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Communications in Soil Science and

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The determination of boron in soil

extracts, plant materials, composts,
manures, water and nutrient solutions
a
Benjamin Wolf
a
Dr. Wolf's Agricultural Laboratories, Ft. Lauderdale, Florida
Published online: 11 Nov 2008.

To cite this article: Benjamin Wolf (1971): The determination of boron in soil extracts, plant
materials, composts, manures, water and nutrient solutions, Communications in Soil Science and
Plant Analysis, 2:5, 363-374

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 SOIL SCIENCE AND PLANT ANALYSIS, 2(5) 363-371* (1971)

THE DETERMINATION OF BORON IN SOIL EXTRACTS, PLANT MATERIALS,

COMPOSTS, MANURES, WATER AND NUTRIENT SOLUTIONS

KEY WORDS: s o i l t e s t i n g , plant a n a l y s i s , leaf or p l a n t

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Benjamin Wolf

Dr. Wolf's A g r i c u l t u r a l Laboratories

Ft. Lauderdale, F l o r i d a

ABSTRACT

A rapid c o l o r i m e t r i c method f o r t h e determination of boron i n s o i l ex-

t r a c t s , p l a n t m a t e r i a l s , composts, manures, water and n u t r i e n t s o l u t i o n i s

proposed. The method i s r a p i d , r e l i a b l e and c a r r i e d out i n aqueous s o l u t i o n .

A marked advantage i s t h a t boron can be determined i n t h e same s o i l e x t r a c t

or p l a n t m a t e r i a l d i g e s t used f o r determination of other elements.

INTRODUCTION

The rapid method f o r t h e determination of boron i n s o i l e x t r a c t s , p l a n t

m a t e r i a l s and i r r i g a t i o n waters i s d e s i r a b l e when devising p r a c t i c a l programs

of boron f e r t i l i z a t i o n . Boron i s an e s s e n t i a l element f o r p l a n t s and i t s

a p p l i c a t i o n i s complicated since t h e r e i s a r e l a t i v e l y narrow range between

deficiency and t o x i c i t y . Knowing t h e boron content of manures, composts and

n u t r i e n t s o l u t i o n s can also be highly u s e f u l f o r c e r t a i n programs.

My e a r l y work with boron ( 1 , 2) was with t h e q u i n a l i z a r i n method as p r o -

posed by Smith (3) b u t with some minor modifications. L a t e r , I used t h e quina-

l i z a r i n method as proposed by Berger and Truog (4) and continued to use t h i s

363
Copyright © 1971 by Marcel Dekker, Inc. NO PART of this work may be reproduced or utilized in any
form or by any means, electronic or mechanical, including xerography, photocopying, microfilm, and re-
cording, or by any information storage and retrieval system, without the written permission of the pubtisher.
 BENJAMIN WOLF

method u n t i l refinements i n the curcumin method (5) allowed f o r rapid d e t e r -

mination of boron. I modified the curcumin method a s o r i g i n a l l y proposed (5)

for aqueous samples, such as s o i l e x t r a c t s and d i g e s t s of wet-ashed plant

tissues.

Both the quinalizarin and curcumin methods have been far from ideal for

routine rapid analyses. The quinalizarin method has the disadvantage since

strong sulfuric acid i s required and sensitivity i s affected by the water con-
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tent of the acid. I t , as well as the curcumin method, i s affected by the pres-

ence of nitrates. In the curcumin method I was able to overcome the effects of

about 25 ppm nitrate-N(100 pp2m in the soil) by use of a reducing agent proposed

by Premi and Cornfield (6). The presence of larger amounts of nitrates in

soils, as often occurs under intensive cropping, required prior destruction of

nitrates with the danger of losing boron as well as being time consuming. The

curcumin method i s also affected by presence of salts which seems to give a

negative error (7).

