Professional Documents
Culture Documents
Spectrochemical Analysis - Ingle PDF
Spectrochemical Analysis - Ingle PDF
STANLEY R. CROUCH
Michigan State University
7 0 9 8 7 6 5 4 3 2 1
I S B N 0-L3-82b900-7
PREFACE xiii
SPECTROCHEMICAL INFORMATION
SPECTROCHEMICAL MEASUREMENTS 13
OF ETER
5-1 Characteristics of Signals and Noise, 135 5-5 Signal-to-Noise Expressions for Absorption
Noise magnitude, 136 Measurements, 150
Noise types, I36 General equations, 150
Signal characteristics and the signal-to-noise ratio, 0% T noise-limited expressions, 152
137 Signal-shot-noise-limited expressions, 153
5-2 Frequency Characteristics of Signal Processing Signal-flicker-noise-limited expressions, 153
and Readout Systems, 138 Other noise sources, 153
Amplitude transfer function, 138 Direct absorbance readout, 153
System bandpass, 139
Noise in readout signals, 140 5-6 Signal-to-Noise Enhancement Techniques, 154
Frequency-domain filtering, 154
5-3 Noise Sources, 141 Adjustment of analytical and background signal
Quantum, secondary emission, and shot noise, 141 levels, 155
Flicker noise, 144 Photon counting, 155
Other noise sources, 145 Modulation techniques, 156
5-4 Signal-to-Noise Expressions for Emission and Double-beam and dual-channel techniques, 158
Luminescence Measurements, 146 Time-domain filtering, 159
General expressions, 146 Multichannel and multiplex systems, 160
Blank-noise-limited SIN expressions, 148 Problems, 161
Signal-shot-noise-limited expressions, 149
References, 162
Signal-flicker-noise-limited expressions, 149
Dependence of SIN on analytical signal, 149
ET NAL 1
7-1 Sample Introduction and Atomization, 189 7-3 Electronic States of Atoms, 202
Overview, 190 Quantum numbers, 202
Nebulizers, 192 Coupling schemes, 203
Free-atom formation after nebulization, 195 Term symbols, 203
Free-atom formation with discrete sample Selection rules and atomic spectra, 205
introduction, 198 Additional splitting effects, 207
7-2 Interferences in Atomic Spectroscopy, 198 Statistical weights and partition functions, 207
Blank interferences, 198 7-4 Spectral Line Profiles, 209
Analyte interferences, 199 Lifetime broadening, 209
viii Contents
8-1 The Ideal Atomic Emission Spectrometric Transducers and electronic components, 243
System, 226 Computer control, 244
Information desired from emission spectra, 226 8-5 Signal and Noise Considerations, 244
Characteristics of the ideal emission source, 226 Readout signals, 244
Atomizer temperature, 227 Signal-to-noise expressions, 245
8-2 Flame Atomic Emission Sources, 227 Signal-to-noise optimization, 246
Properties of flames, 228 8-6 Performance Characteristics, 247
Sample introduction into flames, 231 Linearity, 247
Atomization and excitation characteristics, 232 Precision, 248
Comparison to the ideal source, 233 Accuracy, 249
8-3 Plasma Atomic Emission Sources, 233 Detection limits, 251
Inductively coupled plasmas, 233 8-7 Methodology and Applications, 251
Microwave plasmas, 237 Identification and selection of analytical lines, 252
DC plasmas, 238 Analytical procedures, 253
8-4 Flame and Plasma Emission Spectrometers, Applications, 253
239 Problems, 255
Wavelength selection, 239
References, 256
9-1 Arc Excitation Sources, 258 Photographic detection for arc and spark emission,
Free-burning dc arcs, 258 267
Other types of arcs, 260 Performance characteristics, 268
Arcs as ideal emission sources, 261 9-4 Methodology and Applications, 269
9-2 High-Voltage Sparks and Other Emission Qualitative and semiquantitative methods, 269
Sources, 262 Quantitative methods, 270
The high-voltage spark discharge, 262 Applications, 270
Miscellaneous excitation sources, 265 Problems, 271
9-3 Instrumentation and Performance References, 271
Characteristics, 267
INTRODUCTION TO M
14-1 Basis of Infrared Absorption, 405 Spectral collections and search systems, 422
Requirements for infrared absorption, 405 Applications, 422
Number of vibrational modes, 405 14-5 Quantitative Infrared Absorption
Group frequencies, 407 Spectrophotometry, 423
Vibrational coupling, 408 Nonlinearities, 423
14-2 Instrumentation, 408 Measurement of absorption, 424
Dispersive I R spectrophotometers, 408 Precision of I R absorption measurements, 425
Fourier transform I R spectrometers, 412 Applications, 426
Nondispersive I R instruments, 415 14-6 Near-Infrared and Far-Infrared
14-3 Sample Preparation Techniques, 417 Absorption, 427
Near-infrared spectrometry, 427
Gas samples, 41 7
Liquid samples, 41 7 Far-infrared spectrometry, 428
Solid samples, 419 14-7 Infrared Reflection and Emission, 429
Microsampling devices, 4I 9 Infrared reflectance methods, 429
Infrared emission spectroscopy, 434
14-4 Qualitative Analysis and Structure
Determination, 420 Problems, 435
Correlation charts, 421 References, 436
SPECTROCHEMICAL TE
AC
References, 556
REPARATION METHOD 56
xiii
xiv Preface
taining high-quality spectrochemical data. Indeed, the common to all optical spectrochemical methods. These
title of the book was chosen to emphasize the important include the nature of spectrochemical information and
interplay of spectroscopy and chemistry in this field. measurements, the methodology of spectrochemical
The breadth of the field of spectrochemical anal- analysis, the instrumentation used in optical spectrom-
ysis makes it necessary to restrict the coverage to "op- eters, and the factors that determine signal-to-noise ra-
tical" spectrochemical methods to allow in-depth cov- tio (SIN) in spectrometry. Some instructors may choose
erage in a reasonable amount of space. The optical to delay the introduction of certain topics in Chapters
spectrochemical techniques covered involve radiation1 3 to 6 until the general concepts and equations are
matter interactions in the UV-visible-IR regions of the modified to apply to the specific spectrometric tech-
electromagnetic spectrum, which can be used primarily niques discussed in later chapters. Chapter 7 introduces
for determination of the concentration of species in the general principles and fundamental equations for
mixtures. Thus techniques such as NMR, microwave atomic spectrometric techniques, while Chapters 8 to
spectrometry, ESR, refractometry, and Mossbauer 11 cover specific atomic spectroscopic methods, includ-
spectrometry, which are used primarily for identifica- ing flame emission, arc, spark, and plasma emission,
tion and determination of structures and properties of atomic absorption, and atomic fluorescence spectrom-
pure compounds, as well as qualitative and quantitative etry. Chapter 12 serves to introduce the basis of mo-
uses of mass spectrometry, are not covered. It is ex- lecular spectroscopy. Specific molecular spectroscopic
pected that at the graduate level these topics would be methods, including UV-visible absorption, IR absorp-
covered in detail in a spectral interpretation course or tion, molecular fluorescence, phosphorescence, and
a molecular spectroscopy course taught by organic and chemiluminescence and molecular scattering tech-
physical chemists, respectively. Radiochemical and X- niques, are presented in Chapters 13 to 16. Chapter 17
ray techniques are not covered in this textbook because deals with those spectrochemical methods, both atomic
they do not conveniently fit into the common frame- and molecular, that are becoming or should become
work of optical methods due to significant differences standard "tools of the trade," such as photoacoustic,
in methodology, instrumentation, and origins of ana- thermal lensing, and laser ionization techniques. Six
lytical signals. appendices contain additional information about statis-
Throughout the book we have attempted to stress tical concepts, optical materials, optical filters, photo-
principles and concepts by presenting them first before multiplier tubes, sample preparation methods, and the
the details of a given topic are introduced. This ap- relationships between the different quantities that are
proach has the advantage that it allows a topic to unfold used to express the strength of transitions.
and expand based upon a strong fundamental frame- The atomic and molecular spectroscopy sections
work. It also allows and, indeed, encourages consid- were written to be used independently of each other.
erable flexibility in the use of the book. Instructors of Thus some instructors may prefer to introduce atomic
broad, survey-type courses can pursue a given topic to spectroscopy prior to molecular spectroscopy because
the depth desired without getting "bogged down" in atomic transitions are simple transitions between purely
too much detail or without the feeling that fundamental electronic states. On the other hand, a good case can
principles and concepts are being skipped if the cov- be made for the opposite order because many students
erage of a topic is shortened. Sufficient depth is given are more familiar with molecular absorption spectros-
in each topic, however, for the book to be useful in copy and the quantitative treatment of this topic is
more specialized courses that might stress one or more somewhat easier than the treatment of atomic absorp-
of the major topics presented, such as atomic spec- tion, which requires a discussion of line profiles and
troscopy or molecular spectroscopy. A second impor- atomization. We feel that the flexibility gained by mak-
tant aspect of our approach is an emphasis on the quan- ing these sections independent more than offsets the
titative, mathematical relationships that relate the signals small amount of redundancy required.
measured and the signal-to-noise ratios obtained in Although the book has been written for a full-
spectrochemical methods to physical, chemical, and in- semester course in optical spectrochemical methods, it
strumental factors. This approach gives the user a quan- can be readily abridged for a one-quarter course. This
titative feel for the way in which an instrumental var- can be accomplished by shortening the coverage of top-
iable, for example, will affect the quality of the data ics that should be review material (parts of Chapters 1
obtained and a knowledge of those variables that are to 4, 7, and 12) and by skipping some sections in later
the most effective in improving or optimizing the meas- chapters on specific techniques or even complete chap-
urement. ters (e.g., Chapters 9, 16, or 17).
The book is divided into four major sections. The We have attempted to made the figures as in-
first six chapters cover the fundamental aspects that are formative as possible by writing explanatory captions.
Preface xv
These extended captions provide a useful second level Fahey, Cecilia Yappert, and Paul Kraus assisted in de-
of explanation that expands on or complements the veloping materials, reading and commenting on the text,
textual information. Each chapter ends with a number and checking the answers to problems. Lucy Ingle, Sara
of illustrative questions and problems and a selected Ingle, and Jeff Louch contributed by checking the ac-
list of references. curacy of equations during proofing of the galleys.
A course in modern spectrochemical analysis should Our colleagues contributed in many direct and
include a parallel laboratory. Because of the specialized indirect ways. Many colleagues including Ed Piepmeier,
equipment available at most universities and because Mike Schuyler, Chris Enke, Joe Nibler, Jim Holler, Jim
many universities do not have graduate laboratories, O'Reilly, Gary Hieftje, Gary Horlick, and Ray Barnes
we have not attempted to include laboratory experi- served as responsive sounding boards as we developed
ments in this text. ideas for the book. Others including James Wineford-
ner, Alexander Scheeline, Eric Salin, Adrian Wade,
Acknowledgments Gil Haugen, Gary Christian, Timothy Nieman, Earl
Wehry, and Scott Goode reviewed in detail all or sec-
We would like to acknowledge the assistance and sup- tions of the manuscript and offered many useful com-
port of many people who were involved in the publi- ments that were incorporated (mostly) into the manu-
cation of this work. The production staff at Prentice script. We are especially grateful to our teacher, Howard
Hall, most of whom we never knew, helped us keep on Malmstadt, who provided us with the foundation for
schedule. Especially we wish to thank Elizabeth Perry, many of the concepts in the book.
Nancy Forsyth, Curtis Yehnert, and Dan Joraanstad, Our families and graduate students exhibited
who each served as the chemistry editor during various thoughtful patience over extended periods of time when
stages of the book. Extra special thanks go to the pro- we seemed to be invisible. Many of our students also
duction editor Kathleen Lafferty for her professional suffered through courses based on early versions of the
and understanding guidance during the final stages of manuscript and gleefully identified numerous errors and
publication. Before we became totally addicted to our unintelligible discussions. Finally, we apologize to all
word processors, Patty Ramus, Sherree Bittner, Brandy those who helped that we did not acknowledge specif-
Schuyler, Jenny Harber, Valerie Borst, Dawn Penrose, ically. We know there were many and greatly appreciate
and Debbie Wuethrich helped in typing earlier versions your assistance.
of chapters. Many students including Max Hineman,
Pat Wiegand, Kim Ratanathanawongs, Peter Wentzell, James D. Ingle, Jr.
Tom Doherty , Helen Archontaki, Mark Phillips, Jeff Stanley R. Crouch
We live in an era in which the analysis of chemicals and a quantity related to the result of photon absorption.
materials is of extreme importance. Analytical methods As examples of the latter, the number of ions or a
are used to help monitor the status of human health, quantity related to the kinetic energy produced by ab-
the ingredients in the foods we eat and the water we sorption can be monitored and plotted as a function of
drink, and the quality of the environment. Analysis is wavelength to obtain a spectrum.
also an important step in industrial processes and in the This chapter describes the information that can
development of safe and valuable industrial products. be obtained from spectrochemical studies and the na-
Spectrochemical analysis is one of the major tools of ture of spectrochemical analysis. The use of spectral
analytical chemistry. The applications of spectrochem- data to express analytical information is examined, and
ical methods range from the determination of extremely several criteria for evaluating spectrochemical tech-
low levels of noxious materials in the environment to niques are introduced. The general concepts and basic
the monitoring of major components, such as sodium definitions introduced here are used throughout this
and potassium in biological fluids. These methods thus book as we explore the field of spectrochemical anal-
cover a broad scope of materials, a variety of matrices, ysis.
and a wide range of concentrations.
"Spectrochemical" is a compound word that comes
from spectrum and chemical. A spectrum is a display
of the intensity of radiation emitted, sbsorbed, or scat-
tered by a sample versus a quantity related to photon Spectroscopy is the science that deals with the inter-
energy, such as wavelength or frequency. The term actions of electromagnetic radiation with matter. Sev-
spectrochernical implies that a spectrum or some aspect eral types of interactions are possible. Many of these
of a spectrum is used to determine chemical species and involve transitions between specific energy states of
to investigate the interaction of chemical species with chemical species and are observed by monitoring the
electromagnetic radiation. Spectrochemical methods can absorption or emission of electromagnetic radiation. In
involve a direct optical measurement of the photons these types of interactions it is useful to consider elec-
emitted or transmitted or an indirect measurement of tromagnetic radiation as being composed of discrete
Chap. 1 1 Spectrochemical Information
packets of energy which we call photons. Electromag- lecular spectroscopic techniques. Atomic spectroscopy
netic radiation also has a wave character, and we can deals with spectrochemical phenomena involving free
relate the energy of a photon to its wavelength and atomic species that are usually in the vapor state, whereas
frequency by molecular spectroscopy deals with optical measure-
ments of molecular species in the vapor state, in solu-
tion, or in the solid state.
Photon energies in the optical regions are typically
expressed in units of joules, ergs (1 erg = J), or
where E is the energy in joules (J), v is the frequency electron volts (1 eV = 1.6 x 10-l9 J). The wavelength
(Hz or s-l), h is Planck's constant (6.63 x J s), of radiation in this region is usually expressed in na-
c is the speed of light (3.00 x lo8 m s-I in a vacuum), nometers (1 nm = l o p 9 m), angstroms (1 A =
and X is the wavelength (m). Spectrometry is a more 10-lo m), or micrometers (1 pm = m). One elec-
restrictive term than spectroscopy and denotes the tron volt of photon energy corresponds to radiation with
quantitative measurement of the intensity of electro- a wavelength of 1240 nm.
magnetic radiation at one or more wavelengths with a Because the wavelength is inversely proportional
photoelectric detector. to energy, the wavenumber is often used, particularly
There are several types of radiationlmatter inter- in the IR region. The wavenumber, T, is the number
actions, such as reflection, refraction, diffraction, and of cycles per unit length (usually cm) and thus the re-
some types of scattering that do not involve transitions ciprocal of the wavelength, ? = llA. The wavenumber
between energy states, but rather cause changes in the is usually expressed in cm-l. It is directly proportional
optical properties of the radiation (e.g., direction and to the photon energy,
polarization). These interactions are often related to
the bulk properties of the sample rather than to specific
chemical species. Several analytical techniques are based
on these bulk interactions.
Spectrochemical analysis, in general, deals with The energy or wavelength of the photon deter-
electromagnetic radiation of an enormous range of fre- mines the type of transition or interaction that occurs,
quencies, from the audio frequencies (<20 kHz) to as shown in Table 1-1.
gamma rays (>1019 Hz). Optical spectrochemical anal-
ysis, the primary topic of this book, covers a more re-
strictive range, the near ultraviolet (UV), the visible,
and the infrared (IR) regions, where instrumental re-
quirements are similar, and the materials used for dis-
persing, focusing, and directing the radiation are con- Before getting into the details of spectrochemical meth-
ventional optical materials (glass, quartz, or alkali halide ods and procedures, it is useful to consider some general
crystals). Optical spectrochemical techniques are often aspects which describe the kinds of determinations that
divided into atomic spectroscopic techniques and mo- can be performed and the types of samples that can be
TABLE 7- 7
Regions of electromagnetic spectrum
Wavelength Frequency
range, range, Wavenumber or
Designation A (Hz) energy range Transition
analyzed by spectrochemical methods. In the process, For example, a liter of water (the initial sample) is
we shall define several important terms and concepts obtained from a lake (the bulk material) in order to
which are used throughout this book. determine the mercury content, or a few grams of soil
(initial sample) are acquired from a field (bulk material)
in order to determine the concentration of a pesticide.
Types of Analyses The analytical sample indicates that portion of the initial
sample which is presented to the instrument for spec-
In spectrochemical methods, spectroscopy is used for trochemical analyses. In some cases the initial sample
the identification of chemical species (qualitative anal- is treated prior to the analyses. Such treatment may
p e s ) and for the determination of the amount of a include grinding, heating, dissolution, preconcentra-
particular species (quantitative analyses). At first thought, tion, dilution, separation, addition of a buffer, adjust-
the different goals of qualitative and quantitative anal- ment of the pH, or many other steps. In all cases the
yses would seem to dictate quite different approaches. analytical sample must be representative of the con-
However, to ascertain the presence of a species with centration of the sought-for species in the bulk material.
some degree of certainty, we must have at least a ru- The sampling and sample preparation operations, which
dimentary knowledge of the quantity required. For ex- are the entire set of procedures done to acquire and to
ample, to identlfy the presence of mercury in river water, prepare the analytical sample from the bulk material,
we must determine that the mercury concentration ex- are thus crucial steps in an entire analysis. Errors made
ceeds a certain threshold, decision level. Thus quali- in sampling or sample preparation are carried through
tative analysis can be considered as merely a low-res- the entire process and lead directly to errors in the final
olution type of quantitative analysis, often involving a result (see Section A-2 in Appendix A).
simple binary, yes or no determination of the sought- The species to be determined in the analytical
for species. Although quantitative methods are stressed sample is designated the analyte. There may be several
in this book, it is useful to keep in mind that similar analytes in a given sample, and their concentrations can
considerations of uncertainty in the results, limits of range from the trace level to the major constituent level.
detectability, upper concentration ranges, and error in The term matrix refers to the collection of all the con-
the procedures also apply to qualitative methods. stituents in the sample. The analytical matrix refers
The constituents determined in a spectrochemical specifically to the matrix of the analytical sample which
analysis can cover a broad concentration range. In some may differ from that of the initial sample due to the
cases spectrochemical methods are used to determine substances added or removed in the various sample
major constituents. These are considered here to be treatment stages. The matrix as defined here includes
species present in the relative weight range 1 to 100%. the analyte as well as all the other constituents, which
Minor constituents are species present in the range 0.01 are called concomitants. In trace analyses the analyte
to 1%, while trace constituents are those present in is present in such small amounts that it is convenient
amounts lower than 0.01% (100 pg gW1). to speak of the analyte and the matrix separately. Thus
The size of the sample is also used to classify the for trace analysis we sometimes think of the matrix as
type of analysis performed. A macro analysis is one being composed of the concomitants. In major constit-
carried out on a sample weighing more than 0.1 g. A uent determinations, however, the analyte is a major
semimicro analysis (sometimes called a meso analysis) part of the matrix and this division is no longer helpful.
utilizes a sample size in the range 0.01 to 0.1 g, while For example, in determining iron in steel, the analyte
a micro analysis employs a sample size in the range (iron) is present in such large amounts that it determines
to g. If the sample size is lower than g, to a large extent the bulk properties of the sample. Here
the term ultramicro is sometimes employed. The term the matrix is no longer composed of just concomitants.
ultra-trace analysis is considered to be the determina- In some analyses, information about specific
tion of a trace constituent in an ultramicro sample. chemical species is of interest. Chemical speciation is
concerned with determining the concentration of spe-
cific chemical forms of the analyte (e.g., the amount of
amples metal in a particular oxidation state or the amount of
a drug bound to protein). The nature of the sample
The nomenclature for dealing with the samples used in matrix and the effect of the matrix on a determination
spectrochemical analyses is often confusing and contra- depend upon the chemical interactions among matrix
dictory. Here we shall use the term initial sample to components and between matrix components and the
mean a portion or subset of the bulk material or pop- analyte. Thus the chemical form(s) of the analyte and
ulation about which analytical information is desired. of the matrix components is critical.
Chap. 7 1 Spectrochemical Information
the blank is treated as identically to the sample as pos- of a sample or standard. The blank or reference signal
sible. For example, if the analytical sample is a solution, is defined as the readout signal obtained from meas-
the blank is a solution composed of the same solvent urement of the blank. It includes the background signal
as the sample plus any reagents used to prepare the due to optical signals from the sample container and
sample. The instrumental response from the blank is the concomitants in the blank. The analytical signal is
then subtracted from that of the sample in order to extracted from the total spectrochemical signal. Ideally,
compensate for the effects of concomitants. An ideal it is directly related to the analyte concentration. In
blank can eliminate some types of interference effects emission spectrometry, for instance, the total emission
due to concomitants but cannot compensate for con- signal contains emission components from the analyte,
comitant species that affect the production and meas- the concomitants, and other sources (e.g., the sample
urement of the analyte response. It is difficult to prepare container). The analytical signal is obtained by sub-
an ideal blank because the concomitants and their con- tracting the blank emission signal from the total emis-
centrations are not usually known. A more desirable sion signal. In some automated instruments the readout
approach is to arrange measurement conditions to min- signal may be the analytical signal if the instrument
imize the response due to concomitants. carries out the appropriate modifications internally.
Actual analyses are further complicated because The analytical signal is related to the analyte con-
the sample is almost always confined during measure- centration and other variables by the calibration func-
ment by a container, except for a few in situ measure- tion, f , defined by
ments. In molecular spectroscopy, the container is typ-
ically a glass, quartz, or salt cell. Optical interactions
with the container walls can give rise to additional in-
terference effects. In atomic spectroscopy the container where S is the analytical signal, c, is the analyte con-
is typically a flame, a plasma, or a heated chamber. centration, A is the analysis wavelength, and x, is the
The hot gases produced can emit or absorb radiation magnitude of the ith experimental variable (e.g., tem-
which can also be a potential source of interference. In perature, time). Often the calibration function is con-
some cases the sample matrix can alter the interference sidered only a function of e,.
effects of the container. Since there are many possible The dependence of the calibration function on a
interferences in a spectrochemical procedure, a major specific variable is often given a particular name. For
part of this book deals with their identification, their example, a plot of the analytical signal versus analyte
effects, and their minimization (i.e., ways to compen- concentration, with all the other variables in the cali-
sate for them). bration function held constant, is called the calibration
curve, the analytical curve, or the working curve. Typ-
ically, the calibration curve is obtained from measuring
1-3 EXPRESSIONS
the analytical signals for a series of standards (analyte
INFORMATION
solutions of known concentrations) of different analyte
concentrations. The calibration curve in conjunction
Several different types of information are required in
with the measurement of the analytical signal from a
order to develop, apply, and optimize an analytical
sample is then used to determine the analyte concen-
technique. This section considers how to express spec-
tration in the sample.
trochemical information in a convenient manner so that
The analytical function, g, is the inverse of the
analyte concentration data can readily be extracted or
calibration function:
the dependence of the results on chemical, physical, or
instrumental variables can easily be summarized.
Calibration Data
For example, if S = 10p4ca,c, = 104S.The analytical
The desired result of a spectrochemical analysis is the function provides a direct method for calculating the
concentration of the analyte. It is almost never obtained analyte concentration from the measured analytical sig-
directly as the result of an absolute measurement of an nal for a given set of experimental conditions. In mod-
optical signal, but is obtained indirectly through cali- ern computerized instruments, the calibration function
bration, subtraction of blanks, comparison with stand- is determined by the instrument from measurments on
ards, and other procedures. standards. The analytical function is then calculated and
The total spectrochemical signal is defined as the used to provide the analyte concentration directly from
unmodified readout signal obtained from measurement the measured analytical signal. Such internal calcula-
Sec. 1-3 I Expressions of Analytical Information 7
tions can eliminate operator errors and the tedium as- and are due to transitions between a vibrational and
sociated with using calibration data. rotational level in one electronic state to a vibrational
and rotational level in another electronic state. Thus,
tornic and Molecular Spectra many transitions between two electronic states are pos-
sible and such molecular spectra are called band spec-
A spectrum is a plot of the analytical signal versus wave- tra. In the gas phase, we can observe the vibrational
length, frequency, or wavenumber with all other vari- and rotational structure. In the condensed phases, much
ables held constant. The peaks (lines or bands) are of the structure of molecular spectra is blurred because
characterized by their shape, height (intensity), width, of frequent molecular collisions and level perturbations
and position (wavelength). Usually, the width is ex- due to near-neighbor interactions. As a result, molec-
pressed as the half-width (AX), which is the width in ular electronic spectra of liquids and solutions often
wavelength units at half the net peak height. The half- consist of one or more broad, featureless bands (typi-
width is also called the full width at half maximum cally, 10 to 100 nm wide), where each band is an en-
(FWHM). A spectrum is an essential summary of spec- velope of the multitude of possible transitions between
tral information of any spectrometric technique because vibrational-rotational levels in two electronic states. Vi-
it indicates the wavelength to use for quantitative anal- bration-rotational spectra involve transitions from the
ysis in order to obtain the maximum analytical signal. rotational levels of one vibrational level to the rota-
The analyte spectrum along with spectra of concomi- tional levels of another vibrational level of the same
tants allows the wavelength of analysis to be chosen electronic state and are observed most often in the IR
both to maximize the analytical signal and to discrim- region of the spectrum. The rotational structure of in-
inate against background signals. frared spectra is usually lost in condensed phases. Ro-
The spectrum of the analyte is the basis of qual- tational spectra involve transitions from one rotational
itative analysis since the wavelengths and relative in- level to another rotational level of the same vibrational
tensities of peaks in a spectrum are characteristic of the level of the same electronic state. Purely rotational spectra
analyte. Qualitative analysis through spectral charac- are normally observed in the microwave region of the
teristics is favorable for species which provide "rich" spectrum. Raman spectroscopy also gives information
spectra (spectra with numerous resolvable peaks), so
that these can be "fingerprints" for the analyte(s).
The spectra of atoms in the ultraviolet, visible,
and near-infrared regions arise from purely electronic
transitions of outer-shell (valence) electrons, as dis-
cussed in detail in Section 7-3. Because the quantized
energy levels are relatively far apart, atomic spectra are
narrow line spectra. A transition to or from the ground
electronic level is a resonance transition, and the re-
sulting spectral line is a resonance line. Atomic spectra
are often quite simple because many of the possible
transitions are forbidden transitions. The allowed tran-
sitions can be predicted from quantum mechanics, and
selection rules tell us the requirements for an allowed
transition. Atomic spectral lines have a finite width
(typically, much less than 1 A) even though the tran-
sitions are between two distinct energy levels because
of line broadening due to lifetime effects, the Doppler
effect, and collisions. Line broadening is discussed in
detail in Section 7-4.
Because molecules have quantized vibrational lev-
els and rotational levels in addition to electronic levels, FIGURE 1-3 Vibrational and rotational levels
as shown in Figure 1-3, their spectra are necessarily of molecular electronic states A and B. The vi-
more complicated than the spectra of isolated atoms. brational levels are given by v" and v ' , while the
There are three distinct types of spectra we can observe rotational levels are given by J" and J'. The three
for molecules; these are considered in more detail in arrows show the three types of transitions pos-
Chapter 12. Molecular electronic spectra are observed sible: electronic (x), vibration-rotation ( y ) , and
in the UV, visible, and near-IR regions of the spectrum pure rotation ( 2 ) .
Chap. I 1 Spectrochemical Information
about vibrational and rotational levels through meas- where S is the analytical signal and N is the root-mean-
urement of the inelastic scattering of UV-visible radia- square (rms) noise in the analytical signal. If noise is
tion. what causes the uncertainty in the signal, the signal-to-
noise ratio is the reciprocal of the relative standard
ptirnization of the Response Function deviation (SIN = [RSDIp1).
The magnitude of S or SIB is often simpler to
The influence of the other variables (in equation 1-3) measure, and frequently yields the same optimal ex-
on the calibration function or the analytical signal is perimental conditions as are obtained by optimizing the
also important. The dependence of S on all possible SIN. Maximizing the analytical signal may also maxi-
variables cannot realistically be determined; in practice, mize the SIN or minimize the RSD if a, is independent
only those variables deemed to be critical or which can of the signal magnitude. The SIN may be optimized by
be conveniently measured are considered. The other maximizing the SIB for cases in which o, is determined
variables of interest depend upon the technique and by noise in the background signal.
particular application and can include instrumental var- Knowledge of the calibratioc function allows quan-
iables, such as excitation light source intensity, volume titative information about precision to be obtained. Ap-
element of total sample in the instrument that is actually plication of propagation of uncertainty mathematics (see
probed, and slit width, as well as physical and chemical Appendix A) to equation 1-3 yields
variables such as temperature, pH, ionic strength, re-
agent concentrations, and concentrations of concomitant
species. Results of studies of the influence of experimental
variables are used to choose optimum operating condi-
tions as discussed below.
