [19]. Therefore,if the reactionis assumedto be nonreversible
(which shouldbe the caseat the onsetof the sorption reaction)and first-order in
[Mnz+], a semilogplot of [Mn]1 vs. t will result in a straightline with the slope =
-k/2.303, and intercept,log [Mnz+]o.
However,as mentionedearlier, obtaininga linear plot basedon Eq. [19] is
necessarybut not definitive proof that the reactionis first-order (Sparks,1989).
1294 SPARKSET AL.
1.6..-------------------,
,......,
~
........
1.4
R' =0.998
"""'
I=:
'-'
~
1.2
bJ)
.Q
1.0
y = 1.38 . (8.57 x 1O")x
o.s+----..-----..-----..-------l
20 30 40 50 60
time (ms)
Fig. 43-11. Initial reactionratesdepictingthe first-orderdependence of Mn(II) sorptionasa function
of time for initial [Mn 2+1 o of 25 and 40 11M (from Fendorfet at., 1993).
If the reactionis truly first-order the rate constantshould not vary with the con-
centrationof Mn(n), and the slopeof the semilogplot shouldremainunchanged
with v~rying [Mn2+]. This was establishedand is describedbelow. .
Two initial concentrationsof Mn(n) were used,40 and 25 ~. If Eq. [17]
is correct and elementary,a linear semilog plot of [Mn2+] as a function of time,
independentof initial [Mn2+], should result (Fig. 43-11). The correlationcoeffi-
cient indicatesthat Eq. [19] describedthe datawell assuminga first-order reac-
tion, and the slopesof the two concentrationplots agreenicely. The sorptionof
the higher Mn(II) concentrationresultedin a slightly higher slope.The rate con-
stantsobtainedfor the first-order reactionmechanismwere 3.73 x 10-3 S-1 and
3.75 x 10-3 S-1 for 25 ~ and 40 ~ [Mn2+]o, respectively,with an averagedk1'
=3.74 X 10-3 ± 0.1 x 10-4S-I. In additionto the k! valuesbeingsimilar, the semi-
log plot also should yield an interceptvalue equalto the log [Mn 2+]o; indeedthe
interceptvaluesare in closeagreementwith the known [Mn 2+]o or 24 ~ and 41
~. Consequently,Eq. [17] appearsto be valid for the experimentalconditions
invoked here.
The measuredk{ was then used in combination with Eq. [16] and the
known reactionparametersto calculatek_1'. The calculatedk_t' was 3.04 x 10-4
S-I. After determiningthe kt' and k_t', the time dependenceof Mn(II) sorption
was calculatedand, for further validation of the rate constants,comparedto the
measuredvalues. RearrangingEq. [16] allows the time dependenceof Mn(II)
sorptionto be predicted.
Fig. 43-12 illustrates the measuredand predicted time dependenceof Mn(n)

sorptionon ~-Mn02. The predictedtrendsof Mn(n) sorptionwere in good agree-
mentwith thosemeasured,validatingthe accuracyof the forward andreverserate
constants.
30 r---------------------------------~
Mn(ll)mc(lsLIl'(:(1
MI1(I I) prcdicICI I
20
10
100 180 260 340 420 500 580
time (ms)
Fig. 43-12. The predictedtime dependenceof Mn(II) sorptionon O-MnOz, using the determinedrate
constants,comparedto the measuredsorption rate curve for an initial [Mnz+lo of 25 /lm (from
Fendorfet aI., 1993).
The validation of Reaction [15] as a first-order elementaryreaction indi-

catesthat chemicalkinetics were measured;thus, transportphenomenawere not
rate-limiting. If diffusion processeswere influencing the reaction rate, or if an
incorrect reaction order was assumed,the reaction would not dependsolely on
[Mn2+]. A deviationfrom linearity in the semilogplot of Fig. 43-11 would result
and there would be poor conformity between measuredand predicted Mn(II)
sorptionon o-Mn02 with time (Bunnett, 1986).
Thus, using the approachdescribedabove,onecan measurekl valuesat the
initial stagesof a reaction.With a knowledgeof kl values,integratedrate expres-
sions can then be usedto calculatek+ Furthermore,since chemicalkinetics are
beingdetermined,equilibrium constants(Keqkin) can be obtainedfrom the ratio of
kllk_ l . The Keqkin values may then be comparedto equilibrium constants(Keq),
determinedvia traditional equilibrium approaches,to further test the validity of
the rate parametersobtained(Sparks,1989).
RELAXATION METHODS
Background
As indicated in Fig. 43-1, many soil chemical reactionsoccur on time

scalesof microsecondsand milliseconds.Examplesof thesereactionsinclude ion
association,ion exchange,and sorption phenomena.Batch and flow techniques,
which are suitablefor measurementof reaction times of secondsor greater,are
not suitableto measuresuch rapid reactions(Sparks,1989).
Fortunately,chemicalrelaxationmethodsare availableto measurereaction
rateson millisecondand microsecondtime scales.Transientand stationaryrelax-
1296 SPARKSET AL.
Table 43-2. Relaxationmethodsusedin kinetic studies(from Bernasconi,1976).

Method Time range Method of detection
s
A. Transientmethods 1-1Q-6(1o-B) Spectrophotometric
1. Temperaturejump F1uorimetric
Polarimetric
2. Pressurejump 5-10 x 10-5 (mechani- Conductometric
cal pressurerelease) Spectrophotometric
5 x 10-4 X 10-7 (liquid
shockwave)
3. Electrical field pulse 1O-4_10-B Conductometric
Spectrophotometric
4. Concentrationjump lOB-1OZ (conventional) F1uorometricand many others
1()3-1O-3 (stopped
flow)
B. Stationarymethods
5. Soundabsorption 10-5-10-11 (overall time Powerloss or frequencychange:
rangefor different Resonanceor reverberation(1(J4-106 Hz),
acousticaltechniques) light diffraction (106-1OS Hz), impulseecho
(106-5 x lOB Hz), Brillouin scattering
6. Dielectric dispersion 10-3_10-12 Powerloss,capacitancechange
ation methods,alongwith time rangesthat can be measuredand methodsof reac-

tion(s) detection,are listed in Table 43-2.
The dynamic equilibrium of any chemical systemis defined by both the
physical stateof the systemas well as the chemicalstate.The chemicalparame-
ters which control the equilibrium positioningof the reactioninclude the reactant
and product concentration(the presenceof other reactantspeciesfor alternative
reaction pathways),and in the caseof reactionsin solution or suspension,the
nature of the solvent and ionic strength.The physical parameterswhich deter-
mine the equilibrium positioning include the pressure,temperature,and in the
caseof charged,ionic, or dipolar reactantspecies,the electric field. Any equilib-
rium can be perturbedby altering one or more of the physicalor chemicalattrib-
utes dictating the equilibrium positioning.
Chemicalrelaxationmethodsare basedon the principle that the equilibri-
um of a reactionmixture can be rapidly perturbedby a changein someexternal
factor suchas pressure,temperatureor electric-field strength.The systemis now
at disequilibrium and one then measuresthe relaxation time or the time that it
takesfor the systemto relax from the disequilibrium stateto the final equilibri-
um. If the perturbationis small, all rate expressionsare reducedto first-order
equationsregardlessof reaction order or molecularity.Therefore,the rate equa-
tions are linearized,greatly simplifying determinationof complexreactionmech-
anisms(Bernasconi,1976; Sparks,1989; Sparks& Zhang, 1991).
Temperature-jumpmethods(t-jump), while they have not beenusedin soil
science,havebeenwidely employedto study electrontransfer,enzymecatalysis,
metal complex formation, nucleic acid folding, proton transfer, spin equilibria,
and protein-ligand binding reactions important in chemistry and biochemistry
research.The t-jump methodis applicableover a wide time range(10- 8-1 s), tem-
perature-jumpunits are readily availablecommercially,and t-jump can be com-
bined with concentration-jump(c-jump) e.g., stopped-flow,methods.
Pressure-jump(p-jump) methodshave been the primary transient relax-

ation methodused in soil chemistry. Pressure-jumprelaxation has beenused to
measureadsorption/desorptionand ion exchangekinetics on metal oxides and
clay minerals (Zhang & Sparks, 1989; 1990a,b;Tang & Sparks,1993). The p-
jump method has severaladvantagesover the t-jump technique.Pressure-jump
measurements can be repeatedat faster intervals than with t-jump. With t-jump,
the solution temperaturemust return to its initial value before anothermeasure-
ment can be conducted.This may take 5 min. With p-jump relaxation,one can
repeatexperimentsevery 0.5 min. Longer relaxationtimes also can be measured
with p-jump than with t-jump relaxationmethods.
Concentrationjump (c-jump) methodsinvolve mixing, and thereforemix-
ing time dictatesthe shortestrelaxationtime that can be measured.For very slow
reactions(min or longer) ordinary mixing is sufficient. However,for shortermix-
ing timeson millisecondtime scales,a stopped-flowtechniquemay be used.With
this method,two reactantsare rapidly mixed in a chamber,flow is rapidly stopped
a short distancefrom the mixing chamber,and somephysicalpropertyof the sys-
tem is measuredwith time (Bernasconi,1976).
Of all the transientrelaxation techniques,the electric field pulse method
allows for measurement of the mostrapid reactionrates(10-8-10-4s,Table43--2).
Sasakiet al. (1983) used an electric-field pulse method to study CI and CI04
adsorptionon goethite.
Relaxationtechniquesare capableof revealing the natureof intermediate
stepsin multistepreactionprocesses,as well as alternatereactionpathways.This
is contingenton the condition that the differencein the relaxationtimes associat-
ed with the individual stepsis of suchmagnitudethat their relaxationspectracan
be discernedone from the other. Two relaxationtimes can be distinguishedfrom
one anotherif the slower relaxationtime is at leasttwo times the faster relaxation
time (Knoche & Strehlow, 1979).
On occasion,two or more reactionstepsin a multistepreactionprocesswill
have similar relaxationtimes under a given set of experimentalconditions,such
that the two effectscannotbe separated.In all likelihood, the relaxationtimes for
thesereactionstepswill have a different reactantconcentrationdependenceand
the experimentalconditions can be altered to separatethe two or more effects.
Anotherpossibility would be to changea secondaryexternalparameterotherthan
the forcing function to separatethe two effects. For instance,two relaxations
causedby a pressureforcing function may have a similar concentrationdepen-
dencebut their dependence on temperaturemay not be the same.A pressure-jump
experimentcould then be conductedwith the systemthermostatedat a higher or
lower temperature.Two or more relaxation times which are indistinguishable
under a given set of experimentalconditions can be quite discernableunder
anotherset of conditions.
Pressure-Jump
Relaxation
Since the p-jump relaxation method is perhapsthe most amenabletech-
nique for studying the kinetics of soil chemicalprocesses,particularly for reac-
tions in colloidal suspensions,we will concentrateon the experimentaldetailsof
this method.
1298 SPARKSET AL.
It is not the purposeof this chapterto give an in-depth discussionon the

theory of p-jump relaxation.Thoroughtreatmentscan be found in a numberof
excellentreviews and monographs(Takahashi& Alberty, 1969; Knoche, 1974;
Bernasconi, 1976; Gruenewald & Knoche, 1979; Yasunaga & Ikeda, 1986;
Sparks,1989).
Pressure-jumprelaxationis basedon the principle that chemicalequilibria
dependon pressureas given below
[21]
where ~ V is the standardmolar volume changeof the reactionandp is pressure

(MPa). For a small perturbation,
M(/K = - (~V~p/RT) [22]

Apparatus
A schematicdiagramof a typicalp-jump apparatus,suchas that devisedby
Knoche and Weise (1974) is depictedin Figs. 43-13 and 43-14. The apparatus
consistsof a mechanicalpressureinducingdevice,the p-jump autoclave,andvar-
ious electronic devicesto detect and record the reactantconcentrationchanges
during the equilibrationprocessfollowing a rapid pressurechange.
The p-jump autoclaveis a brasschamberwith two removablecells mount-
ed on its sides,eachcapableof holding approximately1 mL of solution or sus-
pension.Wateris circulatedthroughthe jacketof the autoclaveto maintain acon-
stant temperaturethroughout the measurementperiod.An internally mounted
thermistorprovidesa continuousreadingof the temperatureinside the autoclave.
One cell servesas a sampleholding cell and the other, a referencesolution hold-
ing cell. Both cells haveinternally mountedelectrodeswhich allow measurement
of the electrical conductivity of the cell contents.The referencecell containsan
electrolytesolution of similar conductivity and ionic mobility to that of the sam-
ple. The suspensionsupernatantis often the best suited for this purposesince it
shouldbe nearly identical in chemicalmakeupto the conductingmedium of the
sample. Both cells experiencean identical change in pressure,therefore, the
physicaleffectson the sample'sconductivity during the relaxationare nullified.
The cumulativeeffect of the relaxation on the electrical specific conduc-
tance,(J (in S m-1), of the samplecan be representedmathematicallyas
whereF is the Faradayconstant(96 493 C-1 equiv.), p is the density of the solu-
tion, Zj is the valenceof Ion j, f..lj is the mobility of Ion j (cm2 V-I S-I), and mj is
the molal concentrationof Ionj (Eigen & Demaeyer,1963).The physicaleffects
which might otherwisecontributeto the relaxationeffect are changesin density
(~p) and ionic mobility (~f..lj)' This leavesa relaxationdue only to the chemical
changes(,lm) resultingfrom the pressureinducedperturbation.
Both the sampleand referencecells are sealedwith a plastic membrane
which allows the pressureto be transmittedto the contentsof both cells. Pressure
Fig. 43-13. Schematicdiagramand sectionalviews of the autoclaveof the pressure-jumpapparatus

of Knoche and Wiese (1974): 1, conductivity cells; 2, potentiometer;3, 4O-kHZ generatorfor
Wheatstonebridge; 4, tunable capacitors; 5, piezoelectriccapacitor; 6, thermistor; 7, 10-tum
helipot for tuning bridge; 8, experimentalchamber; 9, pressure-pump;10, brass foil rupture
diaphragm;11, vacuum pump; 12, pressureinlet; 13, heat exchanger,14, bayonetsocket (from
Knoche & Wiese, 1974).
is increasedby pumpingwater into the autoclavewhich is sealedwith a thin strip

of brass foil. When the pressurein the chamberexceedsthat which the brassfoil
can withstand,the foil bursts, releasingthe pressure.The foil is milled to an exact
thickness,allowing a reproduciblebursting pressureto be reachedin the auto-
clave for each repetition of the relaxation time measurement.The time required
1300 SPARKSET AL.
for the pressurein the autoclaveto return to ambientpressureis approximately

60 Jls (Knoche, 1974). Sincethe p-jump must be instantaneouson the time scale
of the relaxation to be measured,this representsthe lower time scale limit for
reactionsthat can be studiedwith this instrument.
The electroniccomponentsof the p-jump apparatuscomprisea Wheatstone
bridge, an analog-to-digital(AD) converter,a computer,and oscilloscope(Fig.
43-13). The two conductivity cells in the autoclaveare connectedto the Wheat-
stone bridge. The total resistanceof the sampleand conductingcontents,repre-
senttwo of the four armsof the Wheatstonebridge. The total resistanceof the ref-
erence-cellarm of the bridge is adjustable bymeansof a potentiometer.This
allows the resistanceof the referencecell and the samplecell to be balancedarti-
ficially. As pressureis increasedin the autoclave,the equilibrium of the chemi-
cal systemin the samplecell is altered,resulting ina changein the ionic makeup
of the supernatant.An imbalancein the resistance(i.e., the conductivity) between
the sampleand referencearms of the Wheatstonebridge results. The sampleis
allowed sufficient time to attain the eqUilibrium dictated by the high-pressure
condition. At this point, the brassfoil bursts,the chemicalsystemrelaxesto the
low-pressureequilibrium, and the balancein resistanceis restoredbetweenthe
two arms of the bridge.
Data Collection and Detection

As the chemicalsystemin the samplecell relaxesto the ambientpressure
equilibrium, the AD convertercapturesthe decayinganalogsignal of the resis-
tanceimbalancein the bridge and convertsit to a digital signal. The captureand
digitization of the resistancemeasurementfrom the bridge is triggered at the
point of pressurereleaseby a piezo-electriccapacitorbuilt into the walls of the
autoclave.Following captureof the relaxationsignal, the digitized information is
fed to the computerfor datastorageand analysis.An oscilloscopeis usedto dis-
play the digitized relaxationspectrumof resistanceimbalanceamplitudevs. time
for eachrelaxationmeasurement.
Severaldigitized relaxationspectracan be fed to the computerfor signal
averagingand data manipulation.The computeralso is equippedwith software
designedto calculaterelaxation times from the exponentialdecay of the bridge
signal. Statistical parametersassociatedwith the relaxation time measurement
also are provided by the program.
The relaxationtime (t) can be monitoredusing either optical or conductiv-
ity detection.Sincethe equilibrium displacementfollowing a p-jump is small, the
very sensitiveconductometricdetectionmethod is usually preferred.Moreover,
for chemical reactionsinvolving a changein solution concentrationof one or
more ionic speciesduring the courseof the reaction,measurementof conductiv-
ity provides a ready meansof monitoring the reaction. For reactionsinvolving
solution specieswhose concentrationscan be measuredby spectrophotometry,
fluorimetry, or polarimetry, the equilibration processcan be measuredby these
means.All that is requiredis that the reactantconcentrationmeasurement be con-
siderably faster than the time required for equilibration so that the exponential
decay of the equilibrium imbalance can be sampled repeatedly.Colorimetric
measurements can be usedbut are generallylessuseful since the time for forma-
1302 SPARKSET AL.
tion of the color producingspeciesis often too long to allow sufficiently rapid
sampling.In consideringa methodof detection,it must be kept in mind that the
pressure-dependence of K is a function of !l. V (Eq. [21D. For reactionsin solu-
tion, !l. V is most pronouncedfor reactionswhich producea concentrationchange
of ionic species andconductivity detectionis an elegantand sensitivemonitoring
tool. For reactionsinvolving nonionic solutes,choosingan appropriatedetector
may be of secondaryconcern.The !l. V of the reactionmay be too small to yield
a measurablep-jump relaxationeffect and anothertechnique,suchas t-jump may
be more appropriate.
Maintenanceof StableSuspensions
The maintenanceof a stable suspensionthroughout the data collection
interval is critical as relaxationtimes for reactionsin aqueoussuspensionswill
vary as the colloid settles.Since relaxationkinetic determinationsare enhanced
by the signal averagingof severalindependentmeasurementsof the relaxation
spectrum,suspensions must remainstablefor an hour or more, dependingon the
tum aroundtime for repeatmeasurements. Slowerreactionswhich requirelonger
periods of time to relax to equilibrium after perturbationwill of courserequire
longer time intervalsbetweenrepeatmeasurements.
Sincesuspensionstability decreaseswith increasingionic strength,a suffi-
ciently low constantionic strengthmust be chosenfor the relaxationstudy. For
the caseof ionic reactions,ionic strengthcan be no lower than that dictatedby
the highestreactantconcentrationfor which a relaxationtime is to be obtained.
For certain solution species,high reactantconcentrationsmay lead to side reac-
tions such as polymerizationand precipitation which may be misinterpretedas
adsorptionreactions.The maximum adsorptiveconcentrationshould not exceed
the level where theseside effectsbecomeappreciable.
There also is a minimum quantity of reactantfor which an adequatemea-
surementof the decayof the equilibrium imbalanceis feasible,dependingon the
methodof detectionchosen.This is further complicatedby the requirement,as
set forth in the developmentof relaxation kinetic theory, that the reactant con-
centrationchangeduring the decayof the perturbationimbalancemust be small
relative to the total reactantconcentrations.Thus in order for the perturbationto
be detectableyet small relative to the total reactantconcentration in solution, a
certain minimum of reactantmust be presentin solution. The extentof perturba-
tion can be adjustedto some extent, however,by aitering the magnitudeof the
forcing function (e.g., the pressuredrop). Ideally the relaxationtime dependence
on reactantconcentrationscan be studied over a wide range such that several
relaxation times whose differencesare statistically significant can be obtained.
Obviously, some considerationsrequire that reactantconcentrationsand ionic
strengthbe low, whereasothersdictate high concentrations.
Finding the rangeof experimentalconditionswhere all theserequirements
are satisfied can be arduous.Many relaxation studiesof colloidal suspensions
which at first appearfeasiblemay prove impractical due to an inability to main-
tain a stablesuspensionfor a periodof time sufficient for accurate relaxationtime
measurement.Thus, prior to measurementof relaxationtimes, it is essentialthat
various combinationsof colloidal particleconcentrations,reactantsolution con-

centrations,and backgroundelectrolyteconcentrationsbe testedso that an opti-
mal rangeof experimentalconditionscan be chosen.
Experimental Detailsof Static Studies

The relaxation times (t) following the perturbationare dependenton the
final (i.e., the low-pressure)equilibrium concentrations.The concentrationsof
the various products,reactantsand intermediatesneed not be known during the
equilibrationphaseof a relaxationtime measurement. One needonly measurethe
ambientpressureequilibrium concentrationsof the various reactionparticipants.
Following determinationof a range of absorbent,adsorptive and back-
ground electrolyteconcentrationsfor which stablesuspensionscan be obtained,
various static measurements must be made on the chemical systemof interest.
Equilibrium reactantconcentrationsof samplesfor which relaxationtimes will be
obtainedshould be determinedby preparingsufficient sampleSo that a portion
can be usedfor relaxationmeasurements and anotherportion can be set asidefor
subsequentequilibrium analysis.Thesesamplescan be analyzedalong with sev-
eral othersto determinethe equilibrium constantsfor the reactionsfrom adsorp-
tion isotherms.This approachprovidesexact measurements of reactantconcen-
trations correspondingto each relaxationtime measured.This should yield pre-
cise datafor plotting of relaxationtimes vs. reactantconcentrationsfor testingof
reactionmechanisms.
Once the various equilibrium aspectsof the chemical system have been
considered,preliminary relaxationstudiesof the systemcan be conducted.First
and foremost is determining whether a measurablerelaxation effect can be
obtained.The various time regionsthat the AD converteris capableof monitor-
ing mustbe probedto determineif more than one relaxationoccurs.This alsowill
allow the researcherto choosethe appropriatetime regionsfor measurementof
relaxationtimes associatedwith eachsampleonce actual datacollection begins.
Once it has beenconfirmedthat one or more relaxationscan be detected,it must
be establishedthat the relaxationsare only obtainedwhen all of the proposed
reactantsare present.All othersampleconstituents,aloneand in all possiblecom-
binations, must be testedto see if a relaxation occurs. Once it has been estab-
lished that the relaxation occursonly in the presenceof the proposedreactants,
the actual relaxationtime measurements can be taken.
Severalapproachesmay be usedto vary the concentrationsof the proposed
reactants,eliciting a changein the relaxationtime(s). The first option would of
coursebe to changethe amountof one or more of the reactantsaddedto the sus-
pension.Another option is to vary the pH of the suspension.In the caseof a vari-
able-chargecolloid, this will alter the distribution of the threeinterdependentpro-
tonation statesthat the amphotericsurfacefunctional groups can assume.This
also is a convenientdevice for adsorptionstudiesof anions of weak acids. The
concentrationsof the undissociatedand dissociatedforms of the adsorptivecan
be adjustedby varying the pH, allowing the determinationof which solution
speciesare active in the adsorptionprocess.
1304 SPARKSET AL.
ExperimentalDetails of Kinetic Studies

Measurementsof a relaxationtime associatedwith a single samplein a p-
jump concentrationseries begins with a measurementof the resistanceof the
samplein its autoclavecell. Once this hasbeendetermined,a referencesolution
of similar resistancecan be mixed and loadedinto the referencecell. The poten-
tiometer on the Wheatstonebridge can then be used to artificially balancethe
resistancesof the two cells. At this point, the AD converteris programmedto
monitor the equilibration processthroughoutthe appropriatetime region previ-
ously determinedin the preliminary studies.
The p-jump autoclaveis thensealedwith the brassfoil andwateris forcibly
pumpedinto the chamber.The pressureis brought to within 0.505 MPa (5 atm)
of the burstingpressureand the sampleis given sufficient time to reachthe high
pressureequilibrium (>5 times the slowestrelaxationtime). A rapid thrust of the
water pump is then applied to burst the foil, returning the pressurein the auto-
clave to ambient pressure.This suddenpressurechangealso will trigger the
recordingof the relaxationover the specifiedtime interval. The capturedrelax-
ation is now readyfor transferto the computerfor signal averagingand analysis.
Oncetransferredto the computer,the relaxationcan be displayedon the oscillo-
scopescreenfor a visualdeterminationof the signal quality. Signalswhich are of
obviously poor quality due to mechanicalor electronicdisturbancescan be dis-
cardedat this point, so that high-quality relaxationspectra willnot be degraded
by the averagingprocess.The averagerelaxationspectraalsocanbe displayedfor
a visual inspection of the signal enhancementwith each additional spectrum
recorded.The relaxationsalso can be qualitatively inspectedfor multiple relax-
ation effects(single, doubleor more relaxations)at this point.
Onceseveralrelaxationspectrahavebeenmeasuredand recorded,thecom-
puter can be used for calculation and statistical evaluationof relaxation times.
This processis initiated by first choosinga portion of the relaxation spectrum
where the relaxationeffect is most pronounced.Basedon this choice, the com-
puterobtains16 equally spacedrelaxationamplitudeswhich will be usedto cal-
culate'to Five 't valuesare calculatedfrom theseamplitudesby first evaluatingthe
first througheighth datapoints, then the secondthrough ninthdatapoints,andso
on. If there is no systematicvariation in the five 't times, it can be assumedthat
the relaxationeffect is a single exponential.A systematicincreasein the 't values
from 'tt through't5 indicatestwo or morerelaxationeffectsaresuperimposed.The
five calculated't values should vary in a random fashion due to experimental
error. The standarddeviation of the 't valuescan be usedas a statisticalevalua-
tion of the precisionof the 't measurement.
If two or more relaxationeffectsaresuperimposed, the relaxationcurvecan
be segmentedand the final part of the curve can be analyzedfor the slower
effect(s)and the first portion of the curve can be analyzedfor the fast relaxation.
The computeris equippedwith softwarethat allows the first threeamplitudeval-
uessampledto be ignoredin calculatingthe five 't values.Thus, 'tt is calculated
from the 4th through 11th amplitudevalues,'t2 is calculatedfrom the 5th through
12th amplitudes,etc. Once the slower relaxationtime has beendetermined,the
first portion of the curveis analyzedfor the fast relaxationtime. If the two effects
are not readily separated,it is possiblethat the fast relaxationcan be detectedin
the next shortesttime region and the slow relaxation in the next longest time
region. However, alteration of the experimentalconditions may be required to
separatethe relaxations.
REFERENCES
Aharoni, C., and D.L. Sparks.1991. Kinetics of soil chemical reactions,A theoreticaltreatment.p.
1-18. In D.L. Sparksand D.L. Suarez(ed.) Ratesof soil chemical processes.SSSA Spec.
Pub!. 27. SSSA, Madison,WI.
Aharoni, e., D.L. Sparks,S. Levinson,and I. Ravina.1991. Kinetics of soil chemicalreactions:Rela-
tionships between empirical equations and diffusion models. Soil Sci. Soc. Am. J.
55:1307-1312.
Amacher,M.e. 1991. Methodsof obtainingand analyzingkinetic data.p. 1~0. In D.L. Sparksand
D.L. Suarez(ed.) Ratesof soil chemicalprocesses.SSSASpec.Pub!. 27. SSSA,Madison,WI.
Bar-Tal, A., M. Eick, D.L. Sparks,and S. Feigenbaum.1990. Analysesof adsorptionkinetics using a
stirred-flow chamber.I. Theory and critical tests.Soil Sci. Soc. Am. J. 54:1273-1278.
Bernasconi,C.F. 1976. Relaxationkinetics. Acad. Press,New York.
Bernasconi,C.F. (ed.). 1986. Investigationsof ratesand mechanismsof reactions.4th ed. JohnWiley
& Sons,New York.
Bleam,w.F., and M.B. McBride. 1986. The chemistryof adsorbedCu(lI) and Mn(lI) in aqueoustita-
nium dioxide suspension.J. Colloid Interf. Sci. 110:335-346.
Bunnett,J.F. 1986. Kinetics in solution. p. 171-250.In e.F. Bernasconi(ed.) Investigationsof rates
and mechanismsof reactions.4th ed. John Wiley & Sons,New York.
Carski, T.H., and D.L. Sparks.1985. A modified miscible displacementtechniquefor investigating
adsorption-desorption phenomenain soils. Soil Sci. Soc. Am. J. 49:1114-1116.
Carski, T.H., and D.L. . Sparks. 1987. Differentiation of soil nitrogen fractions using a kinetic
approach.Soil Sci. Soc. Am. J. 51:314-317.
Chou, L., and R. Wollast. 1984. Study of the weatheringof albite at room temperatureand pressure
with a fluidized bed reactor.Geochim.Cosmochim.Acta 48:2205-2217.
Eick, MJ., A. Bar-Tal, D.L. Sparks,and S. Feigenbaum.1990. Analysesof adsorptionkinetics using
a stirred-flow chamber.II. Potassium-calciumexchangeon clay minerals.Soil Sci. Soc. Am.
1. 54:1278--1282.
Eigen, M., and L. DeMaeyer. 1963.Relaxationmethods.Tech. Org. Chern. 8:895-1054.
Fendorf,S.E., and RJ. Zasoski. 1992. Chromium(III) oxidation by O-Mn02' I: Characterization.En-
viron. Sci. Tech. 26:79-85.
Fendorf,S.E.,D.L. Sparks,J.A. Franz,and D.M. Camaioni.1993.An EPRstopped-flowkinetic study
of rapid metal sorption reactionsin colloidal suspensions.Soil Sci. Soc. Am. J. 57:57-62.
Gardiner, W.e., Jr. 1969. Rates and mechanismsof chemical reactions.John Wiley & Sons, New
York.
Gruenewald,B., and W. Knoche. 1979. Recent developmentsand applicationsof pressure-jump
methods.p. 87-94.In W.J. Gettinsand E. Wyn-Jones(ed.) Techniquesand applicationsof fast
reactionsin solutions.Reidel Pub!., Dordrecht,The Netherlands.
Harter, R.D., and R.G. Lehmann.1983. Use of kinetics for the study of exchangereactionsin soils.
Soil Sci. Soc. Am. J. 47:666-669.
Hodges,S.C.,and G. Johnson.1987. Kinetics of sulfateadsorptionand desorptionby Cecil soil using
miscible displacement.Soil Sci. Soc. Am. J. 51:323-331.
Holdren, G.R., and P.M. Speyer.1987. Reactionrate-surfacearearelationshipsduring the early stages
of weathering. II. Data on eight additional feldspars. Geochim. Cosmochim. Acta
51:2311-2318.
Jardine,P.M., and D.L. Sparks.1984. Potassium-calciumexchangein a multireactivesoil system.I.
Kinetics. Soil Sci. Soc. Am. J. 48:39-45.
Jardine,P.M., L.W. Zelazny,andJ.e. Parker.1985. Mechanismsof aluminumadsorptionon clay min-
erals and peat. Soil Sci. Soc. Am. J. 49:862-867.
Ikeda,T., J. Nakahara,M. Sasaki,andT. Yasunaga.1984. Kinetic behaviorof alkali metal ion on zeo-
lite 4A surfaceusing the stopped-flowmethod.J. Colloid Interf. Sci. 97:278--283.
Klimes, N., G. Lassmann,and B. Ebert. 1980.Time-resolvedEPR spectroscopy.Stopped-flowEPR
apparatusfor biological application.J. Mag. Res. 37:53-59.
Knoche,W. 1974. Pressure-jumpmethods.p. 187-210.In G.G. Hammes(ed.) Investigationsof rates
and mechanismsof reactions.3rd ed. JohnWiley & Sons,New York.
1306 SPARKSET AL.
Knoche, W., and H. von Strehlow. 1979.Data captureand processingin chemical relaxation mea-
surements.p. 137-142.In W.J. Gettins and E. Wyn-Jones(ed.) Techniquesand applications
of fast reactionsin solution. Reidel Publ., Dordrecht,The Netherlands.
Knoche,W., and G. Wiese.1974.An improvedapparatusfor pressure-jumprelaxationmeasurements.
Chem. Instrum. 5:91-98.
Lasaga,Ae. 1981. Rate laws of chemical reactions.p. 1-68. In AC. Lasagaand R.I. Kirkpatrick
(ed.) Reviews in mineralogy. Kinetics of geochemicalprocesses.Vol. 8. Miner. Soc. Am.,
Washington.De.
McBride, M.B. 1987. Adsorption and oxidation of phenolic compoundsby iron and manganese
oxides. Soil Sci. Soc. Am. J. 51:1466-1472.
Miller, D.M., w.P. Miller, and M.E. Sumner.1986. Kinetics of silicic acid sorption by goethiteusing
a flow-through cell. p. 169. In Agronomy abstracts.ASA, Madison,WI.
Miller, D.M., M.E. Sumner,and w.P. Miller. 1989.A comparisonof batch-and flow-generatedanion
adsorptionisotherms.Soil Sci. Soc. Am. 1. 53:373-380.
Morgan,J.J.,andW. Stumm.1964. Colloid-chemicalpropertiesof manganesedioxide. J. Colloid Sci.
19:347-359.
NumericalAlgorithms Group. 1984. NAG Fortran library manual,Mark II. NAG, Oxford, England.
Ogwada,R.A, and D.L. Sparks.1986a.A critical evaluationon the use of kinetics for determining
thermodynamicsof ion exchangein soils. Soil Sci. Soc. Am. J. 50:300-305.
Ogwada,R.A., and D.L. Sparks.1986b.Kinetics of ion exchangeon clay mineralsand soil. I. Eval-
uation of methods.Soil Sci. Soc. Am. 1. 50:1158-1162.
Ogwada,R.A, and D.L. Sparks.1986c. Kinetics of ion exchangeon clay mineralsand soil. II. Elu-
cidation of rate-limiting steps.Soil Sci. Soc. Am. 1. 50:1162-1164.
Patrick, W.H., Jr., B.C. Williams, and 1.T. Morgan. 1973. A simple system for controlling redox
potentialand pH in soil suspensions.Soil Sci. Soc. Am. Proc. 37:331-332.
Phelan,P.I., and S.Y. Mattigod. 1987. Kinetics of heterogeneouslyinitiated precipitationof calcium
phosphates.Soil Sci. Soc. Am. 1. 51:336-341.
Randle,K., and E.H. Hartmann.1987. Applicationsof the continuousflow stirred cell (CFSC) tech-
nique to adsorptionof zinc, cadmiumand mercury on humic acids. Geoderma40:281-296.
Sadusky,M.e., and D.L. Sparks. 1991. Anionic effects on potassiumreactionsin variable charge
Atlantic CoastalPlain soils. Soil Sci. Soc. Am. J. 55:49-56.
Sasaki,M., M. Morlya, T. Yasunaga,and R.D. Astumian. 1983.A kinetic study of ion-pair formation
on the surfaceof a (X-FeOOH in aqueoussupensionsusing the electric field pulse technique.
J. Phys. Chern. 87:1449-1453.
Schnabel,R.R., and D.J. Fitting. 1988.Analysis of chemical kinetics data from dilute, dispersed,
well-mixed flow-through systems. Soil Sci. Soc. Am. J. 52:1270-1273.
Seyfried, M.S., D.L. Sparks,A Bar-Tal, and S. Feigenbaum.1989. Kinetics of Ca-Mg exchangeon
soil using a stirred-flow reactionchamber.Soil Sci. Soc. Am. J. 53:406-410.
Sivasubramaniam, S., and O. Talibudeen.1972. Potassium-aluminum exchangein acid soils. I. Kinet-
ics. J. Soil Sci. 23:163-173.
Skopp, J. 1986. Analysis of time dependentchemical processesin soils. 1. Environ. Qual.
15:205-213.
Skopp,J., and D.L. McAllister. 1986. Chemical kinetics from a thin disc flow system:Theory. Soil
Sci. Soc. Am. J. 50:617-623.
Sparks,D.L. 1985. Kinetics of ionic reactionsin clay mineralsand soils.Adv. Agron. 38:231-266.
Sparks,D.L. 1986.Kinetics of reactionsin pure and in mixed systems.p. 83-178.In D.L. Sparks(ed.)
Soil physicalchemistry.CRC Press,Boca Raton,FL.
Sparks,D.L. 1989. Kinetics of soil chemicalprocesses.Acad. Press,New York.
Sparks,D.L., and P.M. Jardine.1981. Thermodynamicsof potassiumexchangein soil using a kinet-
ics approach.Soil Sci. Soc. Am. J. 45:1094-1099.
Sparks,D.L., and P.M. Jardine.1984. Comparisonof kinetic equationsto describeK-Ca exchangein
pure and in mixed systems.Soil Sci. 138:115-122.
Sparks,D.L., and1.E. Rechcigl. 1982. Comparisonof batchand miscible displacementtechniquesto
describepotassiumadsorptionkinetics in Delawaresoils. Soil Sci. Soc. Am. J. 46:875-877.
Sparks,D.L., L.w. Zelazny,and D.C. Martens.1980. Kinetics of potassiumdesorptionin soil using
miscible displacement.Soil Sci. Soc. Am. J. 44:1205-1208.
Sparks, D.L.,and P. Zhang.1991.Relaxationmethodsfor studyingsoil chemicalprocesses.p. 61-64.
In D.L. Sparksand D.L. Suarez(ed.) Ratesof soil chemicalprocesses.SSSASpec.Publ. 27.
SSSA,Madison,WI.
Stach, J., R. Kirrnse, W. Dietzsch, G. Lassmann,Y.K. Belyaeva, and I.N. Marov. 1985. Ligand
exchangereactionsbetweencopper(II)-and nickel(II)-chelatesof different sulfur- and seleni-
um-containingligands. VI. Kinetics of ligand exchangereactionsstudied by stopped-flow
ESR. Inorg. Chim. Acta 96:55-59.
Takahashi,M.T., and R.A. Alberty. 1969. The pressure-jumpmethods.p. 31-55. In K. Kustin (ed.)
Methodsin enzymology.Vo!' 16. Acad. Press,New York.
Tang, L., and D.L. Sparks1993. Kinetics of calcium-potassiumexchangeon montmorillonite using
pressure-jumprelaxation.Soil Sci. Soc. Am. J. 57:42-46.
Toner, C.V., IV, D.L. Sparks,and T.H. Carski. 1989. Anion exchangechemistry of middle Atlantic
soils: Chargeproperties andnitrate retentionkinetics. Soil Sci. Soc. Am. J. 53:1061-1067.
van Olphen, H. 1977. An introduction to clay colloid chemistry. 2nd ed. John Wiley & Sons, Inc.,
New York.
van Riemsdijk,W.H., and EAM. de Haan.1981.Reactionof orthophosphate with a sandysoil at con-
stantsupersaturation.Soil Sci. Soc. Am. J. 45:261-266.
van Riemsdijk, W.H., and AM.A van der Linden. 1984. Phosphatesorption by soils: II. Sorption
measurementtechnique.Soil Sci. Soc. Am. J. 48:541-544.
Walker, w.J., C.S. Cronan,and H.H. Patterson.1988.A kinetic study of aluminum adsorptionby alu-
minosilicateclay minerals.Geochem.Cosmochim.Acta 52:55~2.
Yasunaga,T., and L. Ikeda. 1986.Adsorption-desorptionkinetics at the metal-oxide-solutioninterface
studiedby relaxationmethods.ACS Symp. Ser. 323:230.:-253.
Zasoski,R.G., and R.G. Burau. 1978. A techniquefor studying the kinetics of adsorptionin suspen-
sions. Soil Sci. Soc. Am. J. 42:372-374.
Zhang, P.c., and D.L. Sparks. 1989. Kinetics and mechanismsof molybdate adsorption at the
goethite/water interfaceusing pressure-jumprelaxation.Soil Sci. Soc. Am. J. 53:1028-1034.
Zhang, P.c., and D.L. Sparks. 1990a. Kinetics and mechanismsof sulfate adsorption/desorption on
goethiteusing pressure-jumprelaxation.Soil Sci. Soc. Am. J. 54:1266-1273.
Zhang, P.c., and D.L. Sparks. 1990b. Kinetics of selenateand seleniteadsorption/desorption at the
goethite/water interface.Environ. Sci. Techno!. 24:1848-1856.
Zhang, P.c., and D.L. Sparks. 1993. Kinetics of phenol and aniline adsorptionon an organo-clay
(HDTMA-montmorillonite). Soil Sci. Soc. Am. J. 57:340.:-345.
Published 1996
Chapter 44
Equilibrium Modeling in Soil Chemistry
S. V. MATTI GOD
AND JOHN M. ZAeHARA, Battelle, Pacific Northwest National Laboratory,
Richland, Washington
Over the past decade,chemical equilibrium models have becomeindispensable