These disadvantages have prompted me to examine over the years a number of

other boro^i methods. This examination, although superficial in some cases, in-

cluded the following methods: chromatrope (8), Spadns (9), waxoline purple or

ahcoquinone (10), carmine (11) and the potentiometric determination as the tetra-

fluoroborate (12). None were found of advantage because they were either too

time consuming, lacked sensitivity, or necessitated the use of strong acids.

Recently, an automated method using azomethine-H has been proposed for

determination of boron in plant materials (13). The method i s rather sensitive,

subject to l i t t l e interferences and works in an aqueous medium. The authors (13)

prepared their own azomethine-H, a procedure which i s somewhat time consuming.

The availability of a commercial product prompted me to examine i t s possibilities

for determining boron in soil extracts, plant materials, composts, manures,

irrigation waters and nutrient solutions. Suitable methods were developed

and these procedures are described in this paper.

361*
 DETERMINATION OF BORON

MATERIALS AND METHODS

A. Reagents-General:

1. Buffer-masking agent: d i s s o l v e 250 gins of ammonium a c e t a t e ,

25 gms of the tetrasodium s a l t of ( e t h y l e n e d i n i t r i l l o ) t e t r a -
a c e t i c acid and 10 gms of the disodium s a l t of n i t r i l o t r i a c e t i c
acid i n 400 ml of water and slowly add 125 ml of a c e t i c acid.
2. Azomethine-H: d i s s o l v e 0.9 gm of azomethine-H and 2 gms of
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ascorbic acid i n 10 ml of water with g e n t l e heating i n a water

bath, and make up t o 100 ml with water. I f t u r b i d , i t should
be heated again i n the water b a t h , making up t o the o r i g i n a l
volume a f t e r cooling. I t i s claimed t h a t t h i s reagent w i l l
keep f o r 14 days i f stored i n a r e f r i g e r a t o r (13).

3. Standard boron s o l u t i o n (100 ppm B): d i s s o l v e 0.5716 gms b o r i c

acid i n water and make up t o 1 l i t e r .

B. Reagents-Soil:
1. Morgan's Universal E x t r a c t i n g Solution (14): dissolve 100 gms of
sodium a c e t a t e i n l e s s than 1 l i t e r of water, adjust the pH with
a c e t i c acid t o 4.8 and make up t o 1 l i t e r .

2. Activated Charcoal Darco G-69: used t o decolorize waters a s

well as s o i l e x t r a c t s .

3. Soil boron standard ( 2 ppm B): take 2.0 ml of the boron standard .
(100 ppm) and d i l u t e t o 100 ml with Morgan's e x t r a c t i n g s o l u t i o n .
C. Reagents-Plant M a t e r i a l s , Composts and Manures:

1. S u l f u r i c a c i d , concentrated
2. Sucrose

3. Hydrogen peroxide, 30%

4. Standard boron s o l u t i o n (4 ppm B): take 1.0 ml of the general
boron standard (100 ppm) and d i l u t e with 25 ml of the wet ash
blank ( d i r e c t i o n s f o r preparing the wet ash blank a r e given

365
 BENJAMIN WOLF

under procedures f o r wet ashing p l a n t m a t e r i a l s , composts and

manures).

D. Reagents-Water and N u t r i e n t Solutions:

1. Standard boron s o l u t i o n (4 ppm B): take 4.0 ml of the general

boron standard (100 ppm) and d i l u t e t o 100 ml with water.

All reagents should be c.p. a n a l y t i c a l grade wherever p o s s i b l e . The

p r a c t i c a l grade f o r tetrasodium s a l t of EDTA and disodium s a l t of NTA i s

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sufficient.

All water solutions should be stored in plastic or low boron glass. Corn-

ing brand, alkali resistant, lime glass, Vycor brand, or silica i s satisfactory.

Even pyrex can be used for short duration storage if soaked overnight in strong

HC1.