Measurement conditions are optimized by maxi- From equation 1-5 the effect of variations in a given
mizing (or minimizing) a response function, which is an variable (magnitude of a for the ith variable) can be
optimization criterion tailored to a specific application. determined. Thus, in general, the magnitude of a var-
Often the response function is the analytical signal, and iable can be adjusted so that the product of the slope
conditions are adjusted to maximize its magnitude. Since of the calibration function with respect to the variable
the variable values may affect the concomitant and sam- (aflax,) and the uncertainty in the variable a, are small.
ple container spectral signals differently than the ana- This is often accomplished by adjusting the magnitude
lyte spectral signal, another useful response function is of the variable to a value where the analytical signal is
the signal-to-backgroundratio (SIB). Here S is the an- independent of the variable (dfldx, = 0). In general,
alytical signal from the analyte and B is the sum of lowering the background signal without decreasing S
background spectral signals from other species or the (maximization of SIB) will improve precision if fluc-
sample container. Changing the value of a variable to tuations in the background signal determine a, since
double S can make it more difficult to measure S if the the absolute standard deviation in the background sig-
background signal increases an order of magnitude. nal usually increases with the magnitude of the back-
The foregoing two response functions are not nec- ground signal.
essarily adequate because they do not take into account The dependence of the calibration function on a
the precision with which the analytical signal can be variable is graphically expressed as a plot of S vs. the
measured. Increasing the magnitude of S or SIB through variable. It is becoming more common with sophisti-
changing the value of an experimental variable can de- cated instrumentation (e.g., graphic capabilities) to col-
grade the quality of the measurement if the relative lect and display multidimensional data. For example,
uncertainty in the magnitude of S is increased. Thus it in molecular fluorescence the analytical signal depends
is often best to optimize the precision of a measurement on both the excitation and emission wavelength, so that
by minimizing the relative standard deviation (RSD) a three-dimensional plot can be made with the x and y
for measuring the analytical signal, where RSD = axes representing the two wavelengths and the z axis
a,/S and a, is the standard deviation in measuring the representing the magnitude of the analytical signal. De-
analytical signal. The standard deviation is a quanti- spite the difficulty we have in constructing such plots,
tative measure of the uncertainty in a quantity due to they can be extremely informative in choosing favorable
random variations in the factors which affect the quan- operating conditions.
tity from one measurement to the next, and is discussed The single factor at a time approach to optimi-
in detail in Appendix A. zation described above does not always yield the op-
Sometimes the precision is optimized by maxi- timum conditions if the variables interact with each other
mizing a quantity called the signal-to-noise ratio (SIN), (i.e., the optimum value for one variable depends on
Sec. 1-4 I Evaluation Criteria in Spectrochemical Techniques
the magnitude of another variable). In the simplex op- niques requiring minimal sample preparation are pre-
timization approach several experimental variables are ferred.
changed simultaneously, and a systematic search for the
optimum response is made. The simplex is a geometric Automation and Multiple-Species Capability
figure that is moved across the response surface by a
pescribed set of rules until it reaches the optimum Automatio'n of spectrometric instruments has increased
response or fails. Simplex optimization is simple enough dramatically due to the introduction of microcompu-
to be done with the aid of a hand calculator and is well ters. Automation, the performance of tasks without op-
suited for computer-controlled systems where response erator assistance, can free the operator of tedious tasks
measurements, calculations, and variable adjustments and increase precision due to more reproducible per-
can all be made very quickly in real time. Use of this formance of steps formerly requiring operator skill. Nu-
and other mathematical optimization techniques (e.g., merous tasks can be automated, including the selection
steepest descents) is increasing. of preprogrammed instrumental variable values for a
given analyte, measurement of spectral signals, con-
struction of calibration curves, and presentation of an-
1-4 EVALUATION CRITERIA I N alytical and statistical information. Further steps which
SPECTROCHEMICAL TECHNIQUES may be automated include sample preparation, cleanup,
and introduction. Since some techniques are more suit-
ractical Considerations able for automation than others and because only cer-
tain automated instruments are commercially available,
Many factors must be considered in choosing a specific the selection of a specific spectrometric technique may
spectrometric technique for a given application. First, depend on the degree of automation desired. Auto-
there are the practical considerations of cost, sample mation is particularly needed for situations involving
size required, simplicity, portability, and robustness. unattended analysis of samples at night or at remote
Cost considerations must take into account not only the locations. The new generations of microprocessor-con-
initial costs of the instrument, but also the special fa- trolled instruments have introduced error checking and
cilities (e.g., power) required to install the instrument, instrument diagnostic features not previously available.
the maintenance costs, and the operating cost (e.g., We can expect to see more instruments with built-in
gases, reagents, technician salary). The amount of sam- intelligence to assist the operator in optimization, in-
ple required is critical in situations, such as clinical ap- strumental operation, and complex data interpretation.
plications, where sample sizes are limited. The simplest These have been termed "expert" systems.
instrument and procedure which fulfill the requirements Multiple-species analysis is more convenient with
of the method are often preferred to reduce the prob- automated instrumentation. Multiple-species analysis
ability of error and downtime. Simplicity also reduces can be carried out by some instruments which measure
the operator skill required. Portability and robustness each analyte in a sequential manner or by others which
are important where an instrument must repeatedly be measure all analytes simultaneously. Simultaneous
moved between sampling sites with minimal setup time analysis can be implemented by measurement of n an-
or where conditions are harsh, as on an oceanographic alytes in each of n identical, but separate, analytical
research vessel. samples, where all samples are processed separately but
Speed of analysis is important since it affects the simultaneously in n parallel channels or by simultaneous
cost per analysis and the sample throughout, which may measurement of n spectral signals from a given analyt-
be critical in routine anabsis situations involving large ical sample. Simultaneous analysis is faster than se-
numbers of samples. The speed of an analysis is affected quential analysis and requires less sample for the situ-
not only by the actual measurement time for the sample, ation where n spectral signals are measured from one
but also by the time necessary to obtain the calibration analytical sample.
curve and the time needed to prepare the analytical
sample. Often sample prepdration (e.g., dissolution of interferences and Selectivit
biological and geochemical samples) is the most time-
consuming step and the step most demanding of tech- The accuracy of all spectrometric techniques can be
nical skill. Techniques which are more prone to inter- degraded by interferences. An interferent is a substance
ference effects often require additional cleanup or present in the analytical sample which affects the mag-
matrix modification steps to remove or suppress inter- nitude of spectral signal measured for the analyte. Thus
ferences before the actual measurement step. Thus, the choice of a spectrochemical technique depends on
overall, where speed of analysis is paramount, tech- the expected interferences and their concentrations in
1 Chap. I i Spectrochemicat Information
Concentration
freedom from interferences.
FIGURE 1-4 Calibration curve illustrating the
concepts of sensitivity, detection limit, and dy-
Figures of Merit namic range. The calibration sensitivity (m) is the
slope of the calibration curve at a particular con-
The characteristics of a spectrochemical technique for centration or the derivative of the calibration
a given analyte are indicated by several figures of merit, function at a particular concentration [dSlac),].
such as accuracy, precision, sensitivity, detection limit, The detection limit (DL) indicates the lowest an-
and dynamic range. The accuracy indicates how close alyte concentration that can be measured with a
specified degree of certainty and is often defined
the measured analyte concentration is to the true an-
by D L = 2sbkimor D L = 3sbklm,where s, is
alyte concentration in the sample and is normally ex- the standard deviation of the blank measurement.
pressed as the relative percent error (i.e., a 1% error The dynamic range is defined by D L and c,, where
indicates that the measured concentration is within 1 % c, is the maximum concentration that can be
of the true analyte concentration). The accuracy de- measured before the calibration curve deviates a
pends on the analyte concentration, the precision, and specific amount from the extrapolated linear por-
interference effects. tion of the plot.
The precision, usually expressed as a percent RSD,
indicates the reproducibility of repetitive measurements
of equivalent analyte solutions. Averaging of repetitive
measurements can be used to improve precision, and to two or three times the standard deviation of a blank
thus the accuracy, if random errors are limiting rather measurement (33 to 50% RSD). It will be discussed in
than systematic errors due to interferents or other fac- more detail in Section 6-4, but is indicative of the lowest
tors. An inherently high-precision technique can be faster analyte concentration that can be reported as being
since repetitive measurements are not required. detected with a specified degree of certainty. The de-
The precision depends upon the analyte concen- tection limit is a critical figure of merit since a spectro-
tration and can depend upon the sample matrix and metric technique cannot be used without preconcentra-
interferences. As indicated in Section 1-3, instrumental, tion steps if the analyte concentration in the analytical
physical, and chemical variables are often selected to sample is below the detection limit. In fact, the analyte
optimize precision. The analyte concentration in sam- concentration should normally be higher than 10 times
ples is often adjusted by dilution or preconcentration the detection limit to obtain reasonable precision (5%
to be in the range yielding best precision. Thus a plot RSD or less). The choice between two spectrochemical
of precision (RSD) vs. analyte concentration is useful. techniques both with detection limits well below the
The concepts of sensitivity, detection limit, and analyte concentrations to be measured is based on other
dynamic range are illustrated in Figure 1-4. Sensitivity criteria. Often variables are optimized for the best de-
may have several meanings, as discussed in greater de- tection limit at the expense of other criteria, such as
tail in Section 6-4. It usually indicates the response of precision at higher concentrations. Detectability is a term
the instrument to changes in analyte concentration and that denotes the ability to provide low detection limits.
is expressed as the slope of the calibration curve or the Dynamic range can be specified as the concentra-
change in analytical signal per unit change in analyte tion range or analytical signal range over which the
concentration. This definition is the primary definition analytical curve is linear or the calibration slope is con-
of sensitivity used throughout this book. Sometimes stant. It is usually defined at the lower end by the de-
sensitivity refers to the ability to distinguish concentra- tection limit and at the upper end by an analyte con-
tion differences and thus takes into account the preci- centration where the analytical signal deviates a specific
sion of measurements. relative amount (e.g., 5%) from the extrapolated linear
The detection limit (DL) is typically defined as the portion of the curve or where the slope deviates a spe-
analyte concentration yielding an analytical signal equal cific relative amount from the slope in the linear por-
Chap. 1 I References 11
1-1. Express 4000 A in nm, as a frequency, as a wavenum- 1-7. A certain optical transition occurs in the visible region
ber (cm-I), and as an energy in joules, ergs, and elec- of the spectrum at 530 nm. Find the energy of the
tron volts. transition in J and in eV. What is the wavenumber of
1-2. Emission and luminescence techniques both involve the transition?
the measurement of the emission of photons from ex- 1-8. A 0.08-g sample is analyzed for a constituent present
cited species. What makes these techniques different? at approximately 5 mg g-'. Classify the analysis in
1-3. Why are real blanks not ideal blanks? terms of constituent amount and sample size.
1-4. What is the difference between SIB and SIN? 1-9. A certain determination follows a calibration function
1-5. In a photoluminescence spectrometry, the photolu- S = 1.5 x 104c, where S is the analytical signal and
minescence signal is usually proportional to the exci- c is the molar concentration. Find the analytical func-
tation source intensity. If the lamp intensity is pro- tion for this determination.
portional to the fourth power of the lamp temperature, 1-10. You have a "30-cm ruler," which is in fact 28 cm long.
to what degree must the RSD of the lamp temperature Briefly discuss (a) measurement precision, and (b) its
be controlled to ensure that the RSD of the photo- accuracy.
luminescence signal due to lamp temperature fluctua- 1-11. The RSD of a measurement limited by noise is 0.50%.
tions is 1% or less? What is the SIN?
1-6. Why does high precision not necessarily ensure high 1-12. Calculate the detection limit for a determination if the
accuracy? blank standard deviation is 0.010 V and the slope of
the calibration curve is 2.0 V fiM-'.
1. IUPAC, Compendium of Analytical Nonzenclatnre, Per- The following monographs or series include chapters on op-
gamon Press, Oxford, 1978. This compendium contains tical spectrochemical methods of analysis and serve as excel-
many definitions of terms and figures of merit. lent general reference sources.
Chap. I 1 Spectrochemical Information
C. N. Reilley, R. W. Murray, and F. W. McLafferty, 7. R. Belcher, L. Gordon, and H. Freiser, eds., Analytical
eds., Advances in Analytical Chemistry and Instrumen- Chemistry, Pergamon Press, Oxford, 1961-present.
tation, Wiley-Interscience, New York, 1960-1973. 8. T. Kuwana, ed., Physical Methods in Modern Chemical
P. J. Elving, I. M. Kolthoff, et al., eds., Treatise o n Analysis, Academic Press, New York, 1978-present.
Analytical Chemistry, Wiley-Interscience, New York, 9. D. Glick, ed., Methods of Biochemical Analysis, Wiley-
1959-present. Interscience, New York, 1954-present.
G. H. Morrison, ed., Trace Analysis: Physical Methods, 10. J. D. Winefordner, ed., Trace Analysis: Spectroscopic
Wiley-Interscience, New York, 1965. Methods for Elements, Wiley-Interscience, New York,
P. J. Elving, I. M. Kolthoff, and J. D. Winefordner, 1976.
eds., Chemical Analysis, Wiley-Interscience, New York, 11. J. D. Winefordner, ed., Spectrochemical Methods of
1941-present. Analysis, Wiley-Interscience, New York, 1971.
C. L. Wilson, D. W. Wilson, and G. Svehla, eds., C o m - 12. G. D. Christian and J. B. Callis, eds., Trace Analysis:
prehensive Analytical Chemistry, Elsevier, Amsterdam, Spectroscopic Methods for Molecules, Wiley-Intersci-
1959-present. ence, New York, 1986.
CHAPTER 2
Spectrochemical
Measurements
When we discuss the quantitative determination of a trochemical methods are classified according to the types
particular species by spectrochemical methods, we are of spectral information produced, and the principles of
considering the measurement of its concentration using absorption, emission, luminescence, and scattering are
some form of spectroscopy. Before going further it is introduced. It is shown that optical information can be
important to be clear about what the measurement selected by its wavelength, its time or temporal behav-
process is and how it applies to spectrochemical pro- ior, its spatial position, or by several combinations of
cedures. A measurement can be defined, in general, as selection criteria. The final section describes how op-
the determination of a particular characteristic of a sam- tical information representing chemical information can
ple in terms of a number of standard units of that char- be converted into a number (measured) that is related
acteristic. A chemical concentration measurement thus to concentration.
consists of determining how many standard concentra-
tion units (molarity, pg rnLP1, etc.) of a particular spe-
cies are present in the sample. Reference to a standard 2-1 COMPLETE SPECTROCHEMICAL
is implicit in this definition of measurement and it is an MEASUREMENT
important part of a spectrochemical determination.
Spectrochemical measurement systems differ from others, Spectrochemical determination of the concentration of
such as electrochemical systems, in that during the the analyte in a sample involves many steps, including
measurement process the chemical information is car- acquisition of the initial sample, sample preparation or
ried, or encoded, as an optical signal. treatment to produce the analytical sample, presenta-
This chapter first considers the complete spectro- tion of the analytical sample to the instrument, meas-
chemical measurement process and identifies the gen- urement of the optical signals, establishment of the cal-
eral components of a spectrochemical analysis system. ibration function with standards and calculations,
The various ways in which intensities of optical signals interpretation, and feedback. The spectrochemical
are expressed in the radiometric and photometric sys- measurement process can be conceptualized as shown
tems of units are explored so that we can be quantitative in Figure 2-1. The initial sample, obtained from the
about how concentration information is encoded. Spec- bulk material, is often treated prior to its presentation
14 Chap. 2 / Spectrochemical Measurements
Analytical
sample
meters, strip-chart recorders, and digital displays as in and the photometric system. The radiometric system is
digital voltmeters, printers, or CRT terminals. employed almost exclusively in modcrn spectroscopy
In order for the number obtained as a result of and will be used throughout this book. The photometric
spectrochemical measurement to be related to the con- system, based on the response of the human eye, is still
centration of the desired species, it is apparent that the used to describe photodetcctors and several other 6p-
system must be calibrated (standardized) in some man- tical components. The photometric system is thus in-
ner. If the conversion factors or transfer functions of troduced here and compared to the radiomctric system.
all the functional elements in Figure 2-1 were known
exactly, were stable, and were known functions of con-
centration, the measurement system could be calibrated
at the factory to read directly in concentration units and
then checked only occasionally. However, in many The radiometric system of units is based on the actual
spectrochemical instruments in use today, conversion radiant energy emitted by a source or striking a receiver
factors change with time and matrix and may not be (e.g., optical transducer) and is preferred in the Inter-
known exactly. Hence it is usually a requirement that national System ot Units (SI). The basic quantity in this
standards of known concentration be introduced reg- system is the radiant energy Q in joules (J).
ularly for calibration purposes. These standards are taken
through the entire process, if possible, to evaluate the
total system response. The calibration results are often Basic Definitions. In the radiometric system there
expressed as a working curve or as the analytical func- are general quantities used to describe radiation, quan-
tion as described in Chapter 1. In computer-controlled tities used to describe sources, and quantities that deal
systems these calibration data can be manipulated and with the receiver. Table 2-1 lists and defines the basic
stored so that the processing system can produce con- radiometric quantities, symbols, and units. The terms
centration data directly. radiant intensity, emittance, emissivity, and radiance re-
The control system in Figure 2-1 is an integral part fer specifically to radiation from a source and the vol-
of the measurement process. In the past a human op- umes, areas, and solid angles are those of the source.
erator has been the most common controller. However, lrradiance and exposure are used to describe the re-
we are observing more and more instrument functions ceiver and its area. In some cases more than one symbol
being placed under the control of microcomputers. This is given for a quantity in Table 2-1. The first symbol is
frees the human operator for the tasks of experimental that accepted by the International Union of Pure and
design, data interpretation, quality assessment, and often, Applied Chemistry (IUPAC) and used in this book.
sample preparation. The symbols in parentheses are sometimes used and are
The final results of a spectrochemical analysis are included here for complctcness. The units given for
often part of a larger feedback control system, in which radiometric quantitics are practical units rather than
the analytical data are used to indicate the status of a strict •˜I units, which use meters instead of centimeters.
process or the quality of a product. Thus if the infor- Most sources and receivers have dimensions on the or-
mation obtained is outside the limits set for a process der of centimeters and practical units are thus easier to
or product, corrective action must be taken so as to visualize.
reduce the error signal. Even human health care can The qualltities given in Table 2-1 are in gcneral
be viewed as a large feedback control system in which functions of spectral position (wavelength, wavenum-
analytical data are an integral part. Just as in an in- ber, frequency, etc.) in that they are usually employed
dustrial process, the data are used to monitor the status to rcprcscnt the magnitude o l the quantity over some
of the paticnt and to determine the type, intensity, and spectral interval. Often they represent the cun~ulative
timing of any corrective actions that are needed. Spec- magnitude of the quantity over the wavelength interval
trochemical measurements can thus be an extrcmcly from 0 to A. If the nlodificr "total" is employed, as in
important part of such control loops. total radiance, it implies the radiance over the wave-
length interval from 0 t o m .
It is often useful to consider these radiometric
quantities within small spectral intervals. For convcn-
iencc, radiometric quantities per unit spectral interval
In spectrochcmical systems the chemical information is evaluated at :I particular spectral position are called
almost always cncoded as the magnitude of optical sig- aaantities and given a subscript A. if wavelcngth
nals at particular wavclengths. There arc two systcms is used, i if wavenumber is used, and v if frequency is
that describe optical intcnsitics, the radiomctric systcm used. The spectral radiar~ceH, i s t h e radiance per unit
16 Chap. 2 / Spectrochemical Measurements
TABLE 2-7
Radiometric system
General
Radiant energy Q Energy in the form of radiation J(ergs)
(Radiant) energy density U Radiant energy per unit
volume
u = aQ
- dv
J~ m - ~
'Since the radiant flux @ may vary in space and direction, the solid angle 0, the volume V, and the area A in the defining equations must bc small enough to give
meaningful local values. If not, more complicated relationships apply (see Optical Radiatiorz Measurements. Vol. 1 , Radiometry, F. Grum and R. J . Becherer,
Academic Press, New York, NY, 1979). The radiance, irradiance, and emittance are related to the radiant energy density by B = Uc/4?i, E = Uc, and M = Uc,
where c is the speed of light. For a Lambertian surface (constant B independent of viewmg direction), B = M h .
wavelength interval (i.e., per nm) and is given by The total radiance B from a source is related to
its spectral radiance by
Occasionally, one will encounter radiometric terms be characterized by the total radiant power cD that it
with units of photons per second or Einstein's (Avo- emits in all directions. In this case, however, it is more
gadro's number of photons) per second instead of watts. useful to use the radiant power per unit solid angle (the
Because the energy of a photon is just hv, these spectral radiant intensity), which is given by
quantities can be directly converted to watt units if the
frequency is known. A subscript p is often used to de- I = -4<P7 ~ (2-7)
note units of photons s-l so that B , , is the spectral
radiance in photons s-l ~ m sr-l - ~nm-l and B , , =
B,lhv. if the source emits equally in all directions (i.e., is iso-
Geometric Factors. To deal with several of the
radiometric quantities it is important to define the geo-
metric factors of solid angle and projected area that are
used. Figure 2-2 illustrates the plane angle, the solid
angle, and the units used to express these terms. The
unit of solid angle, the steradian (sr), is defined in a
manner similar to that of the unit of angle, the radian.
Since the area of a sphere of radius r is 4nr2, a sphere
subtends 4.rrr2/r2= 4n = 12.57 sr. The concept of pro-
jected area that is used in the definition of radiance is FIGORE 2-3 Projected area. The projected area
illustrated in Figure 2-3. is the effective area of a two-dimensional surface
as seen from a point, 0. If the observer looks
Uses of Radiometric Terms. The radiometric along the normal (N) to the surface (a), the ef-
quantities defined in Table 2-1 are used throughout this fective area is the same as the true area. As the
book, but a few examples are given here for illustrative angle 0 between the line of observation and the
purposes. The specific quantity that is used depends on normal increases the projected area decreases (b)
until at 90" (c) the effective area is zero. The
the situation. In most spectroscopic situations one is projected area A, is then A, = A cos 0. The
eventually interested in the radiant power that is inci- radiance of a source is the radiant power per unit
dent on a receptor (e.g., an optical transducer). Con- solid angle per unit projected area, B = (aWaR)
sider, for example, a point source with dimensions that x (aQldA,) = aZ@l(a@aA cos 0). The solid angle
are small compared to the distance (d) from the source R viewed from 0 is 0 = A,ld2 = A cos @I@,
to the receptor of projected area A,. The source could where d is the distance from 0 to the surface.
Chap. 2 / Spectrochemical Measurements
Area viewed,
A; projected area
Source of
projected area
A1
Photometric System
Note that equations 2-8 and 2-9 both predict the familiar
result that the intensity incident on the receiver is in- The photometric system is a relative system based on
versely proportional to the square of the distance from the apparent intensity of a source as viewed by the
the source. average bright-adapted human eye. The quantities in
In the example shown in Figure 2-4, the source the photometric system have meaning only in the visible
has a significant area and it is usually best to use the region (350 to 770 nm) of the spectrum. The basic unit
source radiance. Also in this figure, only a fraction of the photometric system is the lumen (lm). It is de-
AiIA, of the total source projected area is viewed by fined in terms of the standard candle, which is called a
the receptor. This could occur, for example, because candela. The luminous intensity from a candela is $ of
an aperture between the source and receptor limits the the luminous intensity of a blackbody radiator (see
area viewed. If the source emits radiant power @ equally Chapter 3) of 1 cm2 area at the solidification temper-
in all directions, the source radiance B is ature of platinum (2042 K). A source of 1candela emits
1 lumen per steradian.
Photometric quantities exist which correspond to
each of the radiometric quantities as shown in Table
2-2. The same symbols are used for photometric quan-
tities as for radiometric quantities. To avoid confusion
The radiant power G iincident on the area A, of the when both systems are employed or conversions are
receptor is the source radiance times the solid angle
being made, radiometric quantities should carry the
viewed times the area viewed, or subscript e for energy, while photometric quantities
should carry the subscript v for visible.
Conversions between the two systems are made
with the aid of the following equation, which relates
TABLE 2-2
Photometric and radiometric quantities
Radiometric Photometric Definition of
quantity and units quantity and units photometric quantity
the spectral luminance to the spectral radiance: primarily with the four major classes. The basic rela-
tionships between the quantities measured and concen-
tration are developed in this section and used through-
out the book.
Here V(h) is called the spectral luminous efficiency. It
is a dimensionless factor that depends on wavelength Emission Spectroscopy
and varies in the range 0 to 1 over the visible region
(see W. G. Driscoll and W. Vaughan, eds., Handbook In emission spectrochemical methods the sought-for in-
of Optics, McGraw-Hill, New York, 1978, pp. 1-4). formation is the radiation emitted by the analyte due
At 556 nm a light-adapted human eye is at its peak to nonradiational excitation. We further restrict emis-
efficiency. At this wavelength there are 680 lm W-l. sion spectroscopy in this book so that only techniques
At 510 and 610 nm, the spectral luminous efficiency has that involve excitation by collisional (thermal) processes
dropped to 0.5. Total photometric quantities are ob- are considered emission spectrochemical methods. For
tained by integration of the appropriate spectral quan- example, sodium atoms are excited in a flame by col-
tity as shown for luminance in the following equation: lisional processes and emit characteristic radiation. The
excitation can also be partly electrical as in a direct-
TABLE 2-3
Classification of spectrochemical methods
Class Quantity measured Examples
electrical,
Absorption Spectroscopy
or chemical
energy
In absorption methods the sought-for information is the
magnitude of the radiant power from an external source
that is absorbed by the analyte species. Here absorption
FIGURE 2-5 Emission and chemiluminescence of radiation is accompanied by excitation of the analyte.
(bioluminescence) methods. In (a) the addition For absorption to occur, the frequency of the incident
of thermal, electrical or chemical energy causes
nonradiational excitation of the analyte and emis-
radiation must correspond to the energy difference be-
sion of radiation in all directions (isotropic emis- tween the two states involved in the transition as shown
sion). The energy changes that occur during ex- in Figure 2-6. For many conditions the absorption of
citation (dashed lines) or emission (solid lines) radiation follows Beer's law (see Chapter 3 for deri-
are shown in (b). The energies of states 1 and 2 vation):
are usually relative to the ground level and often
abbreviated E, and E,, respectively. A typical
spectrum is shown in (c).
The Einstein transition probability A,, is discussed fur- FIGURE 2-6 Absorption methods. Radiation of
ther in Chapters 4 and 7 and in Appendix F. incident power Q, can be absorbed by the analyte
Equation 2-15 shows that the radiant power of producing a beam of diminished transmitted power
emission is proportional to the excited-state population @ (a) if the frequency of the incident beam, v,
density and thus to the analyte concentration through or v, corresponds to energy difference, E, or E,
equation 2-14. Even if the emission source is not in (b). The spectrum is shown in (c).
Sec. 2-3 / Spectrochemical Methods
2
Luminescence 21 = hv2, = hc/X2,
@L 1 For low absorbances (abc < 0.01), the luminescence
E2 = hv2 = hc/h2 radiant power is directly proportional to the analyte
El = hv, = hclh, concentration and to the radiant power incident on the
0
sample @., Luminescence methods then involve meas-
urements of @, to obtain the analyte concentration c.
These methods are discussed further in Chapters 11
and 15.
on the order of the wavelength of the incident radiation formation produced is useful or desirable. In analytical
is often called Debye scattering. Here the scattered ra- procedures the selection step allows us to separate the
diation is of the same frequency as the incident radia- analyte optical signal from a majority of the potential
tion, but the angular distribution of the scattered ra- interfering optical signals. The selection process is not
diation, unlike Rayleigh scattering, is not uniform. perfect, however, and we must be aware of the limi-
Scattering from still larger particles is often termed Mie tations of the instruments and components used. The
scattering. Large-particle scattering (Debye or Mie) can vast majority of analytical techniques select the desired
be used to determine particle sizes and is important in information based only on its wavelength. As we shall
turbidimetry and nephelometry where suspended par- see, there are many additional criteria which can be
ticles are the scatterers. employed to improve selectivity. The use of time dis-
Brillouin and Raman scattering are forms of ine- crimination, position discrimination, polarization, mul-
lastic scattering which involve a change in the frequency tidimensional information, and chemical selectivity are
of the incident radiation. Brillouin scattering results but a few of the possibilities.
from the reflection of radiant energy waves by thermal
sound waves, whereas Raman scattering involves the Wavelength Selection
gain or loss of a vibrational quantum of energy by mol-
ecules. These methods are discussed further in Wavelength selection in spectrochemical instruments
Chapter 16. can be based on absorption or interference filters, spa-
The equations describing the dependence of the tial dispersion of wavelengths, or interferometry.
observed scattering signal on experimental parameters Wavelength selectors which disperse the spectral com-
are complex, as shown in Chapter 16. However, as for ponents of the optical signal spatially are the most com-
photoluminescence techniques, the scattering signal is mon, and some of the major configurations are shown
proportional to the incident radiant power. in Figure 2-8. The entrance slit defines the area of the
source of radiation that is viewed. The dispersive ele-
ment can be a prism which spatially separates wave-
2-4 SELECTION OF OPTICAL lengths by refraction or a grating which disperses light
INFORMATION based on diffraction. The image transfer system pro-
duces an image of the entrance slit on the focal plane.
Sorting out all the optical information that might be As shown in Figure 2-8, the name of the dispersive
produced in the encoding step is a major step in a spec- wavelength selector depends on the arrangement of ap-
trochemical measurement. It is a rarity that all the in- ertures or slits in the focal plane where the spectrum is
of the analyte as the radiant power of emission cell, a composite total luminescence signal EtL is ob-
some spectrochemical methods the excitation tained:
source and sample container are an integral unit, as in
e nebulizer-burner used in flame emission and the
action cell used in chemiluminescence. where EL is the analyte luminescence signal, EE is the
When the analytical sample is present in the sam- analyte thermal emission signal, and E,, is the blank
e cell, a total or composite signal E, is obtained. This signal. The blank signal is composed of the dark signal
la1 signal is the sum of analytical signal EE, the dark Ed, the background emission signal E,,, a scattering
Ed, and the background emission signal EbE. To signal E,, due to scattered source radiation, and a back-
ct the analytical signal, a second measurement is ground luminescence signal EbL:
ed to obtain the sum of the dark signal and the
ackground emission signal. This second measurement
ally made by replacing the analytical sample with
ank that is ideally identical to the analytical sample Analyte and background emission in the UV-visible
except that the analyte is missing. Thus the analytical region are usually significant only in atomic spectros-
obtained as follows: COPY.