tools for the soil chemist.They are now routinely usedto computeaqueousspe-
ciation as a basisfor interpretingchemicalreactivity (Le., adsorptionand ion ex-
change)and nutrient/contaminantbioavailability. Chemicalequilibrium models
are a mainstay in experimentaldesign and data interpretationof soil chemical
studiessuch as computingthe aqueousconcentrationat which precipitationwill
commencein a multi-ion solution or determiningwhich solid phasesof a given
elementare in solubility equilibrium with the aqueousphase.Adsorptionmodels,
which integrateaqueousspeciationreactionswith surfacecoordinationreactions,
are a necessaryaspectof all modemstudiesof soluteadsorptionreactionson soil
material.
Severalexamplesof equilibrium model applicationsinclude,studiesof soil
solution phasespeciation(Emmerichet aI., 1982; Spositoet aI., 1982; Lighthart
et aI., 1983; Baham& Sposito,1984; Murray & Lewis, 1985; Lewis, 1986; Dud-
ley et aI., 1987; Jardine& Zelezny, 1987; Alva et aI., 1988; Simard et aI., 1988;
Luther & Dudas,1992), influenceof solution speciationon ion exchange/adsorp-
tion (Mattigod et aI., 1979,1985; Bowmanet aI., 1981, 1982; Spositoet aI., 1983;
Elrashidi & O'Connor, 1982), and the effects of speciationon precipitationand
dissolutionphenomena(Mattigod, 1983; Sullivan et aI., 1986a,b;Inskeep& Sil-
vertooth, 1988; Hollis et aI., 1988). Computedand measuredsolution speciation
data also have beenused in interpretingobservedplant uptakeof nutrients and
potentially toxic elements(Pavan& Bingham, 1982a;Pavanet aI., 1982b,1984;
Bingham et aI., 1983, 1984, 1986; Cramer& Lauchli, 1986). Thesestudiessug-
gest that computationand measurements of chemicalspeciationin soil systems
hasbecomean importantaspectof soil chemistry.
In this chapterwe examinethe basisof equilibrium modelingand describe
briefly severalcodesused by soil chemists.Examplesare provided of equilibri-
um computationsthat are relevantto soil systems.We briefly discussthe concept
of model validation within the context of soil chemical studies and evaluate
future needsto improve equilibrium modelsusedby soil chemists.
SegoeRd., Madison,WI 53711,USA. Methods of Soil Analysis: Chemical Methods, Part 3-SSSA
Book Seriesno. 5.
1309
1310 MATIIGOD & ZAeHARA
MATHEMATICAL BASIS OF CHEMICAL EQUILIBRIUM MODELS
The goal of chemicalequilibrium models isto computethe equilibrium dis-

tribution of componentsof a systemof interest.For soil systems,the objectiveis
to computethe equilibrium speciationin solution, solid, and if appropriatethe
gaseousphases.Suchcomputationswould includevarioustypesof reactionssuch
as hydrolysis, complexation, dissolution/precipitation,ion exchange,adsorp-
tion/desorption,and oxidation/reduction.Thesereactionscontrol not only speci-
ation within eachphasebut also masstransferbetweenphases.
Typically, the chemical equilibrium models are formulated using mass
actionandbalanceconstraintsfor eachcomponent.For instance,the massof each
Componenti in a soil systemcan be representedby
m,{Tot) =1:m,{solution) + 1:m,{solid) + 1:m,{gas) [1]
Where, 1:m,{solution) representsthe sum of massof ith componentspeciesthat

are present insolution phase,1:m,{solid) the solid, and1:m,{gas) the gasphasefor
a metallic soil componentthat is presentin solution and solid phases.Therefore,
the total massof sucha componentin eachof thesephasesis the sumof the mass
of this component ineachof the species.
[2]
whereA/Z+ representsunboundionic species,1: MaLb is the sum of massof vari-

ous complex speciesof the metal ion with various free and protonatedligands
that are in the system,and l:MML is the sum of massmetal ion that is associated
with any mixed metal-ligandspecies.Similarly, the massof metallic elementthat
is presentin the soil solid phaseis composedof variousspecies.
The 1:(MaLc) representsthe masspresentas solid associatedwith a ligand,

1: a(Mp'tl-J-d) representsthe massof M presentin mixed solids, and1: M(ads)
representsthe massadsorbed,and on exchangeablesites of solid surfaces.For
eachof the dissolved,solid, and adsorbedconstituentswith which the metal ion
is associatedwith, the massaction providesrelationshipsbetweenthe concentra-
tions of free metal, free ligand and the complex forms. Theserelationshipsare
expressedas conditional constants(thermodynamicconstantscorrectedfor spe-
cific ionic strengthconditions). The combinationof mass action relationships
with a massbalanceequationfor eachcomponentresultsin a set of linear equa-
tions. The mathematicalsolution of theselinear equationsyields the equilibrium
concentrationsof eachspeciesof every componentin the system.Obviously, the
validity of predictedspeciationdependsmainly on the inclusionof the correctset
of dissolvedand solid speciesfor eachcomponentand accurateconditionalcon-
stants.
The mode describedabove for computingthe equilibrium compositionis
generallyreferredto as the "equilibrium constant"approachand is the mostcom-
EQUILIBRIUM MODELING IN SOIL CHEMISTRY 1311
mon method used in many models. However, the equilibrium compositionof a

systemalso can be computedby combining the massbalanceequationfor each
componentwith the chemical potential for relevant speciesand by minimizing
the Gibbs free energy of the system. This techniqueof solving the speciation
problemis known as the "free energyminimization" method.A good description
of this approachis providedby Waite (1989). This approachto solving the equi-
librium composition is less common becausechemical potentials for various
speciesare not as reliable as equilibrium constantsand also not as readily avail-
able (Nordstrom& Ball, 1984).
MODELING METAL-LIGAND-SURFACE INTERACTIONS
Metal-OrganicLigand Interactions
Somefractions of the dissolvednaturalorganicmatter(DOM) in soil solu-

tions are difficult to characterizebecauseof their complexity. Dissolvednatural
organicmatterin soil solutionsconsistsof variouslow molecularweight organic
acids, sugars,and other compoundsas well as higher molecularweight humic
substances.However, the types of functional groups presentin DOM and their
chemistry of interactionswith protons and metals are well established.These
functional groupsconsisttypically of carboxylic, phenolicand alcoholic hydrox-
yl, quinoneand ketonic carbonyl, amino, and sulfhydryl moieties.Experimental
dataalso suggestthat DOM may form complexesof varying stoichiometrywith
the samemetal. A fundamentalproperty of metal complexationbehaviorof the
larger molecularweight componentsof DOM, noticeably humic substances,is
their binding site and binding energyheterogeneity(de Witt et aI., 1990).Because
such complexationmay significantly influence metal speciationin soil systems,
various modelshave beenformulatedto include this importantcategoryof inter-
action. Somerecentexamplesof humic complexationmodelswere developedby
Tipping et al. (1990), Susetyoet al. (1990), and Bartochetet al. (1992).
Early attemptsat modeling theseinteractionswere classifiedas "mixture"
models(Sposito,1981a).The goal of theseclassesof modelswas to simulatethe
functional group chemistryof DOM with respectto its complex formation with
metals. The earliest attempts(Morel et aI., 1975) used a mixture of equimolar
concentrationof six organic acids to model the DOM behavior.Another "mix-
ture" model developedby Mattigod and Sposito(1979) includes a combination
of aliphatic, aromatic,and amino acids that contain the sametypes of functional
groupsthat exist in DOM. In the "mixture" model, the concentrationof eachacid
is selectedsuchthat the calculatedproton titration curve for the mixture matches
the measuredproton titration curve of the DOM. The rationalefor this approach
was that this criterion simulatedbetterthe interactionof DOM functional groups
with dissolvedmetals.An excellentdiscussionof thesemixture modelswas pro-
vided by Sposito(1981a).
A betterapproachto simulatingmetal-DOM interactionsis providedby the
"quasiparticle"models.A quasiparticlemodel was developedinitially by Sposi-
to (1981a)using hypotheticalnoninteractingligandswhich very closely matched
the metal-binding behavior of real DaM. This "Scatchard" quasiparticle

approachprovided a mathematicaldescription of an "average" metal reaction
with DaM thus avoiding the intractableproblem of an actual molecular level
descriptionof DaM-metalinteractions.Therefore,eachconditionalstability con-
stant in the model representsa set of complexesthat includes different ligands
and a rangeof stoichiometries.A numberof quasiparticlemodelswere compared
by Cabanisset al. (1984) as to their effectivenessin describingmetal-DaMinter-
actions. Detailed descriptionsand critiques of these quasiparticlemodels also
were provided by Sposito (1986). An improved Scatchardquasiparticlemodel
was recentlydevelopedby Spositoand Blaser(1992) to describebetterthe metal-
DaM interactions.Someof thesequasiparticlemodelshavebeenincorporatedin
a few of the geochemicalequilibrium modelssuchas SOILCHEM, MINTEQA2,
and GEOCHEM-PC.
Adsorption Models
Adsorption, or the accumulationof solutesat the solid-waterinterface, is
one of the important processesin soil-water systems.Adsorption, which arises
from the coordinative propertiesof solid surfacesinfluences soil solution and
groundwatercomposition,and is fundamentalto: (i) nutrient retention and sup-
ply, and (ii) attenuationof macro-and microcontaminants.Adsorption has been
a focus for researchin soil chemistry,environmentalchemistry,andgeochemistry
for years(Stumm & Morgan, 1981; Morel, 1983; Sposito,1984, 1989). Histori-
cal perspectiveson soil chemistry researchrelated to adsorptionwere provided
by Harter (1986).
The adsorption of inorganic ions or organic species by soil surfaces
involves the formation of a complex with a surfacefunctional group [inner- or
outersphere;seeSposito(1984) or Brown (1990) for discussionof this concept],
or retentionas a chargecompensatingspeciesin the diffuse ion swarmsurround-
ing chargedsoil particles(Sposito,1989). Binding strength(i.e., the free energy
of reaction)generallydecreasesin the order inner sphere>outersphere>diffuse
ion. The most important surfacefunctional groupsin soil include the following:
(i) hydroxyl groupson AI, Fe, Mn, or Si on soil oxides with AI and Si exposed
on clay mineral edges;(ii) siloxanecavitieson the surfacesof 2:1 layer silicates
with fixed negative charge arising from isomorphic substitution; (iii) carboxyl
and phenolatesiteson soil organicmatter; and (iv) partially coordinatedsiteson
the surfacesof carbonatesand sulfates.Thesesurfacesites have beenimplicated
in the adsorptionof a wide rangeof both inorganicandorganiccationsand anions
in soil. Significant researchhas been devotedto: (i) identifying the selectivity,
thermodynamics,and kinetics of thesesites for different adsorbates;(ii) deter-
mining the relative contributionsof thesedifferent types of sites to soil adsorp-
tion processes;and (iii) developingmodelsto describecomplex formation reac-
tions on single solid surfaces.
Complex formation on mineral surfacescan be describedas a chemical
reaction betweenan aqueousspeciesand surfacefunctional group leading to a
surface complex. For example, anion (Ll-) and cation (M"'+) adsorption to
hydroxylatedsurfacesites (SOH) on iron or aluminum oxides, or layer silicate
edgesleadingto either an inner- or outer-spherecomplexare written as follows
EQUIUBRIUM MODELING IN SOIL CHEMISI'RY 1313
SOH + M"'+ =SOM(m-l) + W (inner sphere) [4]

SOH + M"'+ =So-- M"'+ + W (outersphere) [5]
SOH + LI- =SL(/-l)- + OH- (inner sphere) [6]
SOH + W + L 1- =SOH/ - LI- (outersphere) [7]
Similarly, cation exchangeto fixed chargesites on layer silicates (X) is often
describedas follows
[8]
In Eq. [8], C and M are the exchangingcationswith stoichiometriesof m

and n, respectively.The chemical identity and molecular structure of surface
complexeson mineral surfacesand soil colloidal material(i.e., reactionproducts
on the right-handsideof Reactions[4] to [7]) are not well established.Definitive
chemicalinformation on thesespeciescan only be obtainedby in situ surface
spectroscopy(e.g.,Hayeset aI., 1987; Chisholm-Brauseet aI., 1990),and useof
this techniqueremainsan active currentresearcharea(Brown, 1990).
Surface complexation and cation exchangereactions, as well as those
involving carboxylandphenolatesiteson naturalorganicmatter,areof greatgen-
eral concernto those desiring to model adsorptionreactionsin soil. Because
adsorption reactions are described utilizing eqUilibrium constantsand mass
action/massbalanceequations,their solution(Westall, 1980) is compatiblewith-
in geochemical codes calculating aqueous speciation (Le., MINEQL,
GEOCHEM, MINTEQ). It is thereforecommonto link aqueousspeciationand
adsorptioncalculations.Suchlinkage is crucial becausecoordinativesiteson sur-
facescompetewith aqueousphaseligandsfor adsorbateions, and different aque-
ous speciesof a given ion may exhibit different adsorptionbehavior(e.g., Bow-
man & O'Connor, 1982; Benjamin & Leckie, 1982; Chairidchai & Ritchie,
1990).Furtherdiscussionsof adsorptionmodelsin the subsequent subsectionand
examplecalculationsin later sectionswill focus on cation exchange,and surface
complexation.
CationExchangeModels
Models for cation exchangethat are relevantto soils have beenreviewed
by Sposito(1986a)andSposito(1989),while molecularandthermodynamiccon-
siderationswere discussedby Sposito(1981). A good review of cationexchange
reactionsin soil may be found in Talibudeen(1981).
The most commonly employedmodel for computerizedcation exchange
calculationshasbeenthe ideal solutionmodel.It representsthe mostsimpleform
of the more general,regularsolution model (Sposito,1989).In the ideal solution
model, the rational activity coefficientsfor surface speciesif) are unity, and the
activity of the surfaceexchangecomplexesare equalto their mole fractions.
For the reactiondefined by Eq. [8], therefore,the following relationships
hold with respectto the thermodynamic(Kex) and conditional (Kc) equilibrium
constants
1314 MATIIGOD & ZACHARA
and
[9]
wherex representsmole fraction and componentswithin parenthesesare single

ion activities. Under the conditionsof ideal exchangefMXm =fcxn = 1 and Kex =
Kc. The mole-fractionbasedconditionalequilibrium constant(Kc) in Eq. [9] is the
Vanselow selectivity coefficient (Kv; Sposito, 1977). Ideal solution behavior is
commonly used as a first assumptionin modeling cation exchangebehavior in
soil becauseit avoids the complicationssurroundingthe estimationof rational
activity coefficientsfor the adsorbedspecies,which can be complex (seeSposi-
to, 1981). Unlike the Davies or Debye-Huckelequationsfor solution species,
thereare no generalized,simple equationsto estimatethe rational activity coeffi-
cientsfor the surfaceexchangecomplex(Sposito,1989).While the ideal approx-
imation appearsvalid for selectedspecimensmectites(see Sposito& Mattigod,
1979; Fletcher & Sposito, 1989), a compositional dependenceof Kc is often
observedin soil materials(i.e., nonideal behavior) as a result of the complex
polyfunctional natureof soil cation exchangers.The ideal approximationis best
suitedto narrow rangesin exchangerphasecompositionand to exchangingions
of like charge.
The ideal solutionapproachcan be usedfor cation exchangecalculationsin
multicomponent speciation codes such as GEOCHEM, MINTEQA2, or
MINEQL using half-reactionsdescribing the formation of a complex with an
aqueousligand representingone mole of chargedsurfacewith unit valence(X-),
(Shaviv & Mattigod, 1985; Fletcher& Sposito,1989; Allison et aI., 1991). Half-
reactionsfor Reaction[8] are definedas follows
[10]
[11]
The equilibrium constants forEqs. [10] and [11] (Kcxm KMXm ) are definedbelow
in concentrationterms of molality (or molarity) as used in speciationcodes.In
these,single ion activity coefficientsfor the ion exchangecomplexes(CXm MXm)
are definedas unity, consistentwith the assumptionof ideality
K'cxn = [CXn]/(cn+)[x-]
and
K MXm = [MXm]/(M"'+)[X-] [12]
where bracketsrefer to concentrationand parentheses refer to single ion activity.

As shownby Fletcherand Sposito(1989),Kcxn andKMXm (in concentrationterms
of molarity or molality) are relatedto the conditional equilibrium constant(i.e.,
Kc or K y , in concentrationtermsof mole fraction) by the following relationship
Kv =(K' MXmn/KCXnm)([MX]
m + [CXn ](mon) [13]
The half-reactionapproachrequiresa self-consistentdata basefor solutes

involved in the simulationthat are referencedto a commonreaction,such as the
formation of NaX or caX2 (seefor exampleFletcher& Sposito,1989, and their
attendantdatabasefor montmorillonite).The choiceof the referencereactionand
its log K is arbitrary, but the value of the log K must be high enough(Le., >10) to
limit computedconcentrationsof free X-. Alternatively, NaX or caX2 may be
selectedas componentsif the programFITEQL is applied(to be discussedlater).
The half-reactionapproachhas been used to model macro-ionexchangein soil
(Shaviv & Mattigod, 1985; Krupka et aI., 1988), and macro-ion/tracemetal
exchangeon montmorillonite(Fletcher& Sposito,1989), smectiticsoil clay iso-
lates (Zacharaet aI., 1992), kaolinite (Schindleret aI., 1987), and kaolinitic soil
clay isolates(Cowan et aI., 1992).The latter two applicationsrequiredan addi-
tional exchangereactionwith H+ to accountfor the pH variant cation exchange
capacity.The half-reactionapproachhasgreat flexibility for describinga variety
of ion exchangeprocessesrelevantto soil. It has,for example,beenusedto model
cation andanion adsorptionon calcite by assumingthat the relevantsurfacereac-
tions were adsorbateexchangefor surfaceCa and HC03, respectively(Cowanet
aI., 1990; Zacharaet aI., 1991).
SurfaceComplexationModels
Surfacecomplexationmodels(SCM) utilize massaction and massbalance
equations,analogousto aqueousphasecomplexationequilibria, to describethe
formation of surfacecomplexesinvolving the proton and sorbateions with SOH.
Particlesurfacechargeand electricalpotentialare treatedas consequences of the
surfacecomplexationreactions onSOH sites.Thesemodelsare applicableto soil
surfacessuch as aluminum, iron, manganese,and silicon oxides, as well as the
edgesof 1:1 and 2:1 phyllosilicates whichcontain broken bond aluminol and
silanol sites(White & Zelazny,1988). Equilibrium constantsfor the surfacereac-
tions within SCM differ from those in the aqueousphasein that they are modi-
fied with a correctionterm for the electrostaticenergyof the interfacethat effec-
tively representsan activity coefficient ratio for the surfacespecies(Goldberg,
1991a). These models, which conform to the Langmuir isotherm, include the
well-known ConstantCapacitance(CCM), Diffuse Double Layer (DLM) and
Triple Layer Models (TLM). The different SCM differ in their considerationof
interfacialstructureand statedrelationshipsbetweensurfacepotentialand surface
charge.As noted by Westall (1980), the modelsare mathematicallydegenerate,
and conform to a set of comparablesimultaneousequationsthat are amenableto
numeric solutions (Dzombak & Morel, 1987). The readeris directed to various
recentanalysesof thesemodels(Westall & Hohl, 1980; Morel et ai., 1981; Spos-
ito, 1984; Sposito, 1989, 1990; Davis & Kent, 1990; Dzombak & Morel, 1990;
Hayes et aI., 1991; Goldberg, 1991a)for details on their molecularhypotheses,
similarities, and differences.Some of the attributes of these models including
number of adjustableparameters,types of surface complexesconsidered,and
their ability to deal with ionic strengthaffectsare summarizedin Table 44-1. Of
these,the DLM is generally consideredto be the least and the TLM the most
complex.
Table 44-1. Selectedattributesof surfacecomplexationmodels.

Typesof
surface Ionic
Adjustable complexes strength
Model parameters allowed effects Considerations§
OLM Nt>t K., IL, Ksi Inner-sphere Through GCSG:j: Restrictedto lower [
(gWl)
CCM Nt> K., IL, C" K.i Inner-sphere Restrictedto fixed I Restrictedto higher[
(:5:0.01)
TLM Nt> K., IL, C" C2, Inner- and outer- Through GCSGi Applicable to variable[,
Kcat, KAm Ksi sphere and electrolyte suffersconstraintproblems
t Nt = site density; K_ = equilibrium constantfor

SOH = So- + W; K. = equilibrium constantfor
SOH + W =SOH·;Ksi =equilibrium constant(s)for cationor anion adsorbatebinding (e.g., Reac-
= =
andKcat> KAn TLM equilibrium constantsfor outersphereelec-
tions 1-4); C" C2 capacitances;
trolyte binding.
:j: GCSG = Gouy-Chapman-Stern-Grahame charge potential relationship: -00 = ad = -0.1174 ~[
sinh(zFljld/2RT).
§ Hayeset al. (1991).
The SCM have found wide application in the descriptionof the acid-base
propertiesand adsorptionbehaviorof inorganic and organic cations and anions
on a variety of single phasesorbentsrelevant to soils including iron and alu-
minum oxides and 1:1 and even2:1 phyllosilicates(see comprehensiverecent
reviews by Davis & Kent, 1990; Goldberg, 1991a). The successof SCM in
describingconditionsresultsfrom the validity of the assumedsurfacecomplexa-
tion process(i.e., conformationto massaction and massbalancelaws) and wide
flexibility in terms of adjustableparameters,rather than from the mechanistic
accuracyof the interfacial structureassumedby any of the SCM. Severalauthors
(Westall & Hohl, 1980;Hayeset al. , 1991) havedemonstratedthat variousSCM
are equally capable of fitting acid-basetitration data, although different but
nonuniquevaluesfor analogousparameterswere requiredby the variousmodels
(Dzombak& Morel, 1987).Indeed,this ambiguity hasled someto caution(John-
ston & Sposito, 1987; Goldberg, 1991) that surface complexation modeling,
alone, cannotbe usedto supportmechanisticconclusionsregardingsurfacespe-
ciation or interfacial structureand that their strengthlies more as a heuristictool
to gain chemical insight (Charlet & Sposito, 1989) and visualize the effects of
complexsimultaneousequilibria.
Certain stepsare normally taken in the application of a SCM to a single
phase sorbent/sorbatesystems(Fig. 44-1). Parameterestimation is the most
involved part of the procedure.It is critical to recognizethat the adjustablemodel
parameters(Table 44-1) are poorly constrainedand show great interdependence
(Dzombak& Morel, 1987). Site density(i.e., numberof reactivehydroxyl sites/
m2 of surface,Nt) has beendeterminedby crystallographiccalculations,tritium
exchange,and adsorptionmeasurements (seeJames& Parks,1982,for methods;
and Davis & Kent, 1990for a summaryof valuesrelevantto soil minerals).Com-
mon valuesreportedfor soil mineralsare summarizedin Table 44-2.
Equilibrium constantsfor surface reactionsare determinedsuccessively
from material balancedatawherethe reactivity of a single componentcan be iso-
lated (i.e., H+, M"'+, or LI-), beginningfirst with ionization reactionsand subse-
Select SCM
DLM
CCM
TLM
OIher
Select or Measure N,
Crystallography
Trttium Exchange
Adsorption
Acld-8_ Tltntion Data

Detannine 1<... K.• Ke... K••
(TLM) Parameter
Graphical Extrapolation
Numenc Optimization
Estimation
Detannine Ks. from Single

Soluta Sorption Data with
fixed K•• K. C. Ke... K..
ITLM)
Base on Chemically defensible
Surface Species
Predictions of Absortbate
Intaractlons under Different
Conditions
Variations in I, pH, [L,.[ or [Mm.]
Effects of Complexing Ligand and
Competing Ions
Differenl Solids Fig. 44-1. General steps in the application of
SCM.
Table 44-2. Density of surfacefunctional groupson oxide and hydrousoxide minerals.t

Range Proton Proton
Mineral of site acceptor donor
phase densities Reference groups groups Reference
sites/nm2 - sites/nm2-
a-FeOOH 2.6-16.8 James& Parks.1982; 4.4 6,7 Sposito,1984
a-Fe203 5-22 James& Parks,1982
Ferrihydrite 0.1-0.9 Dzombak& Morel,
molesper 1990
sole of Fe
TI0 2 (rutile) 12.2 James& Parks,1982 2.6 4.2 James& Parks,1982
Ti02 (rutile and 2-12 James& Parks,1982
anatase)
a-Al(OH)3 2-12 Davis & Hem, 1989 Sposito,1984
t-Al 20 3 6-9 David & Hem, 1989
Si02 (am) 4.5-12 James& Parks,1982 0 all James& Parks,1982
Kaolinite 1.3-3.4 Fripiat, 1964 0.35 1.0 Sposito,1984
t From David andKent (1990)
quently with single ion adsorptiondata (all using consistentvaluesof Nt and the
capacitancetermswhere appropriate).Equilibrium constantsfor the surfaceion-
ization reactionsare estimatedfrom alkalimetric titration datausing both graphi-
cal and numeric procedures.An excellentsummaryof valuesreportedfor these
constants(K+ andK_) for different soil mineralsand SCM may be found in Gold-
berg (1991). Nonlinear least squaresfitting techniques(i.e., FITEQL, Westall,
1982a,b)are gaining wider acceptancefor objective parameter estimation to: (i)
avoid the approximationsof graphicalextrapolationwhich bias the pKa of K+ and
K_ (Koopal et aI., 1987; Dzombak & Morel, 1987), and (ii) provide error esti-
matesfor the optimizedconstantsbasedon experimentaluncertainty.The numer-
ic procedureis the method of choice for fitting experimentaluncertainty. The
numeric procedureis the methodof choice for fitting adsorptionconstants(Ks;)
for sorbatespecies;the numberof sorbatesurfacecomplexesshouldbe limited to
the smallestnumberthat are chemically reasonableand that describethe experi-
mental data. Surfacespeciesfor the sorbatemust be carefully selectedto be con-
sistent with the sorbateaqueousspeciationand spectroscopicdata on sorbate
chemical bonding and nearestneighbors(see for example Hayes et aI., 1987;
Sposito, 1988). Equilibrium constantsmay be extractedfrom single data sets,
multiple datasets,and multiple datasetsvarying in ionic strength,eachwith dif-
fering statistical implications. A thoughtful review of parameterestimationfor
SCM may be found in Dzombak and Morel (1990); while Hayes et ai. (1991)
demonstratedthe sensitivity of the surfaceequilibrium constantsfor the DLM,
CCM, and the TLM to fixed valuesof Nt and capacitanceterms.
The SCM have seenincreasingapplicationin the descriptionof ion reten-
tion in soil and aquifer materials. These applicationsare summarizedin Table
44-3 for anionsand Table 44--4 for cations.The generalobjectivein thesemodel
applicationshasbeento testwhetherexperimentalsorptiondatais consistentwith
a hypothesizedseriesof massaction equationswith respectto the sorbateand the
proton. Conformanceof model calculationsand experimentaldata may be taken
as evidencethat the postulatedreactionsequence(i.e., massaction equations)is
an acceptablechemicalmodel of the system.It neednot, however,be representa-
tive or chemicalflexibility to fit data. Poor agreementis generallytaken as evi-
dence that other reactionsof different stoichiometryor intensity, on eHher the
sameor different sorbents,are important,or else that the intrinsic characteristics
of the soil functional groups in terms of concentrationor acid-baseproperties
have not beenproperly described.
Soil materials, however, provide challengesfor the application of SCM
becauseof their complex polyfunctional character.For example,while the mea-
surementof the surface area of a single phasesorbentis relatively easy,deter-
mining the surfaceareaof the sorbingsoil constituent(an input parameterneed-
ed for SCM with electrostaticterms) is difficult. A similar analogyholds for the
acid-basepropertiesof the sorbingsurfaceand its intrinsic affinity for the sorbate.
It is well establishedthat potentiometrictitrations of soil materials(the basisfor
determinationof K+, K_) may be compromisedby dissolution andotherreactions
that consumeprotons (Parkeret aI., 1979). Additionally, mineral mixtures may
assume complex surface properties that influence their surface charge and
impactionretentionas a result of differential dissolutionand adsorptionbehavior
g
c::
~
Table 44-3. Applicationsof SCM to anion adsorptionon soils and heterogeneous
natural sorbents.
~
::
Citation Sorbate Solid Model Nt S K± Ksi
Anions
c;1
Goldberg& Sposito,1984 PO]- 44 soils CCM qmaxt Packingarea:j: Aluminum, iron Averageof individual optimized
~
oxides§ values Z
-
15 soils EGME surfacearea rJ'J
Goldberg& Glaubig, 1986 B(OH)4' CCM qmaxt Optimizedwith Ksi Optimizedwith KJK_
Spositoet aI., 1988 SeOi- Surfacesoil CCM qmax Packingarea(empirical Aluminum, iron oxides§ Ksl> K s2' Ks3 optimizedon one ~
partitioning into two soil over differentpH ranges ~
sites ==
Goldberg& Glaubig, 1988a SeOj- Surfacesoil CCM qmax N2 gassurfaceareaof Optimized with Ksi Optimized with K+/K_ ~
(0-7.6 cm, soil
entisol) ~
Goldberg& Glaubig, 1988b AsOl- Surfacesoil CCM qmax Nz gassurfaceareaof Aluminum, iron Optimized ~
(0-7.6 cm, soil oxides§
entisol)
Zacharaet aI., 1989 CrOl- C-horizon TLM Optimized ~S~ From aluminum-goethite From Al-goethite
material
(ultisols)
t qmax = sorbateadsorptionmaxima.
:j: S = qrnax N Aa whereN A is Avagadro'snumberand a is the sorbatepackingdensity.
§ Averageof averagevaluesreportedfor iron and aluminum oxides-K+ (int) =107.35; K_ (int) = 10-8/ 95.
~ ~ = differencein Nz surfaceareabeforeand after extractionwith citrate-dithionite-bicarbonate.
....
~
....
I,C
....
c.>
N
natural sorbents.
Table 44-4. Applicationsof SCM to cation adsorptionon soils and heterogeneous =
Citation Sorbate(s) Solid Model Nt S K± Ksi
Mouvet & Bourg, 1983 Cu, Zn, Cd, Pb, River sediment NEM Acid-base t Extrapolationand
Ni, Ca, Mg titration regression:j:
Charlet& Sposito,1987 Na oxisol TLM Acid-base NR## titration, Extrapolation Extrapolation
titration ion retention
Loux et ai., 1989 Pb, Zn, Ni, Cd, SandyWisconsin DLM Extraction~ 600 m2/g From arnorphous§ From amorphous§
Cu aquifer Fe203° nH 20 Fe203° nH20 Fe203° nH20
sediment
Osaki et ai., 1990 Fe(III), Co, Zn Freshwaterand CCM Acid-base Extrapolatedfrom Acid-base Extrapolationfrom
estuarine titration experimentaldata titration experimentaldata
sediments
Payne& Waite, 1991 U Weatheredschist TLM Extraction# 700 m2/g From amorphous Fit to experimental
"ultisol like" Fe203° nH20 Fe203° nH 20 data
Cowanet ai., 1992 Cd Ultisol clay Multisite NEM a) surfaceareatt t From kaolinite, From kaolinite,
fractions [kaolinite b) extraction aluminum- aluminum-
aluminum- goethite goethite
goethite
Zacharaet ai., 1992 Cd Mollisol-smec- TLM for Surfacearea~~ N2 surface From aluminum From aluminum and
tite clays smectiteedges area and silicon oxide silicon oxides
t Surfaceareanot requiredin the NEM.
:j: Authors noted the following relationshipthat was usedto estimatebinding constantsfor Pb, Ni, Ca, and Mg-log X Brurface =0.945 log x BfydrOlysis + 5.6.
§ constantsfrom Dzombakand Morel (1990).
~ Nt = Fe(ex)[g/kg)°moles sites/g-Fe203° nH20(am) where Fe(ex) is from NH20H ° HCl extraction.
# Sameas footnote ~ but ammoniumoxalateusedas extractant.
~
tt Nt.kaolinite =Ns.kaolinite ° N2 surfacearea,where Ns is the reportedhydroxyl site density on kaolinite. 8
:j::j: sameas Footnote~ exceptCDB extractionused. =
§§ TLM linked with ion-exchangehalf-reactionapproachfor multisite/multireactioncalculations. Ro
~~ Sameas Footnote:j::j: but the site density of smectiteedgesNs.smectitewas used.
## Not requiredin this application.
~
(Anderson& Benjamin, 1985, 1990; Lovgren et aI., 1990). Consequently,SCM

have beenapplied to soils with assumptionsregardingthe natureof the sorbing
surfaceand its properties.It is often assumedthat (i) iron/aluminum-oxidesor
layer silicatesare the primary sorbentsin soil, and (ii) that the acid basebehav-
ior of thesesoil sorbents(K+, K_), or functional groups,can be approximatedby
their single-phaseconstants(Tables44-3 and 44--4). Total reactivesite concen-
trations(Nt) havebeenestimatedfor anionsfrom sorptionmaxima(qmax) at lower
pH (Table 44-3). A different procedurehas been usedfor cations(Table 44--4)
becauseprecipitation of hydroxide or carbonatesolids may confound sorption
maxima measurements at the higher pH values that are required.Total reactive
site concentrationsfor cationshave beendeterminedby estimatingthe mass/sur-
face areaof the sorbing phasein the soil by extractionor surfaceareameasure-
ment and equatingthis directly to the site densitiesreportedfor well-studiedref-
erencephases(e.g., Fe203• nH20, or KGa-1).
THERMOCHEMICAL DATA
Data Sources
All geochemicalequilibrium models that use the equilibrium constant

approachto predict the equilibrium composition rely on thermochemicaldata
basesconsistingof equilibrium constantsfor aqueouscomplex species,solubili-
ty productsfor solid phases,and the redox potentialsfor variousoxidation/reduc-
tion reactions. These data are generally tabulatedfor zero ionic strength and
ambient temperatureconditions (25°C). Typically these are experimentaldata
obtained from the standardcompilations of equilibrium constants(Martell &
Smith, 1976) or computedfrom listings of standardchemicalpotentialsfor vari-
ous species(Wagmanet aI., 1982). Some of the equilibrium models also may
include estimatedthermochemicaldata for known aqueousand solid speciesfor
which reliable experimental data is unavailable (Mattigod & Sposito, 1977;
Langmuir, 1979).Typically, the geochemicalequilibrium models contain data
basesthat containequilibrium constantsfor hundredsof reactions.In many cases
thesedata may not be internally consistentor reliable. Therefore,it is important
for the model user to verify that the subsetof thermochemicaldata for reactions
of interestare both reliable and consistent. Verificationof thermochemicaldata
setscan be conductedby following the generalstrategiesdescribedin detail by
Nordstromand Munoz (1985).
Nonideality Corrections
The thermochemicaldata basesfor equilibrium models contain data that
are relevant to zero ionic strength. Becausereal soil systemshave finite ionic
strengths,the constantsin the databaseshave to be correctedto reflect this non-
ideality. The nonideality correctionfor eachthermodynamicequilibrium constant
is conductedby using activity coefficientsfor dissolvedspeciesinvolved in that
specific reaction.For instance, the thermodynamicequilibrium constantfor a
reaction
Ca2+ + HCOj" =CaHCOj [14]

is representedby
K =(CaHCOj)/(Ca2+)(HC03) [15]
and the quantities within parenthesesrepresentsthe activities of dissolved

species.The activities of eachof thesespecies(a;) is related to their respective
concentrations(m;) and activity coefficients(Yi) by the relationship
[16]
Thereforefor Reaction[15]
K = (CaHCOj)/(Ca2+)([HCOj"D =
[CaHCOj]YcaHCOi[Ca2+]'Yca[HCOj"]HCOJ [17]
where the quantitieswithin the bracketsrepresentconcentrations.The thermody-

namic equilibrium constantis relatedto the activity coefficientsby the relation-
ship,
K =Kc YCaHCO/YCaYHC03 [18]

Where the conditional constantKc = [CaHCOj]/[Ca2+][HCOj"]. Therefore the
nonideality correction for K for this reaction involves computing the activity
coefficientsof eachof the relevantaqueousspecies.Typically theseactivity coef-
ficients for ionic species(Yi) are computedby the Daviesequation
log Yi =-AZ7[ {/1I2j(1 + /112)} - 0.31] [19]
WhereA is a constant,Zi is the valenceof the species,and I is the ionic strength

of solution (~0.5 mollL) calculatedby the relationship,
/ =1/2 I. m;Zr [20]
Where mi is the molar concentrationof each ionic species.The activity coeffi-

cients of neutral aqueousspecies are computed by the expression(Sposito,
1981b)
log 'Yi =0.11 [21]
For solutions of ionic strengthsexceeding0.5 mol/L, extendedforms of the

Debye-Huckelexpressionsuch as the Pitzer equation are used to compute the
activity coefficients(Pitzer, 1973).
Temperature Corrections
Both temperatureand pressureaffect the eqUilibrium. Small pressurevari-
ationsthat occur within soil systemsdo not cause significant
changesin equilib-
rium. Changesin temperaturehowever,causemeasurablechangesin equilibrium.