SAMPLE PREPARATION

A. Soil:

Measure 12.5 ml of s o i l and 1/4 teaspoon of a c t i v a t e d charcoal

into a 50-ml f l a s k or b o t t l e , add 25 ml of Morgan's e x t r a c t i n g

s o l u t i o n and shake f o r 5 minutes. F i l t e r through a Whatman #1

f i l t e r paper. The f i l t r a t e should be clean and c o l o r l e s s . I f

colored, add 1/4 teaspoon of a c t i v a t e d c h a r c o a l , shake and f i l t e r

immediately. I f t u r b i d , t h e f i l t r a t e should be centrifuged t o

obtain a c l e a r supernatant.

B. Plant M a t e r i a l s , Composts and Manures:

Weigh 0.3 gm of d r i e d , ground (passing 40 mesh) m a t e r i a l i n t o

200 x 25 mm Vycor tube marked t o hold 15 ml. Add 2 ml of concen-

t r a t e d s u l f u r i c acid and r o t a t e t h e tube so t h a t a l l t h e m a t e r i a l

i s moistened by the acid. Care should be exercised t o avoid work-

ing t h e m a t e r i a l up t h e s i d e of t h e tube. Let stand f o r 15 minutes.

Add 1 ml of 30% hydrogen peroxide. The contents a r e mixed by r o t a -

t i n g , slowly a t f i r s t , u n t i l well mixed. Place a small funnel, 40

366
 DETERMINATION OF BORON

mm d i a . a t t h e top and 45 mm i n stem l e n g t h i n t h e tube. Put the

tube i n a hole of an aluminum block, approximately 65 mm deep with

openings of 26 mm d i a . x 55 mm deep. Set the block on a hot p l a t e

in a fume hood. Set the temperature a t 500° C and heat u n t i l dense

fumes a r e produced f o r about 5 minutes. Remove t h e tube from t h e

hot p l a t e , cool and add another ml of 30% hydrogen peroxide. Heat

again u n t i l fumes a r e given off f o r about 20 minutes. Remove t h e

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tube and cool. I f the c o n t e n t s a r e not c o l o r l e s s , add 5 drops of

30% hydrogen peroxide, heat again u n t i l fumes a r e produced f o r 20

minutes. Remove from the hot p l a t e and cool. I f the contents are

s t i l l not c o l o r l e s s , t h e p r o c e s s of adding 2 drops of hydrogen peroxide

and h e a t i n g f o r 20 minutes a f t e r fumes a r e produced i s repeated u n t i l

a colorless digest i s obtained.

Once the contents are colorless, dilute with water to the 15 ml

mark. Mix and f i l t e r through a Whatman #40 paper.

Wet-ash blanks are prepared by digesting 0.3 gm sucrose in the

same manner as the test materials, diluting and filtering as des-

cribed above.

C. Water and Nutrient Solutions:

Samples should be free of suspended matter and colorless. If

turbid, centrifuge; if colored, add 1/4 teaspoon activated charcoal

either before centrifuging (if turbid) or before filtering if colored

and free of suspended matter.

PROCEDURE FOR BORON DETERMINATION

A. Soil:

P i p e t t e 4 ml of the s o i l e x t r a c t i n t o a photometric t u b e , add

1 ml each of the buffer-masking reagent and of azomethine-H i n t h a t

order. Immediately s t i r , l e t s e t f o r 1 hour and read a t 430 mu.

36T
 BENJAMIN WOLF

Sample readings a r e compared with s e r i e s of s t a n d a r d s (0, 0.125,

0.25, 0.5, 1.0, and 2.0 ppm boron) prepared a t the same time and

t r e a t e d i n t h e same way as the s o i l e x t r a c t s . These s t a n d a r d s a r e

prepared by d i l u t i n g t h e s o i l 2 ppm boron standard with t h e appro-

p r i a t e amount of Morgan's e x t r a c t i n g s o l u t i o n . The e x t r a c t i n g s o l u -

t i o n alone serves as the 0 ppm boron standard.