The analyte luminescence signal EL can be ob-
tained with two measurements only if the analyte emis-
ere E,, is the blank signal given by Ed + EbE. If sion signal EE is small compared to EL, which is often
esired, the dark signal can be obtained separately by the case. If EE is significant, subtraction of the blank
locking all radiation from reaching the radiant power signal gives a measured analyte luminescence signal
onitor. The background emission signal could then EL that differs from EL as shown in the equation
e obtained from E,, - Ed. In many instruments the
lank solution is used to adjust the readout device to
read zero by suppression of the blank signal. This es-
To obtain the true analyte luminescence signal EL when
blishment of the zero position is still, however, a
E, is significant, the excitation source must be turned
easurement of the blank signal.
off. Then the two measurements indicated in equation
2-22 are made to obtain E,. Subtraction of EE from
otoluminescence Spectrometry
EL gives the true analyte luminescence signal. In some
he instrumentation used in photoluminescence meth- cases it is possible to eliminate the measured contri-
s is illustrated in Figure 2-11. Here an external source bution from analyte emission optically or electronically.
electromagnetic radiation excites the analyte (pho- For example, if the excitation source is modulated and
toexcitation). The analyte concentration is optically en- alternating-current (ac) amplification is used, the ac
as the luminescent radiant power @ ,, which is luminescence signal can be distinguished from the dc
red with the radiant power monitor. The emission emission signal. Often the blank measurement is used
wavelength selector that views the luminescence of the to set the zero position of the readout device.
sample is typically placed to collect radiation at 90" with
respect to the excitation axis. Other geometries, such
as front surface and 18O0,are used in special situations
Chapters 11 and 15). In some cases only one of A typical absorption spectrometer is illustrated in Fig-
avelength selectors is necessary. ure 2-12. The absorption spectrometer is essentially
When the analytical sample is placed in the sample identical to the luminescence spectrometer of Figure
2-11 except that the source, sample cell, and transducer transmittance (100% T) signal obtained with the shutter
are all on the same optical axis. This permits the meas- open and the blank (reference) in the sample container.
urement of the transmitted radiant power. The shutter The 0% T signal EOt is composed of any back-
allows the user to block the radiation source in order ground emission EbE and dark current Ed as shown by
to obtain the dark signal. Usually, only one wavelength the equation
selector is required.
Absorption measurements can be made in two
ways: The transmittance T can be displayed by the read-
out device and the absorbance A calculated manually
When the blank is in the sample container and the
from A = -log T; or the logarithmic conversion can
shutter open, the measured total reference signal E,,,
be done electronically or with computer software and
called the 100% T signal, is composed of the reference
the absorbance A displayed by the readout device. Both transmission signal Er, the 0% T signal, and any back-
of these readout schemes are used, although direct ab- ground luminescence EbL:
sorbance readout is becoming the more common.
luminescence techniques, analyte and background tainer. The voltage EA is then given by
are usually significant only for atomic spec-
trometry. The measured transmittance T' is then used
to calculate an absorbance A' = -log T' which is an
approximation to the true absorbance A = -log T .
The voltages El, and E, are logarithmically related to
Es and E, as follows:
DirectAbsorbanceReadout. As part of the sig-
nal processing electronics, many modern absorption Es Er
spectrometers have provision for obtaining and dis- El, = - k' log - and E, = - k' log -
k k"
playing absorbance directly. The true absorbance A is
given by
where k is a constant reference voltage. Often El, is
set to zero on the readout device so that E, is read out
directly as EA.
Note that in the two-step absorbance measure-
ment scheme, a measurement is not made with the light-
where EA is the voltage proportional to the analyte source shutter closed (0% T ) since A would be infinity.
absorbance and k' is a logarithmic conversion factor in Thus (Ed +EbE) must be negligible compared to Es
volts per absorbance unit. and Er or electronically or optically set to zero by other
The voltage EA and hence A are found from two means. Also, EE + +
EbL EL must be negligible so
measurements: First a reference logarithmic voltage or that Es = E,, and E, = E,,; otherwise, the measured
zero absorbance voltage E, is obtained with the shutter absorbance A' only approximates the true absorbance A.
open and the blank in the sample container; then a We will make use of these equations in later chap-
sample logarithmic voltage E, is obtained with the shut- ters as we explore the factors influencing the accuracy
ter open and the analytical sample in the sample con- and precision of spectrochemical methods.
PROBLEMS
2-1. Consider the spectrometer below, which can be used diant power incident upon the sample and radiant power
for emission, luminescence, or absorption measure- monitor, respectively. The following measurements
ments. Note that the radiant power monitor can be were made with either an analyte solution (a) or an
placed in two positions in which it is equidistant from ideal blank (b), where C = closed shutter and 0 =
the sample, and that shutters 1 and 2 control the ra- open shutter.
Shutter 1
Monochromatic Radiant
radiation source Sample power monitor
Position A
Radiant
power monitor
Position B
Chap. 2 1 Spectrochemical Measurements
- - - --
Consider a collection o f atoms in thermal equilibrium
Position at 3000•‹C.What fraction o f the atoms are in the first
of radiant
monitor Shutter I Shutter 2 Sample Signal (V) excited state i f the energy differencebetween the ground
and the first excited state corresponds to 400 nm? As-
A C 0 a 0.15 sume that the statistical weights o f the ground and first
A C 0 b 0.10 excited states are the same and that higher excited
A C C b 0.07 states are not significantly populated.
A 0 0 b 3.20 The atomic density o f an excited state is 4.00 x 101•‹
A 0 0 a 1.50 atoms cmW3.Calculate the radiant power o f emission
B C 0 a 0.15
from a volume element of 1.00 mm3 o f this excited
B 0 0 a 0.40
B 0 0 b 0.12 state in photons s-I if the Einstein transition coeffi-
cient is 1.00 x 10' s-I.
What is the difference between a monochromator and
In this experiment, scattering and chemiluminescence a polychromator?
are insignificant. Calculate or indicate the following An ordinary 3.0-V flashlight bulb draws roughly 0.25
signals: Ed, Em, Eb,, E,, El, EL, Em, En, Em Es,En A o f current and converts about 1.0% o f the electrical
T , A , T',A ' . power into light ( A ;= 550 nm). I f the flashlight beam
A point source emits 25.13 W . Express the intensity initially has a cross-sectional area o f 10 cm2:
as a radiant intensity. (a) How many photons are emitted per second?
A n extended source is spherical in shape with a radius (b) How many photons are found in a cubic meter o f
o f 2.00 cm and emits 12.57 W from 399.5 to 400.5 nm. the beam?
Determine the spectral radiant emittance, the spectral (c) What is the radiant emittance (W ~ m - o~f the )
radiant intensity, and the spectral radiance at 400 nm. beam as it leaves the flashlight?
A beam o f 632.8-nm photons from a He-Ne laser strikes A 1.0-mW laser has a beam diameter o f 2 mm. Assume
a detector area o f 5.0 mm2. The laser has a flux o f that the beam has negligible divergence and calculate
3.18 x l O I 5 photons s-I. the radiant energy density ( J ern-;) o f the laser beam.
The 4p level o f the Na atom is 3.75 eV above the 3s
(a) What is the laser radiant power in watts?
ground level. What wavelength o f radiation (in nm)
(b) What is the laser irradiance at the detector?
would be required to excite Na from the ground level
(c) Express the answer for part ( b )in terms o f photons
to the 4p level? What frequency is required? What is
s-I rather than watts.
the energy o f this transition in joules?
W h y are electronic, vibrational, and rotational spectra For the copper atom, there are resonance lines at 324.7
observed in different wavelength regions? and 327.4 nm. What is the energy difference in eV
Calculate the absorbances corresponding to transmit- between the two excited states involved in these two
tances o f 0.100, 0.0316, and 0.0100. transitions?
A n extended source emits 2.00 W ~ m sr-' - nm-'
~ at A sample is illuminated with 1.0 pW o f radiation and
300 nm. Calculate the spectral radiant power impin- transmits 90% o f the incident radiation. Calculate the
gent on a 1.00-cm2 receptor that is 2.00 m away in W maximum value o f the luminescence radiant power
nm-I and photons s-' nm-' i f a 1.00-mm-diameter that could be observed.
aperture is placed at the source. What is the incident The irradiance on a receptor 50 cm from a source with
radiant power in watts over 10.0 nm centered at 300 a projected area o f 0.010 cmZ is 2.0 FW ~ m - Cal- ~ .
nm i f the source radiance is constant over this region? culate the radiance o f the source.
1. F . Grum and R . J . Becherer, Optical Radiation Meas- 3. F. E. Nicodemus, ed., Radiometry: Optical Resource Let-
urements, vol. 1, Radiometry, Academic Press, New York, ter, American Institute o f Physics, New York, 1970.
1979.
4. E . L. Dereniak and D. G . Crowe, Optical Radiation De-
2. W . G . Driscoll and W . Vaughan, eds., Handbook of tectors, Wiley, New York, 1984. Includes a chapter on
Optics, McGraw-Hill, New York, 1978. A very useful
radiometry.
handbook with a good deal o f information about optical
principles and optical materials. Contains a chapter on 5 . H. T . Betz and G . L. Johnson, "Spectroradiometric Prin-
"Radiometry and Photometry" ( J . F . Snell) that is quite ciples," in Analytical Emission Spectroscopy, vol. I , pt.
practical. 11, E. L. Grove, ed., Marcel Dekker, New Y o r k , 1971,
Chap. 2 / References
pp. 323-381. Presents a good introductory discussion of 7. J. Wilson and J. F. B. Nawkes, Optoelectronics: A n In-
the principles of radiometry. troduction, Prentice-Hall, Englewood Cliffs, N.J., 1983.
6. Electro-Optics Handbook, Technical Series EON-11, RCA Includes a brief discussion of the radiometric system.
Corp., Lancaster, Pa., 1974. Has a good discussion of the 8. IUPAC, Compendium of Analytical Nomenclature, Per-
radiometric and the photometric systems. gamon Press, Oxford, 1978.
We have seen in Chapter 2 that there are four or five ments are introduced, and the nonidealities which must
basic components that are present in optical spectrom- be considered in their use are described. Filters, prisms,
eters. In addition to excitation sources for emission, and gratings are commonly employed for wavelength
external radiation sources for luminescence and ab- selection purposes. These components are first consid-
sorption, sample containers, wavelength selection de- ered as optical elements and then they are combined
vices, and radiant power monitors, optical spectrome- with lenses and mirrors to form complete wavelength
ters contain a number of optical elements for collimating, selection systems. The chapter ends with a discussion
focusing, and directing electromagnetic radiation. In of Fabry-Perot, Michelson, and other interferometers,
this chapter we consider the optical components and devices that are used in nondispersive wavelength se-
systems used in spectrochemical instruments. Radiation lection systems.
sources and radiant power monitors are described in
Chapter 4, while the remaining components of spectro-
chemical instruments are discussed in later chapters 3-1 BASIC OPTICAL RELATIONSHIPS
dealing with specific methods.
Prior to considering optical elements in detail, it There are several basic laws in optics from which many
is necessary to review several important optical rela- other important relationships are derived. The conser-
tionships. Thus this chapter begins with a discussion of vation law, the laws of reflection and refraction, and
the conservation law, and the laws of reflection, re- the absorption law are used throughout this chapter in
fraction, and absorption. Then the phenomena of in- our discussion of optical components and spectrometric
terference and diffraction are explored. The nature and systems.
production of polarized light are discussed in some de-
tail. Devices that can amplitude modulate a radiant The Conservation Law
energy beam are then discussed. These modulators can
be mechanical, electro-optical, acousto-optical, or mag- A basic principle of wave motion states that when a
neto-optical devices. The mirrors and lenses that are wave strikes a boundary between two media, a portion
used for imaging purposes in spectrochemical instru- of the wave is reflected, a portion is absorbed, and a
sic Optical Relationships
is transmitted into the new medium. This is The permittivity of a medium is related to that of
often known as the conservation law. Consider radiant vacuum (free space) by E = &E,. where iK, is the
energy of wavelength A to be incident on a boundary or relative permittivity
between two media. The fraction of the incident radiant meability of a material
energy lost by absorption at the interface or surface is
, a@).The fraction is the dimension
e is the spectral re- Thus the index of refraction can also be written as
ransmitted into the
total internal reflection. Many reflex cameras, binoculars, Equation 3-11 shows that the flux (radiant power) de-
and telescopes use totally reflecting prisms for this reason. creases exponentially with increasing distance through
Some refractometers, which measure refractive index, are an absorber.
based on total internal reflection. This phenomenon is The absorption coefficient k has units of cm-l.
also the basis for fiber optics transmission of light, as Its magnitude depends on the wavelength of the inci-
discussed in Section 3-3.
TABLE 3- 1
The Absorption Law Assumptions of the absorption law
derf radnation, the nature of the absorber, and the con- be carefully considered in spectrochemical applications.
certraeaon of the absorber. In atomic spectroscopy, the The absorption coefficient should be small enough that
a b ~ o law ~ isg most
~ ~often~ used in the form shown in attenuation by the material is slight. The absorption
e q 3-11, ~where k~ is understood
~ ~to be concentra-
~ coefficients of common optical materials are plotted vs.
t,,, dependent. In molecular spectroscopy, however, wavelength in Appendix 8 . In the UV region quartz
mon to state explicitly the concentration de- (natural silica) or synthetic silica is used. Glass is used
peridence of the absorption coefficient by expressing k in the region 320 nm to 2 km, while halide salts are
g = k'c, where k t is a new absorption coefficient employed in the infrared and vacuum UV region.
In;3Ppendentof concentration. Thus equation 3-11 be-
= cD,ePk bc. Most often in absorption spec-
we measure the transmittance T = @/@, or -2 INTERFERENCE,
the absorbance A = -log T. Thus AND POLARIZATION OF
ELECTROMAGNETIC WAVES
E = E, + E2 (3-17)
= E , , sin (wt + 6,)f E,,,sin (ot + 41,)
With trigonometry it can be shown that the composite
wave is of the form
E = E, sin (ot + +) (3-18)
Equation 3-19 shows that the square of the am- where x, and x2 are the distances from the sources of
plitude of the composite wave is given by the sum of the two waves to the point of observation and h is the
the squares of the amplitudes of the individual waves wavelength in the medium. If the two waves are initially
plus an interference term, 2E,,E,, cos (6, - 6,).Since in phase at the sources ($,, = $,,), or come from the
the irradiance of the resulting wave depends on the same source by two different routes, we can write
square of the amplitude (Equation 3-16), the irradiance
also depends on this interference term. In fact, if we
define 6 = $, - h,, constructive interference (maxi-
mum irradiance) will occur when 6 = 0, k271, 1 4 ~ ,
and so on. Or we can write the condition for construc- The refractive index of the medium is y = h,/h, where
tive interference as 6 = liz2rnn, where m is 0, 1. 2, and h, is the wavelength in vacuum. Thus
so on. Similatly, destructive interference (minimum ir-
radiance) occurs when 6 = k 71, 1 3 7 and ~ ~ so on. The
resultant electric field vector E for in- and out-of-phase
conditions is shown in Figure 3-4.
The phase difference 6 can arise from a difference
in pathlength or a difference in initial phase: where h(0PL) is the difference in optical pathlength
of the two waves. Constructive interference occurs when
6 = m271 or
Sec. 3-2 I Interference, Diffraction, and Polarization of Electromagnetic Waves 37
where m is 0, ? 1, -+2, and so on. Thus, constructive parallel. The resulting diffraction pattern observed at
interference occurs when the optical pathlength differ- a long distance or with a lens is termed Fraunhofer
ence is an integral multiple of the wavelength. Similarly, diffraction. Rays from the slit that reach Po (Figure
destructive interference occurs when A(0PL) = 3-5a) all have identical optical pathlengths. Since they
(2m + 1)Xo/2. We will use this important result in de- are in phase in the plane of the slit, they are still in
scribing a variety of important optical effects. phase at Po and give rise to a central maximum of ir-
radiance. Now consider two rays that reach P, on the
terference
Lens or
We may define optical interference as the interaction large
Slit distance
of light waves to yield a resultant irradiance that is
different from the sum of the component values, as
predicted by equation 3-19. Two waves for which the
initial phase difference, +,, - b02, is zero or constant
for a long time are said to be coherent. Coherence is
thus a condition for achieving interference. Waves emit-
ted by separate sources are incoherent with respect to
Screen
one another. For this reason we normally observe in-
terference phenomena when light reaching the point of (a)
interference comes from a single source by two different
paths. Another condition is that the interfering waves
ust have very nearly the same frequency composition.
This again is readily achieved if the radiation comes
from a single source. Finally, although we have consid-
ered the sources to be polarized, normal unpolarized
light also produces interference for reasons that are
discussed later in this section.
Interference of light waves can be produced in
several ways. In wavefront-splitting devices, portions
of the primary wavefront are used to emit secondary
waves that produce the interference pattern. The classic
Young two-slit interference experiment is an example.
In amplitude-splitting devices, the primary wave itself
is divided into two segments which travel different routes
before recombining and interfering. An example of this
type is the Michelson interferometer discussed in Sec-
tion 3-7. Interference effects are also produced by thin
films and by plates of various thicknesses. The Fabry-
Perot interference filter and Fabry-Perot interferome-
ter are examples of multiple-beam interference devices;
both are discussed in Section 3-7.
iffraction
screen (Figure 3-5b). The ray originating at the top of travels a distance b is approximately its original width
the slit must travel a distance x = (W12) sin 0 farther W plus b times the angular width AIW. For large enough
than the ray from the center of the slit. According to b, we can neglect the original width and W' = bhlW.
our earlier discussion, we will get destructive interfer- A plot of the irradiance of a diffraction-limited beam
ence when this pathlength difference is A12. Thus a min- upon striking a screen is shown in Figure 3-6.
imum irradiance occurs when (Wl2) sin 0 = A12 or Now consider multiple slits as shown in Figure
W s i n 0 = A. 3-7. The difference in OPL between adjacent rays is
We can locate another of the minima by dividing x = d sin 0, where d is the distance between slits.
the slit into four parts as shown in Figure 3-5c. Now Maxima in irradiance occur when this distance is an
any two adjacent rays will destructively interfere when integral multiple of the wavelength or
(Wi4) sin 0 = A12 or W sin 0 = 2A. By extension, the
general formula for diffraction minima is
half-width (width at half maximum intensity) of the FIGURE 3-6 Fraunhofer diffraction at a single
beam in this case is said to be diffraction 1imited.The slit. The angular half-width is = XIW. The irra-
width cannot be reduced to zero unless W is infinite or diance of the second maximum is about 4.4% of
A is zero. The total beam half-width W' after the beam that of the principal or central maximum.
Screen or
film
Slits
(a)
Large N
u,
FIGURE 3-7 Multiple-slit diffraction from six slits separated by distance
d. In (a) light rays reaching one point P on a screen are shown. If the
path difference x = d sin 0 is an integral multiple of a wavelength, the
waves are in phase at P. In (b) the irradiance pattern at the screen is
shown for N (number of slits) = 2 , 4 , and large. For large N each principal
maximum has the shape of a single slit pattern.
Sec. 3-2 / Intederence, Diffraction, and Polarization of Electromagnetic Waves
e number of slits increases, each diffraction max- of the resultant depends, of course, on the amplitude
urn grows in irradiance and becomes narrower. The and phase of the two superimposing waves. If, however,
ing occurs because with multiple slits only a slight the electric field vectors of the waves are mutually per-
in angle away from that corresponding to a max- pendicular, the resultant wave may or may not be lin-
imum leads to almost total destructive interference. Dif- early polarized. If the waves are of equal amplitude and
fraction gratings can be considered to consist of many orthogonal, superposition can lead to plane-polarized,
closely spaced slits. Such gratings are considered in de- elliptically polarized, and circularly polarized radiation;
tail in Section 3-5. which of these is obtained depends on the phase dif-
ference between the two waves, as discussed below.
Linear Polarization, Let us consider in more de-
tail two waves with mutually perpendicular electric field
e have seen that electromagnetic radiation is a trans- vectors. If the two waves have a phase difference that
verse wave and have discussed the nature of plane- is zero or an integral multiple of k 27r, they are in phase,
arized or linearly polarized light. We will now con- and the resultant wave is linearly polarized, as shown
r what occurs when we have two plane-polarized in Figure 3-$a. We can equally well reverse the process
light waves of identical frequency moving through the and resolve any plane-polarized wave into two mutually
same region of space in the same direction. If the elec- perpendicular plane-polarized components. These two
tric field vectors of the two waves are aligned with each orthogonal components are often denoted the parallel
er, they will simply combine to give a resultant wave (11) or 7r component and the perpendicular (1)or a
is also linearly polarized. The amplitude and phase component.
If the two orthogonal waves have a phase differ- Normal Light. Light from common sources (fil-
ence that is an odd-integer multiple of kn, they are ament lamps, the sun, arc lamps) is emitted by nearly
180" out of phase, but again combine to give a resultant independent radiators (atoms and molecules). Each ra-
that is linearly polarized. In this case the plane of vi- diator produces a polarized wave train for a short time
bration is rotated from that of the in-phase case (not (= s). The light propagating in a given direction
necessarily by 90"). consists of many such wave trains whose planes of vi-
bration are randomly oriented around the direction of
propagation. As a result of the random superposition
Circular Polarization. An interesting case arises of independent polarized wave trains, no single result-
if the two waves orthogonal waves are 90" out of phase. ant state of polarization is observable, and natural light
As shown in Figure 3-8b, such a case gives rise to cir- is referred to as unpolarized light. Unpolarized light can
cularly polarized light because the resultant traces out be described as two orthogonal plane-polarized waves
a circle. Note in Figure 3-8b that the resultant vector of equal amplitude with a phase difference between
(when the x component lags the y component by 90") them that varies randomly in time.
rotates clockwise. The resultant wave is said, therefore,
to be right circularly polarized. If, on the other hand, Optical Rotatory Dispersion. For some sub-
the x component were to lead the y component by 90", stances, the characteristics of optical phenomena such
the resultant electric field vector would rotate counter- as absorption, refraction, and reflection depend on the
clockwise. The resultant in this case is said to be left polarization of the incident radiation. Substances that
circularly polarized. rotate the plane of vibration of plane-polarized radia-
If we were to add two equal-amplitude, in-phase, tion are termed optically active. These include aniso-
but oppositely polarized circular waves together, the tropic crystals and liquids or solutes in solution that can
resultant would be a plane-polarized wave. Hence a exist as enantiomers (e.g., chiral molecules). In the
plane-polarized wave can be considered to consist of latter case, optical activity is observed if one of the
equal-amplitude left and right circularly polarized waves. enantiomers is in excess.
The two oppositely polarized components are often called The rotation of plane polarized light by an opti-
the 1 and d components, where d (right-handed or dex- cally active substance can be viewed as being due to
trorotatory) refers to clockwise rotation and 1 (left-handed the different propagation rates of the d and 1 compo-
or levorotatory) refers to counterclockwise rotation from nents, as shown in Figure 3-10a and b. The propagation
the point of view of the observer looking toward the velocities differ because of the different refractive in-
propagation vector. dicies for the d and 1 components, qd and ql, respec-
tively.
The rotation in degrees (a),also termed the op-
Elliptical Polarization. If the two superimpos- tical rotatory power, is given by
ing, plane-polarized waves have a phase difference be-
tween O" (linear polarization) and 90" (circular polari-
zation), the resultant traces out an ellipse and the
radiation is said to be elliptically polarized. In actuality,
linearly and circularly polarized light are simply special where b is the pathlength and X is the wavelength of
cases of elliptically polarized light where the phase dif- the incident radiation. The difference in refractive index
ference is 0" or 90". The various polarization configu- for the two polarization components (q, - q,) is de-
rations for different phase differences are illustrated in noted the circular birefringence. It is often only on the
Figure 3-9. order of However, with a 10-cm pathlength, op-
tical rotations of tens to hundreds of degrees are often Normally, b is given in decimeters. The quantity [a]is
observed. For a d-rotatory compound, the rotation and termed the specific rotation if c is in g mL-I and has
circular birefringence are positive; these quantities are units of degrees per gram per milliliter per decimeter.
negative for a 1-rotatory compound. The rotatory power If the concentration is expressed in molar units, [a]is
nds not only on the compound, wavelength, and called the molar rotation with units of degrees per mole
length, but also on the temperature, and for so- per liter per decimeter. For pure liquids, c in equation
ns of an optically active solute, the solvent and the 3-26 is replaced by the density (p).
nalyte concentration. In polarimetry, the rotation is measured at one
The rotation can be normalized to a particular wavelength. The temperature and wavelength are com-
athlength and concentration as follows: monly specified by adding a superscript and subscript
to a. Standard wavelengths include 589 nm (the Na D
a line) and 546.1 nm (the green Hg line). In spectropo-
[a] = -
bc larimetry, the dependence of the rotation on wave-
42 Chap. 3 1 Optical Components of Spectrometers
length is measured and is termed the optical rotatory where the difference in molar absorptivities is denoted
dispersion (ORD) . the circular dichroism. The units of [8] are often in
degrees centimeter squared per decimole and the factor
Circular Dichroism. Circular dichroism (CD) of 3300 applies if b is expressed in decimeters and c is
depends on the difference in molar absorptivities for in molarity units. A circular dichroism curve or spec-
the d and 1 components, E, and E,, respectively, by trum is a plot of the molar ellipticity versus wavelength.
optically active materials. If an absorbing sample of an In most cases, circular dichroism and optical ro-
optically active compound is illuminated with plane- tation occur simultaneously as illustrated in Figure
polarized radiation, the differential absorption results 3-10e. The angle of rotation (a)in degrees is taken as
in one of the circularly polarized components being the angle between the plane of polarization of the in-
absorbed more strongly than the other component (see cident beam and the major axis of the emerging ellip-
Figure 3-10c). Thus the transmitted radiation is ellip- tical beam. The rapid change in the CD and ORD near
tically polarized. The eccentricity of the ellipse can be the maximum of an absorption band is termed the Cot-
characterized by a quantity termed the ellipticity (0) in ton effect. Usually, the sign of the optical rotation changes
degrees. It is a measure of the magnitude of CD and is near the maxima of absorption bands. The ellipticity
related to the length of the major and minor axes of can also vary in sign in the vicinity of absorption bands.
the ellipse as shown in Figure 3-10d. The ellipticity Thus a CD curve can exhibit both positive- and nega-
cannot be directly measured as the optical rotation. tive-going bands.
Rather the absorbance with incident radiation that is
circularly polarized in the d direction (A,) and the ab- Production of Linearly Polarized Light, A de-
sorbance with 1 circularly polarized radiation (A,) are vice that produces polarized light from unpolarized light
separately measured. The ellipticity is calculated from in known as a polarizer. A linear polarizer produces
these absorbances and the equation plane-polarized light. Polarizers are based on one of
four phenomena: absorption (linear dichroism), reflec-
0 = 33 (Al - Ad) (3-27) tion, scattering, or linear birefringence (double refrac-
tion). The familiar Polaroid sheet contains dichroic ma-
The molar ellipticity ([el) is given by terial that absorbs the component of unpolarized light
perpendicular to the polarizing direction. The trans-
mitted light is plane polarized. The transmitted intensity
through a second polarizing sheet positioned as shown
-.
components are equal, p = p, + p,. Curves b and c (a)
of Figure 3-2 illustrate how the IT and a reflectances u component
behave for different angles of incidence. At a certain I
n component
angle of incidence, known as Brewster's angle 0,, the Incident ray I No reflected
(polarized) I
flectance of the IT component becomes zero. From
e expression for p, above, this occurs when
+ 82 = 90". From Snell's law the necessary angle
ccurs when tan 8, = %/ql. At Brewster's angle,
= tan-l(q2/ql), the reflectance for the IT component
is zero so that the reflected beam is plane polarized with
its plane of vibration at right angles to the plane of
incidence (Figure 3-12a). A plane-polarized incident
eam with its plane of vibration in the plane of incidence
will undergo no reflections at Brewster's angle (Figure
3-12b). At angles other than Brewster's angle, the re-
nce for the u component is greater than the IT
onent (see Figure 3-2, curve c). This is also true FIGURE 3-12 Polarization by reflection. In (a)
for reflection at metallic surfaces. the incident unpolarized beam consists of a com-
Polarized light is also produced by scattering. The ponent perpendicular to the plane of the propa-
gation vector that is perpendicular to the interface
sky appears blue in daytime because short-wavelength
surface (a-component) and a component lying in
light is most readily scattered. Sunlight scattered at right the plane of incidence (n component). At a cer-
angles is fully polarized; all sunlight is partially polar- tain angle of incidence O,, known as Brewster's
ized. We shall examine the scattering of polarized light angle, the reflected light is completely polarized
and the polarization of light by scattering in more detail (only the a component is reflected; the trans-
in Chapter 16. mitted light is partially polarized). In (b) a po-
Optically anisotropic crystals have optical prop- larized incident beam is fully transmitted a t
erties that are not the same in all directions. Polarized, Brewster's angle. Since the indices of refraction
monochromatic rays travel through the crystal with ve- depend upon wavelength, Brewster's angle is also
locities that depend on the state of polarization and the wavelength dependent.
direction of propagation. An exception to this occurs
along a particular axis known as the optic axis where law at the crystal surface and propagates as though the
monochromatic rays travel with the same velocity re- crystal had a refractive index which is independent of
gardless of the direction of polarization. The optic axis direction (the ordinary refractive index qo). The second
is a property of the crystal and is determined by the beam, called the extraordinary ray, does not obey Snell's
crystal structure. Birefringcnt crystals, such as calcite, law; it propagates with a velocity determined by the
show different indices of refraction for the orthogonal extraordinary refractive index qe, which varies with the
polarization states except for rays that propagate along direction of propagation. Advantage can be taken of
the optic axis. the two indices of refraction to separate the two beams;
If unpolarized light is sent into a crystal of calcite thus birefringent crystals can be used as polarizers. Sev-
at an angle to the optic axis, two beams emerge which eral polarizing prisms based on this principle are dis-
have their planes of vibration at right angles to each cussed in Section 3-5, and values of qo and q, are tab-
other. One beam, called the ordinary ray, obeys Snell's ulated in Appendix B for several important crystals.
44 Chap. 3 1 Optical Components of Spectrometers
ordinary ray. Since the two rays move through the crys-
tal at different velocities, a phase difference is intro-
duced. The resultant wave is a superposition of the
extraordinary and ordinary waves. If the thickness of
the crystal is such that a 90" phase difference results,
the plate is known as a quarter-wave plate; a half-wave
plate introduces a 180" phase difference. Circularly po-
larized light is most often produced from normal, un-
polarized light by a series combination of a linear po-
larizer and a quarter-wave plate. A half-wave plate can
change right circularly polarized light to left, and vice
versa.
Rhomboids (rhombs) are also used as retarders
and have the advantage of being essentially achromatic.
FIGURE 3-13 Fresnel rhomb. Internal reflec- Their use is based on the different phase shifts that
tion introduces a phase difference between the occur for the parallel and perpendicular components
two perpendicular components of a linearly po- during internal reflection. The phase difference de-
larized wave. At an incidence angle of 54.6", the pends on the angle of incidence and is essentially in-
phase shift is 45". In the Fresnel rhomb, two such
reflections are used to produce a 90" phase shift, dependent of wavelength. The Fresnel rhomb, shown
which yields circularly polarized light. in Figure 3-13, is commonly used to produce circularly
polarized light.