Generally,if the temperatureof a systemvarieswithin ± lOoC of the ambienttem-
perature(25°C), the temperaturedependence of the equilibriumconstantmay not
exceedthe uncertaintiesof the value listed for the ambienttemperature.Predict-
ing equilibrium at temperaturesbeyondthis rangerequirescorrections tobe made
for equilibrium constantslisted for 25°C. The temperaturedependenceof an
equilibrium constantis relatedto otherthermochemicalparametersby the expres-
sion
olnK/OT = (-l/R1)(oMr/01) + (Mr/R'f2) + (l/R)(M.s%1) [22]
where l!J{0, llSo, R, and T are the standardenthalpy,standardentropy, gas con-

stant,and the temperatureof reaction,respectively.For narrow rangesof temper-
aturesthe first and the third termson the right-handside of Eq. [22] becomeneg-
ligible quantitiesand the resultingexpressionis the well-known Van't Hoff equa-
tion. Use of this simplified expressionfor larger temperatureranges leads to
appreciableerrors becauseit ignoresthe temperaturederivativesof the standard
enthalpyand standardentropy of reactions.
ThereforeintegratingEq. [22] resultsin an expression(Eq. [23]) (Mattigod
& Kittrick, 1980) that includes the temperaturedependenceof these thermo-
chemicalparameters.
InKr =(Tr/T)lnK298 + [(llSO 29s1RT)(T - Tr)] -

[(l/R1) (LlCOp)OT] + (l/R) LlCJ1)OT [23]
whereLlCop is the heatcapacityof reaction.The temperaturedependenceof reac-

tions can be more accuratelyevaluatedif the standardentropy and heat capacity
for datafor all reactantsand productsare available.Mattigod and Kittrick (1980)
provided a detaileddescriptionand examplesfor evaluatingtemperaturedepen-
dent equilibria for soil systems.
GEOCHEMICAL EQUILffiRIUM MODELS USED

IN SOIL CHEMISTRY
Thereare a plethoraof geochemicalequilibrium computercodescapableof

predictingequilibrium speciationfrom inputs of measuredtotal concentrationsof
various components.A detailed history, description, and pedigree of various
codesbeing usedin various geochemicaldisciplineshave beenreviewedby sev-
eral authors(Nordstrom& Ball, 1984; Waite, 1989; Bassett& Melchior, 1990, p.
2-14). However,only a limited numberof suchcodeshavefound widespreaduse
amongsoil chemists.The principal reasonfor such selecteduse of certain codes
appearsto be the modificationsand improvementsthese codes contain that make
them particularly suitable for equilibrium calculations involving soil systems.
Following are brief descriptionsof someof thesecodes.
GEOCHEM
This equilibrium code was developedas a modified version of another

equilibrium code REDEQU (Mattigod & Sposito,1979). The GEOCHEM dif-
fered from REDEQL2 in including: (i) additional thermochemicaldata that are
relevantto soil systems,(ii) a methodfor describingcation exchangereactions,
and (iii) a mixture model and a quasiparticlemodel to predict metal-DOM inter-
actions. Other important types of interactionssuch as dissolution/precipitation,
and oxidation/reductionalso were includedin this code.
Additional characteristicsof this code included ways to account for
metastability of solid species including mixed solids, corrections for ionic
strength, and the capability to predict changesin speciationas a function of
changingpartial pressuresof CO2, N2, and O2 (via redox reactions)in the soil
gaseousphase.
The GEOCHEM usesthe eqUilibrium constantapproachto solvespeciation
problems.The set of nonlinearalgebraicequationsresulting from massbalance
for all the componentsprovideseqUilibrium speciationin dissolved,solid, and
adsorbedphases,respectively.The codeoffers sufficient flexibility for the userto
define a specificproblemby appropriatemodification of a numberof input para-
meters.Since its inception GEOCHEM has beena useful tool in understanding
and interpretingexperimentalobservationsrelatedto solid and solution specia-
tion and their effects on adsorption/ionexchange,nutrient uptakeby plants and
to estimate the effects of changing important parameterssuch as pH, ionic
strength,redox potential, and concentrationof selectedcomponentson specia-
tion.
Currently thereare two versionsof GEOCHEM that can be run on person-
al computers.One of theseversionsretainsthe data input format of the original
versionthat can be editedor changedusing an ASCII editor. The secondperson-
al computerversion of GEOCHEM (GEOCHEM-PC V 2.0) includes both an
interactiveand data file mode of data input, and was developedspecifically for
use in soil chemistryproblemsrelatedto plant nutrients(Parkeret aI., 1995). In
addition to the mode of data input, other maJor modifications in this version
include a thermodynamicdatafile that hasbeencustomizedby including a num-
ber of dissolvedspeciesrelevantto plant nutrition, and the eliminationof adsorp-
tion/ion exchangesubroutines(Parkeret aI., 1987). A numberof minor modifi-
cationsalso are included to enhancethe utility of this personalcomputer(PC)
version.
SOILCHEM
The SOILCHEM was developedby Sposito and Coves (1995) with the
same data base structure and computationalbasis as GEOCHEM. However,
SOILCHEM differs from GEOCHEMprincipally by: (i) building the input file in
an interactivemode; (ii) calculating theactivity coefficientsof dissolvedspecies
in soil solutionsof ionic strengthapproaching2 mol dm-3; (iii) computingmetal-
DOM interactions through the improved Scatchardquasiparticlemodel; (iv)
using activities (PH) insteadof concentrationsof protonsto improve the compu-
tational precision;and (v) incorporatingthe CCM (one of the SCM's) to predict

adsorptionreactions.A compiledMacintoshPC versionof this codealso is avail-
able from the authors(Sposito& Coves,1990).
MINTEQ
This codewas formulatedby Felmy et ai. (1984) combiningthe computa-

tional format of MINEQL (Westall et aI., 1976), and a data baseupdatedfrom
WATEQ3 (Ball et aI., 1981). The MINTEQ also usesthe equilibrium constant
approachto solve the speciation problems. The MINTEQ differs from other
codesdescribedhere by allowing equilibrium computationsto be madeat tem-
peraturesother than 298 K. This task is accomplishedby using the Van't Hoff
equationor analyticalexpressionsrelatingthe equilibrium constantsas a function
of temperature.However,as discussedpreviously,useof the Vant'Hoff equation
for computing equilibria over large temperatureranges should be avoided.
Adsorptionand ion exchangereactionscan be computedby a numberof options
that have beenincorporatedin this code.Theseinclude the use of a distribution
coefficient (Kd), a Langmuirian adsorptionmodel, a model basedon the Fre-
undlich isotherm,a simple ion exchangemodel basedon selectivity coefficients
for major cations,and SCM'ssuchas the ConstantCapacitance approach and the
TCM for adsorption.A PC (IBM compatible)versionof MINTEQ also hasbeen
compiled(Petersonet aI., 1987).
Another PC version of MINTEQ is the MINTEQA2/PRODEFA2 code
(Allison et aI., 1991).In this code,an interactivepreprocessor,PRODEFA2eases
input file preparationwith extensiveediting capabilities.The MINTEQA2 has
three significant features that distinguishesit from MINTEQ. These include,
implementationof the model of Dobbs et ai. (1989) to simulate humate-metal
interactions,the ability to perform calculationsat multiple pH values(pH scans),
and improvedcapabilitiesto computeion exchange.
MINEQL+
This PC software(version3.00) wasdevelopedat EnvironmentalResearch

Softwareas a userfriendly version of the chemicalequilibrium code, MINEQL
(Westall et aI., 1976). This programincludesan user interface,an extendedther-
modynamicdata basefrom MINTEQA1, and an on-screenmultiple run manag-
ing system.Adsorption phenomenacan be addressedwith anyoneof the three
surface complexationmodels (CC, generalizedDLM, and TLM). One of the
uniquefeaturesof this softwareis that the output canbe regulatedand displayed
graphically. The MINEQL+ has beenset up so that the usercan easily manipu-
late and customizethe attachedthermodynamicdatabase.
C-SALT
This recently developedchemical equilibrium model was designedby

Smith et ai. (1995) to predict speciationin solutionsof ionic strengthexceeding
0.5 moVL. The nonideality correctionsare computedby Pitzer equations.This
1326 MATIlGOD & ZACHARA
Basic Sorption da..
TOTM _ 10·'
Ilaction
sorbed
01
t
adsorpllon
denslly
pH __
FITEQL Modeling
IlaII grw:
TOTM TOTM K+, K., KSI _ TOT SOH _ TOT SOH +/. s
FIDeM FleeM K+, K., TOT SOH _ KSI -~ KSI ./. S
loglH'1 pH
Fig. 44-2. Exampleof FlTEQL applicationto ion adsorptiondataon soil.
model containsonly a limited numberof componentsand is designedto predict

the sequenceof formation of evaporitemineralsin soil systems.
FITEQL
The FITEQL model developedby Westall and Morel (1977) and Westall
(1982a,b)is a nonlinearleast squaresoptimization programthat can be usedto
adjust parametersin a chemicalequilibrium model to fit experimentaldata.The
codeis basedon the Gaussmethod(Gaizer,1979; Dzombak& Morel, 1990),and
is flexible with many potentialusesin soil chemistry.The adjustableparameters
may take various forms including equilibrium constantsfor solubility, surface
and solution complexation,and ion exchangereactionsas well as total concen-
trations of componentssuch as surfacesites (X-, SOH). The program FITEQL
and other such programs(e.g., NONLIN; Felmy, 1989) offer a methodologyto
estimate equilibrium constantsand their associateduncertainty (Dzombak &
Morel, 1987, 1990).The FITEQL may be usedto optimize parametersor to per-
form equilibrium calculationswith fixed equilibrium constantsand component
concentrations(Westall, 1982a).
The FITEQL containsfour adsorptionmodels(CCM, DLM, TLM, and the
StemLayer model), and can readily be formulatedto perform ion exchangecal-
culationsvia the half-reactionapproach.The Daviesconventionis employedfor
single-ionaqueousphaseactivity coefficients.The FlTEQL hasbeenusedwide-
ly for the calculationof KJK. of mineral solids from alkalimetric titration data,
and sorptionconstantsfor surfacespecies,K si . A typical applicationof FITEQL
to soil chemicalexperimentaldata is shown in Fig. 44-2 for the adsorptionof a
metalcation (M) by soil material.Adjustableparameterssuchas TOTSOH or Ksi
for multiple surface speciesmay be optimized separatelyor simultaneously,
althoughexperiencehasshownthat FlTEQL convergenceis lesslikely for more
than two adjustableparameters.The FITEQL will not convergeusing the TLM if
KJK. andKeatlKAn are adjustedsimultaneously,becausethe reactionsare inter-
dependent.A thoroughand insightful discussionof the useof FITEQL for extrac-
tion of equilibrium constants forsurfacecomplexationreactionsmay be found in
Dzombakand Morel (1990).A listing (Table 44-5) showssomesampleapplica-

tions of FITEQL to complexationand adsorptionproblemsrelevantto soil.
The FlTEQL providesoutputon the goodnessof fit of the optimizedmodel
parametersto the experimentaldata and the standarddeviationof eachadjusted
parameter.An overall varianceof approximatelyone is indicative of good fit,
with a rangeof 0.1 to 20 beinggenerallyacceptable(Westall, 1982a,b;Dzombak
& Morel, 1990). The variance(Vy) is dependenton user input estimatesof the
absolute[sCabs)]andrelative[s(rel)] experimentalerrorassociatedwith eachtype
of input data(e.g., pH, free metal concentration,total metal concentration,etc.).
Error estimatesrecommendedby Dzombakand Morel (1990) for applicationof
FITEQL to acid-basetitration dataof solids and metal ion sorptionexperiments
are summarizedin Table 44--6.
EXAMPLES OF EQUILIBRIUM COMPUTATIONS
SpeciationCalculationwith GEOCHEM·PC
All examplesof speciationcalculationswereconductedusingGEOCHEM-

PC Version2.0 (Parkeret aI., 1995).Typical examplesof speciationcomputations
will be illustratedfor a systemof sevencationiccomponents(Ca, Mg, Na, K, Cu,
Zn, and H) and six anionic components(C03, S04' CI, P04, N03, and OH). The
total concentrationsof the componentshave beenselectedto representa typical
soil solution composition.The examplesare intendedto show different types of
equilibrium computationsrangingfrom simple solution speciationto more com-
plex redox speciationand naturalorganiccomplexation.
Solution PhaseSpeciation
One of the commonproblemsin soil chemistryinvolves computingspeci-
ation of dissolvedspecies.The neededdata for suchcomputationsare the total
concentrationsof all the componentsof interest.As a rule for a soil system,all
the major cationic and anionic componentsalways should be included because
thesecomponentscontributesignificantly to the ionic strengthand speciation.If
equilibrium computationsinclude trace components,it also is necessaryto
include all tracecomponentsthat are presentin concentrationscomparableto the
tracecomponentsof interest.Becausespeciationinvolves equilibria betweenall
components,omitting componentswill fail to provide a completeand accurate
picture of speciation.
The total concentrationsof componentsusedin theseexamplesare shown
in Table 44-7. The datainput and outputformats are not describedherebecause
they are describedin the manualsof eachof the equilibrium codes.Typically, the
output may include: (i) a table that echoesinput data,the initial guessesfor ionic
strengthand free ionic concentrations,the numberof metals,ligands, complexes
and solids involved in the computation;(ii) a list of relevant thermochemical
data, numberof iterations;(iii) computedionic strength,free and total concen-
trationsof eachcomponent;the chargebalanceof the solution; (iv) a tabulation
of stoichiometryand concentrationsof various complexes;(v) free concentra-
§
Table 44-5. Examplesof FITEQL applicationrelevantto soil chemistry.

Adsorption
Citation Sorbent Solute model Use of FITEQL
Fish & Morel, 1985 Aquatic fulvic acid H+, Cu Nat Optimized stability constantsand discreteligand concentrations
Zacharaet aI., 1987 Fe203·nH20 Cr01-, HCOj", S01-, Ca, TLM Multicomponentadsorptioncalculations
Na
Schindleret aI., 1987 Kaolinite Cu,Cd,Pb CCM and half- OptimizedK.i for metal binding to edgeSOH, and K Mex for cation
reaction exchange
exchange
Goldberg& GJaubig,1988b Surfacesoil As04 CCM OptimizedKsi for severalAs surfacespecies
Zacharaet aI., 1989 Ultisol subsoil Cr04 TLM Optimized SOHT for hypothesizedsoil aluminum-goethitesorbent
Dzombak& Morel, 1990 Fe203·nH20 Many metal cations/anions DLM K± for Fe203• nH20, extensiveself-consistentdatabasefor K.i
Zacharaet aI., 1990 CaC03 Ni, Zn, Cd, Co,Ba, Sr Optimized half-reactionconstants(KMex) for exchangeagainstCa
Half-reaction
(Kcax)
Goldberg,1991 FeOOH Various anionic sorbates CCMandTLM OptimizedK si , use of Vy 2 to distinguishbetweenpotentialinner-
and outer-spherecomplexationmechanisms
Hayeset aI., 1991 FeOOH H+ TLM,CCM Sensitivity of K± to different input values,error estimates,data
groupings
Ronngrenet aI., 1991 ZnS H+, Zn2+ CCM Optimized log~oh 10g~_Il' log~101§
t NA =not applicable.
:j: Vy = SOS/DF
§ Formationconstantsfor the generalequilibria pH< + qZn2+ + r(-SZn) =HpZnq(-SZn)p'+Zq)+ ~
8
t:I
Ro
~
~
Table 44-6. Error estimatesfrom acid-basetitration dataand sorption data(from Dzombak& Morel,
1990).
Measurementt S[rel]:t: S[abs]§ Remarks
XH 0.05 0.0 ±0.02 pH unit
XH 0.10 0.0 ±0.04 pH unit
TH 0.01 0.01 x min TH
XM 0.05 0.0 ±0.02 pM unit
TM 0.01 0.01 x min TM
XA 0.05 0.0 ±0.02 pA unit
TA 0.01 0.01 x min TA
t H = hydrogenion (W); M = cation; A = anion.

:t: SH[rel] =0.05 usedfor pH measurements
in sorptionexperiments;SH[rel] =0.10 usedfor pH mea-
surementsin titration.
= =
§ For b log", Sb[abs] 0.434Sa[ref]; seeWestall (1982a)or Skoogand West (1982, p. 75-80).
tions of componentsand the total concentrationof complexesbetweeneachmetal

and ligand; (vi) a table that lists free and complexconcentrationsas the percent-
age of total concentrationof eachcomponent;(vii) tablesof interaction intensi-
ties, and capacities(Sposito & Mattigod, 1980); and (viii) a list of conditional
constantsfor variousspecies.
In this example,speciationwas computedin the solution phaseassuming
that the systemwas closedregardingtransferof massbetweenthe solution phase
and the solid and gas phases.Selectedoutput from GEOCHEM-PC,described
under (i), (iii), (iv), (v), (vi), and (viii) from this computation are tabulated
(Tables44-8 to 44-13).The first sectionof the output(Table 44-8) lists the input
data and the numberof metals,ligands, complexes,and solids that are included
in the computations.In this example,99 complexesresulting from the interac-
tions betweenseven metals and six ligands were considered.No solids were
included because it wasspecifiedthat the computationsmust consideronly aque-
ous speciation.
The secondtabularoutput(Table 44-9) lists the total andthe free ionic con-
centrations,the free ionic activities of eachmetal and ligand, and the remainder
values.Theseremaindervaluesare computedas the differencebetweenthe total
input concentrationsand the sum of computedconcentrationsof all speciesof
eachcomponentdivided by the total input concentration.This value must be less
than 10-4 for the computationto be convergent.Additional data (Table 44-9)
includes thecomputedionic strength,the chargebalance,and the alkalinity of the
solution.
Table 44-7. Total concentrationsof componentsusedin illustrative computations.t

Component pCT Component pC,
Ca 2.43 2.59
Mg 3.22 3.00
K 3.00 2.70
Na 2.38 4.66
Cu (II) 5.76 2.60
Zn 4.90
t pH = 7.90; pCT = -log (molar concentration).
Thble ~. Listing of input dataandcomputationalconditionsfor GEOCHEM-PC.

The conditionsfor the different casesare:
Metal Codeno. GUESS Case1
- -
Ca 1 3.500 2.430
Mg 2 3.700 3.220
K 4 3.500 3.000
Na 5 3.000 2.380
eu2• 9 6.000 5.760
Zn 12 6.000 4.900
Ligand Codeno. GUESS Case1
C03 1 4.000 2.590
S04 2 3.000 3.000
Cl 3 3.000 2.700
P04 9 5.000 4.660
Fixed pH 7.900
t All simple solid phasesare disallowedfor this run. Maximum iterations= 50; Convergencecriteri-
on =1.oooE-04.
The concentrations(-log molar) and the stoichiometryof complexesare

listed in Table 44--10. For instance,the first line of the table lists two Ca-C03
complexes.The stoichiometryof the first complex (1 1 0) indicates that this
complexcontainsone mole of Ca and one mole of C03 but doesnot containany
H or OH. Thereforethe complex is CaC03° and the concentrationis 4.596 (-log
molar). The stoichiometry of the second complex (1 1 1) indicates that this
speciescontains one mole of each of Ca, C03, and H thus identifying it as
CaHCOj. The stoichiometryof hydrolytic speciessuch as CaOH+ are indicated
asl 0-1.
Another tabularoutput(Thble 44--11) lists free concentrations(-log molar)
of eachmetal and ligand (this is the repetitionof datacontainedin Table 44--11)
Thble 44-9. Free metalsand ligandswith ionic strength=1.553 x 10-2 (computed)and fixed pH of
7.900.
-log -log
total -log free
Total concen- Free free Free concen-
Ionst,:j: concentration tration activity activity concentration tration Remainder
Ca 3.715 x 10-3 2.430 2.059x 10-3 2.686 3.395 x 10-3 2.469 1.070x 10-10
Mg 6.026x lQ-4 3.220 3.360x lQ-4 3.474 5.541 x lQ-4 3.256 2.352x 10-1l
K 1.000X 10-3 3.000 8.762x lQ-4 3.057 9.929x lQ-4 3.003 1.319x 10-1l
Na 4.169x 10-3 2.380 3.641 x 10-3 2.439 4.126x 10-3 2.385 2.302x 10-10
Cu2• 1.738x 1~ 5.760 1.955x lo-B 7.709 3.224x 10-8 7.492 1.139x 10-12
Zn 1.259x 10-5 4.900 3.235 x 1~ 5.490 5.335 x 1~ 5.273 6.612x 10-13
C03 2.570x 10-3 2.590 7.863 x 1~ 5.104 1.297x 10-5 4.887 2.401 x 10-9
S04 1.000X 10-3 3.000 4.613 x lQ-4 3.336 7.607x lQ-4 3.119 7.076x 10-1l
Cl 1.995x 10-3 2.700 1.740x 10-3 2.760 1.971 x 10-3 2.705 -1.021 x 10-10
P04 2.188x 10-5 4.660 2.069x 10-10 9.684 6.375 x 10-10 9.196 1.492x 10-11
N03 2.512X 10-3 2.600 2.182x 10-3 2.661 2.473 x 10-3 2.607 1.081 x 10-10
t The solution contains1.315 x 10-2 equivalentsper liter of cationic species,-8.426x 10-3 eq/L of
anionicspecies,andthus hasa computednet chargeof 4.727x 10-3 eq/L. This representsan error
equalto 35.94%of the total chargeof cationicspeciesin solution.
:j: The computedalkalinity for this solution is 2.554x 10-3 eq/L. The predictedpartial pressureof CO2
in equilibrium with this solution is 1.749x 10-3 atm. (pC02=2.757).
S
Table 44-10. Concentrationof solublecomplexes.
Metal Ligand pC, and stoichiometry -e=~
Ca C03 4.596 1 1 0 4.232 1 ~
Ca S04 3.728 1 1 0
Ca Cl 4.794 1 1 0
3:
0
Ca P04 5.170 1 1 1 7.116 1 1 2 5.815 1 1 0 Cl
l'"l
Ca N03 4.695 1 1 0 7.413 1 2 0 t::
Ca OlL 7.433 1 0 -1 14.888 1 0 -2 :z:
~
Mg C03 5.684 1 1 0 5.119 1 1 1
Mg S04 4.615 1 1 0 :z:
til
Mg Cl 5.682 1 1 0
-
0
Mg P04 5.758 1 1 1 7.903 1 2 6.602 1 1 0 t'"
-
Mg N03 4.983 1 1 0 n
Mg OlL 7.021 1 0 -1 15.675 1 0 -2
K C03 7.210 1 1 0 5.866 1 1 1 10.523 2 1 0 §!=
K S04 5.442 1 1 0 12.398 1 1 1 7.955 2 1 0
K Cl 6.118 1 1 0
K P04 10.629 1 1 0 7.084 1 1 9.040 1 1 2
-3
K N03 5.920 1 1 0
K OlL 9.657 1 0 -1
Na C03 6.892 1 1 0 4.947 1 1 9.976 2 1 0
Na S04 4.623 1 1 0 11.679 1 1 6,718 2 1 0
Na Cl 5.300 1 1 0
Na P04 9.810 1 1 0 6.366 1 1 8.521 1 1 2
Na N03 5.701 1 1 0
Na OlL 8.739 1 0 -1
...
~
...
m

Metal Ligand pC, and Stoichiometry
Cu2+ C03 6.119 1 1 0 7.896 1 2 0 6.055 1 1 1
Cu2+ S04 8.650 1 1 0 13.059 1 2 0 15.886 1 1 1
Cu2+ Cl 10.017 1 1 0 13.632 1 2 0 17.737 1 3 0 22.533 1 4 0 11.071 1 1 -1
eu2+ P04 9.793 1 1 1 11.839 1 1 2 7.537 1 1 0 10.113 1 2 0 15.264 1 2 2
Cu2+ N03 9.918 1 1 0 13.435 1 2 0
eu2+ OH- 7.756 1 0 -1 9.210 1 o -3 11.754 1 o -3 15.489 1 0 -4 9.802 2 o -2
Zn C03 5.800 1 1 0 9.577 1 2 0 5.336 1 1 1
Zn S04 6.432 1 1 0 10.840 1 2 0 13.667 1 1 1
Zn Cl 8.398 1 1 0 10.813 1 2 0 15.119 1 3 0 17.814 1 4 0 8.452 1 1 -1
Zn P04 7.474 1 1 1 9.820 1 1 2 7.118 1 1 0 11.593 1 2 0 12.845 1 2 2
Zn N03 7.700 1 1 0 11.116 1 2 0
Zn OH- 6.537 1 o -1 6.591 1 o -2 10.136 1 o -3 14.870 1 0 -4
H+ C03 2.623 0 1 1 4.219 0 1 2
H+ S04 9.185 0 1 1 27.440 0 1 2
H+ Cl 19.061 0 1 1
H+ P04 5.021 0 1 1 5.877 0 1 2 11,683 0 1 3
H+ N03 11.963 0 1 1 ~
8
CI
Ro
~
~
Table 44-11. Concentrationsoffree metal, free ligand, and total metal complexedby eachligand.
Free ligand Free metal C03 S04 CI P04 N03 OH-
Free ligand 4.89 3.12 2.71 9.20 2.61 6.05
Ca 2.47 4.08 3.73 4.79 5.08 4.60 7.43
Mg 3.26 5.01 4.62 5.68 5.70 4.98 7.02
K 3.00 5.85 5.44 6.12 7.08 5.92 9.66
Na 2.38 4.94 4.62 5.30 6.36 5.70 8.74
Cu2+ 7.49 5.78 8.65 9.98 7.53 9.92 7.73
Zn 5.27 5.21 6.43 8.12 6.96 7.70 6.26
H+ 7.85 2.60 9.18 19.06 4.91 11.96
andthe sumof concentrations(-log molar) of all complexesinvolving eachmetal

and ligand. For examplethe sumof the concentrationsof complexesinvolving Ca
and C03 (CaC03': 4.596and CaHCO;: 4.232) is listed as 4.08 (-log molar).
Free concentrationsof each metal and ligand, and the sum of concentra-
tions of complexesof eachmetal and ligand as a percentageof the total concen-
tration also are listed (Table 44-12). The data provided in this tabulation helps
assesspromptly the significantforms of metalsand ligandsat equilibrium. In this
example,we can quickly notice that a major fraction (>90%) of the alkaline earth
and alkali cationsexist in free ionic forms whereasa major fraction of Cu (>95%)
and nearly one-halfof Zn are complexedwith C03• The secondpart of this tab-
ulation shows that nearly all of Cl and N03 (>98%) exist in free ionic forms,
whereasabout 95% of C03, and about 50% of P04 exist in protonatedforms.
About 20% of S04, and 10% of P04 are predictedto be complexedwith Ca. This
tabulationprovidesa helpful summaryof major interactions,while other outputs
provide detailedinformation aboutthe concentrationsand stoichiometryof vari-
ous species.
Finally, the thermodynamicconstantscorrectedfor the ionic strengthof the
solution (conditionalconstants)are tabulated.Here only a part of the tabulation
is reproduced(Table 44-13). Every line includes the identification number of
eachmetal and ligand involved in the interaction,the conditional constantsand
stoichiometryof three solids and six aqueouscomplex species.In this example,
Table 44-12. Primary distribution of metalsand ligands,no solids are allowed.

Component Free metal C03 S04 CI P04 N03 OH
% of total
Ca 91.36 2.26 5.03 0.43 0.23 0.68
Mg 91.95 1.60 4.02 0.35 0.33 1.72 0.02
K 99.29 0.14 0.36 0.08 0.12
Na 98.97 0.27 0.58 0.12 0.01 0.05
Cu(lI) 1.86 95.26 0.13 1.68 1.06
Zn 42.38 49.25 2.94 0.06 0.87 0.16 4.34
Component Freeligand Ca Mg K Na Cu(II) Zn H
C03 0.50 3.26 0.38 0.06 0.44 0.06 0.24 95.05
S04 76.07 18.71 2.43 0.36 2.40 0.04
CI 98.80 0.80 0.10 0.04 0.25
P04 38.22 9.19 0.38 0.38 1.98 0.13 0.50 49.59
N03 98.45 1.01 0.41 0.05 0.08
...~
....
Table 44-13. Computedconditionalconstants.

Metal Ligand Solids Complexes
1 1 .00 0 0 0 .00 0 0 0 .00 0 0 0 2.76 1 1 0 10.97 1 1 1 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
1 2 .00 0 0 0 .00 0 0 0 .00 0 0 0 1.86 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
1 3 .00 0 0 0 .00 0 0 0 .00 0 0 0 .38 1 1 0 .0000 0 .00 0 0 0 .00 0 00 .00 0 0 0 .00 0 0 0
1 9 .00 0 0 0 .00 0 0 0 .00 000 14.34 1 1 1 20.24 1 1 2 5.85 1 1 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
1 57 .00 0 0 0 .00 0 0 0 .00 0 0 0 .48 1 1 0 .27 1 2 0 .00 0 0 0 .000 () 0 .00 0 0 0 .00 0 0 0
1 99 .00 0 0 0 .00 0 0 0 .00 0 0 0 -28.11 1 0 -1 28.11 1 1 -2 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
2 1 .00 0 0 0 .00 0 0 0 .00 0 0 0 2.46 1 1 0 10.87 1 1 1 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
2 2 .00 0 0 0 .00 0 0 0 .00 0 0 0 1.76 1 1 0 .0000 0 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
2 3 .00 0 0 0 .00 0 0 0 .00 0 0 0 .28 1 1 0 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
2 9 .00 0 0 0 .00 0 0 0 .00 0 0 0 14.54 1 1 1 20.24 1 1 2 5.85 1 1 0 .00 0 0 0 .00 0 0 0 .00 00 0
2 57 .00 0 0 0 .00 0 0 0 .00 0 0 0 .88 1 1 0 .0000 0 .00 0 0 0 .00 0 00 .00 0 0 0 .00 0 0 0
2 99 .00 0 0 0 .00 0 00 .00 0 0 0 -11.61 1 0 -1 -28.11 1 0 -2 .00 0 0 0 .00 0 0 0 .00 0 0 0 .00 0 0 0
~
C'l
0
t:l
1(0
N
>
~
~
the first line indicatesthe interactionsinvolving Ca (metal no. 1) and C03 (ligand
no. 1). The identification numbersfor variousmetalsand ligandsare tabulatedin
the manualsthat accompanythe codes.The constantsfor solids are not listed
becausethis specimencomputationinvolved only solution speciation.Two solu-
ble complexes(CaC03° and CaHCOj) are listed with conditional logarithmic
associationconstantsof 2.75 and 10.97,respectively.
Speciationand MassTransfer
In this calculation,all other computationalconditionsare the sameexcept
that the mass transfer between solution and the solid phasewill be allowed
throughprecipitationof solid phasesif the solution is found to be supersaturated
with respectto the secondarysolid phasesincluded in the database.
Thereare two ways to interpretthe resultsthat predict a significantamount
of solid precipitation. First, the systemmay be kinetically supersaturated with
respectto that precipitatedphase.This condition may arise becausethe solid
phasemay thermodynamicallybe the most stable phasein the soil systemand
may form slowly. Becausethe computationalschemeallows the precipitationof
the most stablephase,the resultsmay indicatea significant amountof precipita-
tion. Such a possibility may be examinedby allowing metastablefast-forming
solid phases,and disallowing the precipitationof the most stablephase.Second,
the soil solution may contain colloidal particlesthat are included as part of the
dissolvedphase.It is necessarytherefore to make sure that only the solution
phaseis being analyzed.
In the following example(Table 44-7) any solid is allowed to precipitate
without restrictionfrom the soil solution consideredas a closedsystem.The sec-
tion of the resultingoutput that lists only the percentagedistribution of the com-
ponentsis tabulated.The resultsof computationindicatethat among20 potential
solid phases,the soil solution is supersaturated with respectto calciate(CaC03),
hydroxyapatite [Ca5(P04)30H], malachite [Cu2C03(OH)2], and hydrozincite
[Zn5(C03)z(0H)6](Table 44-14). About 20% of Ca, 50% of Cu, and 5% of Zn
are predictedto precipitateas solids. Becauseno specific conditions have been
imposedon the nature of precipitatingsolids, the most stable phasesare being
precipitatedin this setof calculations.However,the soil solution may be in equi-
librium with solids that precipitatemore rapidly (metastable)than the most sta-
ble solids. In such cases,additionalcomputationsare necessaryto identify these
setsof metastablesolids.
Assuming that the identities and the thermochemical data of these
metastablephasesare known computationscan be made with imposedsets of
metastablephasesto obtain the equilibrium speciation.
CarbonateEquilibrium
Oneof the previousexamples(solution phasespeciation)showedhow car-
bonateequilibrium is computedfor closed systems(Table 44-9). For a closed
system, the measuredtotal dissolved carbonateconcentrationwas used as an
input without specifyingthe partial pressureof CO2. For opensystems,a number
of computationsare run with different partial pressuresof COz. The partial pres-
Table 44-14. Primary distribution of metalsand ligands,solids precipitationallowed.