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B. P l a n t M a t e r i a l s , Composts and Manures:

P i p e t t e 2 ml of t h e wet-ashed d i g e s t i n t o a photometric t u b e ,
add 4 ml of buffer-masking reagent and 2 ml of azomethine-H reagent
in t h a t o r d e r . Stopper the tube and mix thoroughly by i n v e r t i n g 3
times. Let stand f o r 2 hours and read a t 430 mu.

Sample readings a r e compared with a s e r i e s of s t a n d a r d s (0, 0.2,

0.5, 1.0, 2.0, and 4.0 ppm boron) prepared a t the same time and
t r e a t e d i n the same manner as the d i g e s t s . These standards are p r e -
pared by d i l u t i n g the 4 ppm boron p l a n t m a t e r i a l , manure and compost
standard t o 2 ml with the wet-ash blank. The wet-ash blank serves
as the 0 ppm boron standard.

C. Water and N u t r i e n t Solution;

P i p e t t e 4 ml of the c l e a r c o l o r l e s s sample i n t o a photometric

tube, add 1 ml each of the buffer-masking and then the azomethine-H

reagent i n t h a t o r d e r . S t i r immediately and read a f t e r 1 hour a t

430 mu.

Sample readings a r e compared with a s e r i e s of standards ( 0 , 0.25,

0.5, 1.0, 2.0, and 4.0 ppm boron) t r e a t e d a t the same time and manner

as t h e water and n u t r i e n t s o l u t i o n s . These standards a r e prepared by

d i l u t i n g the 4 ppm boron standard with a p p r o p r i a t e amounts of water.

368
 DETERMINATION OF BORON

DISCUSSION

A. Interferences:

Barron, Bohmer and Stanton (13) found t h a t Al, Cu and Fe i n t e r -

f e r e d with t h e azomethine-H method f o r determining B i n p l a n t m a t e r i a l .

The i n t e r f e r e n c e was overcome by u s e of a 0.25 M EDTA (disodium s a l t )

solution. However, I found t h a t i t was inadequate f o r c e r t a i n s o i l s

which were high i n Al and a l s o f o r c e r t a i n garbage composts which were

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q u i t e high i n Fe a s well a s Al. Adding more EDTA s o l u t i o n was n o t

satisfactory since the additional dilution reduced the color intensity

and, therefore, the sensitivity. Increasing the concentration of the

EDTA solution was not practical because of the limited solubility of

the disodium salt of EDTA.

Use of the tetrasodium salt of EDTA was a partial and practical

solution of these problems. Sufficient tetrasodium salt could be

dissolved and added with the buffer to overcome the interference of

Cu, Fe, and Al in most samples. This buffer-masking solution, however,

tended to deteriorate rapidly because of precipitation of salts. The

addition of the disodium salt of NTA gave much longer shelf l i f e as

well as eliminating interferences from metals in certain composts.

Other interferences were also found. When boron was determined

in soil extracts, no buffer was used at f i r s t because Morgan's extract-

ing solution is already buffered at pH 4.8, a suitable pH for the

azomethine-H reaction. However, ammoniacal nitrogen in ranges commonly

found in fertile soils gave a positive error. When large amounts of

ammonium are added with the buffer-masking reagent, the interference

disappeared.

Even small amounts of hydrogen peroxide can interfere with the

test. In the wet digestion of plant materials, composts and manures,

369
 BENJAMIN WOLF

i t i s necessary t o b o i l the s o l u t i o n f o r a t l e a s t 10 minutes a f t e r

adding the l a s t a l i q u o t of hydrogen peroxide i n order t o remove a l l

t r a c e s of hydrogen peroxide.

Stoppering and mixing by i n v e r t i n g 3 times eliminates poor

r e s u l t s t h a t can be obtained with p l a n t m a t e r i a l s and composts.