Production of Circularly Polarized Light.
Circularly and elliptically polarized light can be pro-
duced by retardation of one polarization state in a re-
tarder plate. If a birefringent crystal is cut so that its
optic axis is parallel to its front and back surfaces, an Several types of devices are used to amplitude modulate
incident monochromatic beam normal to the axis will a radiation source (see Section 4-5). Modulation is based
propagate through the crystal as an extraordinary and on mechanical interruption of a light beam or on elec-
-
0' \
Radiant
energy FIGURE 3-14 Types of mechanical
modulators. In (a) a rotating wheel
or disk chopper (often called a toothed
wheel chopper) is illustrated. A high-
quality motor is used to control the
rotation rate of the wheel. The pe-
riodic interruption of the light beam
can also be controlled by rotating
vanes as in (b). In the oscillating or
tuning fork design of (c), vanes are
attached to the tines of a tuning fork.
The tines are driven by electromag-
nets. The resonant frequency is de-
termined by the size, mass, and spring
tension of the tines. Oscillating chop-
pers as shown in (d) are based on a
periodic rotational oscillation of a vane
over a controlled arc (2 to 30'). These
are also electromagnetically con-
trolled resonant devices.
tro-optic, magneto-optic, or acousto-optic phenomena. Because the birefringence and retardation are re-
In this cection, modulators based upon all of these phc- lated to the applied electric field, these E - 0 effects
nomena are described, including mechanical choppers; provide a means to construct an electrically controlled
c~ckels,and Bragg cells; and the Debye-Sears variable retarder or wave plate. The Merr effect is a
modulabr. quadratic electro-optic effect because the induced bire-
fringence is proportional to the square of the electric
field or voltage applied. By contrast, the birefringence
hlanical modulators or choppers provide a con- induced by the Pocke s effect is directly proportional to
ed periodic physical blocking of a radiation beam. the electric field or applied voltage and is a linear elec-
The lnost common mechanical modulators are based tro-optic effect. Crystals which are linearly electro-optic
on a rotating disk or wheel with apertures or vanes as are also piezoelectric; application of an electric field
shown in Figure 3-14a and b. The chopping frequency produces a change in direction, and application of pres-
is determined by the number of apertures and the ro- sure induces a voltage change.
tation rate of the shaft of the motor. The maximum Electro-optic modulators based on the Kerr and
modulation frequency is generally in the range 1 to 10 Pockels effects are shown in Figure 3-15a and b. Note
kHz. Vibrating or tuning fork modulators are based on that the electro-optic "cell" is sandwiched between two
lateral movement of one or two vanes in and crossed linear polarizers. When no electric field is ap-
out of the light path as illustrated in Figure 3-14c. Be- plied to the cell, no radiation is passed. If a voltage is
cause these are resonant devices, a given chopper is applied that induces a 180" phase shift, the plane of
designed to operate at one frequency (10 to 6000 Hz). polarization is rotated 90" and maximum transmission
The modulation waveform depends on the size, shape, is obtained. By applying the appropriate sinusoidally
and configuration of the vanes and can be sinusoidal. modulated voltage, a sinusoidally modulated radiation
types of resonant choppers include oscillating beam is produced. Pockels cells can provide sinusoidal
ers based on torsion rod and taut band designs modulation at frequencies as high as 10 to 100 MI-Iz. If
(Figure 3-14d), which rely on rotation of a vane in and the applied voltage is pulsed, rise times from 1 to 30 ns
our of the light path. are achieved. Kerr cells can be modulated up to about
Choppers can be used for specialized functions by 10 MHz. Pockels cells are usually preferred because
ounting mirrors, refractor plates, gratings, or filters they require 1 to 3 kV for operation, which is 5 to 10
to the vanes. A chopper with mirrored vanes is com- times less than that required for Merr cells.
only used as a chopping beam splitter, as discussed Circular birefringence induced by a magnetic field
in Section 3-4. A mirrored oscillating chopper can be is termed the Faraday effect. This magneto-optic effect
used to sweep a beam across a target or aperture at can be used to construct a Faraday modulator, as shown
high speeds (e.g., up to 105degrees s-I). in Figure 3-1%. The overall design is similar to an
In some applications, it is only necessary to block E - 0 modulator except that a magnetic rather than elee-
or unblock a radiation beam at certain times in an ex- tric field is applied. In this case, the transmitted radia-
erianent (e.g., to measure the dark signal). Beam tion is always linearly polarized and the rotation of the
locking can be accomplished by pulling a vane in an plane of polarization is proportional to the applied mag-
suh of the light path by attaching the vane to a shaft netic field.
whose two rest positions are controlled by a solenoid.
Electromechanically controlled camera-type iris shut-
ters are also available.
The acousto-optic (A-0) effect is concerned with changes
tic an in optical properties that occur when light passes through
ors a transparent medium into which an acoustic wave is
launched. These induced changes can affect the deflec-
Most electro-optic (E-0) modulators are based on the tion, polarization, phase, frequency, or amplitude of
Merr effect or the Pockels effect. Both of the effects the incident beam. The pressure waves cause a periodic
are concerned with the linear birefringence induced in change in the density and refractive index of the me-
certain materials that are subjected to a strong electrical dium. More generally, the change in refractive index
field. The application of an electric field alters the re- caused by pressure (mechanical stress or acoustic waves)
f m active index and propagation velocity lor the ordinary in crystals is termed the photoelastic effect or the
extraordinary rays. If the medium is illuminated
with plane-polarized radiation, the induced phase shift Many A - 0 modulators are based on the Bragg
between the two components results in ellipticallly po- cell design, as illustrated in Figure 3-16a. The moving
larized radiation. stress maxima and minima act like multiple reflection
46 Chap. 3 I Optical Components of Spectrometers
Polarizer
Polarizer
/
! magnetic
field
layers where the separation between layers is the acous- beam can be parallel to the acoustic waves. The medium
tic wavelength in the medium (A,). The incident radia- becomes a moving transmission grating with "refractive
tion beam is aligned to intercept the pressure waves at index grooves" with a groove spacing equal to the acoustic
a particular angle called the Bragg angle for which a wavelength in the medium. This Debye-Sears config-
large fraction of the output beam is periodically dif- uration is commonly used for A - 0 modulators with
fracted into the first order at the acoustic frequency. liquid media as shown in Figure 3-16b. A portion of
The Bragg angle (0,) is given by the incident beam is periodically diffracted to other
angles as regions of low and high refractive index pass
across the incident beam.
In both the Bragg and Debye-Sears configura-
tions, the acoustic wave is usually generated with a
If the optical pathlength in the acoustic medium piezoelectric transducer that is in contact with the
is sufficiently thin [b < (As)214A], the incident radiation acoustic-wave supporting material. The transducer's
Sec. 3-4 / Imaging and Beam Directing Optics
RF generator
Piezoelectric
crystal , '
Diffracted
radiation
-u
Stop
resonant frequency is determined by the piezoelectric In the discussion above we have stressed primarily
constant of the material used (usually lithium niobate). the application of modulators as an external component
A given A - 0 modulator can only be used at a few set used in conjunction with a continuous source. Many of
frequencies (the fundamental and harmonic frequen- these E - 0 and A - 0 devices are also used inside laser
cies). The bandwidth is controlled by the acoustic ve- cavities for Q-switching and mode locking (see Section
locity in the medium. For Bragg cells, the efficiency 4-3). In addition, some of these A - 0 devices are used
with which the incident beam is diffracted into the first as electrically controlled polarizers (i.e., to rotate the
order is controlled by the radio-frequency (RF) power plane of linearly polarized light or to produce elliptically
plied and the sixth power of the refractive index. or circularly polarized light). In some applications it is
though glass or quartz can be used for crystal media. necessary to determine the difference in the optical sig-
TeO, is often the preferred material because of its high nal observed for different states of polarization (i.e.,
refractive index (q = 2.2). Sinusoidal modulation in parallel vs. perpendicular components or d vs. 1 com-
the range 3 to 100 MHz is common, with rise times of ponents). Here electrically controlled polarizers are used
4 to 30 ns. as polarization modulators.
Some A - 0 modulators use mechanical stress to
induce birefringence in isotropic materials. The differ-
ence in retardation of the ordinary and extraordinary -4 IMAGING A
rays is proportional to the stress applied. Thus an OPTICS
acoustically driven crystal can be used as a variable wave
Pate, where the stress is applied perpendicular to the As discussed in Chapter 2, an optical spectrometer con-
lght beam. The crystal can be placed between two cross tains an image transfer system to produce an image or
olarizers to construct a modulator similar in design to set of images of the entrance aperture on the focal
E - 0 modulators (Figure 3-15). plane. In addition, imaging optics are frequently used
48 Chap. 3 I Optical Components of Spectrometers
vertex (convex mirror). ith this convention equation Mere f is positive for concave mirrors (R < 0) an
3-29 applies equally we1 convex as well as concave negative for convex mirrors (X > 0). The mirror for-
mirrors. mula can then be written
If the object is sufficiently far from the mirror to
ensure parallel light ( S , -+ a,1/S, -+ O), the image is
t a distance -R/2 from the vertex V , called
length f of the mirror. Thus
Figure 3-18 shows the action of several spherical
mirrors. Note that in the case of the convex mirror
(Figure 3-18c), the image is virtual (S2 < 0, R > 0,
FIGURE 3-18 Focal points for spherical mirrors. In (a) a parallel beam
of light strikes a concave mirror and produces a real image at the focal
point. In (b) a parallel beam makes an angle with the mirror axis and
produces a focus in the focal plane (dashed line). In (c) a convex mirror
is shown; the focal point is virtual and the focal length is negative. Placing
a source at the focal point of a concave mirror (d) produces a collimated
beam. The region of the mirror directly behind the source is masked by
the source. In (e) an off-axis mirror is used to avoid having the source in
the light pai.h.
5 Chap. 3 I Optical Components of Spectrometers
f < 0) as it is constructed from the intersection of pro- When an extended object (object size significant
jected rays. compared to S,) is used, we can locate the image by
The mirror formula indicates that moving the point the method shown in Figure 3-19a. The lateral mag-
source 0 from infinity toward the mirror or decreasing nification m = -do, where i and o are the height of
S,, increases S,, and the image moves away from the the image and object. respectively. and the minus sign
focal point toward the center of curvature. If the object indicates the image is inverted. The two similar right
is at the center of curvature, the image must also be at triangles, shown in Figure 3-19. must have proportional
that point. As the object approaches the focal point, sides. Thus ilo = S,/Sl and m = - S,IS,.
the image approaches infinity. Thus a point source of Table 3-2 summarizes the types of images that can
light placed at the focal point produces a parallel or be obtained with concave mirrors. Note that when the
collimated beam as shown in Figure 3-18d and e. object is located closer to the mirror than the focal
point, the reflected rays are divergent, and the image
is a virtual, erect (noninverted) image.
TABLE 3-2
Images formed by spherical concave mirrors and thin convex lenses
Image
0
Figure 3-22 has been moved close to the lens such that
distance S , is quite small. If the distance S, is small, the
paraxial rays from 0 form a virtual image of 0 at dis-
tance S; (S; < 0). The distance S; is now the object
Biconvex Meniscus Biconcave distance for the second surface. Since we have assumed
convex the lens to be thin, there is no need to modify this
distance for the lens thickness. If we use -S; as the
object distance for the second surface, the image will
be formed at distance S2. Or
Note that for the biconvex lens of Figure 3-22, R1 > 0 120 mm. If the object is placed at 300 mm from the
and R, < 0. Here the sign convention, compatible with lens, the image position can be found from equation
that of mirrors, is that R is positive if the center of 3-37 as US, = 11120 - 11300, from which S, = 200
curvature of the surface is to the right of the vertex and mm. If S, = 200 mm, S, = 300 mm. For this lens the
negative if to the left. The focal length f is related to points 200 mm and 300 mm represent conjugate points.
the index of refraction of the lens and its radii of cur- If SI = 240 mm (2f), S, = 240 mm. If S1 = m, S, = f.
vature by Thus a parallel incident beam of light will form an image
at the focal point. Similarly, a point object placed at
the focal point will produce a parallel or collimated
beam.
The image of an extended object can be located
This follows from equation 3-35 since f is the image graphically by the method illustrated in Figure 3-23.
distance if the object is at infinity or f = S, if S, = m. The lateral magnification m is the ratio of the image
Thus the lens formula, equation 3-35, can also be ex- height to the object height. Just as with mirrors, m is
pressed as given by rn = -S,IS,. The types, locations, orienta-
tions, and sizes of the images obtained with thin convex
lenses are the same as those for concave mirrors and
are shown in Table 3-2.
As an example, let us calculate the focal length In addition to the simple lenses presented here,
of a planoconvex lens of radius of curvature 60 mm and cylindrical lenses are also used. These are used to con-
index of refraction 1.5. Let us assume that light is in- vert a point image into a line image or to change the
cident on the planar surface so that R, = x and height of an image without changing its width. Cylin-
R, = 60 mm. From equation 3-36 we have llf = drical surfaces are difficult to produce and they are
(1.5 - 1)(0 - 11- 6O), from which we find that f = available only in planar-cylindrical shapes (planar con-
(a) (b)
FIGURE 3-23 Extended objects imaged by biconvex lenses. In (a) graph-
ical location of the image is shown. Ray 1 is parallel to the optical axis
and passes through the focal point. Ray 2 passes through the focal point
and emerges parallel to axis. Ray 3 is undeviated. In (b) rays from the
top of the arrow illustrate the lateral magnification rn = - S,IS,.
Sec. 3-4 1 Imaging and eam Directing Optics
,ex or planar concave). Aspherical lenses are also avail- erture) or an adjustable diaphragm is used to restrict
able to reduce certain aberrations. the rays that form the image. This has the effect of
Reflective losses with lenses can be substantial limiting nonparaxial rays and improving image quality.
-
unless they are coated. For example, a typical glass lens
can suffer 8% reflection loss. By coating the lens with
a thin layer of a material of index of refraction inter-
A lens with a rear aperture stop is shown in Figure 3-
24. In many optical systems an additional element, called
a field stop, limits the size of the image, as shown in
mediate to that of air and glass, reflective losses can be Figure 3-24. In a camera, the field stop is the edge of
greatly reduced. With a lens such antireflective coatings, the film itself.
designed for normal incidence, can reduce reflective The image of the aperture stop formed by ele-
losses to = 3.5% from = 8%. Antireflective coatings ments between the object and the aperture stop is called
are discussed in Appendix B. il. It is the smallest opening that can
be seen at the object point. The image of the aperture
Lens Stops. Any element that determines the stop formed by elements between it and the image is
amount of radiant energy reaching the image is called il. It is the smallest opening that can
erture stop. In the simple lenses discussed thus be observed at the image point. The entrance pupil
far, the size of the lens itself limits the amount of light controls the cone of light entering the optical system,
accepted. In other cases a separate fixed opening (ap- while the exit pupil controls that leaving it.
Figure 3-25 illustrates how two lenses can be ar-
ranged in such a way that a larger cone of light comes
from on-axis object points than from those that are off-
axis. This is a process known as vignetting.
by an aperture stop, and S, is its distance from the the optical element, the image is formed in the focal
source. The irradiance of the image El in the focal plane plane at a distance f from the optical element (S, = f).
at S, is just the total radiant power of the image divided With S2 = f and the relationship m = - S,lS,, equation
by its area, A,. If we assume there are no reflection or 3-40 becomes
absorption losses in the optical system, El is just
The effective area of the optical element is just A = The F-number of the optical element (Fin), is flD, so
~ r ( D l 2 ) where
~, D is the effective or limiting diameter.
The ratio ASIA, in equation 3-39 is just llm2, where m B,nl 4
is the magnification, if there is no field stop. Thus equa- E. = -
(Fin)',
tion 3-39 becomes
Beam Splitters
Saggital
plane
\
Tangential
plane
Circle of least
confusion I\
Principal
Tangential ray
1
Sagittal
plane
i
Optical
system
ally front-surface coated. Different coating thicknesses also used to make beam splitters of higher efficiency
are used to produce various ratios of transmitted to and a wide range of beam intensity ratios. The effi-
reflected beam intensities. For example, the typical half- ciencies and beam splitting ratios are often wavelength
silvered mirror might transmit 30% of the incident ra- dependent. The second surface of mirror-type beam
iation and reflect 30%. The remaining 40% is lost splitters can be antireflection coated (see Appendix B)
through absorption. Metallic coatings a r e useful to minimize the multiple reflections that can produce
throughout the UV and visible regions and into the unwanted "ghost" beams. Beam splitters are often used
near-infrared region (> 1 pm). Dielectric coatings are in reverse to produce beam combiners.
58 Chap. 3 1 Optical Components of Spectrometers
Reflected
beam 1 Ghost
reflection
Front
Side
view
1 I Beam view
Transmitted
beam
Metal
frame I Membrane
Membrane
A pellicle beam splitter is made of a durable elastic biners. For example, the fundamental infrared beam
membrane stretched over an optically flat metal frame from a Nd:YAG laser (1.06 pm) is often combined with
and bonded to the edges as illustrated in Figure 3-31b. visible harmonics or with a visible dye laser beam by
the membrane is made from optical-grade using a dichroic mirror.
nit lose and has a thickness of only 7 pm. An Another type of beam splitter is based on bifur-
pellicle beam splitter reflects 8% and trans- cated fiber optics bundles as described below.
mits 92% of the incident radiation throughout the vis-
ible and near-infrared regions. Coatings are used to vary
the ratio of the reflected to transmitted beam intensi-
ties. Standard coatings provide ratios of (33167%, Fiber optics from the communications industry are being
40/40%, and 50150%). Pellicles are of high efficiency used in and with spectrometers to transfer light between
they do not produce multiple beams. various points. The construction of a fiber optic is shown
The beam splitters described above all produce in Figure 3-32. A light ray entering the core at angle 8,
beams that are spatially separated, but temporally over- will be totally internally reflected if sin 8; q 2 i q , . Since
g. In some applications it is desirable to split a sin 0; = cos 0 , and cos2 0 , = 1 - sin2 el, the condition
beam into two spatially separated beams at dif- for total internal reflection is 1 - sin2 8 , e (rlz/91)' or
ferent times. The double-beam in-time spectrometer ql sin 0 , r ( q : - ~ $ ) l / The
~ . entrance angle 8 , is related
escribed in Section 4-6 is an example. For such ap- to 0 , by Snell's law ( q , sin 0 , = q , sin O,), so total in-
lications, the chopperibeam splitter illustrated in Fig- ternal reflection occurs when q, sin 8, i (-qf q $ ) l f 2 .
-
ure 3-31c is often used. The chopper wheel is usually There is thus a maximum value of 8 , for which to-
e by cutting open slots in a circular mirror. tal internal reflection occurs given by q , sin 0, =
Many other types of beam splitters are used. Sev- (q: - q:)1/2.Rays that are incident at larger angles
eral of these separate an unpolarized beam into its two are only partially reflected at the core-clad interface
mutually perpendicular components. We have already and soon pass out of the fiber. The numerical aperture
encountered such a polarizing beam splitter in Section NA is defined as 9, sin 0,, or
3-2, where an optically anisotropic crystal, such as cal-
'te, was shown to produce an ordinary and an extraor- NA = qo sin 8, = ( q f - ~2)"~ (3-46)
inary ray. Polarizing prisms, used as beam splitters,
are described in Section 3-5. Another polarizing beam The NA represents the cone of light accepted by the
litter is often referred to as the "pile-of-plates" po- fiber. The Fin of a fiber optic is related to the NA by
izer. This type of beam splitter is based upon polar- Fin = 1/(2 tan 8,). Core diameters are typically 50 to
ization by reflection, as described in the preceding sec- 600 pm and the refractive index of the clad is typically
tion. The pile-of-plates polarizer is illustrated in Figure
3-31d. This type of beam splitter can be made with glass I Clad q,
plates for use in the visible region, with quartz plates
Core q 1
for the ultraviolet region or with alkali halide plates for So
use in the infrared region. A crude beam splitter of this
type can be made from 10 to 12 microscope slides. Fiber optic
Another type of b splitter is a wavelength-
(a)
selective beam splitter o roic mirror. Such mirrors
are made from multilayer, nonabsorbing films. (See
Section 3-5 for a description of interference filters based
on thin films.) A hot mirror reflects infrared radiation
while transmitting visible radiation as illustrated in Fig-
ure 3-31e. A cold mirror reflects visible radiation and
transmits infrared radiation. There are many uses of
dichroic mirrors. Many sources used in the visible also
produce intense infrared radiation which can cause
heating problems. A dichroic mirror can be used to FIGURE 3-32 Fiber optic. The fiber optic in (a)
direct the unwanted infrared radiation away from the consists of a core of refractive index -q, and a clad
sample. In movie projectors, dichroic mirrors are often of refractive index qz,where q2 < ql. Normally.
used to send the infrared radiation to the back of the additional jackets surround the clad to provide
projector away from the film. Like other beam splitters, physical strength. In (b) the various angles are
dichroic mirrors can be used in reverse as beam com- defined.
60 Chap. 3 I Optical Components of Spectrometers
0.02 to 0.2 less than the core. This gives a value for the 3-5 FILTERS, PRISMS, AND GRATINGS
NA in range 0.1 to 0.5.
Fiber optics are experiencing greater use in spec- In the majority of spectrochemical techniques, the op-
troscopy for several reasons. Because they are me- tical signals produced by the encoding process are sorted
chanically flexible, light can be transmitted over curved by means of wavelength so that the information con-
paths. Thus fiber optics can replace several mirrors in cerning one component or a set of components can be
directing light between two points in a spectrometer isolated and measured. Wavelength selection is achieved
(e.g., from the source to the wavelength selector or in simple, low-resolution systems by filtering the radia-
from the wavelength selector to the detector). Fiber tion (attenuating all but the desired wavelengths) and
optics can transmit light over long distances (e.g., 500 in higher-resolution systems by dispersing it (spreading
m), which allows remote monitoring in hazardous en- out the wavelengths spatially) into a spectrum. This
vironments since the more delicate components (de- section considers the optical components used in wave-
tectors, monochromators, signal processing electronics) length selection. Dispersive spectrometer systems are
can be far from the monitoring site. Over long distances, discussed in Section 3-6, while systems based on inter-
the absorption and scattering losses in the fiber can be ferometers are discussed in Section 3-7.
significant.
A single fiber optic cannot transmit an image be-
cause the rays from different parts of the object are
scrambled by multiple internal reflections. Bundles of Filters are used to pass a band of wavelengths (bandpass
fine glass, quartz. or plastic fibers can, however, be filters) or to block wavelengths longer or shorter than
used for image transmission if the fibers are small enough some desired value (cutoff filters). The most widely
in diameter that each fiber transmits rays from a small used filters in spectroscopy are based on absorption or
area of the object. Some typical uses of fiber optic on interference.
bundles are illustrated in Figure 3-33.
Absorption Filters. Filters based on absorption
by colored glasses, crystals, sintered materials, solu-
tions, and thin films are widely used. Filters made from
these materials are simple to use, inexpensive, and rel-
atively insensitive to the angle of incidence of the in-
coming radiation.
Bandpass filters are characterized by plots of their
spectral transmittance [T(X)]vs, wavelength, as shown
in Figure 3-34a. The characteristics of interest to the
user are shown in the figure. The transmission char-
acteristics of several colored glass filters are shown in
Appendix C. Typical values of the full width at half
maximum (FWHM) range from 30 to 250 nm. Bandpass
filters made with an organic dye impregnated on gelatin
and sandwiched between glass plates (Wratten filters)
are also commercially available. Liquid filters are often
made from colored chemical solutions contained in glass
Photocathode cells.
Cutoff filters contain substances that absorb all
Focal planed-I the radiation at wavelengths shorter than a given cutoff
(short-wavelength cutoff) or all the radiation at long
(cf
wavelengths (long-wavelength cutoff). The transmit-
FIGURE 3-33 Some applications of fiber optic tance of a short-wavelength cutoff filter is illustrated in
bundles. In (a) a bifurcated, randomized bundle Figure 3-34b. Cutoff filters can be colored glasses, the
is shown. This can be used as a beam splitter or
Wratten type, organic dyes in polymeric hosts, or thin
to send light down one path and have it return
along the other. In (b) one end of the bundle is films.
circular, while the other is rectangular to match
a spectrometer slit, for example. In (c) a fiber Interference Filters. Interference filters are
optic face plate is shown. This allows a flat image constructed so that the rays from most wavelengths that
to be transferred to a curved surface such as the strike the filter suffer destructive interference, while
active surface of a detector. only rays within a small wavelength band experience
Sec. 3-5 I Filters, Prisms, and Gratings
Glass Glass
Sem~transparent
silver film
(4
FIGURE 3-34 Transmittance of bandpass (a) and
short-wavelength cutoff (b) filters. Wlth the I Dielectric I
bandpass filter (a) the wavelength of maximum
transmittance A,, the maximum transmittance T,
and the full width at half maximum height Ah or
FWHM are illustrated. For the cutoff filter (b)
the cutoff wavelength A, is the wavelength where
T(h)f = 10, 1 or 0.1%. Wavelength h, is the
wavelength at which the transmittance reaches 90
or 99% of its maximum value T,,. Wavelength h,
is the wavelength at which T(h), = 0.5 T,.
constructive interference and are passed. Two types are
widely used: the single-layer or Fabry-Perot type and
e multilayer dielectric type.
The Fabry-Perot type of interference filter is il-
lustrated in Figure 3-35. The thin dielectric is a material (b)
of low refractive index such as quartz, CaF,, MgF,, FIGURE 3-35 Fabry-Perot interference filter:
ZnS, ThF,, or sapphire. Waves that reach points g and (a) cross section; (b) exploded view showing spac-
e in phase undergo constructive interference at the lens ing d between reflective surfaces and angles. The
focal point. As shown in the figure caption, the con- lens in b is equidistant from points g and e. Ray
dition for constructive interference is 1 travels the path a + h e , while ray 2 travels
the path a-.bbf+g. The two waves will be in
phase (constructive interference) at points g and
e if A(0PL) is an integral multiple of the wave-
In many uses of interference filters, the incident length. Because of the law of reflection
beam is normal to the plane of the filter, instead of ab = bf = fg. Thus the condition for constructive
interference is 2abq - be = mh. Since cos 8' =
skewed as in Figure 3-35. In these cases 0 = O", so
dlab, tan 8' = 1 bgld, sin 0 = belbg, and sin 0'
sin 0 = 0. The central wavelength passed by the filter
= sin 01y, this can also be expressed as 2dyIcos
A, can be written 0' - 2 d q sin 8' tan 0' = 2 d y cos 8' = mh. Since
cos 8' = [I - (sin 01y)2]112,the condition for
constructive interference in terms of the angle of
incidence is 2 d ( y 2 - sin2 8)' I 2 - mh.
62 Chap. 3 1 Optical Components of Spectrometers
Vitreous quartz
ference filters is frequently employed when they serve Flucrite
as wavelength selection devices in dye laser cavities.
Filters are commercially available in which the d spacing
varies along the length of the filter. Such wedge filters
are also available in circular styles in which d varies
with the filter rotation.
The transmission curve of a typical Fabry-Perot
interference filter is shown in Figure 3-36 and additional
information is given in Appendix C. Higher-order bands
24-
Polarizing Prisms. Polarizing prisms are made interfere at different angles. Gratings are either de-
of birefringent materials. They depend on the different signed for transmission of the incident radiation or for
indices of refraction for the ordinary and extraordinary reflection. Modern spectrometers invariably use reflec-
rays to separate these rays. With some polarizing prisms tion gratings.
only a single polarized beam emerges; in others, called Spectroscopic gratings are in actuality replicas of
polarizing beam splitters, both rays emerge. The earliest master gratings. Contemporary master gratings are ruled
polarizing prism (1828) was the Nicol prism, but it has by a diamond ruling tool operated by a ruling engine.
long since been replaced by more efficient devices; two The tool rules thin aluminum films that are evaporated
of these are shown in Figure 3-40. If the two prism onto optically flat glass. Machine-ruled gratings are ex-
sections in the Glan-Foucault prism are cemented to- tremely difficult to manufacture and, as a consequence,
gether and the interface angle changed, the device is few master gratings are produced. Replica gratings are
called a Gllan-Thompson polarizer. produced by vacuum deposition of aluminum on a ruled
master and subsequent coating of the A1 layer by an
Diffraction Gratings epoxy-type material. After polymerization the replica
is separated from the master. Today, replica gratings
A diffraction grating is a plane or concave plate that is are produced that are superior to the master gratings
ruled with closely spaced grooves. The grating acts like of only 15 to 20 years ago.
a multislit source when collimated radiation strikes it. Holographic gratings have also become practical
Different wavelengths are diffracted and constructively in recent years. These are made by coating a flat glass
from which
Specular
reflection
Normal
Diffracted
I
I
Incident
I
I
Monochromators are the most widely used dispersive
instruments. They isolate a small wavelength band from
a polychromatic source. Monochromators consist of a
dispersive element (prism or grating) and an image
transfer system (entrance slit, mirrors or lenses, and
exit slit). Within the monochromator an image of the
FIGURE 3-43 Echelle grating. Echelles are rel- rectangular, or sometimes curved, entrance slit is trans-
atively coarse gratings (typically 100 grooves ferred to the exit slit after dispersion of the wavelength
mm-l) with large blaze angles. The steep side of components of the incident radiation.
the groove is employed so as to achieve large In this section grating monochromators are em-
diffraction angles. They are typically used in very phasized because there are few prism systems being
high orders, as discussed in Section 3-6. made at this time. However, the general principles pre-
sented apply equally to both types of monochromators.
A grating monochromator based on the Czerny-Turner
mula (equation 3-50) with respect to wavelength. For configuration shown in Figure 3-44 is used as the model
a constant angle of incidence a , D, in rad nm-l is given for the following discussion because it is a common
?'Y design; many other configurations are available.