Component Free metal C03 S04 CI P04 N03 OH
% of total
Ca 73.81 1.38 4.38 0.36 0.57
18.53t 0.98*
Mg 92.29 1.21 4.35 0.36 1.77 0.02
K 99.31 0.11 0.38 0.08 0.12
Na 99.01 0.21 0.61 0.12 0.05
Cu(II) 1.22 47.12 0.09 0.71
50.85§
Zn 45.86 40.17 3.42 0.07 0.18 4.85
5.46~
Component Free ligand Ca Mg K Na Cu(II) Zn H

C03 0.36 1.99 0.28 0.04 0.33 0.03 0.20 69.94
26.79 0.02 0.01
S04 78.17 16.26 2.62 0.38 2.53 0.04
CI 98.93 0.67 0.11 0.04 0.26
P04 0.04 0.01 0.06
99.89
N03 98.60 0.84 0.43 0.05 0.08
tCaC03·
0 H.
* Cas(P04)3
§ CU2C03(OHh·
~ Zns(C03)z(°H)6·
sure at which CO2 either barely dissolvesinto or degassesout of solution is the

equilibrium partial pressure.Open system computationsindicate that the soil
solution is in equilibrium with the partial pressureof CO2 at 2.89 (PC02).
Oxidation/ReductionReactions
Redox reactionsshould only be included in equilibrium computationsif
there are indicationsthat both the speciesof a redox couple exist in significant
concentrations.The type of dataneededfor redox computationsincludeeither: (i)
the concentrationsof eachspeciesof eachredox couple,or (ii) the total concen-
trations of both speciesof each redox couple and the electron activity. Typical
availabledatafor redox speciesconsistsof total concentrationswith no measured
electronactivity.
If the availabledata are of type (i), i.e., the measuredtotal concentrations
of each redox couple are available,the data input is similar to other previously
describedspeciationproblems.For Type (ii) data, the input must include both
redox species,with total measuredconcentrationbeing assignedto one of the
species,while the other speciesis assigneda nominal total molar concentration
of 1 x 1Q-8. Additionally the input must include all relevantredox couples.The
electron activity either measuredor estimatedmust be specified. The electron
activity (pe) is a function of redox potential(Eh) and the relationshipis specified
(Stumm & Morgan, 1981) as
pe = Eh (measuredin millivolts)/59.155 (mv) [24]

Table 44-15.Total concentrationsof componentsusedas input for redox calculations(with pH = 7.90

and pe =-4.(0).t
Component pCT Component pCT
Ca 2.43 C03 2.59
Mg 3.22 S04 3.00
K 3.00 S(II) 8.00
Na 2.38 Cl 2.70
Cu(II) 5.76 P04 4.66
Cu(I) 8.00 NH3 2.60
Zn 4.90 N03 8.00
t Redox couples:SOJS- 2, NHy'N03, Cu+/Cu2+.
In soil systems,the electronactivity may rangefrom -7 to +14 (Sposito& Mat-

tigod, 1980). Typically in soils, the redox potentials exhibit seesaweffects
(Bartlett, 1986);thereforemeasuredpe may not be in equilibrium with redox cou-
ples. The input data for a typical computationinvolving redox reactionsare tab-
ulated below (Table 44-15). Three redox couples,S041'S-2, NHYN03, Cu+/Cu+2
were included and the measuredtotal concentrationsof both speciesof a redox
couple(Type ii data)were assignedto one of the redox couples.The redoxpoten-
tial was set at -237 mV correspondingto a pe value of -4.00.
The resultsof the redox computation(Table 44-16) show that at specified
pe, all of Cu(II) will be reducedto Cu(I) and precipitatedas CU2S, A major frac-
tion of reducedS (95.5%)also is predictedto precipitateall of dissolvedZn in the
Table 44-16. Primary distribution of metals and ligands, redox reactionswith solid precipitation
allowed.
Component Free metal C03 S04 S(II) Cl P04 NH3 N03 OH
Ca 74.71 1.39 4.44 0.36

18.12t 0.98:1:
Mg 93.96 1.23 4.44 0.36 0.02
K 99.44 0.11 0.38 0.08
Na 99.07 0.20 0.60 0.12
Cu(II)
Cu(I)
100.0§
Zn
100.~
Component Free ligand Ca Mg K Na Cu(II) Cu(I) Zn H

C03 0.36 2.00 0.29 0.04 0.33 69.08
26.19t
S04 75.60 16.29 2.64 0.37 2.44
S(II)
6.53§ 93.97.
Cl 98.92 0.67 0.11 0.04 0.26
P04 0.01 0.06
99.89:1:
NH3 6.59 93.41
N03
t CaC03·
:I: Cas(P04hOH.
§ CU2S,
.ZnS.
Table 44-17. Input datafor computingspeciationusing the mixture model (with pH =7.90).
Component peT Organiccomponent peT
Ca 2.43 Citrate 4.14

Mg 3.22 Salicylate 4.27
K 3.00 Phthlate 3.97
Na 1.04 Arginine 4.49
Cu (II) 5.76 Ornithine 4.36
Zn 4.90 Lysine 4.36
C03 2.59 Valine 4.36
S04 3.00 Maleate 3.97
Cl 2.70 Benzene-sulfonate 4.36
P04 4.66
N03 1.05
form of ZnS. Even though the model predictionsfor this pe indicatethat NH3 is
the dominantspecies,observationsof soil systemsindicate that under reducing
conditions (flooded soils) the dominant speciesthat result from N03 reduction
are gaseousN2 and N20, with minor amountsof NH3 and organic N (Reddy et
aI., 1978). Speciationof the nonredoxspeciesdo not significantly differ from the
results of previous examples where solid precipitation was allowed (Table
44-14).
Natural Organic Interactions

In this final exampleof speciationcalculationswe will compareresults
obtainedfrom the Mixture Model (MM) and the ScatchardQuasiparticleModel
(SQM) (Sposito,1981a)with identical input data.The concentrationsof all inor-
ganic componentswere the samein both cases,and the dissolvedorganic acid
concentrationsin both models were set to 226 g of C/m3• Becausethe SQM
model is relevantfor an ionic strengthof 0.1 mol!L, comparableionic strengths
were usedin both models.The input datafor thesemodelsand the organiccom-
ponentsin the MM that are selectedto simulate the behaviorof natural fulvic
material are listed in Table 44-17. Similar organiccomponentsusedin the SQM
are listed as FULl and FUL2 in Table44-18.The concentrationsof organiccom-
ponentsin MM and in the SQM werecomputedas describedby Sposito(1981a).
The resultsof outputsfrom the MM and the ScatchardQuasiparticleModel
are listed in Tables44-19 and 44-20. Thereare no significant differencesin spe-
ciation of Ca, Mg, K, and Na predictedby both models.However,totally differ-
ent speciationpatternsof Cu are predictedby thesemodels.The MM predictsthat
Table 44-18. Input data for computingspeciationusing the Scatchard Quasiparticle

model (with pH
=7.90).
Component peT Component peT
Ca 2.43 C03 2.59
Mg 3.22 S04 3.00
K 3.00 Cl 2.70
Na 1.04 P04 4.66
Cu (II) 5.76 N03 1.05
Zn 4.90 FULl 3.73
FULZ 2.73
Table 44--19. Primary distribution of metals and ligands, mixture model for organic interactions,
solids precipitationallowed.
Organic
Component Free metal C03 S04 Cl P04 N03 OH interactions
% of total
Ca 80.80 1.02 1.39 0.23 13.66 1.93
0.98t
Mg 69.05 0.62 0.94 0.15 28.18 1.04
K 96.54 0.09 0.18 0.05 3.13
Na 98.06 0.18 0.37 0.09 1.27 0.04
Cu(II) 0.02 0.47 99.50
Zn 42.24 25.31 0.91 0.03 3.47 2.78 25.23
t Ca4(P04)JOH.
Table 44-20. Primary distribution of metalsand ligands,quasiparticlemodel for organicinteractions,

solids precipitationallowed.
Organic
Component Free metal C03 S04 Cl P04 N03 OH interactions
% of total
Ca 77.05 0.98 1.33 0.22 13.03 6.42
0.98t
Mg 68.61 0.62 0.94 0.15 28.01 1.67
K 96.54 0.09 0.18 0.05 3.13
Na 98.03 0.18 0.37 0.09 1.27
Cu(II) 2.00 51.61 0.04 0.16 0.78 0.92
44.48:j:
Zn 49.98 29.95 1.08 0.04 4.11 3.29 11.53
t Cas(P04)JOH.
:j: CU2C03(OH)z.
almost all dissolvedCu (99.5%) is complexedwith organics,whereasthe SQM

predictsthat almost the samefraction (96%) of Cu is complexedand precipitat-
ed with carbonate.Also, significantly different speciationof Zn is predictedby
thesetwo models. For instance,the MM predictsup to 25% of total Zn bound
with organics,whereasthe SQM estimatesthat about 12% of Zn is complexed
with organics. These large discrepanciesindicate that the organic interaction
modelsusedhere should be usedwith caution.
Adsorption Models
Calculationswith FITEQL. Two examplesof adsorptioncalculationswill
be presentedusing the generalizedequilibrium and optimization code FITEQL.
The data to be modeledinclude: (i) Cd adsorptionto a clay-sizedisolate «2.0
!lm) from a Mollisol subsoil and, (ii) CrOi- adsorptionon an Ultisol subsoil and
its clay fraction. Disk copies of the input files used in these examplesmay be
obtainedfrom the secondauthor. The input files will not be fully discussed,the
reader is directed to the FITEQL Version 2.0 manual (Westall, 1982a,b)for a
line-by-line descriptionof the input format.
The FITEQL providesthe following output: (i) input data for verification,
(ii) serial data and estimatedstandarddeviations,(iii) valuesof adjustablepara-
100 .. , I!b o Experiment
¢rIJ • Model 1
BO <> Model2
1J
Q)
OIl
<>
•
• • Model3
.0 OIl <>
0
'"
60
.;
[)
il
E
Q)
!!
Q)
40 Il/I •
a. ~ 00
•
• .-
20
0
4 6
pH
Fig. 44-3. Predictiveandbest-fit modelcalculationsof Cd sorptionto a smectiticsoil clay isolatewith

both ion exchangeand edgecomplexationreactions.
metersat eachiteration (if any), (iv) descriptionof chemicalequilibrium includ-

ing total concentrationanderror in the materialbalanceequation,(v) negativelog
free concentrationof all speciesat eachtitration point, and free concentrationof
all speciesat all titration points, (vi) descriptionof surfaceelectrostaticterms at
equilibrium, (vii) a listing of the experimentaldata, (viii) estimatesof standard
deviationin the experimentaldata,(ix) detailsof the optimizationprocedure,(x)
error propagationdata,and (xi) termsin weightedsumof squaresand derivatives
for the normal and optimizationmatrices.
The following representsgeneralguidancepoints for performing adsorp-
tion modelingin soil materials.
1. Model building must proceed from simple to increasingcomplexity
(i.e., multiple surfacespecies).The degreeof complexity shouldbe ade-
quateto describethe data.
2. Chemically realistic sorbentsand surface speciesshould be selected.
Wheneverpossible these choices should be basedon direct analysis
(e.g., mineralogicalor spectroscopic).
3. Optimizationof the chosenmodelshouldbe conductedon aminimal set
of selectedparametersthat are justifiable. Predictivecalculationsmust
be constrainedto demonstrateapplicability under different chemicalor
experimentalconditions.
4. Equilibrium constants fromdifferent SCM are not interchangeable.
Appropriatestepsnecessaryfor successfulSCM modelingof cationic and anion-
ic adsorptionis illustratedas a flowchart (Fig. 44-1).
Cation Adsorption.The adsorptiondata to be modeledare shown in Fig.
44-3 and involve the fractional adsorptionof Cd2+ (10-6 mollL) on a smectitic
soil clay isolate (2.3 gIL) in a NaCI04 electrolyte(I =0.1 mollL) as the pH was
incrementally raised from approximately4.5 to 8.5. The sorbentwas isolated
from the BC horizon of a Mollisol by sedimentationandcontainedsmectite asthe
dominantmineral phaseand a minor amountof mica. Prior to performingadsorp-
tion studies,the isolate was treatedwith dithionite-citrate-bicarbonate(DCB) to
removefree iron oxidesand dilute hydrogen-peroxideto oxidize residualreduc-
tant, and then dialyzed againstdeionizedwater. The resulting sorbentcontained
someresidualorganicmatter(<1%). Additional information regardingthis sor-

bent may be found in Zacharaet at. (1992) and Zacharaand Smith (1994).
To model the datait is assumedthat Cd adsorptionis dominatedby cation
exchangeto fixed chargesiteson the basalplaneof the smectiteand ionizedcar-
boxylate siteson organicmatter(X-) and coordinationto amphotericedgesites
(AlOH and SiOH) on the smectitecrystallites.It is further assumedthat the pro-
ton (KJK+) and solute(Ksj, KCat and KAn for TLM) associationreactionson the
edgesitesare equalto thesesamereactionson the pure componentoxides(Le.,
silica and alumina).The basic tableaufor this adsorptionproblem is shown in
Table 44-21. Activity coefficients for aqueousspeciesare computedvia the
Davies equationthrough Component170 (by convention).Edge-sitereactions
are describedwith the TLM with an inner-spherecomplex for Cd on AlOH
(AlOCd+) and an outer-spherecomplex for Cd on SiOH (SiO--Cdz+); note an
alternateinner-spherereaction for Cd binding to SiOH (SiOCd+) is shown in
parentheses. Cation exchangeis describedwith half-reactionsfor the formation
of individual cationexchangecomplexes(NaX, CdXz). Component270 is usedin
this and othercomparableproblemsas a dummy componentfor the surfacecon-
centrationof the sorbate (inthis caseCd). Component270facilitatesconvergence
of FITEQL during parameteroptimization(Le., log Ks, total site concentrations,
etc.) in adsorptioncalculations.
A representativeFITEQL input file is shownin Table 44-21. This file can
be usedto exactly reproducethe calculationlabeledModell (a predictivecalcu-
lation using FITEQL in Fig. 44-3, or speciesor site concentrationsto be dis-
cussedsubsequently).The input file includesserial datafor pH (componentno.
50 input as W concentrationin moVL; Le., pH-log gamma),the adsorbedcon-
centrationof Cd (in molelL) at the input pH values[componentno. 270; i.e., CdT
(moVL) • fraction sorbed],and Xi (in molelL) at eachof the input pH values
[componentno. 3; CECNa measuredat eachpH (moVg) • sorbentconcentration
(gIL)]. The equilibrium constantsfor the exchangecomplexes(log KNaX =13.8;
log K CdXZ =31.2) havebeenselectedto yield nonpreferencecation exchangeon
X- (i.e., KNa ~ Ked =1) and to maintainfree X- at low concentration(e.g., <10-9
moVL). The exchangeconstantsare in units of moles/liter,and, for heterovalent
exchange,require a concentrationadjustmentterm for expressionin the more
common units of mole fraction (see Fletcher& Sposito, 1989; Zacharaet aI.,
1993). The total concentrationsof edgeAlOH and SiOH (in mol/L) have been
estimated(for this representativefile) using: (i) the simplifying assumptionthat
the externalsurfacearea (measuredin multipoint Nz adsorption)representsan
approximationof the edgesurfacearea(in mZ/g) and (ii) the site densities(Le.,
molesof sites/mz) for edgeAlOH and SiOH on montmorillonite as reportedby
White and Zelazny(1988).
Adsorption calculationsperformed with this and modified versions of
Table 44-21 are shown in Fig. 44-3. Although it is not shown, the readermay
verify that AlOCd+ and CdXz are the dominantsurface species that contributeto
Cd sorption given the assumptionsand constantsused in the calculation. The
agreementof the model calculationsto the experimentaldatacan be improvedif
the log K for CdXz is increasedto 31.65 (Model 2, an optimizedvalue),suggest-
ing preferentialexchangeof Cd over Na. Optimizing on the concentrationsof
g
Table 44-21. Input data FITEQL modeling of cadmiumsorption on soil smectite.
H+ Act AIOH SiOH X- Ca2+ Na+ CIO" Cd2+ Ads PSI 0 PSI B PSI D logK B-matrix
50 170 2 1 3 4 7 60 270 160 161 162
Cd2+ -10.08 170, 2.0
CdOW -1 2 1 -20.36 170, 0.5
CdOH£ -2 2 1 -33.0 170, 0.0
AIOH 1
AIO- -1 -1 1 -1 0 0 -11.5 170, 0.0
AIOHt 1 1 1 1 0 0 5.7 170, 0.0
AIO-Na+ -1 0 1 1 -1 1 0 -9.2 170, 0.0
AIOH 2+-CIO" 1 2 1 1 1 -1 0 7.9 170, 0.0
AIOCd+ -1 3 1 1 1 0 0 -2.7 170, 0.0
SiOH 1 170, 0.0
SiO- -1 -1 1 -1 -6.95 170, 0.0
SiO--Na+ -1 0 1 -1 1 0 -6.6 170, 0.0
SiO--Ca2+ -1 3 1 -1 2 0 170, 0.0
SiO--Cd2+ -1 3 1 1 1 -1 2 0 -10.0 170, 0.0
SiO--CdOH -2 2 1 1 1 -1 1 -11.7 170, 0.0
(SiOCd+)' (-1) (3) (1) (1) (1) (1) (0) (0) (-7.91) 170, 0.0
NaX 1 1 1 13.8 170, 0.0
CsX2 4 2 1 1 31.2 170, 0.0
caX2 4 2 1 30.6 170, 0.0 c;')
-~
0
t [Cdr = 10-6 mol/L, I = 0.1 mol/L as NaCI04, sorbent= 2.3 gIL]. t:I
f(o
~
~
100 + +
o o --.
80 [)
+
~co 60 §
C
e
Q)
40
Q)
a.
20
Q GI
7 10
pH
Fig. 44-4. Optimized model fit to chromatesorption dataon the U1tisol clay isolate with total con-
centrationof reactiveFeOH surfacesites asthe optimizedparameter.
edge sites (Model 3) producesa better fit to the data at higher pH, but poorer
agreementover the pH rangeof the experiment;the optimized edgeconcentra-
tions (1.4 x 10-4 mollL) are approximately10-fold lower than the valuesinput in
Tables44-21 (1.7 x 10-3 mollL). The readermay be thejudgeasto whetherthese
changes(optimized log KCdX2 and AIOH T) are justifiable given the assumptions
in the precedingparagraphand the known propertiesof soil smectites.
Under no circumstancesis the agreementof the model to the dataquantita-
tive, underscoringthe difficulty in modeling adsorptionreactionsin soil. Other
factors or reactionsexist in this casethat are importantto Cd sorptionon the soil
smectite.In spite of thesefailings, the seriesof reactionsused in FITEQL pro-
vides a qualitative description of the trend in the adsorptiondata that may be
improveduponby the reader.Input file (Table44-21) may be modified to testthe
sensitivity of the model calculationsto site concentrationand equilibrium con-
stant estimates.
Anion Adsorption.The secondexampleinvolves the modelingof CrOi-
(10-6 mollL) adsorptionto an Ultisol subsoil (BC horizon of the Cecil soil) and
its clay fraction which containsgoethite, hematite,gibbsite, and kaolinite. The
examplecalculationis basedon the assumptionsthat: (i) AI-goethite is the dom-
inant sorbentfor CrOi- in the soil (seeZacharaet aI., 1989), and(ii) the soil alu-
minum-goethiteexhibits comparableacid-baseproperties(KJK_), and CrOi-
binding constants(Ksj) to specimenAI-goethite with similar mole-percentage AI
(seeAinsworth et aI., 1989).The examplecalculationwill utilize FITEQL to: (i)
estimatethe concentrationof FeOH sites in the soil isolate «2.0 !lm) through
optimization (curve fitting) on the experimentaladsorptiondata(Fig. 44-4) and
(ii) predictionof CrOi- adsorptionon the whole soil materialin the absenceand
presenceof sOi- as a competitiveion (Fig. 44-5).
The input datadescribingthe adsorptionproblemis given in Table 44-22.
It is assumedthat the ratio of FeOH to AIOH sitesin the soil goethiteis equalto
the Fe/AI ratio in a citratedithionite-bicarbonateextractof the soil clays(=0.22),
and that the surfaceareaof the soil goethiteis equal to the differencein the N2
surfaceareameasuredbeforeand after DCB extraction.The CrOi- sorptionreac-
tions arewritten asouterspherecomplexationreactions,consistentwith the noted
100 o CP
[!J!!J 6~:;
80
$~
+ [IH
• []
.
o
CP-Modell
CP-Mode12
u
Q) 8+ + 8 ., ~
.,
CP/S04
-eg 60
~ 1;1
CP/S04-Modell
q,.,
"E + CP/S04-Model 2
Q) +
~ 40
Q)
"-
.,0 0
20
,." e· (!).B 0
o
4 10 11
pH
Fig. 44-5. Predicted(Modell) and best-fit (Model 2) calculationsof chromatesorptionon the Cecil
soil without and with sulfate.
sensitivity of CrOi- adsorptionon iron-oxide to both ionic strength and SOi-

concentration(Rai et aI., 1986; Zacharaet aI., 1987, 1989) and recentmodeling
of such phenomenologcal responseson iron-oxides(Hayeset aI., 1988). Compo-
nentsnos. 170 and 270 are usedcomparablyto the previousexample.The adsorp-
tion constantsfor the FeOH and AlOH sites have beenderived from adsorption
studieswith single phaseoxide sorbents(Zacharaet aI., 1987; Ainsworth et aI.,
1989).
Serial data is input for log [W] (componentno. 50) and adsorbedCrOi-
(componentno. 270) at the input pH values.The optimization is performediter-
atively; optimizing first on FeOHT (FOH Component1) then readjusting the
AlOH T concentrationin the input file (AOH, Component2) to maintainthe Fe/Al
concentrationratio in the DCB extract(0.22), and reoptimizingon FeOHT again.
This procedure,if repeatedapproximatelyfour times, yields an optimizedvalue
of FeOHT that changeslittle with further iterations.The resultingmodel parame-
ters are unableto describethe more gradualslope to the sorptionedgebelow pH
7 (Fig. 44-4), but otherwiseresult in good agreementto the experimentaldata.
The SOS/DFis 13.2, which is dependenton the error estimatefor no. 270). The
computedsite concentration(FeOH + AlOH) is 4.15 x 10-5 mollL for the exper-
iment, or 6.64 x 10-6 mol/g of clay.
Theseoptimized hydroxyl site concentrationsfor the soil clay can be used
to estimatethe site concentrationsfor the 50 gIL soil suspensionsin Fig. 44-2
accordingto the relationship
SOHsoil =SOHclay(mol/g)• soil concentration(gIL) •
massfraction clay in soil.
Implicit in this relationshipis the assumptionthat the clay fraction controls the
sorption propertiesof the whole soil. The predictedCrOi- adsorptionusing the
optimizedsite concentrationfrom aboveand the binding constantsof crO]- and
SO]- for aluminum-goethitequalitatively agreewith the experimentaldata(Fig.
44-5). The predictededgesare generallysteeperin slope than the experimental
t'!l
10
<::
Table 44-22. Input data FITEQL modeling of CrOj- sorption to Ultisol soil clay isolates.
...t::
H+ Act AIOH FeOH Na+ NOj" SO]- CrO]- Ads PSI 0 PSI B PSID logK B-matrix =
e
<::
50 170 2 10 11 12 60 270 160 161 162
CrOj· 1 ==
1 4 1 6.51 170, 0.5 0
==
HCr04" I:'
H2Cr04 2 6 1 5.56 170, 0.5
NaCr04" 4 1 0.70 170, 0.5 ...Z~
Cr0201- 2 6 2 14.56 170, 2.0 C'l
SOj- 4 170, 2.0 ...z
HS04" 4 1 1.99 170, 0.5 fI:J
0
FeOH 1 170, 0.0 ...
t'"'
FeO- -1 -1 1 -1 0 0 -10.8 170, 0.0 ~
FeOH2 1 1 1 1 0 0 4.2 170, 0.0
FeO--Na+ -1 1 -8.9 t'!l
=
-1 0 1 0 170, 0.0
FeOH2-NOj" 1 2 1 1 1 -1 0 6.1 170, 0.0 ...==
FeOH2-S 0i- 1 5 1 1 -2 0 11.6 170, 0.0 ~
FeOH2-H S04" 2 6 1 1 -1 0 16.3 170, 0.0 e;
FeOH2-CrOi- 1 5 1 1 -2 0 9.8 170, 0.0
FeOH2-H Cr04" 2 6 1 1 1 -1 0 19.4 170, 0.0
AlOH
AIO- -1 -1 -1 0 0 -12.0 170, 0.0
AlOH2 1 1 1 0 0 6.3 170, 0.0
AIO--Na+ -1 0 -1 1 0 -9.9 170, 0.0
AIOH2-NOj" 1 2 1 -1 0 8.3 170, 0.0
AlOH2-CrOi- 1 5 1 1 -2 0 9.4 170, 0.0
AlOH2-HCr04" 2 6 1 1 -1 0 15.9 170, 0.0
AIOH2-Soi- 1 5 1 1 -2 0 11.6 170, 0.0
AIOH 2-SOj- 2 6 1 -1 0 16.3 170, 0.0
....
Ul
-""
1346 MA1TIGOD & ZACHARA
data,exhibiting higher sorption at low pH and lower sorption at high pH. How-
ever, the model correctly describesthe direction in shift of the CrOl- edgethat
occursin the presenceof sol-, althoughthe absolutemagnitudeof the competi-
tive effect is not well describedbelow pH 7. The agreementof the model to the
datais sufficient to testvarioushypothesesregardingthe soil sorptionreaction(s),
such as competitiveion, electrolyte,and pH effects.
The disparitiesbetweenexperimentand prediction in Fig. 44-5, given the
good fit to the adsorptiondata on the soil clay isolate in Fig. 44-4, indicate that
the calculation assumptions(Le., specimenaluminum-goethiteis a quantitative
surrogateof the soil adsorbent)are an oversimplification,and possibly an inac-
curaterepresentationof, the soil sorption process.The importanceof other con-
tributing sorbentsand the possibleexistenceof heterogeneous sorptionsiteswith
nonuniformadsorptionenergiesis thereforeimplied. For example,the adsorption
constantsfor CrOl- and sol- can be optimized sequentiallyusing the soil data
(holding all other parametersconstant)to give a more gradualslope to the pre-
dicted adsorptionedgesthat more closely agreewith the experimentaldata(Fig.
44-5, Model 2 calculations).The optimized constants,which should be viewed
as curve fitting parameterswithout mechanisticsignificance,are KCr04 = 12.5,
K HCr04 = 16.7, KS04 = 9.5, and K HS04 = 12.5 (log values).Incorporatingadsorp-
tion reactionsfor kaolinite (e.g., Zacharaet aI., 1987) along with that for alu-
minum-goethitecan improve the predictionsfor the soil (yielding more gradual
predictededges),but lead to a more complicatedcomputationalproblemwith an
additionalset of assumptionsregardingthe propertiesof the soil kaolinite.
VALIDATION OF EQUILIBRIUM MODELS
Geochemicalequilibrium codesare capableof making detailedpredictions

of elementalspeciationin soil systems.Becauseof increasinguse of speciation
codesin a wide variety of studies,it is essentialthat the predictionsmadeby these
codesare reliable and accurate.The processof comparingpredictedand mea-
sured speciationis defined as validation. Thereforevalidation is viewed as an
integratedtest of accuracywith which a geochemicalmodel and its thermody-
namic databasesimulateactualchemicalprocesses(Jenne& Krupka, 1984).The
criterion for validation can either be subjectiveor quantitative,thereforethe cri-
terion needsto be specifiedfor comparingmodel predictionsand the experimen-
tal observationsof a natural systemat a selectedlevel of detail.
Either a quantitativeor a SUbjective criterion can be used for validation.
The quantitativecriterion consistsof either a deviative or statistical basis. The
deviative criterion definesa deviation coefficient D, that is a measureof differ-
encebetweena predictedvalue P and a measuredvalue M expressedas
D = (P-M)IP [25]
The deviation coefficient (D) should be less than the validity coefficient (V) for
validation. The validity coefficient is selectedby the model useron the basisof
establishedconstraintssuch as regulatorystatutes.
ATTAINED 'Pi
Fig. 44-6. The levels of elementalspeciationpredictedby equilibrium models.
The statisticalcriterion for validation involves the use of appropriatesta-

tistical teststo comparethe predictedand measuredquantitiesat a specifiedlevel
of confidence.The statisticalcriterion thereforeis the most rigorous and objec-
tive validation criterion. Thomaset al. (1991) discussand explain the use of var-
ious statisticalmethodsfor validating different kinds of predictive models.
Methodsof Validation
Geochemicalequilibrium modelsare designedto predict speciationat the

highest level of detail (concentrationof all speciesfor which thermochemical
data is available),whereasvalidation of thesemodelsonly can be accomplished
at a selectedlevel of detail (measureableconcentrationof speciesor groupsof
species).Various levels of predictionsthat can be madeby the equilibrium mod-
els are graphicallyshown (Fig. 44-6). However, detailedpredictionsat the level
of individual speciescan only be made if reliable thermochemicaldata for such
speciesare available. Validating models at the highest level of detail involves

increasinglycomplex measurements and cost. Also, the current state of knowl-
edge in analytical techniquesprecludesdetailed validation at the individual
specieslevel for a majorityof species.A detailedreview of methodsandschemes
of validation for geochemicalequilibrium models was provided by Mattigod
(1995).
Becauseof limitations in analytical techniques,validation of geochemical
equilibrium codesat the highestlevel of detail cannotbe achieved.Also if vali-
dation of a code is conductedfor a selectedsystemundera specifiedset of con-
ditions, the code may not be valid for different setsof conditions and systems.
Thereforegeochemicalequilibrium codescan only be conditionally validated.
Measurableparametersthat are usedin validating model predictionsfor a
soil systeminclude the concentrationsof free ions, complex species,the proton
activity, the electron activity if applicable,and the types and amountsof solid
species.The qualitative or subjectivevalidation at the simplest level of detail
would measuringpartitioningof a componentbetweensolid and aqueousphases.
A more detailedlevel would include the validation of predictedchargeforms of
all componentsnamely cationic, anionic and neutral aqueousspecies.Several
analytical schemeshave beenproposedto separateand measurethe concentra-
tions of aqueousspeciesbasedon their charge.Ion sensitiveelectrodes(ISE) can
be usedto measuredirectly andvalidatethe free ionic concentrationsor activities
in the aqueousphase.If a redox couple is dominantin a systemof interest,the
predictedconcentrationsof redox species ata measuredelectronactivity can also
be validated.
The types and quantitiesof major solid phasespeciescan be either direct-
ly or indirectly validatedby using analytical or solubility methods.Existenceof
someminor and trace element-bearingsolid speciesalso can be validatedquali-
tatively. Even though the major adsorbedcomponentscan be validated,thereare
no reliable techniquesto determinethe typesof adsorbedspecies.However,sev-
eral spectroscopictechniquesare currently beingdevelopedto determinethe spe-
ciation on adsorptionsurfaces(Brown, 1990).
A Schemefor Validation
Appropriatemethodsand criteria can be selectedto conditionally validate
equilibrium models.A proposedscheme(Fig. 44-7) includesa stepwiseselection
of detail and criteria for validation. According to this scheme,the validation at
the lowest level of detail (Level III) consistsof measuringthe partitioning of
componentsbetweensolutionandsolid phase.A higherlevel of validation(Level
II) includes measurements of aqueousspeciationin addition to Level III mea-
surements.Highestlevel of validation(Level I) accordingto this schemeincludes
all feasiblemeasurements of aqueousand solid phasespeciation.
Even thoughthereare a few examplesof limited and conditionalvalidation
studies(seeexamplescited in Mattigod, 1995), equilibrium modelsusedin soil
chemistry have not been systematicallyvalidated. Therefore it is necessaryto
conduct validation studiesof geochemicalequilibrium models and incorporate
the resultsas part of the model documentation.
Fig. 44-7. A proposedvalidation schemefor equilibrium modelswith levels of validation keyed to

the numberand complexity of measurements.
MODELING LIMITATIONS
Equilibrium Conditions
The goal of equilibrium modeling is to predict the speciationof compo-

nentsand their distribution of massbetweenphasesin a given system.Therefore
equilibrium conditions are assumedto prevail for all categoriesof interactions
such as hydrolysis, complex formation, ion exchange/adsorption, and dissolu-
tion/precipitationreactions.
The assumptionof equilibrium conditionsfor aqueousphasereactionssuch
as hydrolysisand complexformation is usually valid becausea majority of these
reactionsare known to occur very rapidly (Wilkins & Eigen, 1965; Pankow &
Morgan, 1987). Many studiesof ion exchangeand adsorptionin soils haveshown
that these reactions also attain equilibrium within several hours. While these
adsorptionreactionsare intrinsically rapid, many reactionsin soils show slower
kinetics due to diffusion of sorbing speciesinto the intersticesof soil minerals.
Available data on the kinetics of dissolution/precipitationreactionsof minerals
indicate that attainmentof equilibrium may range from a few hours to several
months. Clearly the time scale is an important factor in predicting equilibrium
speciation.Becauseof rapidity of reactions,predictionsbasedon equilibrium for
speciationof all componentsin aqueousand adsorbedphaseswould be reliable.
If equilibrium predictions are neededthat include solid phasespeciation,then
knowledgeis neededof the kineticsof dissolution/precipitation reactions of com-
1350 MATI1GOD & ZAeHARA
ponentsof interestso that a time scalecan be specifiedfor the predictionsto be

reliable.
ThermochemicalData
The reliability of predictions using equilibrium models dependssignifi-

cantly on including the effectsof all prominentinteractionsin a systemof inter-
est.It is necessarytherefore,to identify the relevantreactionsand incorporatethe
thermochemicaldatafor thesereactionsin the predictive model. Such complete
information and data are not available for all interactions.For instancethere is
someevidencethat aqueousmixed metal and mixed ligand complexesmay exist
undercertainconditions.Becausethe identitiesandthermochemicaldatafor such
speciesare unknown,the significanceof theseinteractionson soil solutionchem-
istry cannotbe evaluated.
Thereis considerabledatato suggestthe existenceof surface-coprecipitat-
ed or solid solution phasesin soils and sediments.The surfacecoprecipitated
phaseshave different compositionsthan the substratesand exist in metastable
equilibrium with ambient aqueousphase.Therefore the compositionsand the
limits of substitutionfor all componentsand the thermochemicaldata for these
surfacecoprecipitatedsolid phasesneedto be resolvedfor improving the model
predictions.Due to the potential significanceof thesesolid solution speciesin
defining trace elementinteractionsin soil systems,it is necessaryto include the
contributionof thesereactionsto improve the predictive capabilitiesof equilibri-
um models.For a few known solid solutionsystems,modelshavebeen proposed
to predict the stoichiometryof the solid solution phasesfrom the compositionof
the aqueousphase(Glynn et at., 1990a,b).A practical and useful approachis to
incorporatethesesolid solution modelsinto existing equilibrium codes.
AdsorptionModeling
Modeling limitations can be viewed from two perspectives:(i) intrinsic

limitations of the models themselvesand (ii) limitations to the application of
thesemodelsto complexsoil materials.
Limitations to Models
As noted in previous sectionsof this chapter,the various SCM differ in
their assumedstructureof the interfacial region and whethersurfacecomplexes
with electrolyte ions are allowed. Severalauthorshave comparedthesemodels
(Westall & Hohl, 1980; Morel et aI., 1981; Davis & Hem, 1989; Davis & Kent,
1990; Hayeset at., 1991),with the generalconclusionthat eachof the mostcom-
monly employedcodes(DDLM, CCM, and TLM) is sufficiently flexible to pro-
vide equivalentfits to alkalimetric titration dataand single concentrationadsorp-
tion data at low adsorptiondensities(i.e., adsorptionedges)and single ionic
strengths.Although all the codescan explicitly calculatethe competitiveeffects
of specifically sorbing minor solutes (discrepanciesare normally observed
becauseof sorbatespecific site populations;seeGoldberg,1991 for summaryof
SCM applicationsto competitivesorptionstudies),the modelsvary in their abil-