B. Concentration Ranges:
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1. Soils; Using 1 p a r t s o i l to 2 p a r t s e x t r a c t i n g s o l u t i o n permits

the determination of 0-4 ppm or 0-8 pp2m i n the s o i l , a range

covering inadequate t o t o x i c l e v e l s of B f o r most crops.

2. P l a n t m a t e r i a l s , composts and manures: The 0-4 ppm B range i n t h e

d i l u t e d wet-ash d i g e s t s permits the determination of 0-200 ppm B

on a dry weight b a s i s . For many p l a n t s , t h i s also covers the

range from deficiency to excess.

3. Water and n u t r i e n t s o l u t i o n s : The method i s most accurate f o r

the range 0-2 ppm B but can be used f o r values up to 4 ppm B

which covers the amount of boron normally found in i r r i g a t i o n

water and d i l u t e n u t r i e n t s o l u t i o n s . For more accurate d e t e r -

mination i n the 2-4 ppm B range in i r r i g a t i o n w a t e r s , i t i s

b e s t to use only 2 ml sample d i l u t i n g with 2 ml of water be-

f o r e adding the buffer-masking reagent.

C. Comparison With Other Methods:

B determinations using the azomethine-H method gave r e s u l t s i n

good agreement with the curcumin method and probably more r e l i a b l e

r e s u l t s f o r s o i l s with high n i t r a t e contents. For p l a n t s , the azome-

thine-H r e s u l t s agreed more c l o s e l y with spectrographic analyses than

with the curcumin r e s u l t s . All t h e s e comparisons a r e given i n Table 1.

370
 DETERMINATION OF BORON
TABLE 1
Results of t h e Analysis of Several M a t e r i a l s by t h e Proposed Method a s Compared
With R e s u l t s Obtained by Curcumin or Spectrographic Analysis.

A n a l y t i c a l Method

Material Azomethine-H Curcumin Spectrographic

pp2m pp2m

Soils
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Lakewood sand 0.8 0.7

Parkwood Sandy loam 2.7 2.8

Norfolk sand 0.8 3.2*

Plant Tissue, Compost, Water & Nutrient Solution

ppm ppm ppm

Composite l e a f t i s s u e 23 21 24

Chrysanthemum l e a v e s 108 92 112

(Toxicity symptoms)

Garbage compost 28 25

I r r i g a t i o n water 1.5 1.5

I r r i g a t i o n water 1.9 1.6

Nutrient solution 0.5 0.6

*0ver 200 pp2m of nitrate-N in t h i s s o i l . Nitrates not destroyed.

INTERPRETATION OF RESULTS

A. Soils:

The i n t e r p r e t a t i o n of a B s o i l t e s t depends on crop, duration of

a p a r t i c u l a r B concentration, s o i l t e x t u r e , pH, Ca and K content. For

example, t h e B requirements of crops such as a l f a l f a , clover, b e e t s ,

t u r n i p s , celery, b r o c c o l i , and cauliflower a r e considerably greater

than f o r c i t r u s , cotton, tobacco, corn, and most monocotyledonous

plants. Soil B may be s u f f i c i e n t during i n i t i a l crop growth but l a t e r

371
 BENJAMIN WOLF

TABLE 2

I n t e r p r e t a t i o n of Boron Data i n S o i l Analyses

Soil Boron Rating Evaluation

pp2m

0-0.4 poor B contents i n t h i s range f o r extended

periods will result in deficiencies
for most crops.
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0.5-0.9 fair Probably satisfactory for crops with

low B requirements although additional
amounts of B generally need to be applied.

1.0-2.4 good Satisfactory for most crops but for high

requirement crops, maintenance amounts of
B probably should be applied.

2.4-4.9 high High although toxicity symptoms may not

always be apparent. B additions should
cease, particularly on light soils.

5.0+ toxic Such B levels maintained for an appreciable

time will develop toxicity symptoms.

may become insufficient due to extensive leaching during wet periods or

fixation during dry periods. B uptake by plants i s affected by soil pH,

Ca and K contents. I t s release i s markedly affected by texture, being

much greater in sandy than in clay soils.