As we will see in more detail later, the slits play
D dp
=-=-
lml -
-
sin a + sin p (3-52)
an important role in determining the resolution and
dX d cos 6 h cos p throughput of a monochromator. A slit is formed by
two slit jaws. The slit width is formed by the separation
of the two opposing edges of the slit jaws. Slit widths
f the grating is operated with nearly normal incidence are typically in the range of a few micrometers to several
and small angles of diffraction, cos is only weakly
dependent on A. Thus D, is nearly independent of wave- Entrance
length. For example, for a = 3.73", d = 833.3 nm, and Pr-+:...- slit S,
rn = t 1, cos p varies from 0.985 to 0.845 over the Collimating
range 200 to 500 nm. The angular dispersion for the mirror MI
same grating would vary from 1.22 x rad nm-I
at200 nm to 1.42 x rad nm-I at 500 nm.
fraction. Note in equation 3-48 that D, is directly pro- FIGURE 3-44 Czerny-Turner plane grating
portional to m and to llcos p. monochromator. Incident radiation passes through
the entrance slit S, and strikes the parabolic, col-
limating mirror MI. The entrance slit, at the focal
point of MI, acts as a point source for MI which
-6 DISPERSIVE WAVELEN produces parallel radiation for the dispersion ele-
SELECTION SYSTEMS ment, a grating in this case. The grating spatially
disperses the spectral components of the incident
As discussed in Section 2-2, complete optical instru- radiation. Collimated rays of diffracted radiation
strike the parabolic focusing or camera mirror
ments for wavelength selection fall into several cate-
M,. The dispersed radiation is focused in the focal
gories. This section describes systems based on disper- plane producing entrance slit images in that plane.
sion. The monochromator, which isolates a single Because the parallel rays of a given wavelength
wavelength band at a time, is first discussed and then are incident on the focusing element at a specific
polychromators and spectrographs are presented. Sec- angle, each wavelength is focused to a slit image
tion 3-7 discusses Fabry-Perot, Michelson, and other at a different center position on the focal plane.
interferometers that are finding increasing applications The exit slit S, placed in the focal plane isolates
in spectrochemical measurements. a particular wavelength interval.
68 Chap. 3 1 Optical Components of Spectrometers
millimeters. Some monochromators have fixed, un- Wavelength Selection. To change the wave-
changeable slit widths, while others have adjustable slits. length selected by the monochromator, the dispersive
Slit heights usually vary from 1 to 10 mm. element is rotated to bring a different wavelength band
In most monochromators the exit slit has the same through the exit slit. This is shown more clearly in Fig-
dimensions as the entrance slit so that it can be fully ure 3-45 for a Czerny-Turner grating monochromator.
illuminated by the entrance slit image. Many mono- Here angle @, the takeoff angle, is fixed and the grating
chromators allow both slits to be adjusted in width angle 0 is changed to isolate a different wavelength.
simultaneously. Note that changing 0 changes both the incident angle
We discuss first how different wavelength bands a and the diffraction angle P, as can be seen by com-
are brought to the exit slit before discussing mono- paring Figure 3-45a to Figure 3-45b.
chromator characteristics. From Figure 3-45 we can see that the grating ro-
tation angle 0 is given by 0 = a + @. The takeoff angle
@ is given by @ = p - 0 . Thus the incident angle and
diffraction angle can be written in terms of 0 and @ as
To exit
slit
a = 0 - @ and p = 0 + @. If we substitute these
relationships into the grating formula (equation 3-50)
and use the trigonometric identity l/2[sin ( a + P) +
sin ( a - p)] = sin a cos (3, the result is
\
\N d [sin (8 - @) + sin (8 + @)I = 2d sin 8 cos @ (3-53)
= mA
TABLE 3-3
Angles and dispersion characteristics of a Czerny-Turner monochromator in the
first ordera
I plane
FIGURE 3-46 Linear wavelength drive mecha- FIGURE 3-47 Relationship between angular and
nism. A precision leadscrew L is used to move a linear dispersion. Rays of A, from the dispersion
contact flat F. The grating is attached to sine bar element are parallel to each other and are focused
S of length y . Movement of the flat by distance at point x , by the focusing element. Parallel rays
x pushes contact ball B and rotates the grating of A, strike the focusing element at a different
through angle 0. (The optical axis is parallel to angle as determined by the dispersion element
the leadscrew.) Note that sin 0 equals xly, the and are focused at x,. For a grating monochro-
hypotenuse length y is fixed, and x equals np, mator, Ap is the difference in diffraction angles
where n is the number of turns of the leadscrew of A, and A, with respect to the grating normal.
and p is the pitch. If these terms are substituted For a prism monochromator, Ap = A0, where
into the grating formula 3-53, solving for A gives A0 is the difference in refraction angles for A, and
A = 2dnp cos Wmy. Thus the wavelength is di- A, with respect to a ray undeviated by the prism.
rectly proportional to the number of leadscrew
turns n. It is also possible to construct a linear
wavenumber or cosecant drive. However, mod-
-
Since tan (p, - p,) = tan Ap = Axlf, for small
angles Ap = Axlf and dp dxlf. Thus dxldh =
f(d@W.
ern monochromators usually use a linear wave-
length drive and calculate wavenumber with a For a prism monochromator the reciprocal linear dis-
microprocessor. persion is usually quite wavelength dependent because
of the variation of D, with wavelength. Some prism
Dispersive Characteristics. We have already
monochromator configurations, like the Pellin-Broca,
shown how angular dispersion D, can be calculated for provide dispersion that is wavelength independent.
a prism (equation 3-49) and a grating (equation 3-52). The relatively constant dispersion properties of a
The angular dispersion is the angular separation d p ( d o grating monochromator are iilustrated by the mono-
for a prism) corresponding to the wavelength separation chromator considered in Table 3-3. For 300 nm, D ,
d h . It is a property of the grating or prism itself. For a
from equation 3-52 is 1.26 x 10W3rad nm-l. Radiation
monochromator, the dispersion in the focal plane is of
more interest. The linear dispersion, Dl = dxldA, tells
w far apart in distance two wavelengths are separated
Ap -
of 310 nm would be separated from that of 300 nm by
1.26 x rad. When the grating is rotated to
isolate 400 and then 500 nm radiation, similar calcu-
the focal plane. Figure 3-47 illustrates how Dl is de- lations show D, = 1.28 x and 1.32 x
termined for a dispersion element illuminated by col- rad nm-I, only a small difference. For the grating
limated radiation of wavelengths A, and h,. For the monochromator of Table 3-3 and a collimating focal
configuration shown, the linear dispersion is given by length f of 250 mm, the linear dispersion D, = fD, =
D, = fD,, where f is the focal length of the focusing 250 mm x D,. The reciprocal linear dispersion R,, for
element. The linear dispersion Dl is conveniently ex- a grating monochromator is found by combining equa-
pressed in units of mm nm-I in the UV-visible region tions 3-52 and 3-54. Values calculated from the equa-
of the spectrum. Most often it is the reciprocal linear tions below are shown in Table 3-3 for the 250-mm-
ispersion R, that is specified for a spectrometer. The
focal-length monochromator described there.
reciprocal linear dispersion represents the number of
wavelength intervals (e.g., nm) contained in each in-
terval of distance (e.g., mm) along the focal plane. For
the configuration shown in Figure 3-47,
h cos p
R, =
f(sin a + sin p)
70 Chap. 3 1 Optical Components of Spectrometers
It should be noted from Table 3-3 that although the where W is the slit width. For a monochromator with
reciprocal linear dispersion does not change drastically R, = 1 nm mm-I, a slit width of 100 pm would give
with wavelength as it does for a prism instrument, it a spectral bandpass of
does change with wavelength.
Fin =
f
- t(h) = (1 - /(X - A,)/s,/) for A, - s, 5 A 5 A, + s,
D, t(A) = 0 elsewhere
The solid angle is given by
A0
x - s,
h - l s , 25% overlap
Direction
X - 1 s , 50% overlap
X - 1 s , 75% overlap
X 100% overlap
Percentage of image
A0 - xo
Percentage of image overlap
(slit function)
a) (b)
Slit function of a monochromator with equal exit and
entrance slits. In (a) the slit function is shown as the convolution of the
entrance and exit slit images. In (b) the slit function is shown as a function
of wavelength.
stituted into the foregoing definition of R,,, the result spectral radiance B, and the monochromator through-
put factor Y(h).
is for a plane grating, but Two interesting cases can be considered. If the source
for a concave grating if the width is than a is monochromatic and the monochromator is set to the
certain optimum value. appropriate wavelength [h = A,, t(h) = 11, then To, is
Since the diffraction order is given by m =
constant over the source width and J"; B, dh = B, where
[d(sin a +
sin P)]/A, the theoretical resolving power is the line radiance, In this case
can also be written
metric spectral bandpass. Thus diation is specified according to the relative amount of
radiation passed x nm from a specified laser line. Stray
radiation is usually specified as the percentage of the
total radiation passing the exit slit at a given wavelength
Note that the radiant power passed by the monochro- over a specified wavelength range. Thus
mator for a continuum source is proportional to the slit
width squared, whereas for a monochromatic source it % stray radiation @SR
= -x 100 (3-75)
is proportional to the first power of the slit width. This @o
difference arises because as we open the slits for a con-
tinuum source, we not only increase the source area where @,, is the stray radiant power and a, is the
viewed but also pass more of the source spectral dis- source radiant power passed by the monochromator
tribution. For a monochromatic source, the source line over X, * s. Stray radiation is very difficult to measure
width is much smaller than the monochromator slit width. accurately because it depends on the wavelength, the
Opening the slits only increases the source area viewed spectral bandpass, and the type of source used.
by the monochromator in this case. There are many possible causes of stray radiation.
In the equations above we assume that the en- For example, room light can leak into the monochro-
trance and exit slits are of equal size. that the light image mator or radiation can be reflected internally from walls,
on the entrance slit is of uniform irradiance and larger optics and baffles. Scattering from dust particles, and
than W and H for all W and H, and that the limiting fluorescence from optical materials are also sources of
aperture of the monochromator is filled. This is usually stray radiation. An overlapping grating order, grating
accomplished by using a lens or mirror to focus an image imperfections, and diffraction from slit edges can also
of the source on the slit with the cone of radiation produce stray radiation. With gratings, some stray ra-
entering matching the monochromator Fin. Often 1: 1 diation appears at all wavelengths because there is not
imaging is used, where the optical element is two focal complete destructive interference between diffraction
lengths from the source and entrance slit. In this case, orders. This results in the secondary maxima seen in
the Fln of the lens or mirror should be one-half the Figure 3-7.
Fin of the monochromator. When external optics are Stray radiation can be reduced in several ways.
used, To, in the expressions above is replaced by TopT,, Good instrument design, including appropriate baffles
where T, is the efficiency factor (0 < T, < 1) for the to intercept various reflected and scattered rays, is one
external imaging optics or Top is redefined to include obvious way. Holographic gratings provide much lower
the losses of the imaging optics. If other than 1: 1 im- stray radiation factors than do ruled gratings. Stray ra-
aging is employed, the expressions above are divided diation figures can vary from 0.1% to 10-j%. Typical
by m2, where m is the magnification; the solid angle of values are l o p 3 % for ruled gratings and for
the imaging optics is used as R in this case. holographic gratings. Broad absorption filters are often
If a filter is used in place of a monochromator, employed in front of the entrance slit to narrow the
similar expressions apply and Y (A) = A,RT, T(X)f Here source bandwidth. These can also serve as order-sorting
A,R is the non-wavelength-dependent geometric factor, filters. Double monochromators or two mono-
where A, is the source area viewed and R is the solid chromators in tandem can achieve extremely low stray
angle of the source viewed. The area A, is determined radiation figures. These are often employed in situa-
by the actual projected area of the source or an aperture tions such as laser Raman spectroscopy where a weak
in front of the source and R is determined by the lens spectral line (Raman scattering) must be measured near
or mirror (area and object distance) used to focus or a very strong line (Rayleigh scattered line). Note that
collimate the source radiation on the filter. The product if a single monochromator produces 0.1% stray radia-
T,T(X)f is the wavelength-dependent factor, where T(Alf tion, two such monochromators in tandem would pro-
is the spectral transmittance of the filter and T, is the vide 0.0001% stray radiation. The slit function of a
optical efficiency of optical components used in con- double monochromator is the convolution of two tri-
junction with the filter. angular functions (equal ganged slits).
Stray Radiation. Stray radiation or stray light OpticalAberrations. Let us briefly consider op-
in a monochromator is considered to be any radiation tical aberrations with the Czerny-Turner monochro-
passed that is outside the interval A, ? s. where X, is mator of Figure 3-44. Parabolic mirrors are used for the
the wavelength setting and s is the spectral bandpass. collimating and focusing mirrors to eliminate spherical
In some cases, the range specified above is broadened aberrations. Chromatic aberrations are, of course, ab-
to include a larger wavelength range or the stray ra- sent because the image transfer system uses mirrors and
Sec. 3-6 1 Dispersive Wavelength Selection Systems 7
Main
collimating
mirror
-
I I 1 1 1
wavelength region of several differ-
800 80 I I 1
ent orders, efficiency can be very high
I a B l l b
at many wavelengths instead of only Free 800 400
near the blaze wavelength. Scanning spectral
can be accomplished by rotating the range
grating or by moving the detector
along the focal plane.
ent sf finesse, CF, is defined by C, = shown in Figure 3-57b. For 0 = O",interference maxima
where p is the reflectance of the mirror occur when X = 2dlrn. Just as with a diffraction grating,
coatings (typically, X . 9 ) . Here @, is the maximum we can define the free spectral range AXf as the range
radiant power for a given fringe. of wavelengths for which no overlap of adjacent orders
A plot of the relative radiant power vs. 6 is shown occurs, or AXf = hl(m +
1) == Xlrn, because rn is very
in Figure 3-57a for a monochromatic source. Note that large. Thus
eaks in the relative tr mittance occur at multiples
f the phase difference. measure of the sharpness of
the full width at half maximum in phase
),, which can readily be shown to be
The FWHM in wavelength units (FWHM), is thus In the visible region of the spectrum F for most
Fabry-Perot interferometers is =30. The limitation is
(FWHM), 4 1 c ~ most often not the reflectances of mirrors as implied
(FWHM), = - - by equation 3-80, but the deviation of the mirrors from
4rdX - 2 6X-I
parallelism. Curved mirror systems with thin dielectric
coatings have achieved F values of greater than 1000.
Since 6 near an interference maximum is 2 ~ m we
, can
Today the confocal interferometer is more popular than
write
the plane mirror type. In confocal designs concave
spherical reflectors are used separated by a distance
nearly equal to their radii of curvature. The focal points
of the mirrors are nearly coincident on the optical axis.
The confocal design is much easier to align and use than
The theoretical resolving power is then the plane-parallel type.
- For use as a spectrometer the Fabry-Perot can
achieve remarkable resolution. For A = 500 nm, d =
10 mm, F = 30, equation 3-81 gives R,, = 1.2 x lo6
and (FWHM), = 4 x l o p 4 nm. Note, however, that
The quantity n f l F / 2 in equation 3-79 is called the m = 4 x lo4 and the free spectral range would be
finesse F of the etalon. It is a measure of the number AXf = 0.0125 nm. Thus spectrometric applications of
of resolvable elements in one free spectral range given Fabry-Perot interferometers require order sorting. Often,
by the ratio of the separation between adjacent maxima in fact, a grating monochromator is used as an order
AXf to the FWHM. Thus sorter.
For spectrometric applications a pinhole can be
used to allow radiation to be transmitted to a detector
or order sorter. Wavelength scanning is accomplished
by varying the d spacing or the refractive index of the
gap. Piezoelectric mirror mounts are often used to move
The theoretical resolving power is then written in terms one of the mirrors by applying a voltage to the piezo-
of the finesse as electric device. Alternatively, by changing the gas pres-
sure within the interferometer, the refractive index of
the gap can be changed. Although we have assumed an
air gap (T = 1.00) for simplicity, the exact condition
for constructive interference is A(0PL) = 2dq cos 8.
For arbitrary angles of incidence, R,, = 2d cos 8 FIX. For many gases, q is proportional to pressure over a
Sec. 3-7 / Nondispersive Systems
range. Hence pressure-scanned Fabry-Perot arm, the two beams will be in phase at the detector and
interferometers (constant d) are also common. thus constructively interfere (bright field at the detec-
Because of their complexity and small AXf values, tor). When the movable mirror is adjusted XI4 away
y-Perot systems are employed in applications re- from this position, the two beams are 90" out of phase
ng extremely high resolution. Thus they are used at the mirrors and 180" out of phase when they reach
measure the widths of atomic spectral lines, the widths the detector (dark field at the detector). If the mirror
laser lines and for other demanding applications. is continuously moved at a constant rate, the intensity
abry-Perot interferometers can also be used to de- at the detector alternates sinusoidally between light and
termine refractive indexes of gases. dark for each XI4 movement, as shown in Figure 3-59a.
The ac component of the detector signal S(x) as
a function of mirror displacement is related to the source
elssn Interferometer spectrum Q, QV, or 0, by
- zc
=
S(x) cos (4rrxi;) dx (3-84)
culating the spectrum.
The limits of integration in equation 3-84 indicate
Sec. 3-7 1 Nondispersive Systems
or it can be a multichannel (array) detector. The sample trometer, each resolution element is observed for only
can be placed between the source and the beam splitter a fraction of the scanning time. As discussed in Chapter
or between the beam splitter and the detector. 5 , the multiplex advantage results in a signal-to-noise
advantage for multiplex spectrometers when detectors
are limited by detector noise. As a result, Fourier trans-
Advantages of Fourier Transform Methods form techniques find their major applications in spectral
regions where noisy detectors (see Chapter 4) must be
Fourier transform spectroscopic methods have two ma- used, such as the mid- and far-IR regions. Because of
jor advantages over conventional, dispersive spectrom- the increased throughput they are also used in meas-
eters. First there is a throughput advantage, called Jac- urements of weak sources. Although most applications
quinot's advantage, because no entrance slits are present. have been in mid- and far-IR absorption spectroscopy,
Radiation must, however, be collimated in a Michelson Fourier transform spectrometers have also been used
or common-path interferometer system. Thus a circular in UV-visible absorption, in IR emission, in Raman
aperture and a collimating element are normally used. scattering and in rapid-scanning measurements. Com-
The effective solid angle is often, however, much larger mercial Fourier transform spectrometers are available
than that of a conventional spectrometer. Jacquinot's based on both the Michelson and the common-path
advantage can be a factor of 10 to more than 200. A interferometer. Inexpensive computers and rapid Four-
second advantage, called the multiplex advantage (Fell- ier transform algorithms have made Fourier transform
get's advantage), results because the signals from all spectrometers extremely competitive with dispersive
resolution elements are observed all the time in a mul- systems in the mid-IR and clearly superior in the far-
tiplex spectrometer. In a conventional scanning spec- IR region, as discussed in Chapter 14.
PROBLEMS
Consider a light beam of 579-nm unpolarized radiation The peak absorption coefficient for a molecule with a
in air that is incident a sapphire crystal at an angle of number density of 1.0 x 1016 c r r 3 is 1.00 cm-l. Cal-
50.0" with respect to the normal. Calculate the angle culate the absorption cross section and the molar ab-
of refraction, the reflectance for each polarization sorptivity.
component, and the total reflectance. See Appendix Collimated laser radiation in 488 nm is impingent on
B for refractive index data. a 100-pm-wide slit. A diffraction pattern is seen on a
Consider an F12 lens that is 5.0 cm in diameter. A light screen that is 1.0 m behind the slit. What is the half-
source is placed 10 cm from the lens. Where is the width (FWHM) of the central maximum?
image focused, and what is the magnification? What Calculate the numerical aperture and Fin for a fiber
are the Fin of the system and the solid angle collected? optic with core and cladding refractive indices of 1.50
A monochromator has the following specifications: re- and 1.48. respectively.
ciprocal linear dispersion, 2.5 nm mm - I ; focal length. Consider a Fabry-Perot interferometer with a mirror
0.30 m ; F-number. 6.0: grating size. 44.33 x 44.33 spacing of 5.0 mm and mirror reflectances of 0.95. For
mm; groove density. 1200 grooves per mm. Calculate h = 500 nm and normal incidence, calculate the coef-
the following at 400 nm assuming the first order is ficient of finesse, the order. the FWHM in nanome-
used. ters, the finesse, the theoretical resolving power, and
(a) The angular and linear dispersion. the free spectral range.
(b) The projected limiting aperture diameter and pro-
What performance characteristics of a monochromator
jected limiting area.
are affected when only the grating groove density is
(c) The slit width needed to obtain a 5-nm geometric
changed?
spectral bandpass.
(d) The angle of incidence and angle of diffraction. A ray in air is incident on a block of crown glass
(e) The diffraction-limited slit width, diffraction-lim- (T~,,,, = 1.52) at a 30" angle from the normal to the
ited spectral bandpass, and diffraction-limited re- glass surface. A t what angle relative to the normal will
solving power. the ray be transmitted through the glass?
(f') The radiant power in watts passed by the mon- Locate and describe the image of a safety pin 100 cm
ochromator if the special radiance of the contin- away from a convex spherical mirror having a radius
uum source is 2.00 x 1 0 - V ~ m sr-I - n~ m l with of curvature of 80 cm.
H = 5 . 0 m m , W = 0 . 5 0 m m , a n d T 0 , = 0.40. A thin biconvex lens (q = 1.5) has radii of curvature
Calculate the velocity of light in H,O at 579 nm. of R, = 40 cm and R, = 20 cm.
Chap. 3 1 Problems
(a) What is the focal length of the lens? (a) What wavelength will appear in the first order at
) Locate and describe the image of an object 40 cm a diffraction angle of 30.0"?
away from the lens. (b) At what diffraction angles will 300, 400, 500, and
A point source of radiant intensity 1.0 W sr-I is placed 600 nm appear?
at the focal point of a 1.0-cm-diameter Fl6.0 lens. Cal- (c) Find the diffraction angle for the second order of
culate the following values: 280 nm. What first-order wavelength is overlapped
(a) The focal length of the lens. by the second order of 280 nm?
(b) The solid angle collected by the lens. For the grating in problem 3-20:
(c) The radiant emittance of the collimated beam (ig- (a) What blaze angle would give a blaze wavelength
nore transmission losses). of 300 nm?
A Fabry-Perot interference filter has a d spacing of ) The grating is used in a Czerny-Turner mano-
2.04 x cm and a dielectric with -q = 1.35. chromator with a takeoff angle of 10.0". Find the
(a) What central wavelength is passed in the first or- grating rotation angle 0, the angles of incidence
der? and diffraction a and P, and the angular dispersion
(b) What wavelength is passed in the second order? D, for 300-, 400-, and 500-nm radiation.
(c) If the filter oscillates such that the angle of inci- (c) What percentage change in angular dispersion oc-
dence varies from - 10 to lo0, over what wave- curs over the range 300 to 500 nm?
length range is the first-order wavelength scanned? The monochromator in problem 3-21b has a focal length
A grating has a groove density of 1500 grooves per f of 0.30 m.
mm. If the incident beam strikes the grating at an angle Find the linear dispersion D, and the reciprocal
of 20.0"; linear dispersion R, at 300,400, and 500 nm in the
What diffraction angles will the first order of 300, first order.
400, 500, 600, and 700 nm appear? Find the geometric spectral bandpass and the slit-
What wavelength in the second order o ~ e r l a p the
s width-limited resolution at 300, 400, and 500 nm
600 nm first-order beam? for slit widths of 100, 200, and 500 pm.
What is the free spectral range for the first order What radiant powers in watts are passed by the
at 600 nm? monochromator for the slit widths in part (b) if a
If the grating in problem 3-15 is blazed at an angle continuum source is incident with a spectral ra-
of 15.0", what is the blaze wavelength? diance of 1.50 x 10-I W cm-'sr-l nm-I when
A t what angle would the grating in part (a) need the monochromator is set to 500 nm; the slit height
to be blazed to obtain a 450-nm blaze wavelength? H = 5.0 mm, To, = 0.43, and the monochromator
Fin = 7.0?
The grating in problem 3-15 is operated at 500 nm
) Describe in words why the radiant power through-
in the first order. What is the angular dispersion
put of a monochromator shows a different de-
of the grating?
pendence on slit width for a continuum source than
What is the angular dispersion at 300 nm in the
for a line source.
first order?
(e) What are the theoretical resolving power and the
A thin biconvex lens of refractive index 1.5 and di- diffraction-limited spectral bandpass for the first
ameter of 2.5 mm has radii of curvature of R , = 1 cm order of 500 nm if the grating width is 5.0 em?
and R, = 0.5 cm.
A Michelson interferometer has a mirror driven at 1.5
(a) Find the focal point of the lens. cm s ' . What frequency would the interferogram show
(b) If the object is 2 cm away from the lens, where is
if the source radiation were at (a) 400 nm: (
the image? What is the magnification'?
(c) 10 pm?
(c) If the object is a radiation source with a radiance
of 2.0 x W cm -' s r l and a projected area What distance must the mirror be driven in a Mich-
of 0.10 cm2, what is the irradiance of the image? elson interferometer to separate (a) infrared radiation
(Ignore transmission losses.) at 10.15 and 10.18 pm; (b) visible radiation at 725 and
(d) Whcre should the object be placed so that rays 730 nm?
emerging from the lens are parallel? Characterize the shape of the slit function, and hence
Why are mirrors preferred over lenses for imaging in the shape of a scan of a line source, if the exit slit is
many spectroscopic instruments that must cover mul- twice as wide as the entrance slit, and vise versa.
tiple wavelengths? If lenses must be used, how can At 589 nm, the focal length of a lens is 5.0 cm and
their imaging properties be idealized at least for two -q = 1.544. A t 200 nm, the refractive index is 1.651.
wavelengths? What are the disadvantages of this ap- Calculate the focal length at this wavelength. If the
proach? lens was used at 589 nm for 1 : 1 imaging, where is
A grating has a groove density of 2000 grooves per the image focused at 200 nm3
mm. The incident beam strikes the grating at an angle A t 400 nm, a solution of an optical active compound
of 20.0". has a circular birefringence of 4.0 x 10 and a circular
86 Chap. 3 I Optical Components of Spectrometers
dichroism of 10 L mol-1 cm-1. Calculate the optical 3-29. Calculate the maximum amplitude of the electric field
rotatory power and the molar ellipticity in a 10-cm associated with a beam of irradiance 1 mW ~ m - ~ .
cell. 3-30. A line source is scanned with a monochromator at 5.0
3-28. The angular separation of maxima for a multiple-slit nm min-I. On a recorder. a triangular peak is observed
mask illuminated with 400-nm radiation is 0.10 rad. with a half-width of 1.0 nm with a chart speed of 2.0
Calculate the slit spacing. cm min- l. Calculate the spectral bandpass of the mon-
ochromator.
The following books, chapters, or reviews treat the general and Spectrometers" (H. W. Faust), "Gratings and Grat-
principles and applications of basic optics. ing Instruments" (R. M. Barnes and R . F. Jarrell) and
.Spectroradiometric Principles" (H. T. Betz and G. L.
1. E. Hecht and A. Zajac, Optics, Addison-Wesley. Read- Johnson).
ing, Mass., 1974. An excellent modern optics textbook.
R. C. Elser, "Optical Instrumentation," in Trace Anal-
The illustrations are superb and the discussion is both
ysis; Spectroscopic Methods for Elements, chap. V,
quantitative and qualitative. Highly recommended.
J. D . Winefordner, ed., Wiley-Interscience, New York,
2. M. Young, Optics and Lasers, Springer-Verlag, New 1976.
York, 1984. A fairly mathematical treatment, but clearly
J. F. James and R . S. Sternberg. The Design of Optical
done.
Spectrometers, Chapman & Hall, London, 1969.
3, W. J. Smith, "Image Formation: Geometrical and Phys-
R. A . Sawyer, Experimental Spectroscopy, Dover, New
ical Optics," in Handbook of Optics. W. G. Driscoll and
York, 1963. Includes a good discussion of spectrographs
W. Vaughan, eds.! McGraw-Hill. New York. 1978. At and spectroscopic equipment.
an elementary level, but a good review.
E . J. Meehan, "Spectroscopic Apparatus and Measure-
4. J. A . Meyer-Arendt. Introduction to Classical arzd Mod-
ments," in Treatise on Analytical Chemistry. 2nd ed.,
ern Optics, 2nd ed., Prentice-Hall, Englewood Cliffs, pt. 1, vol. 7, chap. 3, I. M. Kolthoff and P. J. Elving,
N.J., 1984. eds., Wiley-Interscience, New York. 1981.
5. F. A. Jenkins and H . E. White. Fundamentals of Optics, P. R. Griffiths, ed.. Transform Techniques, in Clzem-
4th ed.. McGraw-Hill, New York, 1976.
istry, Plenum Press. New York, 1978. Contains a dis-
6. M. Born and E. Wolf, Principles of Optics: Electro- cussion of interferometers and their design and opera-
magnetic Theory of Propagation, Interference and Dif- tion.
fraction of Light, 6th ed.. Pergamon Press, Elmsford, P. R. Griffiths and J. A , deHaseth. Fourier Transform
N.Y., 1980. Infrared Spectroscopy: Wiley. New York, 1986. Dis-
cusses interferometers and their use in FTIR techniques.
The following are devoted to more specialized topics in optics.
J . E . Stewart, Infrared Spectroscopy: Experimental
7. E. Hartfield and B . J. Thompson. "Optical Modula- Methods and Techniques, Marcel Dekker. New York,
tors," in Handbook of Optics, W. G . Driscoll and W . 1970. Has a good discussion of conventional dispersive
Vaughan, eds., McGraw-Hill, New York, 1978. spectrometer designs as well as interferometer systems.
8. M. Gottlieb, C. L . M. Ireland, and J. M. Ley. Electro- G . A. Vanasse, ed., Spectrometric Techniques. vols. I-
Optic and Acousto-Optic Scanning and Deflection, Mar- IV, Academic Press, New York, 1977-1985. The first
cel Dekker, New York, 1983. two volumes, in particular. discuss interferometers and
9. Diffraction Grating Handbook, Bausch and Lomb, their applications in FTIR methods.
Rochester, N.Y. : 1970.
The references that follow provide additional details on spe-
10. J . Wilson and J. F. B. Hawkes, Optoelectronics: A n cialized subjects.
Introduction, Prentice-Hall: Englewood Cliffs, N.J. 1983.
;
Includes a discussion on optics. 21. R. Bracewell, The Fourier Transfornz and Its Applica-
11. Electro-Optics Handbook, Technical Series EOH-11, tions, McGraw-Hill, New York. 1965. A classic text on
RCA Corp., Lancaster. Pa., 1974. Includes a concise the use of Fourier transforms.
discussion of basic optics and optical components. 22. E. Charney. The Molecular Basis of Optical Activity:
Optical Rotatory Dispersion and Circular Dichroism,
The following references are devoted to the optics of spec- Wiley. New York, 1979.
troscopic instruments.
In addition to the above, various catalogs and guides are
12. E . L. Grove, ed., Analyrical Emission Spectroscopy, vol. available from optics and spectrometer manufacturers. Many
1. pt. 11, Marcel Dekker. New York, 1971. Contains of these provide a wealth of fundamental and practical in-
excellent chapters on "Prism Systems. Spectrographs. formation.