ity to describethe competitiveeffectsof weakly binding soil solution macroions
becauseof their different interfacial hypotheses.The TLM can accountfor these
effects specifically through surface ion pairing, while both the other models
(CCM and DDLM) havedifficulty describingthesephenomena(Charlet& Spos-
ito, 1987; Zacharaet aI., 1989; Cowan et aI., 1990; Davis & Kent, 1990).The
models have not yet beencomparedfor their ability to describeadsorptiondata
over larger rangesin sorbateconcentration,especiallyas surfacesite saturation is
approached.Becausethe modelsoften employ different site densitiesto describe
commondatasets,differencesbetweenmodelsmay becomeexaggeratedat high-
er site occupations,especiallyif electrolytebinding is assumedas in the TLM.
A limitation to all SCM is that they embody Langmuirian adsorption
behavior(i.e., a single adsorptionenergyor log K). As a result, they are unable
to describe, with a single set of adjustableparameters,the more commonly
observedFreundlichisothermbehaviorfor metal constantsthat describesorption
well at low concentrationbut may significantly overpredict sorption at high
adsorptiondensities.Overpredictionmay be lessenedby explicitly incorporating
sorbatesite coveragefactors > 1. There are additional recognizeddeficiencies
specific to eachof the codesthat should be recognized.
1. The CCM requiresa site densitybelow that found by isotopic exchange
or crystallographiccalculationsto achieve a good fit betweensurface
charge/pHrelationshipsat different I.
2. The DDLM greatly overestimatesmeasuredvaluesof the zetapotential.
3. The numerousfitting parameterswithin the TLM lead to constraint
problemsand require a larger databasefor parameterestimation.
Ion exchangemodels basedon the half-reaction approachare limited by their
assumptionof ideality. Soils are complex mixed-exchangersystems,where the
combinedeffectsof multiple equilibria on different phases contribute to the over-
all observedsorption behavior. Sposito (1981, p. 139-142) has shown that the
soil exchangecomplex will behave as a nonideal exchanger, regardlessof
whetherthe individual componentphasesthemselvesbehaveas ideal exchangers.
This leadsto the practical implication that a given suite of self-constituenthalf-
reaction exchangeconstantsmay be valid only over limited sorbateand elec-
trolyte/major ion concentrationranges. These concentrationranges will vary
betweensoils dependingon the identity, concentration,and exchangeselectivity
of the phasescomprisingthe soil exchanger.
Problems in Application
As describedby Davis and Kent (1990) the applicationof adsorptionmod-
els to naturalsorbentssuchas soilsis complicatedby their compositemineral and
organiccharacterand complexsurfacepropertiesresultingfrom weatheringreac-
tions, microbiological activity, and other soil-forming processes.Thesedynamic
processeslead to coatings of secondarymineral phasesand organic materials
whoseproperties aredifficult to characterizeandwhoseidentity and composition
changewith time and depth from the soil surface.As a result of their composite
character,ion sorption in soil reflects the contributionsof various surfacereac-
tions on different functional groups,and mineral phasesand coatings.The con-

tributions of theseindividual reactionsmay be difficult or impossibleto experi-
mentally identify, forcing the modelerto hypothesizethe identity of specificreac-
tions and their dominance.
The compositecharacterof soil complicatesthe measurementof site con-
centrations,reaction constants,and sorbent surface area (as required by the
SCM). Therefore,a major modelingquestionis whetherone should utilize reac-
tion constantsfor specific phases(or functional groups) presumedto dominate
soluteadsorption(Le., iron- or aluminumoxides),or shouldone apply constants
descriptiveof the bulk phasepropertiesof the soil? The answerto this dilemma
dependson the mineralogicalcompositionalcomplexity of the soil material.As
previouslynoted,it is especiallydifficult to determinethe site concentrationsand
their distribution betweendifferent adsorbingphases.While various techniques
have been used to estimatereactive site concentrationson soil materials (Le.,
solutesorptionmaxima and extractableconstituents;Davis & Kent, 1990), none
has met with universal acceptance.The applicationof techniquesemployedfor
oxide materialssuch astitration and tritium exchange are limited by mineral dis-
solution (Parkeret aI., 1979) and complicationsarising from watersof hydration
surroundingchargecompensatingcationson layer silicates.Becauseof the diffi-
culty in measuringsite concentrations,Davis and Kent (1990) recommendedthe
use of a site density of 3.84 Ilmoles/m2 for modeling adsorptionin all natural
materials.
The existenceof nonuniqueadsorptionenergiesalso createserror in the
applicationof adsorptionmodelsto soil. All adsorptionmodels described in this
chapterare discretesite modelswhere specific surfacereactionsare assumedto
exhibit a singleor uniquefree energychange(Le., constantlog K R ). In soils, how-
ever, similar functional groups (Le., FeOH, AlOH, SiOH) may exhibit variable
affinities for the proton and other solutesas a result of attachmentof different
phasesand the existenceof defectand structuraVSUrfaceimpurities. Suchbehav-
iors may be more appropriatelydescribedwith heterogeneous adsorptionmodels
(e.g., Bolt & van Riemsdijk 1987; van Riemsdijk et aI., 1987; Parrish& Perdue,
1989) where a probability density function is derived for important parameters
(Le., K+> K_, and KsJ
Dynamic phenomenasuch as bacterialgrowth, organic matter solubiliza-
tion, mineral dissolution and reprecipitation, and other related reactions that
occurduring soil sorptionor titration experimentsmay complicateapplicationof
adsorptionmodels.Theseshouldbe recognizedas they frequently requireexplic-
it considerationfor successfulmodelingof adsorptionreactionsin soil. For exam-
ple, Andersonand Benjamin(1990) and Lovgren et al. (1990) observedthat Al3+
can precipitate oniron-oxide surfaces,changingtheir acid baseand ion retention
properties.Wieland and Stumm(1992) notedthat the releaseand readsorptionof
Al3+ by kaolinite was an important determinantof its surfacechemistry under
weakly acidic conditions. Solubilized elementssuch as AI, Fe, Ca, and Si may
competewith the target elementsfor surfacecoordinationsites while displaced
ligandssuchasorganicmaterialmay alter solutionspeciationand free metal con-
centrations.
FUTURE TRENDS IN EQUILmRIUM MODELING
Equilibrium modeling is a well-establishedtool for gaining insights into a

number of soil chemical processes.Equilibrium codes are continually being
improved and developedto addresssome of the major limitations in modeling
describedin the previoussection.In addition to the developmentof multicompo-
nent equilibrium codesanotherclassof codesare being developedfor modeling
specific typesof interactionsin soils. A major featureof thesecodesis to model
the interactionsof a limited numberof components.One exampleof sucha code
is C-SALT designed for modeling the evaporite mineral formation in soils.
Another exampleis the CHROMAT code that predictsequilibrium speciationof
Cr in soils (Felmy et aI., 1992).
Another emergingtrend in modelingis the couplingof expertsystemswith
equilibrium models.Expertsystemsareknowledge-based codesthat are designed
to examinedatavalidity, provide guidance,and makedecisionson computation-
al conditions.A prototypeof an expert systemto guide the computationof car-
bonate equilibrium in groundwaterwas described recently by Pearsonet al.
(1990).
One of the importantactivities in modelingwill include the systematicval-
idation of equilibrium codesto confirm and improve the predictability of these
codes.Improved samplingand analytical techniqueswill provide the necessary
impetus forvalidating the equilibrium codes.
REFERENCES
Allison, J.D., D.S. Brown, and KJ. Novo-Gradac.1991. MINTEQA2/PRODEFA2.A geochemnical
assessment model for environmentalsystems:Version 3.0 user'smanual.EPN600/3-91/021.
USEPA
Alva, AK., M.E. Sumner,and AD. Noble. 1988. Effects of fluoride on colorimetric measurement of
solution aluminum. Soil Sci. Soc. Am. J. 52:374-378.
Anderson,P.R., and M.M. Benjamin. 1985. Effects of silicon on the crystallization and adsorption
propertiesof ferric oxides.Environ. Sci. Technol. 19:1048-1053.
Anderson,P.R., and M.M. Benjamin. 1990. Surfaceand bulk characteristicsof binary oxide suspen-
sions.Environ. Sci. Technol. 24:692-698.
Baham,J., and Sposito,G. 1984. Proton and metal complexationby water-solubleligands extracted
from anaerobicallydigestedsewagesludge.J. Environ. Qual. 15:239-244.
Ball, J.W., E.A Jenne,and M.W. Cantrell. 1981. WATEQ3: A geochemicalmodel with uranium
added.U.S. Geol. Surv. Open-FileRep. 81-1183.
Bartlett, R. 1986. Soil redox behavior.p. 179-207.In D.L. Sparks(ed.) Soil physical chemistry.CRC
Press,Boca Raton, FL.
Bartoschat,B.M., S.E. Cabaniss,and EM.M. Morel. 1992. Oligoelectrolytemodel for cation binding
by humic substances.Environ. Sci. Technol. 26:284-294.
Bassett,R.L., and D.C. Melchior. 1990. Chemical modeling of aqueoussystems.Am. Chern. Soc.,
Washington,DC.
Benjamin, M.M.,and J.O. Leckie. 1982. Effects of complexation byCl, S04, and S203on the adsorp-
tion behaviorof cadmiumon oxide surfaces.Environ. Sci. Tech. 16:162-170.
Bingham,ET., G. Sposito,and J.E. Strong. 1984. The effect of chloride on the availability of cadmi-
um. J. Environ. Qual. 13:71-74.
Bingham, ET., G. Sposito,and J.E. Strong. 1986. The effect of sulfate on the availability of cadmi-
um. Soil Sci. 141:172-177.
Bingham, ET., I.E. Strong,and G. Sposito.1983. Influence of chloride salinity on cadmiumuptake

by swisschard. SoilSci. 135:160-165.
Blanchar,R.W., and C.E. Marshall. 1981. Eh and pH measurementin Menfro and Mexico soils. p.
103-128.In R.H. Dowdy et a1. (ed.) Chemistryin the soil environment.ASA Spec.Publ. 40.
ASA, CSSA, and SSSA,Madison,WI.
Bowman,R.S., and G.A O'Connor.1982. Control of nickel and strontiumsorptionby free metal ion
activity. Soil Sci. Soc. Am. 1. 46:933-936.
Bowman,R.S., M.E. Essington, andG.A O'Connor.1981. Soil sorptionof nickel: Influenceof solu-
tion composition.Soil Sci. Soc. Am. J. 45:860-864.
Brown, G.E. 1990.Spectroscopicstudiesof chemisorptionreactionmechanismsat oxide-waterinter-
faces. p. 309-353.In M.F. Hochella andA.E White (ed.) Mineral-waterinterfacegeochem-
istry. Reviewsin Mineralogy 23. Miner. Soc. Am., Washington,De.
Cabaniss,S.E., M.S. Shuman,and B.J. Collins. 1984. Metal-organicbinding: A comparisonof mod-
els. p. 165-179.In C.J.M. KramerandJ.e.Duinker (ed.) Complexationof tracemetalsin nat-
ural waters.Martinus Nijhoff, The Hague,the Netherlands.
Chairidchai,P., and G.S.P. Ritchie. 1990. Zinc adsorptionby a lateritic soil in the presenceof organ-
ic ligands. Soil Sci. Soc. Am. 1. 54:1242-1248.
Charlet, L., and G. Sposito. 1989. Bivalent ion adsorption by an oxisol. Soil Sci. Soc. Am. 1.
53:691-695.
Chalet, L., and G. Sposito. 1987. Monovalent ion adsorptionby an oxisol. Soil Sci. Soc. Am. J.
51:1155-1160.
Chisholm-Brause,C.J., P.A Day, G.E. Brown, Jr., and G.A. Parks.1990. Evidencefor multinuclear
metal-ion complexesat solid/water interfacesfrom x-ray absorption spectroscopy.Nature
(London) 348:528-530.
Cowan, C.E., I.M. Zachara,and e.T. Resch.1990. Solution ion effects on the surfaceexchangeof
selinite on calcite. Geochim.Cosmochim.Acta 54:2223-2234.
Cowan,e.E.1.M. Zachara,S.C. Smith, and e.T. Resch.1992.Individual sorbentcontributionsto cad-
mium sorptionon ultisols of mixed mineralogy.Soil Sci. Soc. Am. 1. 56:1084-1094.
Cramer,G.R., and A Lauchli. 1986. Ion activities in solution in relation to Na+-Ca2+interactionsat
the plasmalemma.1. Expl. Bot. 37:321-330.
Davis, J.A., and I.D. Hem. 1989. The surface chemistry of aluminum oxides and hydroxides. p.
185-219.In G.A Sposito(ed.) The environmentalchemistryof aluminum. CRC Press,Boca
Raton,FL.
Davis, JA., and D.B. Kent. 1990. Surface complexation modeling in aqueousgeochemistry.p.
177-248. In M.E Hochella and AE White (ed.) Mineral-water interface geochemistry.
Reviewsin Mineralogy. Vol. 23. Miner. Soc. Am., Washington,De.
Davis, J.A., and1.0. Leckie. 1978b.Effect of adsorbedcomplexingligandson trace metal uptakeby
hydrousoxides. Environ. Sci. Tech. 12:1309-1315.
DeWit, 1.e.M., W.H. Van Riemsdijk, M.M. Nederlof, D.G. Kinniburgh, and L.K. Koopal. 1990.
Analysis of ion binding on humic substancesand determinationof intrinsic affinity distribu-
tions. Anal. Chim. Acta 232:189-207.
Dudley, L.M., B.L. McNeal, J.E. Baham,C.S. Coray, and H.H. Cheng.1987. Characterizationof sol-
uble organiccompoundsand complexationof copper,nickel, and zinc in extractsof sludge-
amendedsoils. J. Environ. Qual. 16:341-348.
Dzombak,D.A., and EM.M. Morel. 1987. Adsorption of inorganic pollutantsin aquaticsystems.1.
Hydraul. Eng. 113:430-475.
Dzombak, D.A, and EM.M. Morel. 1990. Surface complexation modeling: hydrous ferric oxide.
JohnWiley & Sons,Inc., New York.
Elrashidi, M.A, and G.A O'Connor.1982. Influenceof solution compositionon sorptionof zinc by
Emmerich,W.E., L.J. Lund, A.L. Page,and A.C. Chang. 1982. Predictedsolution phaseforms of
heavy metalsin sewagesludge-treatedsoils. J. Environ. Qual. 11:182-186.
Felmy, AR., D.C. Girvin, and E.A Jenne.1984. MINTEQ A computerprogramfor calculatingaque-
ous geochemical equilibria. NTIS PB84-157148. EPA-600/3-84-032. Nat!. Tech. Inform.
Serv.,Springfield, VA
Felmy, AR., D. Rai, I.M. Zachara,M. Thapa,and M. Gold. 1992. CHROMATTM: The soil chromi-
um attenuationevaluationmodel. Electric PowerRes. Inst., Palo Alto, CA
Fish, W., and EM.M. Morel. 1985. Propagationof error in fulvic acid titration data:A comparison
of methods.Can . J. Chern.63:1185-1193.
Fletcher,P., and G. Sposito. 1989. The chemicalmodeling of clay/electrolyteinteractionsfor mont-
morillonite. Clay Miner. 24:375-391.
Fripiat,1.1. 1964. Propertiesof aluminosilicates.p. 327-358. In Clays Clay Min., Proc. 12th Natl.
Conf.
Gaizer,F. 1979. Computerevaluationof complex equilibria. Coord. Chern. Rev. 27:195-222.
Glynn, P., and E.1. Reardon.1990a.Solid-solutionaqueous-solutionequilibria: thermodynamictheo-
ry and representation.Am. I. Sci. 290:164-201.
Glynn, P., E.1. Reardon,L.N. Plummer, and E. Busenberg.1990b.Reaction paths and equilibrium
end-points in solid-solution aqueous-solutionsystems. Geochim. Cosmochim.Acta
54:267-282.
Goldberg,S., and G. Sposito. 1984. A chemicalmodel of phosphateadsorptionby soils. II. Noncal-
careoussoils. Soil Sci. Soc. Am. 1. 48:779-783.
Goldberg,S., and R.A Glaubig. 1988a.Anion sorption on a calcareous,montmorillonitic soil-Sele-
nium. Soil Sci. Soc. Am. I. 52:954-958.
Goldberg, S., and R.A Glaubig. 1988b. Anion sorption on a calcareous,montmorillonitic soil-
Arsenic. Soil Sci. Soc. Am. J. 52:1297-1300.
Goldberg,S., and R.A. Glaubig. 1986. Boron adsorptionon California soils. Soil Sci. Soc. Am. J.
50:1173-1176.
Goldberg,S. 1991a.Sensitivity of surfacecomplexation modelingto the surfacesite density parame-
ter. J. Colloid InterfaceSci. 145:1-9.
Goldberg,S. 1991b. Use of surface complexation models in soil chemical systems. Adv. Agr.
47:233-329.
Harter, R.D. (ed.). 1986. Adsorption phenomena.van NostrandReinhold, New York.
Hayes,KF., G. Redden,W. Ela, and J.O. Leckie. 1991. Surfacecomplexationmodels: An evaluation
of parameterestimation using FITEQL and oxide titration data. I. Colloid Interface Sci.
142:448-469.
Hayes,K.F., AL. Roe, G.E. Brown, KO. Hodgson,J.O. Leckie, and G.A Parks. 1987. In situ x-ray
absorptionstudy of surface complexes:Seleniumoxyanions on a-FeOOH. Science(Wash-
ington, DC) 238:783--786.
Hayes,KF., C. Papelis,and I.O. Leckie. 1988. Modeling ionic strengtheffects on anion adsorption
at hydrousoxide/solutioninterfaces.J. Colloid InterfaceSci. 125:717-726.
Hollis, I.F., R. Keren, and M. Gal. 1988. Boron releaseand sorptionby fly ashas affectedby pH and
particle size. J. Environ. Qual. 17:181-184.
Inskeep,w.P., and I. Silvertooth. 1988. Inhibition of hydroxyapatiteprecipitation in the presenceof
fulvic, fulvic, and tannic acids. Soil Soc. Am. J. 52:941-946.
James,R.O., and G.A Parks. 1982. Characterizationof aqueouscolloids by their electrical double-
layer and intrinsic surfacechemical properties.p. 119-126.In E. Matijevic (ed.) Surfaceand
colloid science.Vol. 12.
Jardine,P.M., and L.W. Zelezny.1987.Influence of organicanionson the speciationof mononuclear
and polynuclearaluminum by ferron. Soil Sci. Soc. Am. J. 51:885-889.
Jenne,E.A, and KM. Krupka. 1984. Validation of geochemicalmodels.p. 143--152.In B.R. Redal
(ed.) Proc. Conf. Workshopon fundamental geochemistryneedsfor NuclearWaste Isolation.
U.S. Dep. Energy,Washington,DC.
Johnston,C.T., and G. Sposito.1987. Disorderand early sorrow: Progressin the chemicalspeciation
of soil surfaces.p. 89-99.In L.L. Boersma(ed.) Futuredevelopmentsin soil scienceresearch.
ASA, Madison, WI.
Koopal, L.K, W.H. van Riemsdijk, and M.G. Roffey. 1987. Surface ionization and complexation
models:A comparisonof methodsfor determiningmodel parameters.J. Colloid InterfaceSci.
118:117-136.
Langmuir, D. 1979. Techniquesof estimatingthermodynamicpropertiesfor someaqueouscomplex-
esof geochemicalinterest.p. 353--387.In E.A Jenne(ed.) Chemicalmodelingin aqueoussys-
tems. ACS Symp. Ser. 93. Am. Chern. Soc. Washington,DC.
Lewis, B.G. 1986. Application of chemical equilibrium modeling to predict the fate of chemical
species in phosphogypsumleachate. p. 61-73. In Proc. 2nd Int. Symp. Phosphogypsum,
Miami, FL. FIPR Pub. no. 01-037-055.Vol. 1.
Lighthart, B., J. Baham,and V.V. Volk. 1983. Microbial respirationand chemicalspeciationin metal-
amendedsoils. J. Environ. Qual. 12:543--548.
Loux, N.T., D.S. Brown, c.R. Chafin, J.D. Allison, and S.M. Hassan.1989. Chemicalspeciationand
competitive cationic partitioning on a sandy aquifer material.J. Chern. Spec. Bioavail.
1:111-125.
Lovgren, L., S. Sjoberg,and P.W. Schindler. 1990. Acid basereactionsand Al(III) complexationat
the surfaceof goethite.Geochim.Cosmochim.Acta 54:1301-1306.
Luther, S.M., And M.1. Dudas.1992. Porewaterchemistryof phosphogypsumtreatedsoil. J. Environ.
Qual. 22: 103--108.
1356 MATIlGOD & ZACHARA
Martell, A.E., and R.N. Smith. 1976. Critical stability constants.PlenumPress,New York.
Mattigod, S.V., and G. Sposito. 1977. Estimatedassociationconstantsfor somecomplexesof trace
metalswith inorganic ligands. Soil Sci. Soc. Am. J. 41:1092-1097.
Mattigod, S.Y., and G. Sposito. 1979. Chemicalmodeling of trace metal equilibria in contaminated
soil solutions using the computer program GEOCHEM. p. 837-856. In E.A. Jenne (ed.)
Chemical modeling in aqueoussystems.ACS Symp. Ser. 93. Am. Chem.Soc., Washington,
De.
Mattigod, S.V., and J.A. Kittrick. 1980. Temperatureand water activity as variablesin soil mineral
activity diagrams.Soil Sci. Soc.Am. J. 44:149-154.
Mattigod, S.Y., J.A. Frampton,and e.H. Lim. 1985.The effect of ion-pairformation on boron adsorp-
tion by kaolinite. Clays Clay Miner. 33:433-437.
Mattigod, S.V. 1995. Validation of geochemicalequilibrium models.p. 201-218.In R.H. l.oeppertet
al. (ed.) Chemicalequilibrium and reaction models. SSSA Spec.Publ. 42. ASA and SSSA,
Madison,WI.
Mattigod, S.Y., A.S. Gibali, and A.L. Page.1979. Effect of ionic strengthand ion pair formation on
the adsorptionof nickel by kaolinite. Clays Clay Miner. 27:411-416.
Mattigod, S.V. 1983. Chemicalcompositionof aqueousextractsof fly ash: Ionic speciationas a con-
trolling factor. Environ. Tech. Lett. 4:756-761.
Morel, F.M.M., J.e.Westall, e.R. O'Melia, andJ.J. Morgan. 1975.Fateof tracemetalsin Los Ange-
les county wastewaterdishcarge.Environ. Sci. Technol.9:756-761.
Morel, F.M.M. 1983. Principlesof aquaticchemistry.John Wiley & Sons,New York.
Morel, F.M.M., J.G. Yeasted,and J.e. Westall. 1981.Adsorptionmodels:A mathematical analysis in
the framework of generalequilibrium calculations.p. 263-294.In M.A. Anderson,and A.1.
Rubin (ed.) Adsorption of inorganicsat solid-liquid interfaces.Ann Arbor Sci., Ann Arbor,
MI.
Mouvet, e., and A.C.M. Bourg. 1983. Speciation(including adsorbedspecies)of copper,lead, and
zinc in MeuseRiver. Water Res. 17:641-M9.
Murray, L.C., Jr., and B.G. Lewis. 1985. Phosphogypsumwaste anion removal by soil minerals.J.
Environ. Eng. 111:681--698.
Nordstrom,O.K., andJ.W. Ball. 1984.Chemicalmodels,computerprogramsand metal complexation
in naturalwaters.p. 149-169.In C.J.M. KramerandJ.C. Duinker (ed.) Complexationof trace
metalsin natural waters. Martinus Nijhoff, The Hague,the Netherlands.
Nordstrom, O.K., and J.L. Munoz. 1985. Geochemical thermodynamics.Benjamin/Cummings,
Menlo Park, CA.
O'Connor,G.A., e. O'Connor,and G.R. Cline. 1984. Sorptionof cadmiumby calcareoussoils: Influ-
enceof solution composition.Soil Sci. Soc. Am. J. 48:1244-1247.
Osaki, S., T. Miyoshi, S. Sugihara,and Y. Takashima.1990.Adsorptionof Fe(III), Co(II) and Zn(II)
onto particulatesin fresh water on the basisof the surfacecomplexationmodel. I. Stabilities
of metal speciesadsorbedon particulates.Sci. Tot. Environ. 99:105-114.
Pankow, J.F., and J.1. Morgan. 1981. Kinetics for the aquatic environment.Environ. Sci. Technol.
15:1155-1213.
Parker,O.R., W.A. Norvell, and R.L. Cheney.1995. GEOCHEM-PC:A chemicalspeciationprogram
for IBM and compatiblepersonalcomputers.p. 253-270.In R.H.l.oeppertet al. (ed.) Chem-
ical equilibrium and reactionmodels.SSSASpec.Publ. 42. ASA and SSSA,Madison,WI.
Parker,J.e.,L.w. Zelazny,S. Sampath,andw.G. Harris. 1979.Critical evaluationof the extensionof
zero point of charge(ZPC) theory to soil systems.Soil Sci. Soc. Am. J. 43:668.
Pavan,M., adnF.T. Bingham.1982a.Toxicity of aluminumto coffeeseedlingsgrown in nutrientsolu-
tion. Soil Sci. Soc. Am. J. 46:993-997.
Pavan,M., and F.T. Bingham, and P.F. Pratt. 1982b.Toxicity of aluminum to coffee in ultisols and
oxisols amended with CaC03, MgC03, and CaS0402H20. Soil Sci. Soc. Am. J.
46:1201-1207.
Pavan,M., F.T. Bingham,and P.F. Pratt. 1984. Redistributionof exchangeablecalcium, magnesium,
and aluminumfollowing lime or gypsumapplicationsto a Brazilian oxisol. Soil Sci. Soc.Am.
J. 48:33-38.
Payne,T.E., andT.O. Waite. 1991.Surfacecomplexationmodelingof uraniumsorptiondataobtained
by isotopic exchangetechniques.Radiochim.Acta 52153:487-493.
Pearson,F.1., B.S. Jensen,andA. Haug. 1990.Expert systemsto supportgeochemicalmodeling.Am.
Chem.Soc.
Pitzer, K.S. 1973.Thermodynamicsof electrolytesI. Theoreticalbasisand general equations. J. Phys.
Chem.77:268-277.
Rai, D., J.M. Zachara,L. Eary, D.e. Girvin, D.A Moore, C.T. Resch,B.M. Saso,and R.L. Schmidt.
1986. Geochemicalbehaviorof chromiumspecies.EPRI EA-4544. Electric PowerRes. Inst.,
Palo Alto, CA
Reddy, K.R, W.H. Patrick, Jr., and RE. Phillips. 1978. The role of nitrate diffusion in determining
the order and rate of denitrification in flooded soil: I. Experimentalresults.Soil Sci. Soc. Am.
1. 42:268-272.
Ronngren,L., S. Syoberg,Z. Sun, W. Forsling, and P.w. Schindler. 1991. Surfacereactionsin aque-
ous metal sulfide systems2. Ion exchangeand acid/basereactionsat ZnS-H20 interface.1.
Colloid InterfaceSci. 145:396-404.
Schindler,P.w., P. Liechti, and J.e. Westfall. 1987. Adsorption of copper,cadmium,and lead from
aqueoussolution to the kaolinite/waterinterface.Neth. 1. Agric. 35:219-230.
Shaviv, A, and S.V. Mattigod. 1985. Cation exchangeequilibria in soils expressedas a cation-ligand
complexformation. Soil Sci. Soc. Am. J. 49:569-573.
Simard, RR, L.1. Evans,and T.E. Bates. 1988. The effects of additionsof Cac03 and P on the soil
solution chemistryof a podzolic soil. Can. J. Soil Sci. 68:41-52.
Skoog, D.A., and D.M. West. 1982. Fundamentalsof analytical chemistry,4th ed. w.B. Saunders,
Philadelphia,PA
Smith, G.R., K.K. Tanji, J.J.Jurinak,and R.G. Burau. 1995.C- SALT - A chemicalequilibrium model
for multicomponentsolutions.p. 289-324.In R.H. Loeppertet al. (ed.) SSSASpec.Pub!. 42.
ASA and SSSA,Madison,WI.
Sposito,G. 1977. The Gapon and Vanselow selectivity coefficients. Soil Sci. Soc. Am. J.
41:1205-1206.
Sposito, G., and S.V. Mattigod. 1979. Ideal behavior in Na-trace metal cation exchangeon Camp
Berteaumontmorillonite. Clays Clay Miner. 27:125-128.
Sposito,G., and S.Y. Mattigod. 1980. GEOCHEM: A computerprogramfor the calculationsof chem-
ical equilibria in soil solutionsand other natural water systems.Dep. Soil Environ. Sci. Univ.
California, Riverside,CA
Sposito,G. 1981a.Trace metalsin contaminatedwaters.Environ. Sci. Techno!. 15:396-403.
Sposito,G., 1981b.The thermodynamicsof soil solutions.Oxford Univ. Press,New York.
Sposito,G., F.T. Bingham,S.S. Yadav,and C. Inoye. 1982.Traceelementcomplexationby fulvic acid
extractedfrom sewagesludge: II. Developmentof chemical models. Soil Sci. Soc. Am. 1.
46:51-56.
Sposito,G., K.M. Holtzclaw, L. Charlet,e. 10uany,and AL. Page.1983. Sodium-calciumand sodi-
um-magnesiumexchangeon Wyoming bentonitein perchlorateand chloride backgroundionic
media. Soil Sci. Soc. Am. 1. 48:1244-1247.
Sposito,G. 1984. The surfacechemistryof soils. Oxford Univ. Press,New York.
Sposito,G. 1986a. Sorption of trace metals by humic materials in soils and natural waters. p.
193-229. In CRC Crit. Rev. Environ. Control. 16:193-229.
Sposito,G. 1986b.Thermodynamicsof the soil solution. p. 147-178.In D.L. Sparks(ed.) Soil phys-
ical chemistry.CRC Press,Boca Raton, FL.
Sposito,G., J.e.M. deWit, and R.H. Neal. 1988. Selinite adsorptionon alluvial soils. III. Chemical
modeling. Soil Sci. Soc. Am. J. 52:947-950.
Sposito,G. 1989.The chemistryof soils. Oxford Univ. Press,New York.
Sposito, G. 1990. Molecular models of ion adsorptionon mineral surfaces.p. 261-279. In M.F.
HochellaandAF. White (ed.) Mineral-waterinterfacegeochemistry.Rev. Mineral. 23. Miner.
Soc. Am.
Sposito,G., and 1. Coves.1990. SOILCHEM on the MACINTOSH. Chapter14 in Chemicalequilib-
rium and reactionmodels.p. 271-288.In R.H. Loeppertet al. (ed.) SSSASpec.Publ. 42. ASA
Sposito,G., and P. Blaser. 1992. Revisedquasiparticlemodel of protonationand metal complexation
reactions.Soil Sci. Soc. Am. J. 56:1095-1099.
Stumm,w., and J.J. Morgan. 1981. Aquatic chemistry.2nd ed. JohnWiley & Sons,Inc., New York.
Sullivan, PJ., AA Sobek, and 1. Rybarczyk. 1986a. Evaluating mineral dissolution in laboratory
weatheringexperimentsusing leachateequilibria. Soil Sci. Soc. Am. J. 50:251-254.
Sullivan, PJ., S.Y. Mattigod, and AA Sobek. 1986b. Dissolution of iron sulfatesfrom pyritic soil
waste.Env. Sci. Technol. 20:1013-1016.
Susetyo,W., 1.e. Dobbs,L.A Carreira,L. V. Azarraga,and D.M. Grimm. 1990.Developmentof a sta-
tistical model for metal-humicinteractions.Anal. Chem.62:1215-1221.
Talibudeen,0.1981.Cation exchangein soils. p. 115-177.In D.J. Greenlandand M.H.B. Hayes(ed.)
The chemistryof soil processes.Wiley, Chichester,England.
1358 MATI'lGOD & ZACHARA
TIpping, E., M.M. Reddy,and M.A. Hurley. 1990. Modeling electrostaticand heterogenityeffectson
proton dissociationfrom humic substances. Environ. Sci. Techno!.24:1700-1705.
Wagman,D.O., W.H. Evans,Y.B. Parker,R.H. Schumm,I. Halow, S.M. Bailey, K.L. Chumey,and
R.L. Nuttall. 1982. The NBS tablesof chemicalthermodynamicproperties:Selectedvalues
for inorganic and C1 and Cz organic substancesin SI units. I. Phys.Chem. Ref. Data. 11,
Supp!. 2.
Waite, T.D. 1989. Mathematicalmodeling of trace elementspeciationin trace elementspeciation:
Analytical methodsand problems.p. 117-184.In G.E. Batley (ed.) CRC Press,Boca Raton,
FL.
Westall,I.C., I.L. Zachary,and F.M.M. More!. 1976. MINEQL: A computerprogramfor the calcu-
lation of chemicalequilibrium compositionof aqueoussystems.Tech. Note 19. RalphM. Par-
sonsLab., Dep. Civil Eng., Massachusetts Inst. Techno!.,Cambridge,MA.
Westall,I.C., andF.M.M. More!. 1977.FITEQL: A generalalgorithm for the determinationof metal-
ligand complexstability constantsfrom experimentaldata.Tech. Note 19. Ralph M. Parson
Lab., Dep. Civil Eng., MassachusettsInst. Techno!.,Cambridge,MA.
Westall,I.C.1980.ChemicaleqUilibrium including adsorptionon chargedsurfaces.p. 33-44.In M.C.
Kavanaugh,and1.0. Leckie (ed.)Particulatesin water.ACS Adv. Chem.Ser. 189.Am. Chem.
Soc.,Washington,DC.
Westall, I.C., and H. Hohl. 1980.A comparisonof electrostaticmodelsfor the oxide/solutioninter-
face.Adv. Colloid InterfaceSci. 12:265-294.
Westall,I.C., 1982a.FITEQL. A computerprogramfor determinationof equilibrium constantsfrom
experimentaldata.Verso 2.0. Rep. 82-01. Dep. Chem.,OregonStateUniv., Corvallis, OR.
Westall,I.C. 1982b.FITEQL. A computerprogramfor determinationof eqUilibrium constantsfrom
experimentaldata.Verso 2.0. Rep. 82-02. Dep. Chem.,OregonStateUniv., Corvallis, OR.
Westall,I.C., I.L. Zachary,andF.M. More!. 1976.MINEQL: A computerprogramfor the calculation
of chemicaleqUilibrium compositionof aqueoussystems.Tech. Note 18. Dept. Civil Eng.,
Massachusetts Inst. Techno!.,Cambridge,MA.
White, G.N., and L. Zelazny. 1988. Analysis and implicationsof the edgestructureof dioctahedral
phyllosilicates.ClaysClay Miner. 36:141-146.
Wilkins, R.G., andM. Eigen. 1965.The kinetics and mechanismof formation of metalcomplexes.p.
55-66. In R.F. Gould (ed.) Mechanismsof inorganic reactions.Adv. Chem.Ser. 49. Am.
Chem.Soc.,Washington,DC.
Zachara,I.M., D.C. Girvin, R.L. Schmidt,and C.T. Resch.1987.Chromateadsorptionon amorphous
iron oxide in presenceof major groundwaterions. Environ. Sci. Techno!.21:589-594.
Zachara,I.M., C.E. Cowan,R.L. Schmidt,and C.c. Ainsworth. 1988.Chromatesorptionon Na-sat-
uratedkaolinite. Clays Clay Miner. 36:317-326.
Zacham,I.M., C.C. Ainsworth, C.E. Cowan,and C.T. Resch.1989.Adsorptionof chromateby sub-
surfacesoil horizons.Soil Sci. Soc.Am. 1. 53:418-428.
Zachara,I.M., C.E. Cowan,and C.T. Resch.1991. Sorptionof divalent metalson calcite. Geochim.
Zachara,I.M., D. Rai, A.R. Felmy, C.E. Cowan, S.C. Smith, D.A. Moore, and C.T. Resch. 1992.
Aqueouscomplexation,precipitation,and adsorptionreactionsof cadmium in the geologic
environment.EPRI TR-l00751.Electric PowerRes.Inst., PaloAlto, CA.
Zachara,I.M., andS.C.Smith. 1994.Edgecomplexationreactionsof cadmiumon specimenandsoil
derivedsmectites.Soil Sci. Soc.Am. 1. 58:762-769.
Published 1996
SUBJECT INDEX
Absorption coefficient, 379 properties, 551-552
Absorption edge, 380---381, 389, 391, 393 Alkali metal determination, 555-557
Accuracy, 34-35, 40-42, 45 atomic absorption spectrophotometry, 556
Acetate-extractable sulfur, 953 flame photometry, 556
Acetic acid dissolution method, carbonate inductively coupled plasma-mass spec-
determination, 456-458 troscopy, 556--557
Acetic acid-dithizone method, Co determina- ion chromatography, 556
tion, 734 neutron activation analysis, 556--557
Acetylacetone extraction, organic P, 878-881 total alkali elements, 557-559
Acid digestion, 4~3 hydrofluoric acid digestion, 558--559
advantages and disadvantages, 50 x-ray fluorescence, 556--557
methods, 56--58 Alkaline oxidation method, oxidation of total S
oxidation of total S to sulfate, 931 to sulfate, 931
principles, 51-52 Alkaline-earth metal(s), 575-598
properties of mineral acids, 50 availability indices, 585-586
safety, 54-55 mineralogy and distribution in rocks and
sample preparation for Si determination, soils, 576--577
632-634 partitioning in soils, 581-586
Acid dissociation method, carbonate determina- properties, 575-576
tion, 439 soluble, 581
Acid soil extraction method, Alkaline-earth metal determination
extractable/exchangeable alkaline-earth atomic absorption spectrophotometry, 578,
metals, 584 587-589
Acidic functional groups, humic substances, colorimetric method, 597-598
1036--1040 electrothermal atomic absorption, 590
carboxyl groups, 1039--1040 extractable/exchangeable fraction, 582-584
phenolic groups, 1040 acid soil extraction, 584
total acidity, 1036--1038 ammonium acetate method, 583-584
Acid-insoluble nitrogen determination, ammonium chloride method, 584
1185-1186, 1191 flame emission photometry, 590
Activated charcoal, 709 inductively coupled plasma-atomic emission
Active acidity, 493-494 spectrometry, 579-580, 589--590
Active carbonate method, determination of car- ion chromatography, 591-593
bonate reactivity, 465-468 single column method, 591
Activity coefficient, 1322 suppressed gradient method, 591-593
Adams-Evans single buffer method, determina- surface-sulfonated cation resin exchange
tion of lime requirement, 498, 505-508 column, 591
Adsorption analysis, 256 permanganate method, 596--597
Adsorption constant, 1318 titrimetric method, 593-596
Adsorption method, fractionation of humic sub- total analysis, 577-580
stances, 1027-1028 x-ray fluorescence, 578--579
Adsorption models, 1312-1321 Allophane, 350---351
anion adsorption, 1343-1346 Alternating current polarography, 255-256
calculations with FITEQL, 1340---1341 Alumina, 347
cation adsorption, 1341-1343 Aluminon method, Be determination, 597-598
cation exchange models, 1313-1315 Aluminosilicate, 517
limitations, 1350 Aluminum, 517-543
surface complexation models, 1315-1321 amorphous, 530---531
Adsorption stripping voltammetry, 256 exchangeable,492,522-531
Advanced photon source, 386 determination of lime requirement,
Aerated soils, 1255 511-513
Agricultural chemicals, 1071 extractable, 522-531
Alcoholic groups, humic substances, fluoro-, oxalato-, and citrato-Al complexes,
1041-1042 524
Alkali carbonate, fusion with, 52 hydroxy-Al compounds, 478, 524
Alkali metal(s), 551-572 inorganic noncrystalline, 531
mineralogy in soils, 552 interlayer, 524
1359
1360 SUBJECT INDEX
Aluminum (cont.) Amino sugar-nitrogen determination, 1186