Therefore, i t i s important to determine soil B several times

during the growing season and use in conjunction with plant analysis

results.

Despite the many factors which can affect an interpretation of

a B soil t e s t , Table 2 gives useful data for evaluating these tests.

B. Plant Materials:

As with the B soil t e s t , the interpretation of a B plant analy-

sis depends on several factors. The crop, kind of plant tissue and

age have an important bearing upon the evaluation of B plant analysis.

Many papers, and several books and monographs give interpretating

372
 DETERMINATION OF BORON

data f o r B p l a n t analyses. Two p u b l i c a t i o n s which are very u s e f u l

are a book edited by Chapman (15) and a b u l l e t i n by Neubert e t a l .

(16).

Generally, values below 25 ppm B i n dry t i s s u e of r e c e n t l y

matured leaves r e p r e s e n t a deficiency range f o r many dicotyledonous

p l a n t s , whereas the deficiency range for monocotyledonous p l a n t s

usually f a l l s below 10 ppm. The t o x i c concentration v a r i e s con-

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s i d e r a b l y but I have not associated B t o x i c i t y with contents below

about 75 ppm of dried t i s s u e of r e c e n t l y matured leaves. However,

I advise no f u r t h e r a d d i t i o n s of B i f the B content of mature leaves

i s over 50 ppm on a dry weight b a s i s .

C. Manures and Composts:

The amount of B i n manures and composts w i l l r e s t r i c t t h e i r u s e

as well as the crop and r a i n f a l l . With a p p l i c a t i o n r a t e s of about 2

tons of dried m a t e r i a l per a c r e , the B content should not be over 300

ppm when applied for B s e n s i t i v e crops.

D. Waters and Nutrient Solutions:

The i n t e r p r e t a t i o n f o r t h e amount of B found i s dependent on

the B s e n s i t i v i t y of the crop. For B s e n s i t i v e crops, i r r i g a t i o n

waters having l e s s than 0.67 ppm B are c l a s s i f i e d as good. For B

t o l e r a n t crops, the B content can be between 1.0-2.0 ppm and s t i l l

be c l a s s i f i e d as good. For most crops, i t appears d e s i r a b l e t o use

water having B contents l e s s than 2.0 ppm (17).

S a t i s f a c t o r y growth of many p l a n t s i s obtained when the B con-

t e n t of n u t r i e n t s o l u t i o n s i s 0.25-0.5 ppm.

SUMMARY

The rapidity and r e l i a b i l i t y of azomethine-H method as well as i t s adapta-

tion to aqueous solution offers real davantages over a number of colorimetric

methods now being generally used for determining B. The ability to determine

373
 BENJAMIN WOLF
B i n the same s o i l extract and plant t i s s u e digest a s the other e s s e n t i a l nu-

t r i e n t elements i s of r e a l value i n programs of f e r t i l i t y control involving

the use of s o i l t e s t s and plant analyses.

ACKNOWLEDGMENT

The azomethine-H f o r these studies was provided by:

Pierce Chemical Co., Rockford, I l l i n o i s 61105

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REFERENCES

1. B. Wolf, Soil Sci., 48, 41 (1939)

2. B. Wolf, Soil Sci. 50, 209 (1940)

3. G. S. Smith, Analyst, 60, 735 (1935)

4. K. C. Berger and E. Truog, Idus. and Engin. Chem. Analyst. Ed.,

11, 540 (1939)

5. R. B. Grimstead and S. Snider, Analyst, 92, 532 (1967)

6. F. R. Premi and A. H. Cornfield, Analyst 92, 196 (1967)

7. D. E. Williams and J. Vlamis, Com. Soil Sci. and Plant Anal., 1,

131 (1970)

8. A. Stettbacher, Mitt. Lebenom, Hyg. 29., 201 (1938)

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