In addition to the optical components presented in tronic signal processing and reaaout systems most widely
Chapter 3, spectrochemical methods require sources used in spectrochemical methods is presented. The prin-
detectors of radiant power. Radiation sources and ciples of modulation, photon counting, lock-in ampli-
detectors, along with electronic and computer signal fication, and several other signal processing methods
cessing systems, are the major topics of this chapter. are described. The chapter ends by considering com-
cause practical radiation sources for absorption, lu- plete single-channel, multichannel, and multiplex op-
minescence, and scattering measurements are often tical spectrometers and the expressions which govern
compared to blackbody radiators, the laws of blackbody their use in making emission, absorption, and lumi-
radiation are introduced here first. The Einstein prob- nescence measurements.
ability coefficients for emission and absorption are then
explained. Our discussion of specific radiation sources
first deals with conventional line and continuum sources
and their characteristics. Then the laser is introduced,
and the principles and characteristics of several types We are all familiar with the fact that a heated surface
of lasers are considered. Radiation transducers (detec- emits electromagnetic radiation. The energy emitted
tors) convert the radiant power of an incident beam increases, and the spectral distribution of the radiation
into an electrical signal that can be processed by elec- shifts toward shorter wavelengths as the temperature
tronic instrumentation. The operating principles and of the surface increases. This is readily noted by a shift
the characteristics of the most widely used thermal and in the color of the surface from red to blue with in-
photon detectors are considered here, and the array creasing temperature. A ody absorbs all radiant
detectors that are making rapid spectral measurements energy incident upon it regardless of wavelength. None
possible are introduced. of the radiant energy is transmitted and none is re-
Al-ter the optical signal of interest is converted flected. Thus a blackbody has unity spectral absorp-
into an electrical signal, it can be manipulated and proc- tance [ a @ ) = 11 at all wavelengths. Because a black-
essed conveniently by analog andlor digital circuits and body is in thermal equilibrium with its environment, it
microcomputers. A brief review of several of the elec- must emit as much radiation as it absorbs. Being a
88 Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
perfect absorber implies that a blackbody is also the stant. Planck's law was found to be in extremely good
most efficient thermal radiator possible. A blackbody, agreement with the observed data.
in fact, radiates more total power and more power per The spectral radiance B, is related to the spectral
unit wavelength interval than any other thermal ra- energy density by B, = U , c l ( 4 ~ )and
, the spectral ra-
diating source at a particular temperature. Although no diance in wavelength units is B, = B,clX2. Thus two
material is an ideal blackbody, one can approximate a useful alternative forms of Planck's law are
blackbody in the laboratory by a hollow, insulated en-
closure or an oven with a small exit hole in one wall.
Radiation that enters the hole has little opportunity to
escape by reflections so that the enclosure acts nearly
as a perfect absorber. If the oven is heated, it can serve and
as a source and emit radiation through the hole. In
thermal equilibrium, absorption balances emission ex-
cept for the small amount of energy that escapes through
the hole.
where c, = 2hc2 = 1.190 x 1016 W nm4 cm-2 sr-l and
Planck's Law c2 = hclk = 1.438 x lo7 nm K . A plot of Bf: vs.
wavelength is shown in Figure 4-1 for several temper-
With an ideal blackbody, absorption exactly balances atures. Note from equations 4-1 to 4-3 that the spectral
emission. Since absorption occurs at the same rate as
emission, the blackbody must contain an equilibrium
density of radiation. We will call the spectral energy
density of a blackbody radiator UE (J H z - l ) or
Ub, (J ~ m nm-I),
- ~ where the superscript b signifies a
blackbody.
Classical radiation theory does not explain the
spectral distribution of blackbody radiation. Wien pro-
duced an expression that fits the observed spectral dis-
tribution fairly well at short wavelengths, but it failed
at long wavelengths. Rayleigh and Jeans were able to
match the observed data, but only in the very long
wavelength region. Planck's success in 1900 in fitting
the spectral distribution of blackbody radiation proved
to be a turning point in the history of physics, for it
eventually led to the quantum description of radiation
and matter. Planck's model assumed that the walls of
the oven were in thermal equilibrium with the radiation
field inside. He assumed, furthermore, that the atoms
inside behaved as oscillators absorbing and emitting
radiant energy. All oscillator frequencies were possible
in Planck's model, and thus a continuum distribution
was predicted in the spectrum. Planck's final assump-
tion was that each atomic oscillator could absorb or
emit only discrete amounts of energy directly related
to its oscillation frequency. Each energy value had to W a v e l e n g t h , nm
be an integer multiple of the basic quantum of energy FIGURE 4-1 Spectral radiance of a blackbody
hv. Statistical arguments then led directly to Planck's as a function of wavelength for several temper-
radiation law: atures. Note the wavelength shift to the blue and
the increase in spectral radiance as the temper-
ature increases. (Adapted with permission from
J. D. Winefordner, S. G. Schulman, and T. C.
O'Haver, Luminescence Spectrometry in Analyt-
ical Chemistry, Wiley-Interscience, New York,
where h is Planck's constant and k is Boltzmann's con- 1972. Copyright O John Wiley & Sons, Inc., 1972).
Sec, 4- 9 1 Blackbody Radiation
Characteristic Examples
Continuum Sources
TABLE 4-2
Common continuum sources
Window or Approximatea
envelope Wavelength spectral radiance
Type Radiating material material range (W cm-, nm-' s r r l )
-- - - - - - - - - - -
Wavelength, p m
lo-'
-5 Spectral distributions
of some continuum sources: (a) spec-
tral radiances of Nernst glower, glo-
bar, and 900•‹Cblackbody vs. wave-
-
length; (b) spectral emittance of
'5
.;f lo-'
tungsten filament lamp vs. wave-
length; (c) spectral irradiance of D, 5
lamp vs. wavelength measured at 50
cm from source. [(a) Adapted with
,;
permission from W. Y. Ramsey and
J. C. Alishouse, A Comparison of 200 300 400 5 00 600 700
Infrared Sources, Infrared Physics, A, nm
vol. 8, copyright Pergamon Press,
Elmsford, N.Y., 1968, p. 151.1 (c)
94 Chap. 4 I Optical Sources, Transducers, and Measurement Systems
globar causes the rod to heat and emit radiation at lem. In some cases an igniter circuit is used to provide
temperatures exceeding 1000•‹C.The power consump- a short high-voltage (10 to 20 kV) start pulse to initiate
tion is normally higher than that of the Nernst glower. ionization. Some lamps provide a third electrode for
Water cooling is needed to cool the metallic electrodes ignition. The pressure of the gas in an arc lamp can
attached to the rod. The spectral distributions of the vary from a few torr to more than 100 atm; the pressure
Nernst glower, the globar, and a 900•‹Cblackbody are determines the type of discharge and the type of spectral
shown in Figure 4-5a. The globar has slightly better output obtained.
emissivity at short wavelengths (A < 6 pm) than does Hydrogen and deuterium arc lamps (Figure 4-4c)
the Nernst glower. It is less convenient to use and more provide strong spectral continua in the UV spectral
expensive, however, because of the necessity for water region. As can be seen in Table 4-2, these lamps are
cooling. useful to wavelengths as short as 185 nm. This short
Tungsten-filament lamps (Figure 4-4b) are nor- wavelength limit is set by the transmittance character-
mally used in the visible and near-IR regions. Because istics of the quartz envelopes. At wavelengths longer
of their widespread applications, these lamps are avail- than about 370 nm, the spectra (see Figure 4-5c) from
able in a variety of sizes and shapes. Ribbon filament hydrogen and deuterium are no longer continua. Deu-
rather than wire filament types are excellent for spec- terium lamps provide higher intensity than do hydrogen
troscopy because the image of the ribbon can be ori- discharges. Spectrophotometric instruments that op-
ented in the plane of a spectrometer slit. The trans- erate in the UV and visible regions often require two
mission characteristics of the glass envelope limit the sources: a hydrogen or deuterium lamp is used for the
wavelength range to 320 to 2400 nm. as shown in Table UV region, 200 to 350 nm, and a tungsten-filament lamp
4-2 (see also Figure 4-33). Quartz-halogen lamps have is used for the visible region, 350 to 800 nm.
operating lifetimes that are considerably longer than Molecular fluorescence and phosphorescence
normal tungsten lamps. They can be operated at higher spectrometry require high-intensity. continuum sources
temperatures to provide higher intensities than tungsten of UV-visible radiation. The xenon arc lamp shown in
lamps. The added iodine reacts with gaseous tungsten Figure 4-4d is the most common source for molecular
near the quartz wall to form WI, which diffuses to the fluorescence spectrometry. It is available in a variety
hot filament and redeposits tungsten. This regeneration of sizes and shapes. The wattages range from 35 to
process is responsible for the longer lifetime. Because 10,000. The EIMAC lamp (Varian. Inc.) shown in Fig-
of their higher operating temperatures. quartz-halogen ure 4-4e is a xenon arc lamp with the electrodes at the
lamps tend to be less stable than normal tungsten lamps focal point of a parabolic reflector. The beam emerging
unless various feedback stabilization techniques are uti- from this lamp is nearly collimated, which has advan-
lized to control them. tages for certain types of spectroscopy.
In a high-pressure lamp such as a Xe arc lamp,
Arc Lamps. In an arc lamp an electrical dis- the discharge is constricted to a small volume between
charge is sustained through a gas or metal vapor en- the electrodes (typical 2 to 10 mm separation). These
closed in a sealed glass or quartz envelope. The dis- lamps are typically operated with a dc power supply of
charge is initiated and maintained by dc or ac current 20 to 30 V and 2 to 20A. Pulsed xenon lamps and power
through the gap between two electrodes (cathode and supplies are also available; these provide 1 to 10 ps
anode) in the enclosure. Ionization of the vapor within pulses. Figure 4-6 shows that the spectrum from a Xe
the tube is necessary to achieve conduction. Some dis- arc lamp is effectively a continuum with little fine struc-
charges operate with a heated cathode. and the electron ture.
emission from the cathode causes ionization of the gas.
With cold cathode discharges, ions are formed by cosmic Continuum Plus Line Sources
rays, radioactivity. UV radiation, or another external
source. To sustain ionization, a potential difference that Other high-pressure arc lamps produce intense line
exceeds the breakdown voltage of the gas is applied. A spectra superimposed on an intense spectral continuum.
self-sustaining discharge results when positive ions. ac- The high pressure and the arc restriction considerably
celerated by the field, bombard the cathode to produce broaden the atomic lines. The most common of these
secondary electrons. These electrons then collide with lamps are mercury and xenon-mercury arc lamps. Fig-
gaseous molecules or atoms to maintain ionization in ure 4-6 compares the spectrum from a xenon arc to that
the tube. Since this type of discharge gives a negative from a mercury arc lamp. The high-pressure mercury
dynamic resistance, a ballast resistance is necessary to arc contains many broad lines from atomic mercury
prevent burnout. superimposed on a continuum. The spectral radiance is
With all arc lamps, starting the arc can be a prob- high, exceeding 1 W cmp2 nm-l s r r l for the 100-W
Sec. 4-2 I Conventional Radiation Sources
pectral distribution
arc lamps: curve a,
lamp; curve b, 100-
rcury arc lamp. Strong self-ab-
sorption (see Chapter 7) causes the
lack of radiance at 254 nm. (Adapted
with permission from J. D. Wine-
fordner, •˜. G. Schulman, and T. C.
O'Haver, Luminescence Spectrome-
try in Analytical Chemistry, Wiley-
Interscience, New York, 1972. Co-
pyright O John Wiley & Sons, Inc.,
1972.) Wavelength, nm
lamp shown in Figure 4-6. Because of the presence of hot-cathode variety and operate at relatively low volt-
atomic lines, this source is not very useful for molecular ages (105 to 150 V ac). They differ from conventional
absorption spectroscopy, where it is desirable to have fluorescent lamps in that they are made to transmit
a flat spectral distribution; however, it is an excellent ultraviolet radiation. In a normal fluorescent lamp the
source for photoluminescence spectroscopy, especially wall coating absorbs ultraviolet radiation and reemits
if a Hg line corresponds to the wavelength of excitation. visible light.
ercury arc lamps are available that operate from al- Mercury Pen-Ray or Sterilamp types are cold-
ternating current or direct current and in wattages up cathode types that start and operate at higher voltages
to 2500. (200 to 410 V ac operating voltage) than the hot-cathode
Mercury-xenon arc lamps can provide very high variety. These small pencil-shaped lamps are available
spectral radiances, particularly at wavelengths where commercially; most can be obtained with an appropri-
mercury emits. They are available in wattages up to ate transformer and ballast for starting and sustaining
5000. These sources have proven particularly useful in the discharge. The small sizes of these lamps and the
molecular fluorescence spectroscopy, where high inten- ness of the atomic lines made them excellent sources
sity is desirable. The presence of the Xe stabilizes the for wavelength calibration of spectroscopic equipment,
arc compared to an ordinary Mg lamp. particularly in the ultraviolet region.
Anode
Shield
but are usually in the range 10 to 50 mA. High currents
I
can cause appreciable line broadening and self-reversal
(see Section 7-5). Operation at high currents can sig-
nificantly reduce the lamp lifetime. Manufacturers of
atomic absorption spectrometers often recommend op-
I I
Base I erating hollow cathodes at currents substantially lower
than the maximum values. High intensity or boosted
output hollow cathode lamps are also available for many
elements. The intensities of these lamps can be several-
I fold higher than normal HCLs.
Hollow For use with synchronous detection systems, HCLs
cathode lamp can be electrically modulated or a mechanical chopper
can be used. Lock-in detection systems (see Section 4-
5) require 50%-duty-cycle modulation, as is commonly
FIGURE 4-7 Shielded hollow cathode lamp (a) used in atomic absorption spectrometry. (The duty cycle
and power supply circuit (b). The cathode in (a) is the fraction of the time the lamp is on.) With electrical
is a cuplike hollow cylinder made of or lined with modulation schemes the hollow cathode current is often
the element or an alloy of the element to be ex- not turned completely off, to enhance lamp stability.
cited. A simple wire serves as the anode. The For atomic fluorescence spectroscopy dc operation of
envelope is filled with neon or argon at a pressure HCLs does not produce sufficient intensity. High in-
of a few torr. Commercial lamps are available for tensities can be obtained, without exceeding the max-
most elements. Typically, a few hundred volts is imum current ratings, by pulsing the hollow cathode in
applied between the cathode and anode (b). a low-duty-cycle mode. Here the current during the
lamp on-time can be high, but because of a long off-
Hollow cathode lamps can produce extremely nar- time, the average current is maintained below the max-
row atomic lines. When operated properly, the atomic imum rating. Boxcar integration schemes are often used
line widths are only 0.01 to 0.02 A (FWHM). These with pulsed HCLs (Section 4-5).
line widths are narrower than absorption line widths in
flames, as discussed in Chapter 7. A typical spectrum Electrodeless Discharge Lamps. The elec-
from a hollow cathode lamp (HCL) is shown in Figure trodeless discharge lamp (EDL) is a very intense atomic
4-8. line source, often producing line radiances 20 to 50
Unlike most sources, provision is usually made to times greater than those of a hollow cathode lamp.
vary the lamp current in order to vary the line radiance. Because of this EDLs have found use in atomic fluo-
The operating currents of HCLs should not exceed the rescence spectroscopy and in atomic absorption for ele-
maximum current rating specified by the manufacturer. ments such as As, Se, Te, and Hg, where HCL intens-
These maximum values vary from element to element ities are relatively low.
The EDL is basically a sealed quartz tube, as shown
in Figure 4-9, that contains a few torr of an inert gas
and a small amount of the metal (or metal salt) whose
spectrum is desired. The lamp is placed in an intense
RF or microwave field that is directed on the lamp by
an antenna or a waveguide cavity. A Tesla coil is used
to start the discharge by ionizing some of the inert gas
atoms. The electrons produced are accelerated by the
high-frequency component of the field and acquire suf-
ficient energy to ionize other atoms and maintain a
plasma. ( A plasma is an ionized gas.) The heat pro-
duced vaporizes the metal or metal compound, which
is excited by collisions to produce the spectrum. Be-
cause the gas pressure and temperature are low, broad-
ening effects are small, and narrow spectral lines result.
The microwave or R F generator employed with
FIGURE 4-8 Portion of spectrum from a dual- EDLs is typically a 50- to 1000-W supply operating at
element hollow cathode lamp. 2450 MHz. Although lamps are available commercially
Sec. 4-2 1 Conventional Radiation Sources
EDL
Glass ,
wool Quartz
shell
Metal or metai
compound
FIGURE 4-9 Electrodeless discharge lamp. The spectral distribution of an LED is a continuum over a
sealed tube is placed in an intense radio-fre- narrow wavelength range (FWHM typically 30 to 50
quency (RF) or microwave field. nm). They are usually operated at low voltages (e.g.,
5 V) with currents in the range 1 to 260 mA. Their
for some elements, EDLs can be readily produced in major advantage is their small size.
the laboratory. Unfortunately, there is still a great deal Fluorescent lamps are not often used as spectro-
of art in the preparation of stable, reliable lamps for metric sources, but they are commonly used to initiate
general-purpose use. Thermostating the lamp often im- photochemical reactions. Fluorescent lamps are dis-
proves stability. Some EDLs have been operated in a charge devices that produce light by the emission from
pulsed mode. luminescent powders excited by a low-pressure mercury
The use of EDLs in atomic fluorescence spec- arc. The spectral distribution of the light is a function
trometry has declined in recent years due to the avail- of the chemical composition of the phosphor used.
ability of lasers. These increasingly important radiation
sources are discussed in detail in Section 4-3.
Thermal Gradient Lamp. The thermal gradient Standard sources are used to calibrate monochomators,
lamp (TGL) is a relatively new atomic line source that detectors, and secondary sources for absolute radio-
can be constructed for elements of high volatility (As, metric or photometric measurements. The primary
Cd, P, S, Se, Te, Zn). A commercial TGL is shown in standard photometric source is a blackbody constructed
Figure 4-10. Intensities of TGLs are as high as those of from a fused-thoria (Tho,) crucible that is nearly filled
EDLs and the lines produced can be narrower. The with pure platinum. A small cylinder of pure fused thoria
TGL has a much shorter warm-up time than an E D L is placed in the cer?ter of the crucible. The bottom of
of the same element. Because of these factors TGLs the cylinder is packed with powdered fused thoria. The
could replace EDLs in atomic absorption determina- crucible has a small hole in the center from which the
tions of As, Se, and Te, and they could become popular light emerges through a funnel-shaped sheath. The cru-
atomic fluorescence sources for some elements. cible is embedded in powdered fused thoria and is heated
by enclosing it in an induction furnace. The temperature
iscellaneous Sources of the crucible is first raised to well above the melting
point of platinum. Then it cools slowly until the soli-
Light-emitting diodes (LEDs) have been used as sources dification temperature (2042 K) is reached. The lumi-
in fixed-wavelength, molecular absorption spectrome- nance of the hole at the solidification temperature of
ters. They are pn-junction devices that when forward
-
biased produce radiant energy. Gallium arsenide (A,,
900 nm), gallium phosphide (X,, 650 nm), and silicon
- platinum is given the value of 60 lm srP1 cm-,.
The National Bureau of Standards (NBS) has es-
tablished working standards of spectral radiance and
carbide (X, = 580 nm) are commonly used. Mixtures spectral irradiance based on calibrations relative to
of these compounds can be used to shift the wavelength blackbody sources. The standards of spectral radiance
maximum anywhere in the region 540 to 900 nm. The cover the wavelength region of 250 nm to 2.5 pnl. The
98 Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
sources are commercial GE type 30 AlT2417 tungsten frequency to cause stimulated transitions in the system,
ribbon filament lamps centered 8 to 10 cm behind a each stimulated emission generates a photon while each
fused silica window that is 3 cm in diameter. Values of absorption removes a photon. The change in radiant
spectral radiance for these lamps are given as a function flux d@ in the distance dz due to absorption is given
of wavelength. They are intended for use with auxiliary by d@ = @ n,udz, where @ is the transition cross section
optics of specified arrangement. Since these standards in cm2 (see Section 3-1 and Appendix F). The change
are expensive they are used infrequently to calibrate in flux due to stimulated emission in distance dz is d@
other working standards. When operated properly, the = an,@ dz. Thus we can write the total change in ra-
uncertainties in spectral radiance are less than 10%. diant flux in distance dz as
Tungsten-filament, quartz-iodine lamps are issued
as standards of spectral irradiance for the wavelength
region 250 nm to 2.6 Fm. They are commercial GE type
6.6 AlT4QllCL-200-W lamps. Values of spectral irra-
diance vs. wavelength are supplied. The irradiance Equation 4-15 shows that there is a net gain in flux
standards require no auxiliary optics. Again these when n, > n, and a net loss in flux with n, < n,. When
standards are expensive and should be used to calibrate the system is at thermodynamic equilibrium, (n,),, >
other, less expensive, working standards. The uncer- (n,),,, and the material is a net absorber. However, if
tainties in the spectral irradiance standards are also less the upper-level population can be made to exceed that
than 10%. of the lower level (n, > n,), the system behaves like an
The primary standard of length is the wavelength amplifier at frequency v,. Under these conditions, the
in vacuum of the radiation of the orange line of 86Kr atomic or molecular system is called an active medium,
unperturbed by external influences. The accepted and it has undergone population inversion. In the IR-
wavelength is A = 6057.80210, A. The emission is pro- to-visible frequency region, such an amplifier is called
duced in a hot-cathode discharge tube filled with high- a laser for light amplification by stimulated emission of
purity 86Kr at a temperature of 64 K. radiation.
Despite the existence of working standards for As in an electronic circuit, we can make an os-
absolute photometry and radiometry, an accuracy of cillator from an amplifier by introducing positive feed-
5% is difficult to achieve and usually obtained only by back. This can be done with a laser by placing the active
experts. Accuracies approaching 1% are very rare and medium in a resonant cavity (Fabry-Perot cavity) made
realized only in a few types of measurements within a from two mirrors, as shown in Figure 4-11. When the
very few laboratories. Fortunately. there are not very gain of the active medium equals the losses in the sys-
many spectrochemical methods that require even an tem, oscillation begins. If we assume that the major
estimate of absolute intensities because relative intens-
ities are used in most procedures.
- -
4-3 LASER SOURCES %
Output
beam
In spectroscopy laser sources are making it possible to - Active medium
-
Mirror 1 Mirror 2
obtain information that was difficult or impossible to partially
obtain with conventional sources. This section presents transparent
the basic principles of lasers and describes the types of FIGURE 4-11 Laser cavity. In the laser the ac-
lasers that are useful in spectrochemical methods. Some tive medium is positioned between two mirrors.
of the unique characteristics of laser sources are pre- The length of the cavity is distance d ' , while the
sented along with the principles of tunable lasers. Ap- optical pathlength is d. Here d is given by d =
plications of lasers to various types of spectrochemical qb + q1(d' - b ) , where -q is the refractive index
methods are described in later chapters that deal with of the active medium and q' is that of the re-
specific techniques. mainder of the cavity. The electromagnetic wave
travels back and forth between the mirrors being
amplified at each pass through the medium. A
Principles of Lasers
partially transparent mirror allows part of the beam
to pass out of the cavity as the output. Although
Let us consider a collection of atoms or molecules in- plane parallel mirrors are shown, spherical mir-
teracting with an electromagnetic wave that is propa- rors are normally used to minimize diffraction
gating along the z axis. If the wave is of the appropriate losses and improve beam quality.
Sec. 4-3 / Laser Sources
2
losses occur because the mirrors are imperfect reflec-
tors, the condition for oscillation is given by t Fast decay
old population inversion can be obtained by substituting FIGURE 4-12 Three- and four-level lasing
G from equation 4-17 into equation 4-16 and solving schemes. In the three-level scheme (a), atoms
are excited (pumped) in an appropriate manner
for n, - n,. The result is
to the upper level (2), from which they rapidly
decay to an intermediate level (1). If the inter-
mediate level is long-lived compared to the upper
level, the population of level 1 can grow relative
to level 0 until inversion is achieved. In the four-
The population inversion can be achieved by sev- level scheme (b), the atoms are excited to level
eral different schemes. In a two-level system, it is not 3 from which they rapidly decay to level 2. Pop-
possible to achieve inversion since stimulated emission ulation inversion occurs between levels 2 and 1.
and absorption must be balanced. Hence systems with It is easier to achieve the inversion in the four-
level system.
three or more levels are used as shown in Figure 4-12.
The excitation process, called pumping, can be by op-
tical methods with other sources, by electrical methods terminating lasers can be operated only as pulsed lasers.
using an electrical discharge, by chemical reactions, or The maximum useful pump pulse width is on the order
by a rapid adiabatic expansion. Once inversion is of the lifetime of the lasing transition. In a pulsed laser,
achieved, oscillation can be initiated by spontaneous the output appears as a repetitive pulse train or the
emission processes. All lasers, of course, have ineffi- pulsed laser can be operated in a single-shot mode. The
ciencies, so that the output energy is significantly less radiant power output of a pulsed laser, during the pulse,
than the pump energy. Even the most efficient lasers can be very high (kW to > MW) even though the av-
may have overall efficiencies of 20% or less. However, erage power is low. Output characteristics of pulsed
as discussed below, the characteristics of laser radiation lasers can be expressed as output energy per pulse, peak
are remarkably different from those of conventional output power, or average power. For a laser that pro-
sources. duces 5-ns pulses at a repetition rate of 50 pulses s-'
with an energy of 10 mJ per pulse, the peak output
Laser Types power is 10 x Jl(5 x s) = 2 MW. The
average power is 10 x l o p 3 Jlpulse x 50 pulsesls =
There are many different types of lasers available and 0.5 W. The radiant power outputs of CW lasers are
several different operational modes. from milliwatts to watts.
With CW and pulsed lasers the population inver-
Operational Modes. The four major opera- sion cannot exceed the threshold value to any large
tional modes are continuous wave, pulsed, Q-switched, extent because lasing occurs when the threshold is
and mode-locked. In a continous-wave (CW) laser the reached. In a Q-switched laser, often called a giant pulse
output beam is continuous with respect to time. To laser, the quality factor (Q) of the cavity is spoiled and
achieve CW operation, the upper level of the lasing oscillation is prevented until the population inversion
transition must be longer lived than the lower level. If has grown well beyond the threshold value. The cavity
the upper level were to have a shorter lifetime than the is suddenly switched on, and a powerful giant pulse is
lower level, the laser would be self-terminating. Self- obtained. Although there are several ways to accom-
100 Chap. 4 I Optical Sources, Transducers, and Measurement Systems
plish Q-switching, one simple method employs a sat- are shown in Figure 4-13. The lowest order TEM,, mode
urable absorber in the cavity. The saturable absorber is the most widely used because the flux is Gaussian
is often an organic dye in solution. Until the radiation over the cross section of the beam. (Emittance de-
in the cavity builds up to a high intensity, the dye ab- creases from the center of the beam cross section ac-
sorbs the light and prevents laser action. The cavity cording to the Gaussian equation; see Appendix A.) A
thus has high losses, and the population inversion must complete mode specification is of the form TEM,,,,
become very large to overcome these losses. When the where n is the axial mode number. For each TEM,,,
radiance in the cavity reaches a critical level, the ab- mode there are many longitudinal modes each sepa-
sorber becomes bleached (saturated) and transparent rated by a frequency Av, = cl2d. For a gas laser with
to the light. If the population inversion is high at this a 1-m cavity, Av, = 150 MHz. This frequency difference
time, a very intense laser pulse is produced. Laser out- is much smaller than the line width of the atomic tran-
put powers on the order of hundreds of megawatts per sition; hence many modes will oscillate simultaneously.
pulse can be achieved. The Q-switched laser can pro- In mode locking the phases of the oscillating modes
duce a single pulse or a repetitive pulse train. are forced to be correlated or locked to each other
Before we can understand the fourth operational rather than random. This results in ultrashort pulses
mode, it is necessary to consider the mode structure of (picoseconds or less) of very high peak power. Often
a normal laser beam. In a resonant cavity such as that the phase locking is accomplished by placing a saturable
shown in Figure 4-11, standing waves occur when there absorber or an acousto-optic modulator (see Section 3-
is an integral number n of half-wavelengths spanning 3) in the cavity. If n' modes oscillate in the presence of
the optical pathlength d between the mirrors. Consec- a saturable absorber, the cavity loss contains a term
utive longitudinal (axial) modes, each with a distinctive that oscillates at Av', the beat frequency of adjacent
frequency v, = ncl2d, are separated by frequency dif- modes. When the absorber bleaches, pulses are pro-
ferences Av, = cI2d. These longitudinal modes corre-
spond to standing waves set up along the z-axis. In
addition transverse modes, called TEM for transverse
The duration of each pulse t, is t, -
duced in which the phases of the modes are locked.
llAv' =
2d/[c(2n1 + I)]. The temporal behavior of such a Q-
electric and magnetic, can be sustained as well. The switched, mode-locked laser is shown in Figure 4-14.
transverse modes are given the symbol TEM,,, where
m and 1 are the integer number of transverse nodal lines Solid-State Lasers. The first operating laser was
in the x and y direction across the beam. The light a pulsed laser which had as its active medium a ruby
patterns for several modes (observed at the mirrors) crystal. A simplified energy-level diagram of the active
I TEM,
~lashlaip YAG
rod
(a)
power is approximately n' times the average power, FIGURE 4-16 Construction (a) and energy lev-
where n' is the number of locked modes. els (b) of a Nd:YAG laser. The pump transitions
are in the red region of the spectrum, while the
Cr3+ ion in a ruby'laser is shown in Figure 4-15. The laser output is in the near IR (1.06 km).
ruby rod is surrounded by a flashlamp which provides
broadband, pulsed optical pumping. The ends of the important laser material. The active material is shaped
rod are silvered to form a resonant cavity. The ruby as a rod and is optically pumped. The Nd:UAG laser
laser emission occurs at 694.3 nm. The laser pulse is has been operated in both pulsed and CW modes. The
-0.5 ms in duration and has a line width of -0.01 nm. pulsed Nd:YAG laser has a pulse duration on the order
Typical peak output power is 500 kW, but Q-switching of nanoseconds and very high output power at 1.06 pm.
has been used to produce peak powers exceeding 150 It has become a popular laser for pumping tunable dye
MW per pulse. lasers, as discussed later in this section. The lasing scheme
In addition to the ruby laser there are many other is illustrated in Figure 4-16. Mode-locked Nd:glass las-
solid-state lasers. Of these Nd3+ in a glass host or in a ers can produce pulses as short as 1 ps and peak output
yttrium-aluminum-garnet (YAG) host is a parti~ularly powers as high as 1013W.