labile, 525 colorimetric method, 1196--1200
polynuclear species, 524 steam-distillation method, 1188, 1191-1192
in soil acidity, 518, 522 (Ammonia + amino sugar)-nitrogen determina-
soil pH and, 478 tion, 1188, 1191
speciation in solution, 534--538 Ammonia electrode, modification of Kjeldahl
physicochemical separation methods, method, 1108-1110
536--538 Ammonia-nitrogen determination, 1185-1186,
timed spectrophotometric assays, 534-536 1188, 1191
Aluminum determination Ammonium, 1123
atomic absorption spectrometry, 540--541 exchangeable, 1123-1124
atomic emission spectrometry, 540--541 fixed, 1186
exchangeable Al, 526 in Kjeldahl digest, 1100--1102
extraction from mineral soils, 525-527 nonexchangeable, 1123
dilute salt extractable Al, 526--527 (Ammonium +nitrate+ nitrite)-nitrogen deter-
potassium chloride method, 526 mination
extraction from organic matter and organic microdiffusion method, 1143
soils, 527-530 steam-distillation method, 1136
copper chloride method, 529 (Ammonium+ nitrate)-nitrogen determination
sodium pyrophosphate method, 529-530 microdiffusion method, 1143
extraction from poorly ordered phases, steam-distillation method, 1135-1136
530--531 Ammonium acetate method
ammonium oxalate method, 530--531 cation exchange capacity determination,
potassium hydroxide method, 531 1220--1221
sodium citrate-dithionite method, 530--531 exchangeable K determination, 560, 568-569
flame atomic absorption spectrometry, Biichner funnel method, 560
540--541 centrifuge method, 560
flameless atomic absorption spectrometry, extractable/exchangeable alkaline-earth met-
541 als, 583-584
inductively coupled plasma- atomic emission Mn determination, 675-676
spectrometry, 541-542 Ammonium acetate-hydroquinone method, Mn
inductively coupled plasma-mass spectrome- determination, 677-678
try, 541-542 Ammonium bicarbonate-DTPA method. See
in soil solution, 531-534 DTPA-AB method
extraction of samples in laboratory, Ammonium chloride method,
532-533 extractable/exchangeable alkaline-earth
filtration of soil solution samples, metals, 584
533-534 Ammonium determination
lysimeters, 532 ammonium electrode, 1162-1163,
spectrophotometric methods, 538-540 1168-1169
8-hydroxyquinoline-butyl acetate method, Berthelot reaction, 1146--1148
538-540 colorimetric methods, 1146--1148,
pyrocatechol violet method, 540 1152-1155
tiron method, 540 extraction of exchangeable ammonia,
total Al content, 518-522 1129-1131
alternative procedures, 522 Nessler method, 1146
hydrochloric acid-nitric acid-hydrofluoric nonexchangeable ammonium, 1171-1176
acid method, 519-521 salicylate method, 1152-1155
lithium metaborate fusion, 519-520 in soil extracts, 1131-1171
titrimetric methods, 542-543 Ammonium oxalate method
x-ray fluorescence spectrometry, 542 Al extraction from poorly ordered phases,
Aluminum oxide, 351 530--531
Amalgamation, 774 "amorphous" iron oxide determination,
Amide-nitrogen, 1186 649-650
Amino acid, 1185 Mo determination, 728-729
Amino acid-nitrogen determination, 1186 Ammonium pyrrolidine dithiocarbamate-chloro-
colorimetric method, 1193-1196 form method
steam-distillation method, 1187-1188, Cd determination, 761-763
1191-1192 Ni determination, 761-763
Amino sugar, 1185 Pb determination, 761-763
SUBJECT INDEX 1361
Ammonium-nitrogen determination, 1126 hydride-mercury vapor generator, 131-132

microdiffusion method, 1142-1143 soil tests, DTPA-AB method, 825-826
steam-distillation method, 1135 standard reference materials, 794
Analysis of variance, 13-15 total As content
Analytical methods digestion of soils, 813
approval system, 25 in soil solutions and water extracts,
detection limits, 46-4 7 818-819
methods manual, 24 in soils, 812--818
modification, 24-25 Arsenite determination, 820-824
quality assurance, 19-20 Ascorbic acid method, P determination,
range of analysis, 46 908-909
standard, 24 modified, 909-910
standard operating procedures, 22-25 Assessment phase, quality assurance program,
training of personnel, 27-28 20
validation, 24 ASV. See Anodic stripping voltammetry
Angle resolved x-ray photoelectron spec- Atomic absorption spectroscopy
troscopy, 371-372 Al determination, 540-541
Anhydrite, 470 alkali metal determination, 556
Anion(s), total anion concentration, 426 alkaline-earth metal determination, 578,
Anion adsorption models, 1343-1346 587-589
surface complexation models, 1319 atomic emission spectroscopy versus, 66---68
Anion exchange, 1231-1251 Cd determination, 759-761
Anion exchange capacity, 1241, 1243 chemical interferences, 68, 77
Anion exchange resin Co determination, 733
Mo determination, 729-730 costs, 68
P buffering capacity determination, 904-905 Cr determination, 689-691
P determination, 898-899 detection limits, 74-76
Anodic stripping voltammetry (ASV), 247 Fe determination, 658-659
applications, 255 instrumentation, 69-74
chemical analysis, 263-264 atomization systems, 70-72
complexation and speciation, 260-262 light sources, 70
oxidation-reduction reactions, 262-263 optical system, 72-74
soil solutions and natural waters, 260-264 ionization interferences, 68, 77-78
instrumentation, 257 Mn determination, 667, 670, 679
interferences and other problems, 265 Mo determination, 728
principles, 253-255 Ni determination, 759-761
sample preparation, 259-260 nonspectral interferences, 76--78
Anthracene, 338 Pb determination, 759-761
9,10-Anthraquinone, 338 principles, 64-67
Aqua regia, digestion with sample introduction
method, 56--57 flow injection, 88
principles, 51 hydride generation, 86-87
Aqueous extract mercury cold vapor method, 87
preparation, 419 solid samples, 87-88
soil/water ratios of one:one and one:five, 420 sample preparation, 84--86
total dissolved solids, 431-433 soil digests, 84--85
Aqueous solution, electrical conductivity, soil solutions and extracts, 85-86
422-427 sensitivity, 74-76
Archiving, 25-27 Si determination, 6314>33
Arsenate determination, 820-824 spectral interferences, 68, 78-83
Arsenic, 793, 811--826 Atomic concentration, x-ray photoelectron
availability indices, 824--826 spectroscopy, 369-371
chemistry, 811--812 Atomic emission spectroscopy. See also Induc-
Arsenic determination tively coupled plasma-atomic emission
arsenite and arsenate in soil solutions and spectroscopy
water extracts, 820-824 AI determination, 540-541
colorimetric method, silver diethyldithiocar- atomic absorption spectroscopy versus,
bamate method, 814--816 664>8
hydride generation atomic absorption spec- chemical interferences, 68
trometry, 815, 817-818 ionization interferences, 68
1362 SUBJECT INDEX
Atomic emission spectroscopy (cont.) extractable/exchangeable fraction, 582-584

principles, 66-67 acid soil extraction, 584
spectral interferences, 68 ammonium acetate method, 583-584
Atomic fluorescence spectroscopy, 66-67 ammonium chloride method, 584
Attenuated total reflectance FnR spectroscopy, flame emission photometry, 590
270,287,302-308 inductively coupled plasma-atomic emission
Attenuation length, 359 spectrometry, 579--580, 589--590
Audit, 31-32 ion chromatography, 591-593
performance, 32 titrimetric method, 593-5%
system, 32 x-ray fluorescence, 578--579
Audit trail, 26 Bassanite, 470
Auger electron, 359, 361 Batch techniques, 1276-1280
Auger electron emission, 384-385 advantages, 1280
Auger electron spectroscopy, 357 analysis of rate data, 1290--1295
Auger lines, 365 disadvantages, 1279--1280
Auger parameter, 361-362 mixing effects, 1279
Authority, quality assurance program and, 20 for point of zero salt effect, 1245-1248
Automatic titration method, Cl determination, stirred-batch reactors, 1277-1279
840,844--845 tube techniques, 1276-1277
Availability indices Beam line hardware, 385-387
for alkaline-earth metals, 585-586 Beamsplitter, Fourier transform infrared spec-
for As, 824--826 troscopy, 277-280
for B, 609-613 Beer's law, 283-284
for Cd, 752-758 Beidellite, 289
for Co, 734 Benchsheet, 25-26
for Cu, 706-717 Berthelot reaction
for Fe, 653-658 ammonium determination, 1146-1148
for Mn, 672--681 inorganic N determination, 1125
for Mo, 728--731 Beryllium, 575-598. See also Alkaline-earth
for Ni, 752-758 metal(s)
for P, 890--903 availability indices, 585-586
for Pb, 752-758 mineralogy and distribution in rocks and
for S, 949--955 soils, 576-577
for Se, 808--811 properties, 575-576
for Zn, 706-717 soluble, 581
Axial symmetry, 327 Beryllium determination
Azomethine-hydrogen method, B determina- aluminon method, 597-598
tion, 617--618 atomic absorption spectrophotometry, 578,
587-589
Babington style nebulizer, 98--101 electrothermal atomic absorption, 590
Background correction, atomic absorption spec- extractable/exchangeable fraction, 582-584
troscopy, 78, 80-83 acid soil extraction, 584
Backscatter, x-ray absorption fine structure ammonium acetate method, 583-584
spectroscopy, 403,410 ammonium chloride method, 584
Backtitration method, for point of zero salt flame emission photometry, 590
effect, 1248 inductively coupled plasma-atomic emission
Backtransformed function, 405 spectrometry, 579--580, 589--590
Band fitting, Fourier transform infrared analy- ion chromatography, 591-593
sis, 285 titrimetric method, 593-596
Barium, 575-598. See also Alkaline-earth x-ray fluorescence, 578--579
metal(s) Bias, 12, 35, 41-42, 44
availability indices, 585-586 Bicarbonate-extractable sulfur, 953-954
mineralogy and distribution in rocks and Bidentate bond, 348
soils, 576-577 Binding energy, electron, 379
properties, 575-576 Biota, soil, 1011
soluble, 581 Blank sample, 41
Barium determination Blind duplicates, 43-45
atomic absorption spectrophotometry, 578, "Blind" sample, 32
587-589 Blue silicomolybdous acid method, Si determi-
electrothermal atomic absorption, 590 nation, 635--636
SUBJECT INDEX 1363
Boehmite, 347, 350 inductively coupled plasma-atomic emission

Boiling nitric acid extraction method, nonex- spectrophotometry, 761
changeable K determination, 562 sampling and sample preparation, 739-740
Boltzmann's law, 324 selective extraction, 745-752
Borate, 604-605, 607 carbonate phase, 747, 750
Borate ion pairs constants, 605 exchangeable phase, 747, 749
Boric acid, 604-<i05, 607 iron oxide phase, 748, 750
Boron, 603-624 manganese oxide phase, 750
adsorption-desorption by soil constituents, organic matter phase, 747, 750-751
606-607 residual phase, 748, 751
availability assessment, 609-613 soil tests, 752-758
chemistry in aqueous media, 6Q4-{iQ5 DTPA method, 754-755
Boron determination DTPA-AB method, 756
colorimetric methods extractants, 753
azomethine-hydrogen method, 617----018 Mehlich I extraction, 757-758
carmine method, 613----014 Mehlich III extraction, 758
curcumin method, 614-<i 15 one-tenth molar hydrochloric acid method,
2-ethyl-1,3-hexanediol in chloroform 756-757
extraction, 615----016 standard reference materials, 742
extraction methods, 607----013 total Cd content
hot-water extract, 610----011 hydrogen peroxide, hydrofluoric acid,
mannitol-calcium chloride method, nitric acid, and perchloric acid method,
612-613 744-745
saturation extract of soil paste, 611-612 nitric acid-hydrochloric acid digestion
inductively coupled plasma spectrometry, method, 742-743
623-624 nitric acid-perchloric acid digestion
potentiometric methods method, 743-744
mannitol method, 618-620 principles, 740-742
tetrafluoroborate selective electrode Calcareous soil
method, 620----023 inorganic P fractionation, 883-884
total B content, 607-609 organic C determination, 985-988
Bouguer-Beer-Lambert law, 283-284 Calcite, 437
Bromine chemistry, 833-834 Calcite determination, 440
Bromine determination Calcium, 575-598. See also Alkaline-earth
direct potentiometric method, 834---836 metal(s)
ion chromatography, 839 availability indices, 585-586
microdiffusion method, 837--839 mineralogy and distribution in rocks and
neutron activation analysis, 863 soils, 576-577
x-ray fluorescence, 863 properties, 575-576
Brucine method, nitrate determination, 1148 soluble, 581
Buffer capacity, soil, 494, 496 Calcium carbonate, 437, 478-479
Buffer solution, 501 Calcium chloride extraction method
Buffered alkaline solution extraction method, P F determination, 857
determination, 895--897 P determination, 891--893
Buffering capacity, P, 903-905 Calcium determination
Bulk density, mean, 14-15 atomic absorption spectrophotometry, 578,
587-589
Cadmium, 739-763 EDTA method, 593-594
availability indices, 752-758 electrothermal atomic absorption, 590
in soil solution, 752 extractable/exchangeable fraction, 582-584
Cadmium determination acid soil extraction, 584
ammonium pyrrolidine dithiocarbamate- ammonium acetate method, 583-584
chloroform method, 761-763 ammonium chloride method, 584
atomic absorption spectrophotometry, flame emission photometry, 590
759-761 inductively coupled plasma-atomic emission
flame atomic absorption spectrophotometry, spectrometry, 579-580, 589-590
759-760 ion chromatography, 591-593
graphite furnace atomic absorption spec- permanganate method, 596-597
trophotometry, 760 titrimetric method, 593-596
1364 SUBJECT INDEX
Calcium determination (cont.) Catechol, 347

x-ray fluorescence, 578-579 Cation(s), total cation concentration, 426
Calcium oxide incineration method, F determi- Cation adsorption models, 1341-1343
nation, 853 surface complexation models, 1320
Calibration, 45-48 Cation exchange capacity (CEC), 1234, 1241,
Calibration curve, 30, 40, 42, 46 1243
linear, 47-48 ammonium acetate method, 1220--1221
quadratic, 47-48 compulsive exchange method, 1215-1218
Calibration standards, 46, 48 criteria for selection of method
Capacity factor, 227 characterization of soils in "field" state,
soil acidity, 491 1205
Carbo-Erba NA 1500, 974-975 soil classification, 1205-1206
Carbon chemistry, 961-963 at field pH, 1202
Carbon determination. See also Organic matter, historical review, 1202-1205
soil operational definition, 1201-1202
inorganic C, 986 origin, 1201
organic C, 961, 983-1001. See also Organic problems in measurement
carbon determination effect of cation and anion type,
total C content, 961, 963-983 1211-1212
dry combustion method, 963-977 ionic strength effects, 1212-1213
high-temperature induction furnace pH effects and use of buffered solutions,
method, 971-973 1212
instrumental methods, 973-977 removal of entrained electrolyte,
medium-temperature resistance furnace 1212-1213
method, 966-971 soluble salts and carbonates, 1210--1211
wet combustion method, 963-964, soils containing salts, carbonates or zeolites,
977-983 1213-1214
Carbon dioxide, soil, effect on soil pH, 482 unbuffered salt extraction method,
Carbonate, 437-470, 962 1218-1220
chemistry, 437-438 Cation exchange column, surface-sulfonated,
effect on cation exchange capacity measure- alkaline-earth metal determination, 591
ment, 1210--1214 Cation exchange models, 1313-1315
soil pH and, 478-479 Cation exchange reactions, formulation,
Carbonate determination 1206-1209
quantitation of calcite and dolomite, 440 Cation exchange selectivity coefficients,
total carbonate content, 438-458 1206-1210
acetic acid dissolution method, 456-458 CCV sample. See Continuing calibration verifi-
acid dissociation method, 439 cation sample
gravimetric method, 440, 444-448 CD-ROM,27
gravimetric method for loss of carbon CEC. See Cation exchange capacity
dioxide, 455-456 Cerium(IV) method, oxidation of inorganic
manometric method, 439 Cr(III), 694-695
pressure calcimeter method, 440-444 Certification, 28
titrimetric procedure, 448-451 Cesium, 551-572. See also Alkali metal(s)
volumetric calcimeter method, 451-455 chemistry, 551-552
Carbonate equilibria, 1336 properties, 555
extraction of soil water for, 470 Charge analysis, 1231-1251
reaction rates, 469-470 Charge determining ions, 1235-1236
solid phase, 468-469 Check sample, quality control, 28, 34, 36, 38,
Carbonate reactivity, 458-459 42
active carbonate method, 465-468 Chemical equilibrium models, 1309--1353. See
in aggregated soils, 462-463 also Equilibrium models
manometric method, 463-464 Chemical kinetics, 1275
pH stat method, 459-463 Chemical relaxation methods, 1296
Carbon-bonded sulfur determination, 946-949 Chemically modified electrode voltammetry,
Carbonyl groups, humic substances 256
quinone groups, 1042-1043 Chemiluminescence, Cr determination, 691
total carbonyl groups, 1042 Chemisorption, metal ions on mineral surfaces,
Carboxyl groups, humic substances, 1039--1040 349--350
Carmine method, B determination, 613-614 Chlorine chemistry, 833, 839
SUWECT INDEX 1365
Chlorine determination oxidation of inorganic Cr(III) with Ce(IV),

automatic titration method, 840, 844-845 694-695
colorimetric methods, 839-840 oxidation of organic ligands binding Cr(III)
direct potentiometric method, 840, 843-844 nitric acid/hydrogen peroxide method, 694
Hg(II) thiocyanate method, 840, 845-846 nitric acid/vacuum oven method, 694
ion chromatography, 846 precipitation method, 691
neutron activation analysis, 863 soil handling, 684-685
potentiometric titration method, 840-843 in soils and plants, 686
stripping voltammetry, 863 total Cr content, digestion methods, 686
Chlorophenoxyalkanoic herbicide, 341 total Cr(VI) content, 692-693
Chlorostannous acid method, P determination, voltammetry, 691
907-908 Chromium net oxidation test, 695-696
CHROMAT, 1353 Chromotropic acid method, nitrate determina-
Chromate, 683 tion, 1148-1149
Chromatographic bed, 226 CIRCLE cell, 306-309
Chromatography, 225. See also specific kinds Citrate-bicarbonate-dithionite method, total
of chromatography "free" iron oxide determination,
definition, 225-226 647-648
Chromite, 684 Citrate-dithionite extraction method, total "free"
Chromium, 683-697 iron oxide determination, 646-647
chemisorption on mineral surfaces, 351 Citrato-aluminum complexes, 524
chemistry, 683-684 Clay suspension, 307
as redox tool, 695-697 Clay-water interactions, 296
available reducing capacity for Cr(VI), Cobalt, 723-734
696 availability indices, 734
Cr net oxidation test, 695-696 chemistry, 723-725
soil reducing intensity for Cr(VI), Cobalt determination
696-697 acetic acid-dithizone method, 734
total Cr(VI) reducing capacity, 696 atomic absorption spectroscopy, 733
Chromium(III), 683 general principles, 725
labile, 693 total Co content, dithizone in carbon tetra-
oxidation of inorganic Cr(III) with Ce(IV), chloride method, 731-733
694-695 Cold vapor-atomic absorption spectrometry, Hg
oxidation of organic ligands binding determination, 771-784
nitric acid/hydrogen peroxide method, 694 Color, soil, 1261
nitric acid/vacuum oven method, 694 Colorimetric method
oxidizability of inert soil Cr, 696-697 alkaline-earth metal determination, 596-598
Chromium(VI), 683 amino acid-N determination, 1193-1196
available reducing capacity, 6% amino sugar-N determination, 1196-1200
soil reducing intensity for, 696-697 ammonium determination, 1146-1148,
soluble, 686-688 1152-1155
in solution, 693-694 arsenite and arsenate determination, 821-822
total reducing capacity, 696 As determination, silver diethyldithiocarba-
Chromium determination mate method, 814-816
atomic absorption spectrophotometry, B determination
689-691 azomethine-hydrogen method, 617-618
chemiluminescence, 691 carmine method, 613-614
colorimetric method, 691 curcumin method, 614-615
Cr oxidizable by hypochlorite, 692 2-ethyl-1,3-hexanediol in chloroform
Cr(IV) in solution, 693-694 extraction, 615-616
Cr(IV) standards, 689-692 Cl determination, 839-840
digestion of soil materials for, 692 Cr determination, 691
s-diphenylcarbazide method, 686-688, 690 Fe determination, 1,10-phenanthroline
oxidative preparation of samples, 688-689 method, 659-661
exchangeable Cr, 692-693 inorganic N determination, 1125-1126,
extraction of labile Cr(III), 693 1146-1161
gas chromatography, 691 automation, 1150-lJ.52
inductively coupled plasma spectrophotome- Mn determination, 680-681
try, 689-691 periodate method, 671-672
liquid chromatography, 690 nitrate determination, 1148-1149, 1155-1158
1366 SUBJECT INDEX
Colorimetric method (cont.) labile, 704

nitrite determination, 1149-1150, 1158-1161 nonlabile, 704
P determination nonspecific adsorption, 703
ascorbic acid method, 908-909 occlusion, 704
chlorostannous acid method, 907-908 soil properties vs. deficiencies, 704-705
modified ascorbic acid method, 909--910 specific adsorption, 703-704
vanadomolybdophosphoric acid method, uncontaminated soils, 704
907 Copper chloride method, Al extraction from
pH determination, 483---484 organic matter/organic soils, 529
Column, 226 Copper determination
high-performance liquid chromatography, soil tests, 706-707
230-231 DTPA-AB method, 706, 709--711
ion chromatography, 240-241 DTPA-TEA method, 706-709
Column efficiency, 227-228 field calibration, 705-706
Complexation greenhouse evaluation, 705-706
applications of anodic stripping voltammetry, Mehlich I extraction, 711-713
260-262 Mehlich III extraction, 706, 713-715
applications of differential pulse polarogra- sample and extractant preparation, 707
phy, 260-262 total Cu content, hydrofluoric, nitric, per-
Composite samples, 16-17, 19 chloric, and sulfuric acid method,
Compton effect, 150 717-719
Compulsive exchange method, cation exchange Core electronic state, 365
capacity determination, 1215-1218 Corrections in data, 31
Computer system Corrective action, 32, 38, 42, 45
backup procedures, 27 Cost
record keeping, 26 atomic absorption spectroscopy, 68
Concentration-jump relaxation methods, 1297 ICP-AES, 94
Conductance meter, 423, 428 ICP-MS, 94
Conductivity cell/electrodes, 423---425 sampling, 14-15
large cup-type, 427---430 Cottrell current, 251
Confidence limits, 6, 9 Counterion adsorption, 1237-1239
Constant capacitance model, 1315 inner-sphere vs. outer-sphere, 1237-1238
Constant charge, 1201, 1234 solid particle vs. embathing solution,
Constant charge density, measurement, 1238-1239
1250-1251 specific vs. indifferent, 1237
Contamination, 42---43, 55, 141 Stem model, 1238
Continuing calibration verification (CCV) sam- Counting efficiency, XRFS, 173, 188-190
ple, 34, 36 CPMAS-NMR, humic substances, 1049
Continuous flow methods, 1280-1286 Crystal field model, 334
disadvantages, 1280-1282 Crystal monochromator, 386-387, 391,398
equipment, 1280 C-SALT, 1326, 1353
Continuous liquid-liquid extractor, extraction of Curcumin method, B determination, 614-615
organic chemicals, 1075 Cylindrical internal reflectance FTIR spec-
Continuum background correction, 80-81, 83 troscopy, 270,287, 308-309
Control chart, 35-37
Control limits, 29-30, 36-37, 42 Data analysis, x-ray absorption fine structure
Conway microdiffusion method, Br determina- spectroscopy, 411---412
tion, 837-839 Data collection, Fourier transform infrared
Copper, 703-719 spectroscopy, 280-281
availability indices, 706-717 Data management, 30-31. See also Record
available forms in soil, 704 keeping
chemisorption on mineral surfaces, 350-351 calculations, 31
chemistry, 703-704 corrections, 31
deficiency, 703-704 primary data, 30-31
divalent secondary data, 30
complex with organic substances, Data recording, 25-26
345-346 Davies equation, 1322
electron spin resonance spectroscopy, Debye-Huckel equation, 1314
345-346 Debye-Waller factor, 403
exchange on layer silicates, 349 Defensibility, 19, 21, 24-26, 30-31
SUBJECT INDEX 1367
~H-~OH, 1247-1248, 1250 Dissolution methods, 49-63. See also specific

Desorbed species, 1279 methods
Desorption isotherm, 292 errors, 51
Detanning, 697 safety, 54-55
Detection limits sample preparation, 55-56
atomic absorption spectroscopy, 74-76 Dithizone in carbon tetrachloride method, Co
ICP-AES, 114-116 determination, 731-733
ICP-MS, 116 DME. See Dropping mercury electrode
method detection limit, 46--4 7 Documentation, 31
XRFS, 188-190 Dolomite, 437-438
Deuterated triglycine sulfate detector, 277, 279 Dolomite determination, 440
Deuteration technique, sample for Fourier trans- "Double blind" sample, 32, 44
form infrared spectroscopy, 297 Double layer thickness, 1232
Deviation coefficient, 1347 OPP. See Differential pulse polarography
1,2-Dibromoethane (EDB), 302-303 Drainage, 417
Dichromate method, N determination, 1117 DRIFf
Dichromate oxidation method, organic C deter- accessories, 298-300
mination, 988-999 humic substances, 1047-1048
principles, 988-995 "Drop knocker," 257
tube digestion method, 997-998, 1000 Dropping mercury electrode (DME), 247-248,
Walkley-Black method, 993, 995-996, 1000 250,257
Differential pulse polarography (DPP) Dry ashing, oxidation of total S to sulfate,
applications, 252-253 931-932
chemical analysis, 263-264 Dry combustion method
complexation and speciation, 260-262 C determination, 963-977
oxidation-reduction reactions, 262-263 high-temperature induction furnace
to soil solutions and natural waters, method, 971-973
260--264 medium-temperature resistance furnace
instrumentation, 257 method, 96&-971
interferences and other problems, 265 organic C determination, 985-988, 1000
principles, 252 pretreatment, 987-988
sample preparation, 259-260 DTPA method
Differential pulse voltammetry, 247-266 Cd determination, 754-755
Diffuse double layer, 1231-1233, 1315 Fe determination, 654-657
Diffuse layer charge density, 1232 correlation with Fe chlorosis, 656
Diffuse reflectance FTIR spectroscopy, 270, sample preparation, 655--{i56
287,297-302 soil factors influencing, 655
Diffusion, 1295 Mn determination, 677
Diffusion current, 249 Ni determination, 754-755
Dilute acid fluoride extraction method, P deter- Pb determination, 754-755
mination, 894-895 DTPA-AB method
Dilute hydrochloric acid method, Zn determina- As determination, 825-826
tion, 715-717 Cd determination, 756
Dilute salt extractable aluminum, 52&-527 Cu determination, 706, 709-711
Dilution effect, on soil pH, 4 79-480 Fe determination, 657-658
Dilution error, 42 K determination, 570--571
5,5-Dimethylpyrroline 1-oxide, 348 Mo determination, 730--731
s-Diphenylcarbazide method, Cr(VI) determina- Ni determination, 756
tion, 68&-688, 690 P determination, 897-898
interferences, 688 Pb determination, 756
oxidative preparation of samples, 688-689 Zn determination, 706, 709-711
Dipole moment, 274 DTPA-TEA method
Direct current polarography, 248-249, 257
Cu determination, 70&-709
Direct injection nebulizer, 102
Zn determination, 70&-709
Direct potentiometric method
Dumas method, N determination, 1085,
Br determination, 834-836
1087-1089
Cl determination, 840, 843-844
Duplicate, 34
F determination, 857-859
blind,434-45
Direct reader, 103
1368 SUBJECT INDEX
ECEC. See Effective cation exchange capacity spin content of humic substances,
EDB. See 1,2-Dibromoethane 339-340
EDTAmethod structural implications of electron spin
Ca determination, 593-594 resonance, 340
extraction of humic substances, 1017 hyperfine splitting, 325, 328, 336, 338
Mg determination, 593-594 inhomogeneous broadening, 333
EELS. See Electron energy loss spectroscopy instrumentation, 329-331
Effective cation exchange capacity (ECEC), double cavity instrument, 337
523-524, 1202,1221-1222 resonant ("microwave") cavity, 329
Eh-pH diagram, 1258-1259 spectrometer, 329-330
Electric double layer, 1231 vacuum tube oscillator, 329
Electrical conductivity, 417-433 line broadening, 338
aqueous solution, 422-427 nuclear quadrupole interactions, 325, 329
apparatus, 423-424 organic paramagnetic species, 335-337
calculations, 425 paramagnetic metal ions
principles, 422-423 complexes with soil organic and mineral
procedure, 424-425 materials, 341-347
reagents, 424 divalent Cu, 345-346
saturated soil paste, 427-431 divalent Mn, 347
apparatus, 428-430 ferric iron, 343-345
calculations, 429 ion exchange on layer silicates, 349
principles, 427-428 vanadylion,346-347
procedure, 429 precautions, sensitivity, and resolution,
reagents, 428 331-333
waters, 422-427
principles, 323-326
Electrical specific conductance, 1298
quantitative, 349
Electrochemical cell, 248-249, 257-258
resonance condition, 326, 330
Electrode potential, 1256
rigid-limit spectra, 327
Electrodeless discharge lamp, 70
sample preparation, 331
Electromagnetic spectrum, 271
signal linewidth, 338
Electrometric method, pH determination,
spectral parameters, determination and theo-
484--485
retical interpretation, 333-334
Electron donor-acceptor system, electron spin
spin derivatization, 347-351
resonance spectroscopy, 340-341
Electron energy loss spectroscopy (EELS), 378 spin Hamiltonian, 326-327, 334
Electron microscopy, humic substances, 1060 superhyperfine splitting, 325, 328-329
Electron nuclear double resonance (ENDOR) thermodynamic constants and, 351
spectroscopy, 333, 352-353 three-line spectrum, 338
Electron paramagnetic resonance spectroscopy. Electron transfer, 1256
See Electron spin resonance spec- Electron Zeeman effect, 325, 327
troscopy Electronic method, pH determination, 485-488.
Electron paramagnetic resonance-stopped flow See also pH meter
apparatus, 1289-1290 procedure, 487-488
Electron spin echo (ESE) spectroscopy, 333, Electronic spectroscopy, 378
353-354 Electron-spin echo envelope modulation
Electron spin resonance (ESR) spectroscopy, (ESEEM), 353
323-354 Electrophoresis, fractionation of humic sub-
applications, 334-351 stances, 1031-1033
interpretation and comments on data, Electrothermal atomic absorption spectroscopy
337-341 alkaline-earth metal determination, 590
whole soil, 334-335 Mn determination, 670
electron donor-acceptor system, 340-341 Elementary reaction, 1292
electron Zeeman effect, 325, 327 Elson-Morgan method, amino sugar-N determi-
forbidden transition lines, 329 nation, 1196-1200
g-factor, 325, 327-329, 336, 342 Eluents
homogeneous broadening, 333 high-performance liquid chromatography,
humic substances, 335-337, 348-349, 233-234
1050-1052 ion chromatography, 241
interactions with organic chemicals and ENDOR spectroscopy. See Electron nuclear
metal ions, 340-341 double resonance spectroscopy
SUBJECT INDEX 1369
Energy-dispersive spectrometer, 162, 172, Ferric oxide, pretreatment of soils high in, 58
176-179 Ferrihydrite, 639-640
procedure for use, 209-211 Ferrimagnetic cluster, 345
Equilibrium computations, examples, Ferromagnetic resonance, 334
1327-1346 Ferrous iron determination, 1,10-phenanthroline
Equilibrium conditions, 1349-1350 method, 659-661
Equilibrium constant, 1257, 1295 Ferrum reductum, 926
Equilibrium models, 1309-1353 FFT. See Fast Fourier transform
equilibrium constant approach, 1310-1311 Film diffusion, 1279
free energy minimization approach, 1311 Filtration method
future trends in modeling, 1353 Al determination in soil solution, 533-534
mathematical basis, 1310-1311 Al speciation in solution, 536-537
problems in application, 1352-1353 First-order equation, 1296
validation, 1346-1349 First-order reaction, 1291, 1293-1295
methods, 1347-1348 FITEQL, 1315, 1326-1327, 1340-1341
scheme for validation, 1349 FITI routine, 411
Equipment. See Instruments/equipment Flame atomic absorption spectrophotometry,
Error 67-69
in dissolution of solids, 51 Al determination, 540-541
measurement, 12 Cd determination, 759-760
neutron activation analysis, 157-158 Mn determination, 670
sampling, 3-5, 7, 11-12 Ni determination, 759-760
selection, 12 Pb determination, 759-760
XRFS, 185-188 Flame atomization, 70-72
Escape depth, 359 Flame emission photometry, alkaline-earth
ESE spectroscopy. See Electron spin echo metal determination, 590
spectroscopy Flame photometry, alkali metal determination,
ESEEM. See Electron-spin echo envelope
556
modulation
Flameless atomic absorption spectrometry, Al
ESR spectroscopy. See Electron spin resonance
determination, 541
spectroscopy
Flooded soils, 1255, 1258
2-Ethyl-1,3-hexanediol in chloroform method,
Flow injection principles, 100
B determination, 615-616
Flow injection system, atomic absorption spec-
EXAFS spectroscopy. See Extended x-ray
troscopy, 88
absorption fine structure spectroscopy
EXAFSPAK program, 412 Flow methods, 1280-1290
Exchange capacity, mean, 8 analysis of rate data, 1290-1295
Exchangeable aluminum method, determination continuous flow, 1280-1286
of lime requirement, 511-513 fluidized bed reactor, 1286-1287
Exchangeable sodium percentage, 1209 stirred-flow reactor, 1282-1286
Exchangeable sodium ratio, 1209 stopped-flow technique, 1287-1290
Expert systems, 1353 Fluidized bed reactor, 1286-1287
Explosive reaction, 718 apparatus, 1286
Extended x-ray absorption fine structure uses, 1287
(EXAFS) spectroscopy, 380, 384, Fluorescence spectroscopy, humic substances,
391-394 1054-1055
data analysis, 411-412 Fluorescent x-ray relaxation process, 384-385
fine structure analysis of soil materials, Fluoride-titratable aluminum, 543
409-411 Fluorine
structural analysis, 398-409 chemistry, 833, 846-847
surface sensitive, 391 soluble, 856-857
Extra-atomic relaxation energy difference, 362 Fluorine determination
calcium chloride-extractable F, 857
Facility management, 28-29 direct potentiometric method, 857-859
Fast Fourier transform (FFT), 281 fluorometry, 863
FEFF computer program, 409, 411-412 ion chromatography, 863
Feldspar, K analyses, 565-566 neutron activation analysis, 863
Fermi level, 360-361 in solution, 857-863
Ferric iron determination, 1,10-phenanthroline SPADNS method, 859-860
method, 659--561 titrimetric methods
1370 SUBJECT INDEX
Fluorine determination (cont.) potassium bromide pellets, 2%

thorium nitrate in presence of alizarin red selective deuteration methods, 297
S,862-863 self-supporting films, 288--294
thorium nitrate in presence of chrome supported deposits on infrared transparent
azurol S, 860--862 windows, 294-295
total F content transmission methods, 287-288
calcium oxide incineration method, 853 signal-to-noise ratio, 282
decomposition of F-containing samples, single-beam spectra, 280--281
848---850 spectral analysis
separation of F from interfering com- application of Beer's law, 283-284
pounds, 850 band fitting, 285
sodium hydroxide fusion method, baseline correction, 284
850--852 spectral subtraction and difference meth-
steam distillation for separation of F from ods, 284-285
perchloric acid, 853---855 spectral processing
steam distillation for separation of F from choice of reference spectrum, 281-282
sulfuric acid, 855-856 optical saturation and electronic gain,
sulfuric acid method, 852-853 282-283
water-soluble F, 856-857 trading rules, 282
Willard-Winter distillation method, 84 7 Fourier-filtered spectra, x-ray absorption fine
Fluoro-aluminum complexes, 524 structure spectroscopy, 405
Fluorometric method Free acids, in soil, 477-478
F determination, 863 Free energy minimization approach, to equilib-
Se determination, 796, 801-803 rium modeling, 1311
Flux, salts used as, 50 Freundlich isotherm, 1351
Fourier transform function, 399, 405 FTIR spectroscopy. See Fourier transform
Fourier transform infrared (FTIR) spectroscopy, infrared spectroscopy
269-315 FTIR-gravimetric cell, 288, 291
attenuated total reflectance, 270, 287, Fulvic acid. See also Humic substance(s)
302-308 aqueous extraction, 1019
apparatus, 305-306 chemical shift assignments, 1051
applications, 307-308 composition, 1018
crystal materials, 306 electron spin resonance parameters, 1052
sample preparation, 307 oxygen-containing functional groups, 1043
theory, 304-305 Fume hood, 29
cylindrical internal reflectance, 270, 287, Fused discs, XRFS, 198--205
308--309 Fusion agents, 49-63
data collection, 280--281 advantages and disadvantages, 50
diffuse reflectance, 270, 287, 297-302 methods of fusion, 59-62
accessories, 298--300 principles, 52-53
applications, 301-302 safety, 54-55
sample preparation, 300--301 salts used as fluxes, 50
theory, 298
far-IR measurements, 272 Gamma rays
group frequencies of soil constituents, 275 detection, 147-150
humic substances, 1047-1048 detectors, 150--152
instrumentation emission, 146
beamsplitters, 277-280 for neutron activation analysis, 147
detectors, 277, 279 prompt gammas, 142-143
interferometer, 276--277 Gapon coefficient, 1209-1210
low temperature studies, 311-313 Gapon convention, 1207
microspectroscopy, 287, 308--312 Gas chromatography, Cr determination, 691
principles Gel filtration, fractionation of polysaccharides,
infrared absorption, 272-273 1034
molecular energy, 270--271 Gel permeation chromatography
molecular vibrations, 271-272 cleanup of extract of organic chemicals, 1080
vibrational analysis, 274-275 fractionation of humic substances,
Raman spectroscopy versus, 286 1028--1029
sample presentation, 287, 300--301, 307 humic substances, 1056--1057
liquid transmission cells, 296--297 GEOCHEM, 1313-1314, 1324
SUBJECT INDEX 1371
Geochemical equilibrium models, 1323-1327 High-temperature induction furnace method, C