Absorption
Gas Lasers. The helium-neon (He-Ne) laser
shown schematically in Figure 4-17 is the most common
\'
\'
,:\ Relaxation of all lasers. It is a CW laser pumped by an electrical
\\
\\ discharge. The lasing transitions occur between the Ne
\:,
\\
,\'x,:
Nonradiative
transiti\ons
energy levels, while He is added to increase the pump-
\\ \ \
ing efficiency. The helium atoms are ionized to maintain
\'
\.'w the discharge. Metastable levels in H e atoms transfer
Metastable state
energy efficiently to Ne. The dominant laser transitions
are at 632.8 nm, 1.15 pm, and 3.39 pm. The outputs
of many He-Ne lasers as well as other noble gas lasers
are linearly polarized as described in Figure 4-17. Sim-
ple He-Ne lasers made for use as alignment aids are
available without Brewster angle windows and thus with
unpolarized outputs. The He-Ne laser is simple and
inexpensive. The output power is limited, however, to
the range 0.5 to 50 mW because at low currents few
Ground state metastables form, whereas at high currents the He met-
FIGURE 4-15 Energy levels in a ruby laser. The astables ionize.
atomic system consists of absorption bands, a All of the noble gases have been made to lase.
metastable (long-lived) level, and the ground state. The argon ion laser can oscillate at several frequencies,
The ruby system thus behaves as a three-level with the most intense outputs being at 488.8 nm in the
laser system, blue and 514.6 nm in the green. The output power is
102 Chap. 4 / Optical Sources, Transducers, and Measurement Systems
-*
Discharge tube minated by end windows tilted at
Brewster's angle (see Section 3-1).
Thus no reflections occur at the win-
Mirror dows for light polarized with the elec-
tric field vector in the plane of the
figure. The orthogonal component is
Power partially reflected on each pass and
supply. quickly becomes an insignificant part
dc
of the output beam.
usually several watts CW, but A r + lasers have obtained charges. A gaseous mixture such as Ar, F,, and H e
powers as high as 150 W CW. Argon ion and He-Ne when subjected to the discharge creates ArF excimers
lasers are the most reliable lasers at this time. (an excimer is an excited dimer or trimer). A unique
The CO, laser emits in the infrared region at 10.6 property of rare gas halide excimers is that they are
p.m. It is, at present, the most powerful CW laser avail- dissociated in the ground state. This makes it easy to
able; output powers of several hundred kilowatts have maintain a population inversion because the ground
been obtained. The CO, laser can be pumped by col- state of the excimer rapidly dissociates.
lisions with electrons, or by resonant energy transfer Some of the output characteristics of excimer las-
from added N, molecules. The laser output occurs on ers are summarized in Table 4-3. Typically, pulse du-
a series of CO, rotational lines within a vibrational tran- rations are on the order of 10 to 30 ns with repetition
sition. rates as high as 500 Hz.
The N, laser utilizes a vibronic transition of the The excimer laser is inherently tunable over a nar-
nitrogen molecule at 337.1 nm. It is a pulsed laser that row wavelength range (few nm). Tuning has been ac-
is self-terminating and requires a very short current complished by inserting a prism inside the resonant cav-
pulse for pumping. The typical arrangement is the ity. The excimer laser is an important ultraviolet radiation
Blumlein circuit, shown in Figure 4-18. The N, laser is source, especially for photochemical studies. Various
a common pump laser for tunable dye lasers as discussed wavelengths in the UV are available from different me-
later. dia, as shown in Table 4-3. The excimer laser is becom-
Several metal vapors (Zn, Hg. Sn, Pb, Cu, Cd) ing especially useful for pumping tunable dye lasers.
have been made to lase in the visible region, but at Because of its output wavelength at 308 nm and its long
present only the Cd vapor laser is in common use. The gas lifetime (>lo7 shots per gas filling), the XeCl laser
copper vapor laser is now commercially available. The is the most suitable excimer laser for pumping dye las-
He-Cd laser emits at 325.0 and 441.6 nm. It utilizes ers. The excimer laser has relatively poor beam quality.
transitions of cadmium excited by collisions with met-
astable helium.
Excimer lasers are among the newest gas lasers; Dye Lasers. The first liquid laser was success-
the rare-gas halide excimer laser was discovered in 1975. fully operated in 1963, while the first dye laser was
Laboratory-type lasers are pumped by electrical dis- introduced in 1966. Since then a great many fluorescent
dyes (rhodamines, coumarins, fluoresceins, etc.) have can be performed with efficiencies up to 30%; however,
been made to lase at frequencies from the IR into the they add a great deal of complexity to the laser system.
UV region. Energy-level schemes of fluorescent mol- Continuous-wave operation is also readily avail-
ecules are discussed in Section 12-6. Because of fre- able for many dyes pumped with noble gas ion lasers.
quent collisions in solution, the spectra of such mole-
cules are fairly broad. Lasing typically occurs between Grating
the first excited singlet state and the ground state. The I / I
is used to decrease the bandwidth of the output beam. -19 Dye laser pumped by N, laser.
With the Nd:YAG laser, the fundamental frequency The beam expanding telescope increases the
(1.06 pm) is unsuitable for pumping most dyes. Thus number of grooves of the grating hit by the beam
the frequency must be increased by doubling, tripling, and decreases the power density so as to avoid
or even quadrupling with nonlinear optical techniques damage to the grating. Tuning is accomplished
(see later in this section). These conversion processes by rotating the grating.
LE
Typical pulsed dye laser characteristics
The output powers of CW dye lasers can vary from is switched to the "up" position, the internal energy
milliwatts to watts; they are tunable throughout the circulating in the cavity is deflected out of the cavity.
visible region. Figure 4-20 shows two CW dye laser Normally, only a small fraction of the total energy avail-
designs. The simple standing-wave cavity in Figure 4- able in the cavity is coupled to the output by the output
20a is useful for multimode operation or for low-power reflector. Pulse separation can be selected by choosing
single-mode operation. The ring-cavity design in Figure the appropriate cavity dumping rate. Typically, the pulse
4-20b can produce higher powers, but it is more com- separation can be varied from less than 1 ps to greater
plex. Tunable picosecond pulses at repetition rates greater than 1 ms, providing variable repetition rates. Peak
than 1MHz can be obtained with synchronously pumped powers can be as high as kilowatts.
dye lasers (see Table 4-4). In such a laser a mode-locked
argon ion laser is used to pump a dye laser. The cavity Semiconductor Lasers. In the semiconductor
of the dye laser is extended so that the intermode spac- or diode laser, population inversion occurs between the
ing of the dye laser is an integral multiple of the argon conduction band and the valence band of a pn-junction
laser mode locker frequency. With partially transmit- diode. Stimulated transitions of the electrons from the
ting mirrors, the output is a continuous train of pico- conduction band to the valence band are responsible
second pulses with peak powers on the order of 100 W. for laser action, and stimulated emission results from
The synchronously pumped dye laser can be mod- electron-hole recombinations. Since the frequency
ified to achieve wider pulse separations and an order emitted is directly related to the bandgap energy, var-
of magnitude more energy per pulse by a technique ious semiconductor compositions can be used to give
known as cavity dumping. Here the output coupler of different wavelengths. Also, since the lasing occurs be-
the dye laser is replaced by an acoustic-optic deflector. tween bands, these lasers can be tuned over small in-
The deflector behaves like a mirror that lies parallel to tervals. Typical materials are GaAs, which lases at 0.84
the laser beam inside the cavity. Whenever the mirror pm, and lead salt diodes (PbSnTe), which lase in the
Flat
Tuning output
Collimating element(s) mirror
mid-IR region. Infrared lasers can achieve very narrow (bandwidth) of the laser. A laser with a bandwidth Av
line widths (lop6 cm-l) but must be operated at low has a coherence time 7, = 1IAv. Radiation is said to be
temperatures (10 to 20 K). For a given composition went if there is a constant time-independ-
tuning can be achieved by changing the diode current, ent phase difference for the amplitude at two different
which heats the diode. This causes refractive index points.
changes that alter the effective cavity length. Typical The final aspect of laser radiation that makes it
tuning ranges are 20 to 50 cmpl. different from conventional radiation is its high irra-
diance. Typically, lasers can achieve irradiances that
Chemical Lasers. With chemical lasers no op- are 4 to 10 orders of magnitude larger than those from
tical or electrical pumping is necessary. An exoergic conventional sources. This is a direct consequence of
chemical reaction produces molecules in excited states. the power and directionality of the laser. For example,
If population inversion is achieved, lasing can occur on a pulsed dye laser might produce a peak output power
specific transitions. The hydrogen fluoride laser is the of 10 kW in a beam of 40-mm2 cross-sectional area.
best known example; lasing here occurs on vibrational Even if a conventional source were to produce an equiv-
transitions. An electrical discharge is used to produce alent output power, it could not be collected and fo-
hydrogen atoms and fluorine atoms. The chemical re- cused into such a narrow beam without many losses.
action (H, + F + HF" + H) produces excited HF.
Continuous powers of 475 have been achieved at
3 p m Many other chemical systems have been utilized
to produce pulsed or CW lasers. The monochromaticity and high irradiance of laser ra-
diation have led to many new discoveries. Among these
are optical effects which depend nonlinearly on the am-
plitude of the electric field of the laser beam. One of
Laser radiation has significantly different properties from these, optical harmonic generation, is extremely im-
radiation emitted by conventional sources. Laser radia- portant in the generation of new laser frequencies and
tion is highly directional, spectrally pure, coherent, and is discussed here. An entire new branch of laser spec-
of very high radiance. troscopy involves the observation and application of
The directionality of lasers is a direct result of the nonlinear radiationlmatter interactions. Many of these
resonant cavity. Only waves that propagate normal to techniques are discussed in subsequent chapters.
the mirrors oscillate. Under optimum conditions, this When an electromagnetic wave propagates through
gives rise to a radiant energy beam with an angular a dielectric medium, the electromagnetic field exerts
divergence 0 (radians) approaching the diffraction limit forces on the outer, valence electrons of the material.
(0 = ~XITW,,where w, is the beam waist or minimum Normally, these forces are small, and the resulting elec-
spot size). As a consequence laser radiation can be tric polarization P (C mp2) is directly proportional to
accurately transmitted over large distances (e.g., to a the field. Thus we can write
spot on the moon and back). In the laboratory, the
directionality of lasers simplifies the alignment of op-
tical materials and enables samples in remote locations where x is a dimensionless constant called the electris
to be probed. tibility. At extremely high irradiance, we might
Laser radiation is of high spectral purity. Lasers expect this relationship to break down, particularly if
are often called quasi-mom omatic sources or the field becomes comparable to the binding energy of
continuum sources, depen on one's point of the electrons. At field strengths lower than those that
Spectral purity is a result of the resonant interaction of cause such dielectric breakdown of the material, we can
the medium with the pump source, the cavity gain, describe the polarization by
which enhances the central frequency, and the resonant
cavity, which allows oscillation of characteristic fre-
quencies only. In the visible region it is fairly routine where x,, is called the nth-order susceptibility. If the
to achieve linewidths on the order of 0.01 to 0.1 A. electric field of the incident wave is of frequency w and
Laser radiation is also classified as coherent radia- oscillates according to = Em sin w t , the resulting po-
tion. Radiation is said to be te larization is of the form
a given point in space, there is always a constant phase
difference between the amplitude of the wave at two
successive instances in time. The degree of temporal
coherence is obviously related to the monochromaticity
106 Chap. 4 / Optical Sources, Transducers, and Measurement Systems
which can be rewritten as new shifted frequencies. A prism is usually used to dis-
perse the various wavelengths spatially and allow iso-
lation of the desired beam.
Summary
sin 3wt) + ...
Perhaps no invention except the transistor has stimu-
lated the imagination of scientists and the public more
than that of the laser. For spectroscopic applications,
The first term in equation 4-21 is just the normal lasers provide coherent, narrow-bandwidth, high-inten-
linear term than predominates at low fields. The po- sity radiation. Dye lasers have made such radiation tun-
larization that results from this linear term oscillates at able over the visible region and parts of the ultraviolet
the incident frequency w, and the light that propagates and infrared regions. Despite the great advances that
through the medium corresponds to the usual refracted have been made with laser sources, many difficulties
wave. The second term makes a significant contribution must be overcome before they are routinely used in
only at high irradiances. It contains a constant term, many analytical applications. Only gas lasers are reli-
the dc rectification term, plus a cos 2wt term. The light able enough for routine uses. The high-powered Nd:YAG
that propagates because of this term is of frequency 2 0 and excimer lasers are still somewhat difficult to align
with characteristics of monochromaticity and direction- and use. Many lasers have "down periods" that ap-
ality similar to the incident laser beam. The process of proach 50%, which is much too large for routine ap-
generating the light at 2 0 is called second-harmonic plications. Users of such lasers must be optical and
generation. The third term in equation 4-21 contains a electronic experts, familiar with elaborate alignment
component at 3w, and this generates the third harmonic. procedures and methods to extract signals from noisy
A material that is isotropic or has a center of environments. The future should see many advances in
symmetry can produce only odd harmonics. An ani- laser technology, especially in the development of in-
sotropic material can, however, produce even and odd expensive, reliable, and automated systems.
harmonics. Among the most important materials used
in second-harmonic generation are the piezoelectric
crystals potassium dihydrogen phosphate (KDP) and 4-4 OPTICAL TRANS
ammonium dihydrogen phosphate (ADP). Centrosym-
metric materials like calcite have been used to generate The radiant power monitor introduced in Section 2-6
third harmonics. consists of a radiation transducer, signal processing
Harmonic generation is frequently used with dye electronics and a readout system. The purpose of the
laser systems. The output of a Nd:YAG laser (1.06 pm) radiation transducer is to convert radiant power info an
is frequently doubled or tripled in frequency in order electrical signal or to another physical quantity (e.g.,
to produce radiation suitable for pumping organic dyes. heat or resistance) that can readily be converted to an
The outputs of dye lasers are often frequency doubled electrical signal. Regardless of the specific mechanism
to produce tunable UV radiation. Efficient harmonic involved, the characteristics of the transducer (sensitiv-
generation must negate the effects of dispersion (vari- ity, linearity, dynamic range, signal-to-noise ratio, etc.)
ation of 7 with A) by a technique known as phase match- play an important role in determining the accuracy and
ing. precision attainable in spectrochemical methods.
If two laser beams of frequencies w, and w, are Optical transducers fall into three major cate-
incident on a nonlinear medium, sum (w, + w,) gories: thermal detectors, photon detectors, and mul-
and difference (o, - w,) frequencies can be generated. tichannel detectors. Thermal detectors sense the change
A device called an optical parametric oscillator uses in temperature that is produced by the absorption of
such nonlinear mixing to produce tunable coherent ra- incident radiation. The temperature change is con-
diation. verted into an electrical signal by methods that depend
Another technique used to shift the frequency of on the specific transducer. Thermal detectors have a
a laser is the Raman shifter. discussed in more detail nearly uniform spectral response that is determined by
in Chapter 16. Raman shifters use stimulated Raman the absorption characteristics of their coatings and win-
oscillation in gases such as hydrogen. deuterium, and dow materials. They are thus highly useful for direct
methane to provide shifts of several thousand wave- radiometric measurements as well as for spectroscopy.
numbers. A pump laser beam is focused into a gas cell. Photon detectors. on the other hand, respond to inci-
where various nonlinear processes occur to generate dent photon arrival rates rather than to photon ener-
Sec. 4-4 I Optical Transducers
gies. The spectral response of these transducers varies is called the linear dynamic range, while the dyna
with wavelength. A major advantage of photon detec- range usually refers to the total range of incident radiant
tors is their rapid response time, generally submicro- power over which the transducer is responsive.
second, compared to thermal detectors, which have re- The sensitivity of a transducer is often not only a
sponse times that are slower than milliseconds. Photon function of wavelength and incident radiant power, but
detectors can also detect lower radiant powers than may depend on such variables as temperature, bias volt-
thermal detectors in many cases. Multichannel detec- age, and component values. Hence, with some trans-
tors are photographic emulsions, arrays of multiple ducers, it might be necessary to keep several variables
semiconductor detectors, or arrays of thermal detec- constant during an experiment to ensure constant sen-
tors. The elements in array detectors are arranged lin- sitivity. The constancy of Q(X) or R(X) with time is
early or in a two-dimensional grid. known as the stability of the transducer. Stability can
The general characteristics of transducers are pre- be expressed as short term (hours) or long term (days
sented first in order to define many of the terms that or weeks). Often, the stability depends highly on main-
are later used in describing specific detectors. Thermal taining constant the variables upon which Q(X) or R(h)
detectors, photon detectors, and multichannel detectors depend. A long-term change in Q(X) at a constant @
are then discussed. is referred to as degradation. Some transducers exhibit
hysteresis in that the responsivity at a particular power
changes if the incident power is increased and then
brought back to its original value.
Radiation transducers vary widely in their sensitivity,
linearity, spectral response, response speed, electrical Response Speed. Transducers vary widely in
output domain, and noise figures. To evaluate a trans- their ability to detect rapid changes in incident radiant
ducer for a particular application, the characteristics power. Quantitatively, the response time is evaluated
discussed below are commonly used. in terms of the time constant T = 1/(2&), where fc is
the frequency at which R(X) has fallen to 0.707 of its
Sensitivity and Responsitivity. There are sev- maximum value (3-dB point) when a sinusoidal input
eral ways to describe the sensitivity of a detector. The of frequency fc is incident on the transducer. The rise
responsivity K(X) is the ratio of the rms signal output time is the time for the output to rise from 10% to 90%
X (voltage, current, charge) to the rms incident radiant of its final value when an instantaneous (step function)
power @ evaluated at a particular wavelength and in- increase in radiant power is incident on the transducer.
cident power. The sensitivity Q(h) is the slope of a plot
of electrical output X vs. incident radiant power @:
Electrical Output Characteristics. Transducers
also differ in their electrical output domains and their
output impedances. The same transducer can, in fact,
produce outputs in different data domains. The pho-
For photon detectors and for many thermal detectors, tomultiplier tube, discussed in detail below, can pro-
Q(X) and R(A) are wavelength dependent and specified duce an output charge (an analog domain), an output
at a particular wavelength. A plot of Q(h) vs. X or R(h) current (analog), an output pulse rate (a time domain),
vs. 1is called the s ectral response of the transducer. or a specific number of output pulses (digital domain).
The overall functional relationship between the The output impedance of a detector often deter-
output quantity X and the input quantity @ is known mines the type of measurement system to be used.
as the transfer function. It can be expressed by an equa- Impedances can range from megaohms (phototubes) to
tion or by means of a plot. The sensitivity is the slope a few ohms (thermocouples).
of the transfer function, whereas the responsivity is its
magnitude at a given incident power [R(h) = XI@]. Dark Signal Characteristics. The electrical out-
If Q(X) is constant and independent of @, the detector put of a transducer in the absence of radiation is known
is said to exhibit linearity. If a transducer shows linearity as its dark signal. Because of their different response
and its transfer function passes through the origin, the mechanisms, radiation transducers vary widely in their
sensitivity is equal to the responsivity. In many cases dark signal characteristics. Photoemissive detectors,
these terms are thus used interchangeably. Usually, which depend on photoelectrons being emitted from
transducers exhibit linearity over a limited range of in- photosensitive materials, exhibit dark signals due to
cident radiant power. The total range, expressed in thermal emission of electrons. Although dark signals
powers of 10, for which the transfer function in linear can in principle be subtracted from the total signal in
108 Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
the presence of radiation, noise and unidirectional drifts Pneumatic. A pneumatic detector is based on
in the dark signal can become major sources of error. a thin blackened membrane placed in a gas-filled, air-
tight chamber. As radiation strikes the detector, the gas
is heated and expands against another membrane. The
Noise Characteristics. Although we shall delay displacement of the membrane is detected in some types.
a detailed discussion of noise and signal-to-noise ratio In others the capacitance of the membrane serves as a
until Chapter 5, it is useful to define several terms here measure of displacement. A commercial pneumatic cell
related specifically to noise characteristics of trans- is the Golay detector, which is widely used in infrared
ducers. The noise equivalent power (NEP or @), is the spectrometers. The Golay cell is sensitive (see D" in
rms radiant power in watts of a sinusoidally modulated Table 4-5), but tends to be fragile.
input incident on the detector that gives rise to an rms
signal equal to the rms dark noise in a 1-Hz bandwidth. Thermocouple. In the thermocouple a thin
(See Chapter 5 for bandwidth definitions.) The rms blackened strip or flake is connected thermally to the
dark noise a, is thus given by a, = R(h)@,. Since it is junction of two dissimilar metals. Radiation absorbed
a function of wavelength, the NEP is usually specified by the strip causes the junction to increase in temper-
at a particular wavelength. The modulation frequency, ature and a change in thermoelectric voltage is pro-
electrical bandwidth, and detector area should also be duced. The thermocouple detector has uniform spectral
specified. response in the region 1 to 40 pm, reasonable sensitiv-
The detectivity D (W-l) is a measure of minimum ity, and excellent linearity. Thermocouples require no
detectability and is defined as D = l/@,. As with the external bias, have high stability, and have low output
NEP, the wavelength, modulation frequency, band- impedance (5 to 15 0).Since their output voltages are
width, and detector area should be specified. The D often on the order of microvolts, they require large
star (D*), in cm Hz1I2 W-I, is a normalization of de- amplification factors. With modern amplifiers, per-
tectivity to take into account the area and electrical formance is limited by thermal noise (see Chapter 5).
bandwidth dependence. It is related to D by D* = Multiple-junction thermocouples, called thermopiles are
DA112(Af)'12,where A is the detector area in cm2 and also used.
Af is the noise equivalent bandwidth in Hz.
Thermistor Bolometer, The thermistor is made
from an intrinsic semiconductor. As the temperature
Thermal Detectors increases the number of valence-band electrons pro-
moted to the conduction band increases, which in-
The thermal detectors discussed here are listed in Table creases the conductivity and decreases the resistance.
4-5 along with several useful characteristics. Thermal A thin blackened tip allows the absorption of radiation,
detectors are widely used in the IR region of the spec- which heats the thermistor. The thermistor is normally
trum. placed in a bridge circuit with a reference thermistor
TABLE 4-5
Thermal detector characteristics
that is not irradiated. The resistance can be measured tubes), pn-junction devices (photodiodes, phototran-
by a null-comparison technique, or the out-of-balance sistors), photoconductive cells, and photovoltaic cells.
voltage of the bridge can be monitored. Bolometers are The operating principles of each of these detectors are
rugged and exhibit moderate sensitivity and a wide lin- discussed below.
ear range. The thermistor spectral response normally
peaks in the near-IR region. Thermistor detectability Vacuum Phototubes. A vacuum phototube
is limited by thermal noise at frequencies above 20 Hz (PT), sometimes called a vacuum photodiode, consists
and by llf noise (see Chapter 5) at lower frequencies. of two electrodes sealed in an evacuated glass or silica
envelope. A phototube, its circuit symbol, and a typical
Pyroelectric Detector. A pyroelectric detector bias network are shown in Figure 4-21. The photosen-
is typically made from triglycine sulfate (TGS). When sitive cathode can be made from a number of photoern-
placed in an electrical field, a surface charge results issive materials (e.g., Cs,Sb, alkali metal oxides, AgOCs).
from alignment of electric dipoles. When a pulse of Typical bias voltages are 75 to 125 V.
incident radiation heats the TGS, a change in surface Quartz or
charge results (pyroelectric effect), which is related to
the incident radiant power. The output current is pro-
portional to the rate of temperature change of the ma-
terial dTIdt; the detector does not respond to constant
radiant energy levels. The pyroelectric detector is fast
(<1 ms response time) because only charge-reorien-
tation limits the response speed for modulated inputs.
For wavelengths below 2 pm, however, the TGS must
be blackened, which can slow the response. The spectral
response of a blackened detector is fairly flat over the
region 1 to 36 pm. Since the pyroelecric effect is an
integrated volume effect, output signals from pyroelec-
tric detectors are not a function of the spatial or tem-
poral distribution of the input radiation. This makes
them highly useful for colorimetry, radiometry, and in
laser power meters. Linear arrays of pyroelectric de-
(b)
tectors are also available.
FIGURE 4-21 Circuit symbol (a) and bias net-
work (b) for vacuum phototube. The photocath-
ode C consists of a photosensitive material. It is
biased negative with respect to the anode A. Ir-
The photon detectors to be considered here are listed radiation of the cathode causes photoelectrons to
in Table 4-6, where various characteristics are com- be emitted and attracted to the anode. The anodic
pared. Photon detectors can be broadly classified as current ,i is in the direction shown; it produces
photoemissive devices (photomultipliers and photo- a voltage drop E, across load resistor R,.
Linear range
TYpe D* (cm Hzu2W-I 1 R(hIb (decades) Spectral range (nm)' Rise time (ns) Output
Photomultiplier tube 1012- 10'' 10-lo5 A W ' 6 110-1000 1-10 Current, charge
Vacuum phototube los- 10'" l 0 - ~ - 1 0 - ~A W - I 5 200- 1000 1-10 Current
Si photodiode 101"- 1012 0.05-0.5 A W-' 5-7 250-1100 1-10 Current
Photoconductive cell 10'- 10" lo4- lo6 v w - 5 750-6000 50-106 Resistance
change
Photovoltaic cell lo8-10" 100-10h V W ' 3 400-5000 1000 Current or
voltage
SOURCE: W. G. Driscoll and W. Vaughan, eds., Hundbook of Optics, McGraw-Hill, New York, 1978.
Since many diffcrent types of detectors are available in each class, the values given represent ranges for several different types.
bFor the photomultiplier, the value of H ( X ) includes the internal gain; for the photoconductive cell voltage, responsivity assumes a constant current of 1 to 10 mA.
CExtended IR-responsive photoconductors are available (see the text).
110 Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
The photocathode is characterized by a threshold diation are then found from substituting equation 4-23
wavelength 1,;at longer wavelengths, photons are in- into 4-25 and 4-24 into 4-26. This yields
effective in causing photoemission. The threshold wave-
length is related to the photocathode work function E,
by X, = hcIE, = 1240/Ec. Here X, is in nanometers
when E, is in electron volts. The work function rep- and
resents the energy which must be given to an electron iap = Y@R(X) (4-28)
in the Fermi level of a metal to raise it to the potential
energy of the metal-vacuum interface. Because some where the radiant power in watts is @ = @, x hv. If
electrons occupy states higher than the Fermi level at K(X) and R(X) are not constant over the wavelength
ordinary temperatures, there is no abrupt cutoff at X > range incident on the photocathode, these expressions
A,. Typical work functions for pure metals are 2 to 5 become integrals over this range. For typical photo-
eV, but semiconductor materials can have values sub- tubes, the anodic current becomes independent of bias
stantially lower. Indeed, extended long wavelength re- voltage (rl -+ I ) above about 60 V. For most phototubes
sponse is obtained with photosensitive materials which the anode current must be kept lower than to 10-j
show negative electron affinity. A for linearity to be obtained.
Only a certain fraction of the photons with greater In the absence of radiation, a small anodic dark
than threshold energy yield photoelectrons with suffi- current i, is obtained, where in, = qi,, and icd is the
cient kinetic energy to escape the photocathode. This cathodic dark current. At moderate bias voltages dark
fraction is called the quantum efficiency K(h) and is the current can be due primarily to thermal emission of
ratio of the number of photoelectrons ejected to the electrons at the photocathode. When voltages are low,
number of incident photons. Typically, K(h) varies from ohmic leakage can dominate. At low temperatures lu-
0 to 0.5. The rate rcp (s-l) at which photoelectrons are minescence and radioactivity can be the primary source
emitted from the photocathode for monochromatic ra- of dark current. When high voltages are applied, ion-
diation of flux @, (photons s-l) is ization of residual gas and field emission can be addi-
tional sources. Typical values of in, for vacuum pho-
totubes are in the range 10-l2 to 10-l4 A .
This rate can also be expressed as a cathodic current Photomultiplier Tubes. The photomultiplier
icp by multiplying rcp by the electron charge, e = tube (PMT), like the phototube, contains a photosen-
1.6 x 10-19 c . sitive cathode and a collection anode. However, the
cathode and anode are separated by several electrodes,
called dynodes, that provide electron multiplication or
gain. Figure 4-22 shows a typical PMT. The cathode is
The efficiency with which photons are converted biased negative by 400 to 2500 V with respect to the
to photoelectrons is R(h), the radiant cathodic respon- anode. A photoelectron ejected by the photocathode
sivity in A WP1. It is related to the quantum efficiency strikes the first dynode and releases two to five sec-
by R(h) = K(X)e/hv = 8.06 x 10-"(A) x X, where ondary electrons. Each secondary electron is acceler-
X is in nanometers. Typical values of R(X) are given in ated by the field between the first and second dynode
Table 4-6 for vacuum phototubes. and strikes the next dynode with sufficient energy to
A fraction of the photoelectrons emitted are col- release another two to five electrons. Since each dynode
lected at the anode. This fraction q , called the collection down the chain is biased .= 100 V more positive than
efficiency, depends on the bias voltage E , and ap- the preceding dynode, this multiplication process con-
proaches unity if E, is sufficiently high. The arrival rate tinues until the anode is reached. The result is a large
of photoelectrons at the anode ,r and the anodic pho- charge packet of a few nanoseconds' duration at the
tocurrent ,i are thus anode for each photoelectron collected by the first dy-
node. The gain rn (the average number of electrons per
anode pulse) depends highly on the power supply volt-
and age, but is often lo4 to 10'. For example, if m were 106,
each anode pulse would contain an average charge of
1.6 x 10-19 x lo6 C = 1.6 x 10-l3 C. If the pulse
were 5 ns in duration, this would result in an average
Complete expressions for the anodic pulse rate and an- current of 32 FA during the pulse.
ode current in the case of monochromatic incident ra- It is most common to measure the average current
Sec. 4-4 I Optical Transducers 1
Photocathode
imum ratings (typically, <1 FA). The limit of linearity from the dynodes can be a source of dark current in
is caused by space-charge effects in the last dynodes. PMTs. Since the cathodic dark current is multiplied by
Since the electrons emitted by the dynodes are drawn the full gain of the tube, thermal emission from the
from the dynode current (current through the bias re- photocathode (or early dynodes) is often a major com-
sistors), the dynode current should be -100 times ,i ponent of the total dark current. Typically, dark cur-
to prevent bias voltage changes and nonlinearity. Op- rents in PMTs are in the range to 10-l1 A. For
erating a PMT at light levels such that the anode current very low light levels, the dark current can be as large
exceeds the maximum rating can cause fatigue, a loss as the photocurrent, and noise in the dark current can
in sensitivity as a result of reduced secondary emission. limit precision. The thermal dark current it, emitted by
Such sensitivity changes are thought to result from ero- the photocathode is given by it, = C A T 2 e-EclkT,where
sion of the dynode surfaces. Tube fatigue varies widely C is a constant, A is the surface area of the photocath-
with PMT type and among tubes of the same type. Some ode, T is the photocathode temperature, and E, is the
PMTs exhibit hysteresis when exposed briefly to high work function. If the dark current primarily results from
light levels. Hysteresis may result in overshoot or un- thermal emission, then in, = mi,,and the cathodic dark
dershoot by a few percent when levels are changed. current i, = it,. In this case, cooling the PMT can
Hysteresis may be caused by capacitance effects within reduce the dark current. Typically, temperatures of - 60
the tube. to P C are used. If external radioactivity or cosmic rays
Dark current in PMTs arises from effects similar are a significant cause of dark current, shielding can be
to those in phototubes. In addition, thermal emission used. Since the anodic dark current is proportional to
Sec. 4-4 I Optical Transducers
the gain, except at very low or very high bias voltages, verter (see the next section) is used to produce a voltage
there is often an optimum bias voltage which gives the proportional to the incident radiant power.
lowest relative dark current noise. The spectral response of most photodiodes reaches
The limiting source of noise in P a maximum in the near-IR region (0.85 to 1.0 pm).