GEOCHEM-PC, 1312, 1324, 1327-1339 determination, 971-973
GFM spectrophotometry. See Graphite fur- using Fe, Sn, and Sn-coated Cu accelerators,
nace atomic absorption spectrophotom- 972-973
etry HMDE. See Hanging mercury drop electrode
g-factor, 325, 327-329, 336, 342 Hollow cathode lamp, 70
Gibbsite, 347,350 Hollow-fiber membrane suppressor, 238
Glancing angle x-ray absorption fine structure Hot-water method, B extraction, 610-611
spectroscopy, 393 HPLC. See High-performance liquid chro-
Glycine, chelating ligand, 350 matography
Goethite, 289, 639-640 Burnie acid. See also Burnie substance(s)
Gouy-Chapman model, 1231 chemical shift assignments, 1051
Graphical assessment, kinetic rate data, composition, 1018
1290-1291 electron spin resonance parameters, 1052
Graphite crucible, 519-521 oxygen-containing functional groups, 1043
Graphite furnace atomic absorption (GFM) Burnie substance(s)
spectrophotometry, 67-69 chemical composition, 1015
Cd determination, 760 complexation chemistry, 348-349
Mo determination, 727 definition, 1012
Ni determination, 760 insolubility, 1015-1016
Pb determination, 760 interactions with organic chemicals and
Graphite furnace atomization, 71-72 metal ions, 340-341
Gravimetric method major proton resonance, 1050
carbonate determination, 440, 444--448 spin content, 339-340
for loss of carbon dioxide, carbonate deter- spin-labeled, 347
structural implications of electron spin reso-
mination, 455-456
nance, 340
Grazing incident mirror, 387
Burnie substance characterization, 1036-1060
Griess-Ilosvay method
acidic functional groups, 1036-1040
inorganic N determination, 1125
carboxylgroups, 1039-1040
nitrate determination, 1149, 1155-1158
phenolic groups, 1040
nitrite determination, 1150, 1158-1161
total acidity, 1037-1038
g-tensor, 327
carbonyl groups
Gypsum determination, 470-472 quinone groups, 1042-1043
total carbonyl groups, 1042
Halides, 833-867 chemical degradation, 1059
Hanging mercury drop electrode (HMDE), 254 chemical methods, 1036-1044
Harmonic rejection mirror, 391 electron microscopy, 1060
Heat-soluble sulfur, 952-953 electron spin resonance spectroscopy,
Hectorite, 289, 351 323-354, 1050-1052
Hematite, 639-640 fluorescence spectroscopy, 1054-1055
Henderson-Hasselbalch equation, 477 Fourier transform infrared spectroscopy, 296
High-performance liquid chromatography hydroxyl groups
(HPLC), 226, 228-236 alcoholic groups, 1041-1042
analytical columns, 232 phenolic groups, 1041
applications, 236 total hydroxyl groups, 1040-1041
columns, 230-231 infrared spectroscopy, 1045-1047
detectors, 234-236 ketonic groups, 1043-1044
eluents, 233-234 molecular size and shape, 1055-1059
gradient, 229 gel chromatography, 1056-1057
instrumentation, 229-230 light scattering, 1058
isocratic, 229 viscosity, 1058-1059
polar-bonded phase columns, 233 ultracentrifugation, 1057-1058
recorders, 236 nuclear magnetic resonance, 1047-1050
reverse-phase columns, 232-233 physical properties, 1055-1059
separation methods pyrolysis-mass spectroscopy, 1052-1054
adsorption, 231 spectroscopic, 1044-1055
ion exchange, 232 ultraviolet-visible spectroscopy, 1044-1045
partition, 231 Burnie substance extraction/purification,
size exclusion, 231-232 1014-1021
1372 SUBJECT INDEX
Humic substance extraction/purification (cont.) Hydrogen-saturated resin extraction method,

with aqueous solutions, 1017-1021 nonexchangeable K determination,
with EDTA, 1017 562-563
IHSS method, 1018-1019 Hydrologic regime, 1261
with non-aqueous solvents, 1021 Hydrolyzable unknown nitrogen. See HUN
with pyrophosphate, 1017 fraction
sequential extraction, 1020--1021 Hydroxy-aluminum compounds, 478, 524
with sodium hydroxide, 1017 Hydroxyl groups, humic substances
Humic substance fractionation alcoholic groups, 1041-1042
adsorption methods, 1027-1028 phenolic groups, 1041
based on charge characteristics, 1031-1033 total hydroxyl groups, 1040--1041
based on molecular size, 1028-1031 8-Hydroxyquinoline-butyl acetate method, Al
based on solubility, 1024--1028 determination, 538-540
electrophoresis, 1031-1033 Hyperfine coupling constant, 325, 329, 342
gel permeation chromatography, 1028-1029 Hyperfine splitting, 325, 328, 336, 338
with organic solvents, 1026--1027 Hypochlorite, Cr oxidizable by, 692
pH-based, 1024--1025 ICP. See Inductively coupled plasma
salting out, 1025-1026 ICP-AES. See Inductively coupled plasma-
ultrafiltration, 1030--1031 atomic emission spectroscopy
Humin, 1027 ICP-MS. See Inductively coupled plasma-mass
HUN fraction, 1185-1186, 1192 spectrometry
Hydride generation atomic absorption spec- Ideal solution approach, 1314
trometry, 86--87 Ignition method, organic P determination,
arsenite and arsenate determination, 822- 875-876
824 IHSS method, extraction of humic substances,
As determination, 815, 817-818 1018-1019
Se determination, 796, 800--801 Ilkovic equation, 249
Hydride-mercury vapor generator lmogolite, 350
description, 132 Imprecision, 33, 38-40
determination of As, Se, and Hg, 131-132 Inaccuracy, 33
ICP-MS and ICP-AES, 101, 131-132 Incident x-ray intensity, 379
Hydriodic acid-reducible sulfur, 944-946 Indicator electrode, 247, 258-259
Hydrochloric acid-nitric acid-hydrofluoric acid Indifferent ions, 1237, 1246
method, Al determination, 519-521 lndophenol blue method, ammonium determi-
Hydrochloric acid-sulfuric acid extraction nation, 1146--1148
method Inductively coupled plasma (ICP)
Mn determination, 676 generation, 94--95
P determination, 893-894 properties, 95-98
Hydrofluoric acid digestion Inductively coupled plasma-atomic emission
alkali element determination, 558-559 spectroscopy (ICP-AES), 67-69,
Fe determination, 644--645 91-134
method, 56--58 alkaline-earth metals, 579-580, 589-590
principles, 51-52 analysis of soil extracts and digests, 128-131
Hydrofluoric acid modification, of Kjeldahl Al determination, 541-542
method, 1110--1111 As, Se, and Hg determination, 131-132
Hydrofluoric acid-perchloric acid-sulfuric acid Cd determination, 761
digestion method costs, 94
Fe determination, 643--o44 detection limits, 114--116
Hydrofluoric, citric, perchloric, and sulfuric instrumentation
acid method inductively coupled plasma generation,
Cu determination, 717 94--95
Zn determination, 717-719 properties of inductively coupled plasma,
Hydrogen peroxide, hydrofluoric acid, nitric 95-98
acid, and perchloric acid method spectrometer, 102-103
Cd determination, 744--745 interferences
Ni determination, 744--745 correction, 118-121
Pb determination, 744--745 ionization, 117
Hydrogen peroxide, nitric acid, hydrochloric solute vaporation, 117
acid reflux method, 717 unwanted radiation, 117
SUBJECT INDEX 1373
Mn determination, 667--670, 679--680 Infrared spectroscopy

Ni determination, 761 Fourier transform. See Fourier transform
Pb determination, 761 infrared spectroscopy
preparation of stock standard solutions, humic substances, 1045-1047
128-129 Infrared transparent window, 294-295
quality control, 133 In-house standard, 38
sample introduction Initial rate, 1290, 1292-1295
hydride-mercury vapor generator, 101, Initial rate method, 1291-1292
131-132 Inner-sphere complex, 410, 1312-1313
LASER sampling of solids, 101-102 Inositol phosphate determination, 886-889
nebulizer, 98-101 Instantaneous equilibrium model, 1284
sample preparation Instruments/equipment
digestion of organic matter, 127 calibration. See Calibration
dissolution of silicates, 127 current list, 30
grinding soil samples, 125 daily check, 29
soil extracts, 125-126 maintenance, 29
wavelength selection, 104-113 Integrated rate equations, 1290-1295
Inductively coupled plasma-mass spectrometry Intensity factor, soil acidity, 491
(ICP-MS), 91-134 Intensity of photoelectron peak, 370
Al determination, 541-542 Interelemental spectral interference correction,
alkali metal determination, 556-557 118-119
analysis of soil extracts and digests, 128-131 Interferogram, 277-278, 281
costs, 94 Interferometer, Fourier transform infrared spec-
detection limits, 116 troscopy, 276-277
instrumentation Interlaboratory precision, 44-45
inductively coupled plasma generation,
Internal vibrational modes, 271
94-95
Intralaboratory precision, 44
properties of inductively coupled plasma,
Intrinsic surface charge density, 1239
95-98
Ion chromatography, 226, 236-243
special apparatus, 128
Al speciation in solution, 536-538
spectrometer, 104
alkali metal determination, 556
interferences
alkaline-earth metal determination, 591-593
correction, 124-125
single column method, 591
mass discrimination, 122-123
suppressed gradient method, 591-593
non-spectroscopic, 121-122
solids deposition on sampler and skimmer surface-sulfonated cation exchange col-
cones, 121 umn, 591
unwanted ions, 123 applications, 242-243
isotope selection, 113-114 Br determination, 839
preparation of stock standard solutions, Cl determination, 846
128-129 columns, 240-241
problem areas, 93 detectors, 241-242
quality control, 133 eluent-nonsuppressed system, 239-240
sample introduction eluents, 241
hydride-mercury vapor generator, 101, eluent-suppressed system, 237-238
131-132 F determination, 863
LASER sampling of solids, 101-102 inorganic N determination, 1127-1128
nebulizer, 98-101 instrumentation, 237
sample preparation P determination, 911-912
digestion of organic matter, 127 recorders, 242
dissolution of silicates, 127 S determination, 924
grinding soil samples, 125 separation methods, 241
soil extracts, 125-126 sulfate determination, 941
spectral interferences, 93 Ion exchange, layer silicates, 349
Inductively coupled plasma spectrometry, 69 Ion exchange chromatography
B determination, 623--624 fractionation of polysaccharides, 1033-1034
Cr determination, 689--691 inositol phosphate determination, 887
P determination, 910-911 Ion product, of water, 476
Si determination, 631,633 Ion scattering spectroscopy (ISS), 357
Infrared absorption, 272-273 Ionic strength, 1322
1374 SUBJECT INDEX
Ionic strength (cont.) Isotopic exchange, P buffering capacity deter-

effect on cation exchange capacity measure- mination, 904-905
ment, 1212-1213 ISS. See Ion scattering spectroscopy
Ion-selective electrode
for ammonium, 1162-1163, 1168-1169 Judgment sample, 3-4
for Br, 834-836
for Cl, 841 Kaolinite, 349
for F, 857-859 K-edge analysis, 389--390
inorganic N determination, 1125, 1161-1171 Ketonic groups, humic substances, 1043-1044
for nitrate, 1163-1167, 1169--1170 Kinetic data, analysis
for nitrogen oxides, 1167-1168, 1170--1171 graphical, 1290--1291
Iron, 639--660 initial rate method, 1291-1292
availability indices, 653--658 integrated rate equations, 1290--1291
chemistry in soil, 639--651 least-squares technique, 1292
complexation by organic matter, 640--641 using initial rate and integrated rate experi-
ferric ments, 1292-1295
complex with organic substances, Kinetic energy
343-345 electron, 360--361
electron spin resonance spectroscopy, photoelectron, 380, 383
343-345 Kinetic methods
in soil solution, 641 batch methods, 1276--1280
Iron chlorosis, 656 analysis of rate data, 1290--1295
flow methods, 1280--1290
Iron determination
analysis of rate data, 1290--1295
"amorphous" iron oxide, 648--650
relaxation methods, 1295-1305
ammonium oxalate method, 649--650
Kinetic model
atomic absorption spectroscopy, 658--659
dependent on solution concentration, 1284
colorimetric method, 1,10-phenanthroline
independent of solution concentration,
method, 659--661
1285-1286
exchangeable Fe, 651--653
Kinetics, 1275
organically bound Fe, 650--651
Kjeldahl method, N determination, 1085-1087,
sodium pyrophosphate method, 651
1089--1102
selective extraction methods, 645--653
ammonia electrode modification, 1108-1110
soil tests, 653--658
determination of ammonium in digest,
DTPA method, 654--657
1100--1102
DTPA-AB method, 657--658 digestion of sample, 1094-1100
total Fe content, 641--645 hydrofluoric acid modification, 1110--1111
hydrofluoric acid digestion method, permanganate-reduced Fe modification,
644--645 1111-1112
hydrofluoric acid-perchloric acid-sulfuric pretreatment of samples, 1090--1094
acid digestion method, 643--644 regular method, 1103-1108
sodium carbonate fusion method, 642--643 using block digester, 1114-1116
total "free" iron oxide, 645--648 salicylic acid-thiosulfate modification,
citrate-bicarbonate-dithionite method, 1112-1114
647--648 Klystron, 329
citrate-dithionite extraction method, Kubeika-Munk function, 298
646--647
water-soluble Fe, 651--653 "Lab dirt," 685
Iron oxide determination Labile pool, 753
"amorphous," 648--650 Laboratory access, 29
ammonium oxalate method, 649--650 Laboratory control sample, 34, 36, 38, 42
total "free," 645--648 Lande factor, 324
citrate-bicarbonate-dithionite method, Langmuirian adsorption behavior, 1351
647--648 Laser disk, 27
citrate-dithionite extraction method, LASER sampling of solids, 101-102
646--647 Layer silicates, 343
Iron oxide-impregnated filter paper method, P ion exchange, 349
determination, 899--901 Leachate, Hg determination, 784-787
Isoelectric point, 1240--1242 Leaching, 417
Isotope, radioactive. See Radioactive isotopes Lead, 739--763
SUBJECT INDEX 1375
availability indices, 752-758 nature of soil acidity, 493-494

soil solution, 752 neutralization sequence, 494
Lead determination soil properties, 494-496
ammonium pyrrolidine dithiocarbamate- field estimation, 499-500
chloroform method, 761-763 importance, 491-493
atomic absorption spectrophotometry, laboratory protocols, 498-499
759-761 other methods, 513
flame atomic absorption spectrophotometry, soil-base titration methods, 49fr..497,
759-760 499-501
graphite furnace atomic absorption spec- soil-buffer equilibration methods, 49fr..497,
trophotometry, 760 501-511
inductively coupled plasma-atomic emission Adams-Evans single buffer method, 498,
spectrophotometry, 761 505--508
sampling and sample preparation, 739-740 Mehlich single buffer method, 508-510
selective extraction, 745-752 Nommik single buffer method, 510
carbonate phase, 747, 750 Shoemaker-McLean-Pratt single buffer
exchangeable phase, 747, 749 method, 498, 502-505
iron oxide phase, 748, 750 Woodruff single buffer method, 510--511
manganese oxide phase, 750 soil-lime incubation methods, 49fr..497,
organic matter phase, 74 7, 750--751 499-500
residual phase, 748, 751 target pH, 491-492
soil tests, 752-758
Lime response curve, 499-500
DTPA method, 754-755
Limit of quantitation, 47
DTPA-AB method, 756
Lineshape, 285
extractants, 753
Liquid chromatography, 225-243
Mehlich I extraction, 757-758
Cr determination, 690
Mehlich III extraction, 758
high-performance. See High-performance
one-tenth molar hydrochloric acid method,
liquid chromatography
75fr..757
principles, 22fr..228
standard reference materials, 742
tailing, 228
total Pb content
Liquid sample, XRFS, 199-200, 214-215
hydrogen peroxide, hydrofluoric acid,
Liquid transmission cell, sample for Fourier
nitric acid, and perchloric acid method,
transform infrared spectroscopy,
744-745
29fr..297
nitric acid-hydrochloric acid digestion
method, 742-743 Lithium, 551-572. See also Alkali metal(s)
nitric acid-perchloric acid digestion chemistry, 551-552
method, 743--744 extractable, 572
principles, 740--742 properties, 555
Least-squares technique, 1290, 1292 Lithium metaborate flux, 202-204
LECO instruments, 975-976 Lithium metaborate fusion method, Al determi-
LECO S Analyzer, 933, 937 nation, 519-520
L-edge analysis, 390 Lithium tetraborate fusion method, Si determi-
LEED. See Low-energy electron diffraction nation, 629---032
Lepidocrocite, 639--640 Lithium tetraborate-lithium carbonate-lan-
Ligand field calculations, 334 thanum oxide flux, 204-205
Light absorption spectrometry, Si determination Logbook,25-26
blue silicomolybdous acid method, 635---036 Long-term standard deviation, 40
yellow silicomolybdic acid method, 634---035 Loss-on-ignition method, organic matter deter-
Light elements, 386 mination, 1003--1005
Light scattering, humic substances, 1058 Low temperature study, Fourier transform
Light source, atomic absorption spectroscopy, infrared spectroscopy, 311-313
70 Low-energy electron diffraction (LEED), 357
Lime requirement, 491-513, 522-523 Lysimeter, Al determination in soil solution,
analytical considerations, 49fr..499 532
definition, 491-493
estimation from soil pH, 477 Maghemite, 639--640
exchangeable acidity determination, 49fr..497 Magnesium, 575-598. See also Alkaline-earth
exchangeable Al method, 511-513 metal(s)
factors affecting availability indices, 585-586
1376 SUBJECT INDEX
Magnesium (cont.) Mannitol potentiometric method, B determina-

mineralogy and distribution in rocks and tion, 618-620
soils, 576--577 Mannitol-calcium chloride method, B extrac-
properties, 575-576 tion, 612-613
soluble, 581 Manometric method
Magnesium carbonate, 437 carbonate determination, 439
Magnesium determination determination of carbonate reactivity,
atomic absorption spectrophotometry, 578, 463-465
587-589 Mass absorption coefficient, 167, 184
EDTA method, 593-594 measurement, 195
electrothermal atomic absorption, 590 Mass balance equation, 1284
extractable/exchangeable fraction, 582-584 Mass spectrometry. See Inductively coupled
acid soil extraction, 584 plasma-mass spectrometry
ammonium acetate method, 583-584 Mass transfer, 1283
ammonium chloride method, 584 speciation and, 1335-1336
flame emission photometry, 590 Matrix effects, XRFS, 190-197
inductively coupled plasma-atomic emission Matrix modification, 79-80
spectrometry, 579-580, 589-590 Matrix spiking solution, monitoring extraction
ion chromatography, 591-593 of organic chemicals, 1082-1083
titrimetric method, 593-596 MCMFE. See Membrane-covered mercury film
x-ray fluorescence, 578-579 electrode
Magnetic dipole moment, effective, 323 Mean,5-S
Magnetic spectroscopy, 378 on per element basis, 13
Magnetite, 639-640 Mean square among elements within units,
Manganese,665-681 13-14
absorbed or retained, 666 Measurement error, 12
availability indices, 672-681 Medium-temperature resistance furnace
chemistry, 665-667 method, C determination, 966--971
divalent Mehlich I extraction
complex with organic substances, 347 Cd determination, 757-758
electron spin resonance spectroscopy, 347 Cu determination, 711-713
easily reducible, 677-678 K determination, 567
exchange on layer silicates, 349 Mn determination, 676
exchangeable, 665-666, 675-676 Ni determination, 757-758
insoluble precipitates, 666 Pb determination, 757-758
in minerals, 666 Zn determination, 711-713
organic matter phase, 666 Mehlich III extraction
soluble, 665-666 Cd determination, 758
water-soluble, 673-675 Cu determination, 706, 713-715
Manganese determination K determination, 567-568
atomic absorption spectroscopy, 667, 670, Ni determination, 758
679 Pb determination, 758
colorimetric methods, periodate method, Zn determination, 706, 713-715
671-672 Mehlich single buffer method, determination of
inductively coupled plasma-atomic emission lime requirement, 508-510
spectroscopy, 667-670, 679-680 Meinhardt nebulizer, 99
soil tests, 672-681 Meker burner, 59-60
DTPA method, 677 Membrane-covered mercury film electrode
easily reducible Mn, 677-678 (MCMFE), 256
exchangeable Mn, 675-676 "Memory effect," 43
hydrochloric acid-sulfuric acid extraction Mercury, 769-788
method, 676 chemistry, 769-771
water-soluble Mn, 673-675 methylation, 771
total Mn content, 679-680 Mercury cadmium telluride detector, 277,279
colorimetric method, 680--681 Mercury cold vapor method, 87
Manganese dioxide, pretreatment of soils high Mercury determination
in, 58 amalgamation, 774
Manganese electron demand, 695-696 cold vapor-atomic absorption spectrometry,
Manganese oxide, selective dissolution from 771-784
soils and sediments, 678-679 dissolved Hg, 786--787
SUBJECT INDEX 1377
hydride-mercury vapor generator, 131-132 dilute salt extractable Al, 526-527

organic Hg compounds, 787-788 potassium chloride method, 526
standard reference materials, 782-783 MINTEQ, 1313, 1325
suspended Hg, 786-787 MINTEQA2, 1312, 1314, 1325
total Hg content Mixing effect, batch methods, 1279
automated method, 775-782, 787 Mixture model, 1311, 1337-1339
manual method, 775-782, 786-787 Modulation amplitude, 332
preparation of soil digest, 779-780 Molecular orbital calculations, 334
principles, 772-775 Molecular polarizability, 274
in soil extracts, leachates, and water sam- Molecular vibrations, 271-272
ples, 784--787 Molybdenum, 723-734
in soils and sediments, 771-784 accumulation by plants, 724
Mercury electrode, 258--259 availability indices, 728-731
Mercury thin film electrode (MTFE), 254, chemistry, 723-725
258--259 deficiency, 723
Mercury(Il) thiocyanate method, Cl determina- toxicity, 723
tion, 840, 845-846 Molybdenum determination
Metal, surface coordination, 349-350 anion exchange resin, 729-730
Metal ions atomic absorption spectroscopy, 728
interactions with humic substances, 340-341 general principles, 725
structural, 341 graphite furnace atomic absorption spec-
Metal spin probes, 348--351 troscopy, 727
Metal-ligand-surface interactions plasma emission spectroscopy, 728
adsorption models, 1312-1321 soil tests
metal-organic ligand interactions, 1311-1312 ammonium oxalate method, 728--729
modeling, 1311-1321 DTPA-AB method, 730-731
Method blank, 41 total Mo content, thiocyanate-stannous chlo-
Method detection limit, 46-47 ride method, 725-728
Methods manual, 24 Monosaccharides, polysaccharide composition,
Methoxybenzene radical, 338 1060-1062
Methylene blue method, sulfate determination, Montmorillonite, 289-290, 351
927,932-936,940-941 Morgan extraction
Methylmercury, 771 modified, K determination, 570
2-Methyl-2-nitrosopropene, 348 K determination, 569-570
Mica, 289 MTFE. See Mercury thin film electrode
K analyses, 566 Muck soil, 421
Microdiffusion method Multichannel analyzer, neutron activation
Br determination, 837-839 analysis, 152
inorganic N determination, 1127-1128, Multiplet splitting, x-ray photoelectron spec-
1139-1146 troscopy, 369
in absence of nitrite, 1142-1143 Multistage sampling. See Subsampling
in presence of nitrite, 1143
Microelectrodes, pH determination, 488 N-associated radicals, 338
Micromembrane suppressor, 238 Natural organic interactions, equilibrium com-
Microprobe, Raman, 313-315 putations, 1337-1339
Microspectroscopy, Fourier transform infrared Natural waters
spectroscopy, 287, 308--312 anodic stripping voltammetry, 260-264
Microwave digestion, 740 differential pulse polarography, 260-264
advantages and disadvantages, 50 Near infrared diffuse reflectance spectropho-
method, 62---03 tometry, N determination, 1116
principles, 53-54 Nebulizer, 98--101
Microwave energy, efficiency of absorption, 54 Nernst equation, 1235, 1257, 1259
Microwave oven, 62 Nessler method, ammonium determination,
Microwave-power saturation, 333 1146
MINEQL, 1313-1314, 1325 Net total particle charge, 1239
MINEQL+, 1325-1326 Neutral salt extractable iron, 653
Mineral acids, 50 Neutralization sequence, 493-494
Mineral potassium analysis, 563-566 Neutron activation analysis, 141-158
Mineral soils, Al extraction, 525-527 alkali metal determination, 556-557
1378 SUBJECT INDEX
Neutron activation analysis (cont.) total Ni content

calculations, 157-158 hydrogen peroxide, hydrofluoric acid,
equipment nitric acid, and perchloric acid method,
gamma-ray detectors, 150--152 744-745
multichannel analyzers, 152 nitric acid-hydrochloric acid digestion
neutron sources, 150 method, 742-743
errors, 157-158 nitric acid-perchloric acid digestion
halide determination, 863 method, 743-744
instrumental, 141, 155 principles, 740--742
neutron irradiation, 154 Ninhydrin method, amino acid-N determination,
principles 1193-1196
neutron capture cross-sections, 143-144 Nitrate, 1123
neutron induced reactions, 142-143 exchangeable, 1124
nuclear decay and detection of radiation, (Nitrate + nitrite)-nitrogen determination
146-150 microdiffusion method, 1143
production and decay of radioactive iso- steam-distillation method, 1136
topes, 144-146 Nitrate determination
radiation safety, 152-153 brucine method, 1148
radioassay, 155-158 chromotropic acid method, 1148-1149
radiochemical, 156-157 colorimetric methods, 1148-1149,
sample preparation, 153-154 1155-1158
encapsulation for neutron irradiation, extraction of exchangeable nitrate,
154 1129-1131
standards, 153 Griess-Ilosvay method, 1149, 1155-1158
Neutron capture, 142 nitrate electrode, 1163-1167, 1169-1170
Neutron capture cross-sections phenoldisulfonic acid method, 1148
definition, 143 salicylic acid method, 1149
thin target assumption, 143-144 in soil extracts, 1131-1171
Neutron source, 150 Nitrate-nitrogen determination, 1126-1127
Nickel, 739-763 microdiffusion method, 1142-1143
availability indices, 752-758 steam-distillation method, 1135-1136
soil solution, 752 Nitric acid/hydrogen peroxide method, oxida-
Nickel determination tion of organic ligands binding Cr(III),
ammonium pyrrolidine dithiocarbamate- 694
chloroform method, 761-763 Nitric acid/vacuum oven method, oxidation of
atomic absorption spectrophotometry, organic ligands binding Cr(III), 694
759-761 Nitric acid-hydrochloric acid digestion method
flame atomic absorption spectrophotometry, Cd determination, 742-743
759-760 Ni determination, 742-743
graphite furnace atomic absorption spec- Pb determination, 742-743
trophotometry, 760 Nitric acid-perchloric acid digestion method
inductively coupled plasma-atomic emission Cd determination, 743-744
spectrophotometry, 761 Ni determination, 743-744
sampling and sample preparation, 739-740 Pb determination, 743-744
selective extraction, 745-752 Nitrite, 1123
carbonate phase, 747, 750 exchangeable, 1124
exchangeable phase, 747, 749 Nitrite determination
iron oxide phase, 748, 750 colorimetric methods, 1149-1150,
manganese oxide phase, 750 1158-1161
organic matter phase, 747, 750--751 extraction of exchangeable nitrite,
residual phase, 748, 751 1129-1131
soil tests, 752-758 Griess-Ilosvay method, 1150, 1158-1161
DTPA method, 754-755 nitrogen oxides electrode, 1167-1168,
DTPA-AB method, 756 1170--1171
extractants, 753 in soil extracts, 1131-1171
Mehlich I extraction, 757-758 Nitrite-nitrogen determination, 1127
Mehlich III extraction, 758 Nitroalkane, 348
one-tenth molar hydrochloric acid method, Nitrogen determination
756-757 inorganic N, 1123-1176. See also specific
standard reference materials, 742 compounds
SUBJECT INDEX 1379
colorimetric methods, 1125-1126, Optical system, atomic absorption spectroscopy,

1146-1161 72-74
extraction of exchangeable ammonium, Organic carbon determination, 961, 983-1001.
nitrate, and nitrite, 1129-1131 See also Organic matter
ion chromatography, 1127-1128 in calcareous soils, 985-988
ion-selective electrodes, 1125, 1161-1171 calculated from total C determination, 985
microdiffusion method, 1127-1128, comparison of methods, 999-1001
1139-1146 dichromate oxidation method, 988-999
nonexchangeable ammonium, 1171-1176 principles, 988-995
in soil extracts, 1131-1171 tube digestion method, 997-998, 1000
in soils, 1125-1129 Walkley-Black method, 993, 995-996,
steam-distillation methods, 1125, 1127, 1000
1131-1138 dry combustion method, 985-988, 1000
ultraviolet difference method, 1127-1129 pretreatment, 987-988
organic N, 1185-1200 in noncalcareous soils, 985
steam-distillation method, 1187-1196 in soil extracts, 988
total N content, 1085-1117 wet combustion method, 985-988, 1000
pretreatment, 986-987
dichromate method, 1117
Organic chemicals
Dumas method, 1085, 1087-1089
extract cleanup, 1080-1081
Kjeldahl method, 1085-1087, 1089-1102
gel permeation chromatography, 1080
Kjeldahl method, ammonia electrode
extraction, 1071-1083
modification, 1108-1110
bound pesticide residues, 1081-1082
Kjeldahl method, hydrofluoric acid modi-
matrix and surrogate spiking solutions,
fication, 1110-1111
1082-1083
Kjeldahl method, permanganate-reduced
purge-and-trap systems, 1072-1074
Fe modification, 1111-1112
solvent extraction of aqueous samples,
Kjeldahl method, regular, 1103-1108
1074---1076
Kjeldahl method, salicylic acid-thiosulfate solvent extraction of solid samples,
modification, 1112-1114 1076-1080
Kjeldahl method, using block digester, sampling and preservation
1114---1116 liquid samples, 1072
NIR spectrophotometry, 1116 solid samples, 1072
persulfate oxidation method, 1116-1117 Organic contaminants, 1071
Nitrogen oxides electrode, 1167-1168, Organic free radicals, electron spin resonance
1170-1171 spectroscopy, 323-354
Nitrone, 348 Organic matter, soil, 1001-1006
Nitrosoarene, 348 Al extraction, 527-530
Nitrosobenzene, 348 copper chloride method, 529
Nitroxide spin label, 347-348 sodium pyrophosphate method, 529-530
Nommik single buffer method, determination of categorization, 1025
lime requirement, 510 definition, 962, 1011
Noncalcareous soil digestion, 127
inorganic P fractionation, 882-883 effect on lime requirement, 495--496
organic C determination, 985 Fe interactions, 640--641, 650-651
Nonideality corrections, thermochemical data, interactions with paramagnetic metal ions,
1321-1322 341-347
NONLIN, 1326 organic C-organic matter conversion factors,
Normal pulse polarography, 250-251 962
Normalization function, 399 oxidation of ligands binding Cr(III), 694
Nuclear decay, 146 Organic matter characterization. See also
Nuclear magnetic resonance spectroscopy Humic substance(s); Polysaccharides
humic substances, 1047-1050 characterization of polysaccharides,
P-31, P determination, 911 1060-1062
Nuclear quadrupole interactions, 325, 329 density fractionation, 1034---1035
extraction of polysaccharides, 1021-1024
One-tenth molar hydrochloric acid method fractionation of humic substances,
Cd determination, 756---757 1024---1033
Ni determination, 756-757 fractionation of polysaccharides, 1033-1034
Pb determination, 756-757 future developments, 1062-1063
1380 SUBJECT INDEX
Organic matter characterization (cont.) Perchloric acid digestion

principles, 1011-1014 method, 57-58
Organic matter determination P determination, 872
calculation of organic matter content, principles, 52
1001-1002 Performance audit, 32
direct estimation, 1002-1005 Periodate method, Mn determination, 671-672
expression of soil organic matter content, Perkin-Elmer CHN2400, 976-977
1005-1006 · Permanent charge. See Constant charge
loss-on-ignition method, 1003-1005 Permanganate method, alkaline-earth metal
Organic matter extraction, 1014-1024 determination, 596-597
extraction of humic substances, 1014-1021 Permanganate reduced-iron modification, of
Organic mercury compounds, 771, 787-788 Kjeldahl method, 1111-1112
Organic nitrogen determination, 1185-1200 Personal protective equipment, 29
steam-distillation method, 1187-1196 Persulfate oxidation method, N determination,
Organic phosphorus determination, 874-881. 1116-1117
See also Phosphorus determination Pesticide, 1071
fractionation, 884-890 extraction of bound residues, 1081-1082
inositol phosphate fraction, 886-889 pH, soil, 475-489. See also Soil acidity
phospholipid fraction, 889-890 definition of pH, 476-477
Organic soils, Al extraction, 527-530 factors affecting
copper chloride method, 529 carbon dioxide content, 482
sodium pyrophosphate method, 529--530 dilution, 479--480
Organic solvents, fractionation of humic sub- salt content, 480-482
stances, 1026-1027 suspension effect, 482-483
Organic sulfur determination, 943-949. See lime requirement, 491-513
also Sulfur determination significance, 477-479
extraction, 952-955 pH determination
Osmotic pressure, 426 colorimetric methods, 483-484
Outer-sphere complex, 410, 1312-1313 electrometric methods, 484-485
Outlier, 37, 39, 46 electronic measurement, 485-488
Out-of-control value, 36-38 with microelectrodes, 488
Overtiming, 705 with pH paper, 489
Oxalato-aluminum complexes, 524 with test kits, 488
Oxidation potential, 1256 pH effect
Oxidation state, x-ray absorption fine structure on cation exchange capacity measurement,
spectroscopy,381,393-394 1212
Oxidation-reduction reactions on redox reactions, 1257
applications pH meter, 475, 485-487
anodic stripping voltammetry, 262-263 electrode problems, 486-487
differential pulse polarography, 262-263 procedure for use, 487-488
equilibrium computations, 1336-1337 standardization, 485-486
pH paper, 489
Packed-column suppressor, 238 pH stat method, determination of carbonate
Pair production, 150 reactivity, 459--463
Paramagnetic metal ions pH-based method, fractionation of humic sub-
chemisorption on mineral surfaces, 349--350 stances, 1024-1025
complexes with soil organic and mineral Phase shift, x-ray absorption fine structure
materials, 341-347 spectroscopy, 404-405
electron spin resonance spectroscopy, 1,10-Phenanthroline method, ferrous and ferric
323-354 iron determination, 659--661
divalent Cu, 345-346 Phenoldisulfonic acid method, nitrate determi-
divalent Mn, 347 nation, 1148
ferric iron, 343-345 Phenolic groups, humic substances, 1040-1041
vanadylion,346-347 Phenyl-N-t-butylnitrone, 348
ion exchange on layer silicates, 349 Phlogopite, weathered, 345
Parent material, effect on lime requirement, Phonon modes, 271-272
494-495 Phosphate extraction, absorbed selenite, 809
Particle diffusion, 1279 Phospholipid determination, 889-890
Pass energy, 363 Phosphorus, 869--912
Peat soils, 421 availability indices, 890-903
SUBJECT INDEX 1381
chemistry, 869-870 three step model, 369--371

Phosphorus buffering capacity determination, Phyllosilicate, 517
903-905 Pitzer equation, 1322
anion exchange resin, 904--905 Platinum crucible, 57-58
isotopic exchange, 904--905 Platinum electrode, 1255
Phosphorus determination cleaning and testing, 1265-1266
colorimetric methods construction, 1262-1265
ascorbic acid method, 908-909 field installation, 1266-1267
chlorostannous acid method, 907-908 field measurement of redox potential,
modified ascorbic acid method, 909--910 1267-1269
vanadomolybdophosphoric acid method, preparation, 424
907 retrieval at end of field study, 1269
fractionation of inorganic P, 881---884 Pneumatic nebulizer, 98-101
calcareous soils, 883---884 concentric, 99
noncalcareous soils, 882---883 cross flow, 99
fractionation of organic P, 884---890 Podzolization process, 341
inductively coupled plasma spectrometry, Point of zero charge, 1239--1240
910-911 Point of zero net charge, 1240-1242
ion chromatography, 911-912 measurement, 1242-1244
P-31 nuclear magnetic resonance spec- Point of zero net proton charge, 1240-1242
troscopy, 911 measurement, 1249-1250
principles, 906 Point of zero salt effect, 1240-1242
soil tests measurement, 1244--1249
anion exchange resin, 898---899 automatic potentiometric titration,
buffered alkaline extraction, 895---897 1248-1249
calcium chloride determination, 891---893 backtitration method, 1248
dilute acid F extraction, 894---895 batch method, 1245-1248
DTPA-AB method, 897---898 Polar-bonded phase high-performance liquid
hydrochloric acid-sulfuric acid extraction,
chromatography, 233
893---894
Polarizability, 274, 362
iron oxide-impregnated filter paper
Polarography, 247
method, 899--901
alternating current, 255-256
P-32 dilution method, 901-903
differential pulse. See Differential pulse
water or dilute salt extraction, 891---893
polarography
total organic P content, 874---881
direct current, 248--249, 257
acetylacetone extraction, 878---881
instruments and electrode, 256---259
concentrated sulfuric acid-dilute sodium
normal pulse, 250-251
hydroxide extraction, 877---878
Policy statement, of quality system, 20-21
ignition method, 875---876
principles, 874---875 Pollutant metals, 739--763, 769--788
sodium hydroxide, hydrochloric acid, Polygorskite, 290
sodium hydroxide extraction, 876---877 Polymerization process, 341
total P content, 870---874 Polysaccharides
perchloric acid digestion, 872 characterization, 1060-1062
principles, 870---871 hydrolysis, 1061-1062
sodium carbonate fusion method, 871---872 monosaccharide composition, 1060-1062
sodium hypobromite oxidation method, extraction and purification, 1021-1024
873---874 fractionation, 1033--1034
sulfuric acid-hydrogen peroxide-hydroflu- gel filtration, 1034
oric acid digestion, 872---873 ion-exchange chromatography, 1033-1034
Phosphorus-31 nuclear magnetic resonance Population, 1
spectroscopy, 911 Potassium, 551-572. See also Alkali metal(s)
Phosphorus-32 dilution method, P determina- chemistry, 551-552
tion, 901-903 exchangeable, 553, 559--560
Photoelectric effect, 147-149 mineralogy in soils, 552
Photoelectron, 359--360, 382 nonexchangeable,553--554,560-563
binding energy, 358, 364--368 properties, 552-554
kinetic energy, 380, 383 in soil solution, 553
survival pathlength, 383-384 Potassium bromide pellet, sample for Fourier
Photoemission, 358 transform infrared spectroscopy, 296
1382 SUBJECT INDEX
Potassium chloride method, Al extraction from Pulse-height selection, XRFS, 172-173

unbuffered soils, 526 Purge-and-trap system, extraction of organic
Potassium determination chemicals, 1072-1074
ammonium acetate method for exchangeable Pyrocatechol violet method, Al determination,
K,560 540
boiling nitric acid extraction for nonex- Pyrolysis-mass spectroscopy, humic substances,
changeable K, 562 1052-1054
hydrogen-saturated resin extraction for Pyrophosphate method, extraction of humic
nonexchangeable K, 562-563 substances, 1017
mineral K analysis, 563-566
soil tests, 566--571 QA. See Quality assurance
ammonium acetate extractable K, QC. See Quality control
568-569 Quality assurance (QA), 19-48. See also Qual-
DTPA-AB method, 570-571 ity control
Mehlich I extraction, 567 assessment mechanisms, 20, 31-32
Mehlich III extraction, 567-568 external, 32
modified Morgan extraction, 570 internal, 31-32
Morgan extraction, 569-570 in chemical testing, 20
Potassium hydroxide method, Al extraction definition, 21
from poorly ordered phases, 531
in physical testing, 19-20
Potential determining ions, 1235-1236
policy statement, 20-21
Potentiometric method
sampling, 19
B determination
standard operating procedures, 22-31
mannitol method, 618-620
statistical terms, 33-42
tetrafluoroborate selective electrode
training about, 28
method, 620-623
Quality assurance (QA) manual, 22, 24
Br determination, 834--836
Quality control (QC), 19-48. See also Quality
Cl determination, 840, 843-844
assurance
F determination, 857-859
definition, 21
Potentiometric stripping analysis, 256
Potentiometric titration method ICP-AES, 133
ICP-MS, 133
Cl determination, 840-843
for point of zero salt effect, 1248-1249 standard operating procedures, 22-31
Powder, XRFS, 197, 214-215 statistical terms, 33-42
Power saturation effect, 326 training about, 28
Precipitation method, Cr determination, 691 Quality control check sample, 28, 34, 36, 38, 42
Precision, 7, 34-35, 38-40, 44 Quantitation, limit of, 47
of analysis, 38 Quartz, K analyses, 566
interlaboratory, 44-45 Quasipartide model, 1311-1312
intralaboratory, 44 Quinone groups, humic substances, 1042-1043
repeatability and reproducibility, 40 Quinone-hydroquinone system, 340
sources of imprecision, 39-40
Pressed pellets, XRFS, 197-198, 211-214 Radiation safety, 152-153
Pressure calcimeter method, carbonate determi- Radioactive isotopes
nation, 440-444 for neutron activation analysis, 147
Pressure-jump relaxation methods, 1297-1305 nuclear decay, 146--147
apparatus, 1298-1301 production during neutron irradiation,
data collection and detection, 1301-1302 144-146
experimental details Radioassay, neutron activation analysis,
kinetic studies, 1304-1305 155-158
static studies, 1303 Raman microprobe, 313-315
maintenance of stable suspensions, Raman scattering, 273-274
1302-1303 Raman spectroscopy, 270, 313-315
Primary data, 30-31 Fourier transform infrared spectroscopy vs.,
PRODEFA2, 1325 286
Proficiency, 28 principles, 273-274
Proficiency testing, 32, 45 Random sample
Prompt gammas, 142-143 simple, 4-6, 11
Proton transfer, 1256 stratified, 7-9, 11
Pseudo first-order reaction, 1292-1293 Range of analysis, 46
SUBJECT INDEX 1383
Rapid dichromate oxidation method. See properties, 555

Dichromate oxidation method
Rapid reactions, 1283 Safety
"Raw" data. See Primary data dissolution of solids, 54-55
Reanalysis, 38, 45 neutron activation analysis, 152-153
Record keeping XRFS, 179-181
standard operating procedures, 25-27 Salicylate method
training plan, 28 ammonium determination, 1152-1155
REDEQL2, 1324 nitrate determination, 1149
Redox, soil Salicylic acid-thiosulfate modification, of Kjel-
available reducing capacity for Cr(VI), 696 dahl method, 1112-1114
Cr as redox tool, 695-697 Salinity, 417-433, 839
Cr net oxidation test, 695-696 soil, 417
soil reducing intensity for Cr(VI), 696-697 determination, 427-431
total Cr(VI) reducing capacity, 696 diagnosis, 417-433
Redox potential, 1255-1271. See also Platinum soil pH and, 480-482
electrode water, 418
applications of redox measurements, Salt content. See Salinity
1258-1261 Salting out, fractionation of humic substances,
controlled studies, 1269-1271 1025-1026
field measurement, 1267-1269 Sample preparation
reliability, 1269 anodic stripping voltammetry, 259-260
theory, 1256-1258 atomic absorption spectroscopy, 84-86
Redox reaction, pH effect, 1257 differential pulse polarography, 259-260
Redoximorphic features, 1261 electron spin resonance spectroscopy, 331
Reducing capacity Fourier transform infrared spectroscopy,
available for Cr(VI), 696 286-297,300-301,307
total Cr(VI), 696 ICP-AES, 98-102, 125-127
Reduction potential, 1256 ICP-MS,98-102, 125-127
Reference electrode, 247-248 neutron activation analysis, 153-154
Reference material, 28 for total elemental analysis dissolution,
Relative standard deviation, 33-35, 38-39 55-56
Relaxation methods x-ray photoelectron spectroscopy, 372-373
background, 1295-1297 XRFS, 197-200
chemical, 1296 Sampling, 1-17
concentration-jump, 1297 composite samples, 16-17, 19
pressure-jump, 1297-1305 cost, 14-15
stationary, 1295-1296 error, 3-5, 7, 11
temperature-jump, 1296 quality assurance, 19
transient, 1295-1296 sampling plans, 3-11, 19
Relaxation spectra, 1304 subsampling, 12-16
Relaxation time, 1296, 1301 for total elemental analysis dissolution,
Repeatability, 23, 40 55-56
Repeatability value, 40 variation of soils, 1-3
Replicate, 24, 33-34, 39, 41 Sampling error, 12
Replicate sample, 43-45 Sampling unit, 12
Reproducibility, 35, 40 definition, 4-5
Reproducibility value, 40 location, 4-5
Resolution, electron spin resonance spec- S-associated radicals, 338
troscopy, 331-333 Satellites, x-ray photoelectron spectroscopy,
Resonance condition, electron spin resonance 369
spectroscopy, 326, 330 Saturated soil paste
Resonance phenomenon, 323-326 electrical conductivity, 427-431
Reverse-phase high-performance liquid chro- preparation, 419
matography, 232-233 Saturated soils, 1258
Rotational correlation time, 349 Saturation extract
Round robin, 45 B determination, 611-612
Rubidium, 551-572. See also Alkali metal(s) preparation
chemistry, 551-552 apparatus, 419
extractable, 572 calculations, 420
1384 SUBJECT INDEX
Saturation extract (cont.) Shoemaker-McLean-Pratt single buffer method,

principles, 419 determination of lime requirement, 498,
procedure, 420 502-505
reagent, 420 Short-term standard deviation, 40
Saturation percentage, 427 Silica gel, chromatographic, 230
Saturation water percentage, 420 Silicates, dissolution, 127
Scale interval, 4 Silicon, 627-636
Scanning electron microscopy (SEM), 378 chemistry, 627
Scanning force microscopy (SFM), 378 Silicon determination
Scanning monochromator, 103 atomic absorption spectrometry, 631-633
Scanning probe microscopy (SPM), 357 inductively coupled plasma spectrometry,
Scanning tunneling microscopy (STM), 378 631,633
Scatchard quasiparticle model, 1312, light absorption spectrometry
1337-1339 blue silicomolybdous acid method,
Scintillation detector, 170 635-636
Secondary data, 30 yellow silicomolybdic acid method,
Secondary ion mass spectroscopy (SIMS), 357 634-635
Second-order reaction, 1291 principles of analyses, 627-629
Security, of records, 26 standard reference materials, 629
Selection error, 12 total Si content
Selection rules, 274 sample preparation by acid digestion,
Selectivity coefficient, measurement, 632-634
1222-1226 sample preparation by lithium tetraborate
Selenate determination, 805-808 fusion, 629-632
Selenite determination, 805-808 Silver diethyldithiocarbamate method, As deter-
Selenium, 793-811 mination, 814-816
availability indices, 808-811 Simple random sample, 4-6, 11
chemistry, 794-796 SIMS. See Secondary ion mass spectroscopy
Selenium determination Slipping plane, 1241
hydride-mercury vapor generator, 131-132 Slurry, determination of alkaline-earth metals,
sample preparation, 807-808 579-580
selective extraction, 809-811 SMDE. See Static mercury drop electrode
selenite and selenate in soil solutions and Smectite, 290
water extracts, 805-808 Smith-Hieftje background correction, 82-83
standard reference materials, 794 Sodium, 551-572. See also Alkali metal(s)
total Se content chemistry, 551-552
digestion of soils, 798-800 mineralogy in soils, 552
fluorometric methods, 796, 801-803 properties, 554-555
hydride generation atomic absorption Sodium adsorption ratio, 1209
spectrometry, 796, 800--801 Sodium carbonate, 437,479
in soil solutions and water extracts, Sodium carbonate fusion method, 59-60
804-805 Fe determination, 642-643
in soils, 796-803 P determination, 871-81'2
SEM. See Scanning electron microscopy Sodium citrate-dithionite method, Al extraction
Semiquinone radical, 338, 340 from poorly ordered phases, 530-531
Sensitivity Sodium determination
atomic absorption spectroscopy, 74-76 exchangeable Na, 571
electron spin resonance spectroscopy, soluble Na, 571
331-333 Sodium hydroxide method, extraction of humic
Separatory funnel method, extraction of organ- substances, 1017
ic chemicals from aqueous samples, Sodium hydroxide fusion method, F determina-
1075 tion, 850-852
Serial dilution, 42 Sodium hydroxide-hydrochloric acid-sodium
Serial training, 27 hydroxide extraction, organic P deter-
SEXAFS. See Surface sensitive extended x-ray mination, 876-877
absorption fine structure spectros- Sodium hypobromite oxidation method, P
copy determination, 873-874
SFM. See Scanning force microscopy Sodium peroxide, fusion with
Shear plane, 1241 method, 61-62
SUBJECT INDEX 1385
principles, 53 Soil-base titration method, determination of

Sodium phosphate method, Al extraction from lime requirement, 496-497, 499-501
organic matter and organic soils, Soil-buffer equilibration method, determination
529-530 of lime requirement, 496-497, 501-511
Sodium pyrophosphate method, organically Adams-Evans single buffer method, 498,
bound Fe determination, 651 505--508
Soil acidity, 475-489. See also pH, soil Mehlich single buffer method, 508--510
active, 493-494 Nommik single buffer method, 510
Alin,518,522 Shoemaker-McLean-Pratt single buffer
capacity factor, 491 method, 498, 502-505
exchangeable,492,494,496-497 Woodruff single buffer method, 510--511
intensity factor, 491 SOILCHEM, 1312, 1324--1325
lime requirement, 491-513 Soil-lime incubation method, determination of
nature of, 493-494 lime requirement, 496-497, 499-500
nonexchangeable,494 Solid electrode, 259
total potential, 492 Soller slit, 390
Soil biota, 1011 Soluble salts, effect on cation exchange capaci-
Soil classification, 1205-1206 ty measurement, 1210--1214
Soil classification unit, 2 Solution phase speciation, 1327-1335
Soil colloid, Fourier transform infrared spec- Sonication extraction, organic chemicals from
troscopy, 288--294 solid samples, 1077-1078
Soil digest, atomic absorption spectroscopy, SOPs. See Standard operating procedures
Soxhlet extraction, 1077
84--85
SPADNS method, F determination, 859-860
Soil extract
SPC. See Statistical process control
atomic absorption spectroscopy, 85-86
Speciation
Hg determination, 784--787
Al in solution, 534--538
ICP-MS and ICP-AES, 125-126, 128--131
physicochemical separation methods,
inorganic N determination, 1125, 1131-1171
536-538
organic C determination, 988
timed spectrophotometric assays, 534--536
Soil horizon, 2
applications of anodic stripping voltammetry,
Soil morphology, 1261
260--262
Soil pH. See pH, soil
applications of differential pulse polarogra-
Soil properties, effect on lime requirement,
phy, 260--262
494-496
As in soil solutions and water extracts,
Soil redox. See Redox, soil 820-824
Soil reducing intensity for Cr(Vl), 696--697 calculations with GEOCHEM-PC,
Soil salinity. See Salinity 1327-1339
Soil solution mass transfer and, 1335-1336
Al determination, 531-534 Se in soil solutions and water extracts,
extraction of samples in laboratory, 805-808
532-533 solution phase, 1327-1335
filtration of soil solution samples, XANES analysis, 393--398
533--534 Specific adsorption, 1231, 1237
lysimeters, 532 Spike recovery data, 34
alkaline-earth metals in, 581 Spiked sample, 42
anodic stripping voltammetry, 260--264 Spin concentration, 336-337
As determination, 818-819 Spin content, humic substances, 339-340
As species in, 820-824 Spin derivatization, 347-351
atomic absorption spectroscopy, 85-86 Spin Hamiltonian, 326-327, 334
Cd determination, 752 Spin labels, 347-348
differential pulse polarography, 260--264 Spin orbit splitting, x-ray photoelectron spec-
Fe determination, 641 troscopy, 368
K determination, 553 Spin traps, 348
Ni determination, 752 Spin-lattice relaxation time, 326, 332-333, 354
Pb determination, 752 Spin-orbit coupling, 327
Se determination, 804--805 Spin-relaxation time, 354
Se species in, 805-808 Spin-spin interactions, 333
Soil tests. See specific elements Spline function, 399, 401
Soil variation, 1-3 Split samples, 39, 43-45
1386 SUBJECT INDEX
SPM. See Scanning probe microscopy Strontium, 575-598. See also Alkaline-earth
Sputter gun, 364 metal(s)
Standard deviation availability indices, 585-586
long-term, 40 mineralogy and distribution in rocks and
relative, 33-35, 38-39 soils, 576-577
short-term, 40 properties, 575-576
Standard hydrogen electrode, 1268 soluble, 581
Standard operating procedures (SOPs) Strontium determination
analytical methods, 22-25 atomic absorption spectrophotometry, 578,
analytical structure, 22-23 587-589
data management, 30-31 electrothermal atomic absorption, 590
documentation, 31 extractable/exchangeable fraction, 582-584
employee responsibility, 23 acid soil extraction, 584
facility management, 28-29 ammonium acetate method, 583-584
format, 22 ammonium chloride method, 584
instrument and equipment care, 29-30 flame emission photometry, 590
management responsibility, 23 inductively coupled plasma-atomic emission
organization, 22-23 spectrometry, 579-580, 589-590
quality assurance structure, 23 ion chromatography, 591-593
record keeping, 25-27 titrimetric method, 593-596
training program, 27-28 x-ray fluorescence, 578-579
Standard reference material, 34-36, 38-39, 42 Subsampling, 12-16
Static mercury drop electrode (SMDE), 258 optimum rate, 14-15
Static surface charge, 361
Sulfate
Stationary relaxation methods, 1295-1296
oxidation of total S to sulfate, 926, 931-933
Statistical process control (SPC), 35
reduction to sulfide, 932-933
Steam-distillation method
Sulfate determination, 926
inorganic N determination, 1125, 1127,
extraction of sulfate-S, 950-952
1131-1138
inorganic sulfate, 938-943
in absence of nitrite, 1135
ion chromatography, 941
in presence of nitrite, 1135-1136
methylene blue method, 927, 932-936,
organic N determination, 1187-1196
separation of F from perchloric acid, 940-941
853-855 principles, 938-940
separation of F from sulfuric acid, 855-856 Sulfur, 921-955
Stem layer, 1238 availability indices, 949-955
Stem model, 1238 chemistry, 921-925
Stem potential, 1238 speciation, XANES analysis, 393-396
Stem thickness, 1238 Sulfur determination
Stirred-batch reactor acetate-extractable S, 953
apparatus, 1277-1278 bicarbonate-extractable S, 953-954
uses and advantages, 1278-1279 heat-soluble S, 952-953
Stirred-flow reactor, 1282-1286 inorganic sulfate, 938-943
advantages, 1283 ion chromatography, 924
apparatus, 1282-1283 organic S content, 943-949
data analysis, 1286 C-bonded sulfur, 946-949
disadvantages, 1283 hydriodic acid-reducible S, 944-946
distinguishing instantaneous and kinetic reac- oxidation of total S to sulfate, 926, 931-
tions, 1284-1286 933
STM. See Scanning tunneling microscopy acid digestion, 931
Stokes scattering, 274 alkaline oxidation, 931
Stopped-flow technique, 1287-1290 dry ashing, 931-932
apparatus, 1288-1289 soil tests
background, 1287-1288 extraction of organic S, 952-955
electron paramagnetic resonance, 1289- extraction of sulfate-S, 950-952
1290 total S content, 925-938
Stratified random sample, 7-9, 11 automated instrumental methods, 933,
s-triazine, 341 937
Stripping voltammetry, halide determination, principles, 925-928
863 wet chemical methods, 925-933
SUBJECT INDEX 1387
Sulfuric acid Thermochemical data

digestion with limitations, 1350
method, 57-58 nonideality corrections, 1321-1322
principles, 52 sources, 13 21
in soil, 477 temperature corrections, 1323
Sulfuric acid method, F determination, 852-853 Thermodynamic constants, electron spin reso-
Sulfuric acid-hydrogen peroxide-hydrofluoric nance parameters and, 351
acid digestion, P determination, Thermometer, electrical conductivity determina-
872-873 tion, 423
Supercritical fluid extraction, organic chemicals Thin film, XRFS, 199
from solid samples, 1078-1080 Thiocyanate-stannous chloride method, Mo
Superhyperfine coupling constant, 326, 342 determination, 725-728
Superhyperfine splitting, 325, 328-329 Thiosulfate, 922
Supporting electrolyte, 259 Thorium nitrate method, F determination
Suppressed gradient method, alkaline-earth in presence of alizarin red S, 862-863
metal determination by ion chromatog- in presence of chrome azurol S, 860---862
raphy, 591-593 Timed spectrophotometric assay, Al speciation
Surface charge, 1231 in solution, 534-536
charge/potential relationships, 1236-1237 Tiron method, Al determination, 540
constant charge, 1234 Titrimetric method
measurement, 1241-1242 Al determination, 542-543
source, 1234-1237 alkaline-earth metal determination, 593-596
variable charge, 1234-1236 carbonate determination, 448-451
Surface charge density, 1234 F determination, thorium nitrate method
Surface complexation models, 1315-1321 in presence of alizarin red S, 862-863
applications in presence of chrome azurol S, 860---862
anion adsorption, 1319
Total anion concentration, 426
cation adsorption, 1320
Total cation concentration, 426
parameter estimation, 1318
Total chromium(Vl) reducing capacity, 696
Surface functional groups, 1317
Total dissolved solids, 417-433
Surface ionization reactions, equilibrium con-
apparatus, 432
stants, 1318
aqueous extract, 431-433
Surface probing microscopy, 378
calculations, 432-433
Surface sensitive extended x-ray absorption fine
principles, 431-432
structure (SEXAFS) spectroscopy, 392
Surrogate spiking solution, monitoring extrac- procedure, 432
tion of organic chemicals, 1082-1083 waters, 431-433
Suspension effect, on soil pH, 482-483 Total element analysis. See also specific ele-
Suspension postcolumn reaction suppressor, ments
238 dissolution methods, 49----63
Symmetry information, x-ray absorption fine sample preparation, 55-56
structure spectroscopy, 393-396 Total elemental concentrations, 141
Synchrotron, 385-387 Total hydrolyzable nitrogen, 1186, 1190---1191
System audit, 32 Toxicity of elements, 475
Systematic sample, 9-11 Traceability, 21, 45-46
one-dimensional case, 10---11 Trading rules, Fourier transform infrared spec-
two-dimensional case, 10---11 troscopy, 282
Systematic square grid pattern, 11 Training program, standard operating proce-
dures, 27-28
Tamm's reagent, 649----650 Transient relaxation methods, 1295-1296
Target pH, lime requirement, 491-492 Transition metal, paramagnetic. See Paramag-
TEM. See Transmission electron microscopy netic metal ions
Temperature corrections, thermochemical data, Transmission electron microscopy (TEM), 378
1323 Transport phenomenon, 1295
Temperature-jump relaxation methods, 1296 Triple layer model, 1315
Test kit, pH determination, 488 True value, 40-41
Test portion, 33 Tube digestion method, organic C determina-
Tetrathionate, 922 tion, 997-998, 1000
Texture, soil, 1261-1262 Tube techniques, kinetic methods, 1276-1277
effect on lime requirement, 495-496 "Typical" site, 3-4
1388 SUBJECT INDEX
Ultracentrifugation, humic substances, Water salinity, 418

1057-1058 Waters
Ultrafiltration, fractionation of humic sub- electrical conductivity, 422--427
stances, 103(}..1031 Hg determination, 784-787
Ultrasonic nebulizer, 98--101 total dissolved solids, 431--433
Ultraviolet difference method, inorganic N Wavelength-dispersive spectrometer, 161-162,
determination, 1127-1129 174-176, 178--179
Ultraviolet spectroscopy, 291 procedure for use, 206--209
Ultraviolet-visible spectroscopy, humic sub- Wavenumber, 271
stances, 1044--1045 Weathering, effect on lime requirement,
Unbuffered salt extraction method, cation 494--495
exchange capacity determination, Wet combustion method
1218--1220 C determination, 963-964, 977-983
Unpaired electron, 324 organic C determination, 985-988, 1000
pretreatment, 986--987
Valence electron, 365 Wetlands, 1261
Validation of methods, 24 White line, 381, 393
Validity coefficient, 1347 Willard-Winter distillation method, F determi-
Vanadomolybdophosphoric acid method, P nation, 847
determination, 907 "Within-strata" mean square, 8
Vanadyl ion Woodruff single buffer method, determination
complex with organic substances, 346--347 of lime requirement, 51(}..511
electron spin resonance spectroscopy, Work function of the sample, 360
346--347
Vanselow convention, 1207 XAD-8 resin, 1019-1020
Vanselow selectivity coefficient, 1208, XAFS spectroscopy. See X-ray absorption fine
1223-1226, 1314 structure spectroscopy
Variable charge, 1201, 1234-1236 XANES analysis. See X-ray absorption near
Variance of the mean, 5-8, 14-15 edge fine structure analysis
on a per element basis, 13 XPS. See X-ray photoelectron spectroscopy
Variation among units, 13-14 X-ray(s)
Vibrational modes, 271-272 interaction with matter
Vibrational spectroscopy, 269-270, 274-275, absorption, 167-168
378 dispersion (diffraction), 168--169
Vibrational transitions, 272 enhancement, 168
Viscosity, humic substances, 1058--1059 scattering, 168
Vitreous carbon crucible, 62 properties, 163-164
Volatile organic chemicals, extraction, X-ray absorption fine structure (XAFS) spec-
1071-1083 troscopy, 377--412
Voltammetry, 247 absorption edge, 38(}..381, 389, 391, 393
anodic stripping. See Anodic stripping backscatter, 403, 410
voltammetry beam line hardware, 385-387
chemically modified electrode, 256 data analysis, 411--412
Cr determination, 691 electron, symmetry, and chemical environ-
modem techniques, 255-256 ment determination, 393-396
Volume weight of soil, 714, 716 experimental and data analysis, 387--412
Volumetric calcimeter method, carbonate deter- experimental considerations, 387-392
mination, 451--455 extended. See Extended x-ray absorption
fine structure spectroscopy
Walkley-Black method, organic C determina- fluorescence detection, 389-391
tion, 993, 995-996, 1000 Fourier-filtered spectra, 405
Warning limits, 36--37 glancing angle, 393
WATEQ3, 1325 incident intensity loss, 390
Water, ion product, 476 information sources, 412
Water extract interferences, 390
As determination, 818--819 K-edge analysis, 389-390
As species in, 82(}..824 L-edge analysis, 390
P determination, 891-893 oxidation state determination, 381, 393-394
Se determination, 804-805 phase shift, 404--405
Se species in, 805--808 principles, 379-385
SUBJECT INDEX 1389
signal-to-noise ratio, 389 sample preparation

solid-state detectors, 390-391 fused discs, 198--205
special experimental designs, 392-393 liquids, 199--200, 214-215
speciation, 393--394 loose powders, 197, 214-215
structural analysis, 398-409 pressed pellets, 197-198, 211-214
synchrotrons, 385-387 thin films, 199
transmission detection, 389, 391 wavelength-dispersive spectrometer,
white line, 381, 393 161-162, 174-176, 178--179,206--209
X-ray absorption near edge fine structure X-ray photoelectron spectroscopy (XPS),
(XANES) analysis, 380, 392-395 357-374
S and metal speciation in soils, 396--398 analysis, 365
structural analysis, 398-409 angle resolved, 371-372
X-ray detector atomic concentrations calculations, 369--371
gas-filled, 169-170 charge-shifted C(ls) peak, 364
proportional, 169--170 depth profile studies, 371-372
scintillation, 170 fixed analyzer transmission, 363
semiconductor (solid-state), 171-172 fixed retarding ratio, 363
X-ray excitation, 164 information gleaned from, 357-358
X-ray fluorescence spectroscopy (XRFS), instrumentation, 362-364
161-218 detector, 363--364
analysis multiplet splitting, 369
major elements, 200-211 principles, 358--362
minor and trace elements, 211-215 quantitative analyses, 369--371
Al determination, 542 recent developments, 373
alkali metal determination, 556--557 referencing binding energy, 364-368
alkaline-earth metal determination, 578--579 sample preparation, 372-373
applications, 215-218 sampling handling and treatment, 373
Br determination, 863 satellites, 369
detection limits, 188--190 sensitivity factors, 369
energy-dispersive spectrometer, 162, 172, spin orbit splitting, 368
176--179, 209--211 x-ray source, 364
errors, 185-188 X-ray tube, 164-166
instrumental, 187 selection and operation, 166
measurement and statistical, 187-188 X-ray white light, 386
operator, 185-187 XRFS. See X-ray fluorescence spectroscopy
sample representation and preparation,
187 Yellow silicomolybdic acid method, Si determi-
instrumentation, 173--181 nation, 634-635
selection, 179--181
interferences, 190-197 Zeeman background correction, 81--83
background, 190 Zeeman effect, 324
spectral, 191 Zeolites, carbon exchange capacity of soils con-
matrix effects, 190-197 taining, 1213--1214
principles Zero-order reaction, 1291
counting efficiency, 173, 188--190 Zeta potential, 1241
interaction of x-rays with matter, 167-169 Zinc, 703-719
nature and production of x-ray spectra, availability indices, 706--717
163--166 available forms in soil, 704
pulse-height selection, 172-173 chemistry, 703--704
x-ray detection and measurement, deficiency, 703--704
169--172 labile, 704
problems associated with, 162 nonlabile, 704
qualitative analysis, 181-182 nonspecific adsorption, 703
quantitative analysis, 182-188 occluded, 704, 709
reducing matrix effects soil properties vs. deficiencies, 704-705
compensation methods, 193--195 specific adsorption, 703--704
dilution methods, 192-193 uncontaminated soils, 704
linear calibration methods, 192 Zinc determination
mathematical correction models, 195-197 soil tests, 706--707
safety, 179--181 dilute hydrochloric acid method, 715-717
1390 SUBJECT INDEX
Zinc determination (cont.)

DTPA-AB method, 706, 709-711
DTPA-TEA method, 706--709
field calibration, 705-706
greenhouse evaluation, 705-706
Mehlich I extraction, 711-713
Mehlich III extraction, 706, 713-715
sample and extractant preparation, 707
total Zn content, hydrofluoric, nitric, per-
chloric, and sulfuric acid method,
717-719
Zirconium crucible, 61

Methods of Soil Analysis

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Methods of Soil Analysis

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Published 1996

METHODS OF SOIL ANALYSIS

1. MINERALS IN SOIL ENVIRONMENTS. Second Edition. 1989.

2. PESTICIDES IN THE SOIL ENVIRONMENT: PROCESSES, IMPACTS,

3. SOIL TESTING AND PLANT ANALYSIS. Third Edition. 1990.

4. MICRONUTRIENTS IN AGRICULTURE. Second Edition. 1991.

5. METHODS OF SOIL ANALYSIS: PHYSICAL AND MINERALOGICAL

METHODS OF SOIL ANALYSIS: CHEMICAL AND MICROBIOLOGICAL

METHODS OF SOIL ANALYSIS: CHEMICAL METHODS. Part 3. 1996.

Editorial Committee: D. L. Sparks

Managing Editor: J.M. Bartels

Editor-in-Chief SSSA: J. M. Bigham

Number 5 in the Soil Science Society of America Book Series

Published by: Soil Science Society of America, Inc.

ALL RIGHTS RESERVED UNDER THE U.S. COPYRIGHT LAW OF

Second printing 1999.

Soil Science Society of America, Inc.

Library of Congress Cataloging-in-Publication Data

Library of Congress Catalog Card Number: 96-70096

Printed in the United States of America

2 Quality Assurance and Quality Control

3 Dissolution for Total Elemental Analysis

4 Atomic Absorption and Flame Emission Spectrometry

5 Inductively Coupled Plasma Emission Spectrometry

6 Neutron Activation Analysis

7 Elemental Analysis by X-Ray Fluorescence Spectroscopy

9 Differential Pulse Voltammetry

10 Fourier Transform Infrared and Raman Spectroscopy

11 Electron Spin (or Paramagnetic) Resonance Spectroscopy

12 X-Ray Photoelectron Spectroscopy

13 X-Ray Absorption Fine Structure Spectroscopy

14 Salinity: Electrical Conductivity and Total Dissolved Solids

15 Carbonate and Gypsum

16 Soil pH and Soil Acidity

19 Lithium, Sodium, Potassium, Rubidium, and Cesium

20 Beryllium, Magnesium, Calcium, Strontium, and Barium

26 Copper and Zinc

27 Molybdenum and Cobalt

28 Nickel, Cadmium, and Lead

30 Selenium and Arsenic

31 Bromine, Chlorine, and Fluorine

34 Total Carbon, Organic Carbon, and Organic Matter

35 Organic Matter Characterization

36 Extraction of Organic Chemicals

40 Cation Exchange Capacity and Exchange Coefficients

41 Charge Analyses of Soils and Anion Exchange

42 Redox Measurements of Soils

43 Kinetic Methods and Measurements

44 Equilibrium Modeling in Soil Chemistry

The editorial committee expresses its sincerest gratitude to the anonymous

D.L. Sparks, Editor-in-Chief

AL. Page, Associate Editor

P.A. Helmke, Associate Editor

R.H. Loeppert, Associate Editor

Y.O.Aochi Staff Research Associate, Department of Soil and Environmental

Richmond J. Bartlett Professor of Soil and Environmental Chemistry, Department of Plant

J. K. Bartz Chemist, Washington State Department of Ecology, 1315 W. 4th Ave-

Paul M. Bertsch Professor, Advanced Analytical Center for Environmental Science,

John M. Bremner Distinguished Professor Emeritus, Department of Agronomy, Iowa

L.D.Calvin Professor Emeritus of Statistics, Department of Statistics, Oregon

Theodore H. Carksi Research Associate/Supervisor, Agricultural Products, Stine-Haskell

D.LCocke J.M. Gill Professor of Chemistry, Department of Chemistry, Lamar

James G. Crock Analytical Geochemist, Branch of Geochemistry, Analytical Chemis-