5) depends on the level of illumination. For low levels, They are thus very useful for UV-visible and near-IR
the limiting noise is usually traceable to dark current detection. These devices often show excellent linearity
shot noise. At higher illumination levels signal-carried over six to seven decades of incident radiant power.
shot or flicker noise can far exceed dark current noise. Responsivities are typically much lower than those of
Therefore, D* and NEP values, which are calculated photomultiplier tubes (see Table 4-6) because of the
by assuming dark current noise is the limiting noise internal gain of the PMT. However, the simplicity, ex-
source, must be used with caution. Considerable error cellent linearity, and very small sizes of photodiodes
in predicting detection capability from D* values can make them attractive for applications where light levels
result if noise in the signal is ignored. Many photo- are relatively high. They have practically replaced vac-
multiplier manufacturers use the term equivalent noise uum phototubes in all but a few applications. Photo-
ut (ENI) instead of noise equivalent power. It is very diodes are extremely fast transducers. A special type,
similar except that a square-wave-modulated blackbody known as a pin junction, in which the p- and n-type
source is used, and the peak-to-peak value is obtained semiconductor materials are separated by an insulating
instead of the rms value of a sinusoidally modulated layer i, has a subnanosecond response time. The pho-
source. Specification sheets for PMTs are discussed in
Appendix D.
Incident
radiation
(input)
- -
- ---9
Amplified
electron
output
measure weak signals in the presence of strong signals,
blooming can seriously limit the use of the integrating
capability and decrease resolution. Another problem is
that of lag, which is a result of incomplete erasures of
the image during a readout cycle. This leads to a car-
ryover of the image from one integration time to an-
film
other. Lag is also highly undesirable in spectroscopy,
particularly in time-resolution applications. Fortu-
Eb Gain control
nately, lag does not appear to be a significant problem
with diode arrays.
Like diode arrays, silicon vidicons have no inter-
i
nal gain. Their dynamic range is similar (two-three dec-
Diode array
ades). To provide gain for the vidicon, the silicon-in-
tensified target (SIT) is commonly used. In the SIT, a
photocathode is placed in front of the silicon target and
Photocathode ' the photoelectrons are accelerated by a large 7 to 8 kV
(b) potential difference. Here, typically, 1000 electron-hole
FIGURE 4-27 Microchannel-plate-intensified pairs are produced per incident electron. The silicon
diode array. The microchannel plate in (a) con- vidicon target is operated as an electron detector rather
sists of a large number of closely packed hollow than a photon detector. Gains of several hundred can
glass fibers (10 to 40 p m in diameter) with a re- be obtained and the linear dynamic range can be ex-
sistive coating for secondary emission on the in- tended to about four decades. Intensified SIT devices
ner surface. Each fiber acts as an individual elec-
are also available. These have an image intesifier in
tron multiplier with a gain of 10" or 10". In (b) a
photocathode produces photoelectrons which front of the SIT tube and are called ISITs. Lag and
strike the microchannel plate. The amplified elec- blooming remain as problems for these intensified de-
tron packets, each with spatial integrity, strike a vices.
phosphor, which produces visible radiation de-
tected by the array. Charge-Coupled and Charge-Injection De-
vices. Like photodiode arrays, charge-coupled de-
Vidicon Tube. The silicon vidicon uses detector vices (CCDs) and charge-injection devices (CIDs) are
elements that are also reverse-biased photodiodes op- solid-state sensors and are constructed with integrated-
erated in the charge-storage mode. However, scanning circuit technology. With both devices, the charges gen-
electron beam readout techniques are used. Radiation erated by photons are collected and stored in metal-
strikes a silicon target where the charge stored on the oxide-semiconductor (MOS) capacitors. The pixel
pn-junction capacitance is discharged by generation of arrangement for the 512 x 320 CCD array made by
electron-hole pairs. The target is then scanned by an RCA is illustrated in Figure 4-28a; some CCD arrays
electron beam with magnetic deflection coils, and the are as large as 2000 x 2000 (4 x lo6 pixels). Each pixel
charge needed to restore each detector to its initial state consists of a thin conducting electrode and a thin in-
is measured. sulating oxide layer on top of a p type silicon substrate,
In contrast to the photodiode arrays, which in as shown in Figure 4-28b.
spectroscopy have been linear arrays, the silicon vidicon The MOS capacitors are inititially reversed biased
is a two-dimensional array containing about lo5 resolv- by a positive voltage applied to the metallic electrode;
See. 4-5 1 Signal Processing and
specific type of signal processing used depends highly of the optical transducers are in one of the analog do-
on the signal form, its level, and the type of environ- mains. The major exception to this is the photomulti-
ment (i.e., noise sources and the expected noise am- plier tube when it is used in a photon counting system.
plitude). Also, the type of signal processing is depend- In this case, if the desired quantity is the total number
ent on the specific instrumentation used and the format of accumulated counts over some boundary condition
of the data to be presented. The signal processing sys- (i.e., counts per spark discharge or counts per laser
tem can perform various operations, including data con- pulse), the photomultiplier output is directly digital. If
versions (current to voltage, analog to digital, etc.), the desired quantity is the count rate, the photomulti-
amplification or voltage division, mathematical opera- plier output is in one of the time domains (frequency
tions (e.g., integration, logarithmic conversion, and dif- domain). Thus the PMT output can be in any of the
ferentiation), and SIN enhancement. Even with a com- three electrical domain classes (analog, time, digital).
puter, some signal treatment by hardware is almost always In the past completely analog signal processing
necessary prior to acquisition by the computer. Readout and readout systems were employed, as illustrated by
systems vary widely from instrument to instrument, but the general block diagram of Figure 4-30a. Increasingly,
some form of digital readout and, increasingly, graph- the outputs of analog transducers are being converted
ical display are commonly included. to digital form as early in the measurement sequence
We will consider in this section many of the stand- as feasible, as illustrated in Figure 4-30b. Early con-
ard ways in which signals are conditioned, modified, version to the digital domain allows the signal to be
and processed prior to readout. After presenting some treated with highly accurate digital methods or with
general considerations, we consider the principles of software in a microcomputer. Even with "digital" sys-
modulation since many signal conditioning methods are tems, some analog signal conditioning is normally done
discussed. Analog signal processing schemes, including to amplify, filter, or otherwise alter the signal (e.g.,
SIN enhancement techniques, are described next fol- convert current to voltage) to make it suitable for an-
lowed by a consideration of computer data acquisition alog-to-digital conversion. In many cases it is desirable
systems. Digital data processing methods are then briefly to shift the signal away from dc, which is a particular
introduced. A detailed discussion of analog-to-digital noisy region of the frequency spectrum (see Section
conversion methods and computer data processing tech- 5-1). With analog systems, this is accomplished by mod-
niques is beyond the scope of this book. The reader ulation (see below) and ac signal processing. With digital
should consult the references for this chapter for ad- systems, modulationldemodulation or gated detection
ditional information on these methods. The section con- schemes are used prior to conversion to digital format.
cludes with a discussion of the common types of readout For photon counting systems, the scheme illus-
and display systems. trated in Figure 4-31 is used. The readout is the number
of pulses occurring during a preselected counting in-
terval. Because of the high probability of pulse overlap
General Considerations at high count rates, photon counting is used in situations
where irradiances are quite low (e.g., Raman scattering
Electrical signals can be grouped into three data domain and some luminescence methods). Photon counting sys-
classes: analog, time, and digital. The outputs of most tems are discussed in more detail in Chapter 5.
- Pulse helght
dtscnmlnator
Analytical
signal
scattering methods, the sample container would be in- modulation, the sample is alternately presented and re-
serted after the chopper so that the source radiation moved from the excitation light beam or the excitation
would pass through the sample on alternate half-cycles. emission source. In molecular absorption or photolu-
Source modulation in absorption or photoluminescence minescence spectroscopy this can be accomplished by
spectroscopy also allows discrimination against any alternately positioning the sample cell and the reference
emission by the sample in that the resulting ac output cell in the light path. In atomic emission, fluorescence,
of the transducer is referenced to the dc emission level, and absorption spectrometry, the sample introduction
which is present during both half-cycles. In emission system can alternately present sample and blank to the
spectroscopy, the analyte and background emission are atomizer. Sample modulation allows automatic refer-
both carried by the modulation waveform, and modu- encing of sample information to that of the blank, which
lation allows discrimination against the dc detector dark can automatically provide correction for blank inter-
signal. ferences if the blank is ideal. Unfortuately, most sample
In some cases the source is pulsed on and off in modulation techniques operate at only a few hertz, which
a low-duty-cycle mode, as compared to the 50% duty is often not far enough removed from the noisy dc re-
cycle depicted in Figure 4-33. Although the methods gion to achieve much improvement in SIN.
for recovering such signals are quite different, as dis- Wavelength modulation is the repetitive variation
cussed below, signal information is still moved away of the wavelength range observed by the detector. This
from the dc region, and it can be amplified and re- can be accomplished by repetitively scanning a mon-
covered in the presence of noise. ochromator back and forth across a fixed-wavelength
In another form of modulation, called sample range. Normally, it is possible to achieve only a low
modulation frequency with this approach, and the re- Many other characteristics can be used for mod-
fractor plate approach shown in Figure 4-34a to c is ulation. Several of these are described in later chapters.
more commonly employed. Wavelength modulation is
used primarily in atomic spectrometry, where the var- nal Conditionin
iation in wavelength is often very small. Here it is pos-
sible to make measurements on and off an atomic line Usually, the first step in the overall signal processing
to obtain alternately the analyte and the background scheme is to condition the transducer output signal for
signals. Note that wavelength modulation is similar to the circuitry that follows. This often involves data do-
sample modulation in that essentially a sample-blank main conversion and dc or ac amplification (or in some
measurement is made, although no actual "blank" is cases, voltage division). After conditioning, the analog
measured. Wavelength modulation can also be imple- signal can be converted to the digital domain or further
mented by oscillation of the spectrometer entrance slit, processed in the analog domain. Some typical signal
which effectively changes the angle of incidence to the conditioning elements are given in Figure 4-35 along
grating. Oscillation of an interference filter or the split- with their input-output relationships (transfer func-
ting of degenerate lines from a source with a modulated tions).
magnetic field is also employed. In molecular spec-
trometry, wavelength modulation can be used to obtain Current-to- Voltage Conversion. The outputs
a derivative, as shown in Figure 4-34d. Here the wave- from several transducers (phototubes, photodiodes,
length interval varied is small compared to the width photomultipliers) are currents and must be converted
of the molecular band. to voltages prior to analog-to-digital conversion or read
out. Current-to-voltage conversion can be accom- pacitance and provides a low output impedance for con-
plished with a simple load resistor as shown in Figure necting voltmeters, analog-to-digital converters (ADCs),
4-35a or with an operational amplifier (op amp) current and so on. With the load resistor technique, the voltage
follower as shown in Figure 4-35b. In either case the monitoring device is connected directly across the load
output voltage E, is given by resistor, which can be as large as 101•‹R .
Commercial current-to-voltage converters are
available with switch-selectable feedback resistances.
Many can measure currents in the picoampere range.
where i, is the transducer output current (photoanodic These require excellent shielding and grounding and
current i, for a PMT or phototube). the use of very high quality amplifiers. Some systems
In the case of the op amp current-to-voltage con- have an independently variable time constant so that
verter, RL is the feedback resistance R f , and the output the response time can be adjusted; they may even have
voltage is inverted (i.e., E, = - ioRf).In the load re- calibrated offset currents for nulling out dark current.
sistor case, the voltage across the transducer E,, is given
by E,, = E, -ioRL. At high light levels, if i,RL is D@ Amplification. Outputs from voltage-out-
signficant compared to E,, the bias voltage of the trans- put or current-output transducers after current-to-volt-
ducer becomes dependent on the light intensity. With age conversion are often amplified to 0.1 to 10 V prior
the operational amplifier circuit, on the other hand, the to conversion to a digital signal or connection to a read-
anode of the transducer is maintained at virtual com- out device. For millivolt-level signals. the voltage-fol-
mon potential by the negative feedback. Thus E,, is lower with gain shown in Figure 4-35c can provide a
maintained at E, independent of the light level. The high input impedance and gains of up to lo3 without
operational amplifier technique also reduces stray ca- inversion. Computer data acquisition systems, as dis-
Sec. 4-5 1 Signal Processing and Readout Systems
cussed later, often prov~dea rogrammable-gain am- to the counting circuitry. Again fast response is nec-
plifier prior to the ADC. Such plifiers are often volt- essary to avoid pulse pile-up (deadtime) effects. In the
age followers with remotely selectable gains. An inverting discriminator of Figure 4-35e, the user adjusts the
amplifier, such as that shown in Figure 4-35d, can also threshold voltage level. If the input voltage exceeds this
be used, but the input impedance is only R,,. value, the comparator output voltage goes to its neg-
Some transducers, such as thermocouples, pro- ative limit. If the input voltage is below the threshold,
duce microvolt-level signals. Special components, such the comparator is at its positive limit. Some photon
as instrumentation amplifiers and isolation devices, are counting systems have a window discriminator, which
used to amplify these tiny signals. There are some cases allows only pulses with amplitudes between a lower and
in which a voltage divider must be used to attenuate a an upper threshold to be counted. These are made from
signal prior to analag-to-digital conversion or readout. two comparators and a logic circuit.
Simple resistive dividers made from precision resistors Commercial photon counters with high-quality
are typically used. components are available from several instrument com-
The outputs of resistive transducers (thermal de- panies. These can be general-purpose counting units or
tectors) can be conditioned in several ways. The resist- made specifically for a spectroscopic instrument such
ance can be converted to voltage with the inverting as a Raman spectrometer.
amplifier configuration of Figure 4-3%. Here a fixed
input voltage and a fixed input resistor are used, and
the transducer is placed in the feedback loop in place
of resistor R+.The amplifier output is then a constant
times the transducer resistance. If the transducer is sub- Although the trend in recent years has been to do as
stituted for the input resistor, instead, an output voltage much of the processing as possible in the digital domain,
proportional to its condu ) is obtained. In analog signal processing is still widely used. Even with
many cases, the resistive is placed in one computer systems, some analog processing may be done
changes in resist- for reasons of speed or economy. The types of pro-
ance, the off-balance bridge voltage is proportional to cessing considered here are mathematical operations,
the transducer resistance. filtering, ratioing, and SIN enhancement. Several of the
most common analog processing elements are shown in
6 Amplification. The current-to-voltage con- Fgiure 4-36.
of Figure 4-35b and the inverting amplifier of
Figure 4-35d can be ma e unresponsive to dc signals
by placing a capacitor in the input circuit before the op mathematical operation
amp input. The capacitor blocks any dc com ational amplifier circuits. The configuration shown in
the signal and allows only the varying signal to be am- Figure 4-36a is used to perform voltage or current in-
plified. tegration. When integration is to begin, the switch is
A capacitively coupled current-to-voltage con- opened and capacitor G begins charging. At the end of
verter is often used with a modulated source and PMT the integration period the switch is closed to discharge
transducer. The voltage signal at the modulation fre- the capacitor. The output voltage just before the switch
quency can be selectively amplified by means of a tuned is closed is given by the expression in Figure 4-36a.
amplier as discussed later. Integration is used to measure peak area or to improve
the SIN.
Differentiation is used to accentuate spectral de-
tails during a scan or to measure the time rate of change
esistor connected to common of signals in reaction-rate measurements. It can be per-
(e.g., < 100 a), The signal c tioning circuitry is formed by the circuit shown in Figure 4-36b. Here the
usually a fast video voltage amp1 and a discriminator output voltage is proportional to the time derivative of
(comparator) such as that shown in Figure 4-3%. Be- the input voltage. This simple circuit is usually highly
cause of the internal gain of the susceptible to noise, which often has a higher rate of
amplification is needed (10 to 100). change with time than the signal being differentiated.
errors due to pulse overlap, the rise time of the amplifier Hence the frequency response of the differentiator is
should be fast and it s have extended high-fre- often intentionally degraded by putting a small capac-
quency response (>I00 for modest irradiances). itor in the feedback loop and a resistor in the input
The discriminator not only allows discrimination circuit. This effectively makes the differentiator an in-
against small pulses, but it also shapes the signal prior tegrator at high frequencies.
124 Chap. 4 l Optical Sources, Transducers, and Measurement Systems
The circuit of Figure 4-36c is a logarithmic voltage the high-pass filter shown in Figure 4-36e are first-order
amplifier. It can also be a logarithmic current amplifier passive filters. They provide a decrease in gain of 20
if the current input is attached directly to the amplifier dB per decade change in frequency at frequencies lower
inverting input (as in Figure 4-36a). Unfortunately log- or higher than their cutoff frequencies rc = 1/(2nRC)],
arithmic elements (transistors and diodes) are quite respectively. Active filters are based on operational am-
temperature dependent, and the constants shown in the plifiers; they can avoid loading effects and provide gain.
transfer function are strong functions of temperature. Active filters with orders as high as 6 (rolloff rate =
Commercial logarithmic amplifier modules are often 120 dB1decade) are practical. Commonly, a feedback
temperature compensated. These are usually logarith- capacitor (CJ is used in parallel with the feedback re-
mic ratio amplifiers. Such amplifiers are frequently used sistor in the op amp current-to voltage converter (Figure
to convert transmittances to absorbances in spectro- 4-35b) to form an active low-pass filter with time con-
photo~netricinstruments (see Section 2-6). stant 7 = RfCP The use of filters is discussed in more
There are many situations in which an output volt- detail in Section 5-2.
age proportional to the difference (A - B) or the ratio For ac signals, bandpass filters, which transmit a
(AIB) of two input signals A and B is desired (e.g., narrow band of frequencies and reject lower or higher
dual-beam or dual-channel methods). Although these frequencies, are readily obtained or constructed. In-
functions can be implemented in the analog domain, it tegrated-circuit (IC) active filters can provide high-pass,
is increasingly common to use a computer to calculate low-pass, and bandpass outputs simultaneously. In ad-
them. dition, they can be used to make notch filters which
specifically reject a particular frequency band, such as
Filters. Filters are widely used in spectrometric 60 Hz. Bandpass filters can have gain, in which case
systems. The low-pass filter shown in Figure 4-36d and they are often referred to as tuned amplifiers. Such
messing and Readout Systems
Reference
signal
(carrier)
duty cycles. A block diagram of a boxcar integrator is data processing routines. The computer also controls
shown in Figure 4-38. the scanning time and the number of samples averaged.
The boxcar integrator allows the recovery of sig-
nals that are time related to a trigger signal and the Multichannel Averaging. A multichannel av-
rejection of those that are not. A typical application erager acquires a large number of evenly spaced signal
might be the measurement of the fluorescence produced readings across a complete waveform on each signal
by pulsed dye laser excitation. The pumping source for repetition instead of one reading per repetition as in
the dye laser could provide the trigger signal, and a the boxcar integrator. A fast analog-to-digital converter
PMT would be used to measure the fluorescence. The and digital memory are required to digitize and store
PMT output, after current-to-voltage conversion, would each reading. On subsequent signal repetitions, the pre-
be the boxcar integrator input. The system could be set viously stored value for the appropriate channel (mem-
up to integrate the PMT output for the entire laser ory location) is summed with the new value in a fast
pulse. Averaging over multiple laser firings would im-
prove the SIN. Alternatively, a small time slice could
be selected and the boxcar time window scanned to
produce a time-resolved plot of the fluorescence emis-
sion. Gate pulse widths from 100 ps to 10 ms are avail-
able on commercial boxcars.
With boxcar integrators a high-pass filter is typi-
cally used to remove dc signals and low-frequency noise +-+
before they reach the integrator. In some situations it r2
Time
is advantageous to use the dual-channel boxcar ap-
FIGURE 4-39 Dual-channel boxcar approach.
proach illustrated in Figure 4-39. Here, one can subtract A boxcar integrator can be constructed with two
background or even ratio the signal of interest to a channels (two gates and integrators). The ana-
reference signal. lytical signal information is integrated and stored
The signal processing techniques provided by lock- in one channel during time t,. During time t2 the
in amplifiers and boxcar integrators can also be imple- background is integrated. The difference (inte-
mented by digital hardware or by software integration gral during time t, minus integral during t,) is then
techniques. A synchronous photon counting system can calculated; this provides correction fir dc back-
be implemented by using an upldown counter synchro- ground signals and discriminates against low-fre-
nized to a modulated waveform. Here the counter counts quency noise. Many dual-channel boxcar inte-
up during the on-cycle of the modulation waveform and grators also provide a ratioing function so that
down during the off-cycle for background subtraction. the ratio of the channel 1 signal to the channel 2
signal is calculated. This is often used with pulsed
At this time the hardware approaches are much faster lasers to reduce the effect of pulse-to-pulse var-
than software-based techniques. A commercial com- iations in the laser output. Here one photode-
puter-controlled boxcar integrator is available in which tector would monitor the laser output itself, while
analog circuitry is used for high-speed operations and a second would monitor the laser-stimulated
a computer is used to implement several sophisticated process (e.g., fluorescence or transmitted light).
See. 4-5 / Signal Processing and Readout Sysrems
adder, and the new sum is stored. After n repetitions signal is connected to a programmable gain instrumen-
the entire acquired waveform has been averaged over tation amplifier. A sample-and-hold circuit acquires the
the n repetitions. Multichannel averagers are normally analog signal at a particular instant in time and holds
used for acquiring signals that are of lower frequencies it constant for the successive approximations ADC. The
than those acquired by boxcar integrators. If averaging digital data can then be loaded onto the computer bus
is not required, a transient recorder can be used to through a bus interface (tristate buffers).
digitize and store a complete single trace of a waveform. Analog 110 boards are also available for many
One commercial transient digitizer uses a high-speed microcomputers. These include a data acquisition sys-
(5 ns) ADC to digitize the analog information. A com- tem and also one or more output digital-to-analog con-
puter can then manipulate the stored digital data. Av- verters (DACs). Some have on-board memory and even
eraging of multiple repetitions can then be done by the real-time clocks. Prices for these boards can range from
computer. a few hundred dollars to over $1000, depending on the
options selected. Several firms are now selling complete
software packages for laboratory uses. These laboratory
notebook packages can be used for many tasks, includ-
Increasingly microcomputers are being used to acquire ing data acquisition, manipulation, and graphical dis-
and manipulate spectrometric data. A few years ago play.
it was a major task to interface a computer to an ex-
periment. However, today, complete data acquisition
systems and analog input-output (110) boards are com-
mercially available for almost all popular micro- A substantial amount of signal processing is now being
computers. done in the digital domain with computer systems. Al-
A typical data acquisition system is shown in Fig- though the details are beyond the scope of this book,
ure 4-40. The input multiplexer allows 16 analog signals a few of the common manipulations are briefly consid-
to be selected and converted. The multiplexer address ered here.
register selects the channel to be converted. The access Many of the mathematical operations discussed
can be random or sequential. The selected analog input above can be done more accurately with computers than
Sample
and-
Gain hold
programming outuut
computer
bus
--a conversion
Address
in
FIGURE 4-40 Computer data acquisition system. A typical system con-
tains a multiplexer, a programmable gain amplifier, a sample-and-hold
circuit, an ADC, and a sequencer. Systems are commercially available
for most microcomputers. A wide range of ADG types and conversion
speeds are available. Integrating ADCs are relatively slow (ms conversion
times), but average the signai during the conversion time. A sample-and-
hold circuit is not needed with these converters. Successive approximations
converters are relatively fast (microsecond conversion times), but require
an absolutely steady voltage during conversion. The fastest converters are
flash ADCs developed for digital TV. These convert continuously and
have throughput rates in the hundreds of MIHz range. They are relatively
expensive and of lower resolution than other ADC types.
128 Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
with analog circuits. Thus such operations as integra- scanning spectrometers, where recorders are used to
tion, differentiation, ratioing, and logarithmic conver- display the spectrum, provide digital readouts for quan-
sion can readily be performed in the computer. Where titative purposes.
high speed is needed, there is still an advantage to using For the measurement of voltages, readout devices
analog circuits for these operations. Although the com- should have high input impedance to prevent loading.
putations can be quite rapid with a computer, it takes Other characteristics to consider when choosing a read-
some time for acquiring, manipulating, and storing the out device for a given application include linearity, res-
data. olution (smallest difference in signal that can be re-
Signal-to-noise enhancement techniques are being solved), range, response time, stability, and drift.
increasingly implemented with the aid of computers. Monitors used for display with computers can be
Digital filtering methods allow the application of fil- either monochrome or color. Increasingly, graphics dis-
tering functions that are impossible or difficult to im- plays are being used to enhance visualization and to
plement with analog circuits (see Section 5-6). Algo- provide various plots and correlations that are easy to
rithms are readily available for common filtering and inerpret and manipulate. High-resolution color graphics
data smoothing operations. Many commercial spectro- display are especially effective in this regard.
chemical instruments with built-in computers contain
programs for carrying out a variety of filtering and
smoothing functions. 4-6 OPTICAL SPECTROMETERS
Many computer-based systems have software for
taking the Fourier transform or inverse transform of The complete optical spectrometer consists of the op-
acquired data. Such techniques allow one to manipulate tical components discussed in Chapter 3, the radiation
the data in the frequency domain for frequency-domain source, transducer, and electronic instrumentation dis-
filtering (see Section 5-6) or to operate on interfero- cussed in this chapter, and various control systems, sam-
metric data. ple introduction systems, and sample containers that
With computers data can also be readily fit to are specific for each technique. Optical spectrometers
theoretical o r empirical models through nonlinear can be broadly broken into three categories: single-
regression methods. For example, linear and nonlinear channel, multichannel, and multiplexing spectrometers.
calibration curves can be calculated and displayed quickly This section considers these spectrometer types and pre-
with computer-based systems. Similarly, data can be fit sents the readout expressions for quantitative applica-
to standard functions such as exponentials, polyno- tions of emission, absorption, and photoluminescence
mials, and so on. Again we are increasingly seeing such spectrometers.
capabilities being incorporated into commercial instru-
ments. Single-Channel Spectrometers
Single-channel spectrometers are considered here to be
Readout and Display Systems
spectrometers in which information at a single wave-
length setting is obtained or a spectrum is recorded
The readout and display system provides a visual rep-
sequentially (one wavelength at a time). They are di-
resentation of the output of the experiment. In the past
vided into fixed-wavelength systems, conventional scan-
analog readout and display systems were most common.
ning systems, and rapid-scanning systems. Single-chan-
Most of these are voltage-actuated devices. Computer
nel spectrometers can also contain more than one optical
outputs can be displayed digitally or they can be con-
beam, as in a double-beam spectrophotometer.
verted to analog signals by DACs and displayed on
analog output devices. Often, an analog display is used Fixed-Wavelength Spectrometers. A spec-
for a fast observation of the acquired signals, but the trometer can be classified as a fixed-wavelength device
quantitative information is obtained from the digital if it has no scanning mechanism and is intended for
data. operation at a single wavelength even though that wave-
The output devices used in spectroscopic instru- length might be alterable. Thus a filter photometer, an
ments are analog voltmeters (older instruments), digital atomic absorption spectrometer with manual wave-
voltmeters (panel meters), oscilloscopes, strip-chart re- length adjustment, and a manual calorimeter are all
corders, printers, monitors, or terminals. Because of classified as fixed-wavelength spectrometers. These sys-
their improved accuracy, freedom from bias, and lower tems are useful only for quantitative analysis after spec-
chance of reading error, various digital displays are in- tral information has been obtained and wavelengths
variably used in modern instruments unless a continu- chosen from spectral scans made on a separate instru-
ous plot of output signal vs. time is desired. Even many ment.
Sec, 4-6 / CqptiGLtl Spectrometers
Conventional Scanning Spectrometers. Here sample and reference beams, As shown in Figure 4-41,
a scanning mechanism is present, usually a motor, to the beams can pass alternately (double beam in-time)
turn a grating or prism in a monochromator at conven- or simultaneously (double beam in-space) through the
tional scanning speeds (0.01 to 10 nm s- I ) . This sweeps sample and reference cells. In contrast to the dual-chan-
the dispersed spectrum across a fixed exit slit, as de- nel spectrometers discussed below, the same wave-
scribed in Chapter 3. Scanning spectrometers can, of length band is viewed by two detectors (double beam
course, be operated at fixed wavelengths, which makes in-space) or by a single detector (double beam in-time).
them useful for quantitative measurements as well as In double-beam systems the sample beam always passes
for spectral scanning. A scanning spectrometer is more through the sample container, while the reference beam
complex and costly than a fixed-wavelength system, but may or may not pass through the reference container
considerably more versatile. The conventional spec- and hence the blank.
trometer obtains a complete spectrum in minutes; it can In many molecular double-beam spectrometers
only be used on static systems or on systems in which both the sample and the blank signals are acquired to
the spectral properties change very slowly in time. allow on-line compensation for blank signals. For ab-
sorption spectrometers, where the ratio of the sample
Rapid-Scanning Spectrometers. Some single- signal to the reference signal is calculated, the double-
channel spectrometers are capable of obtaining a spec- beam system compensates for source intensity drifts. In
trum rapidly. One approach is to oscillate an optical addition, during scanning, the double-beam system
component in the monochromator (grating or mirror) compensates for the source spectral distribution and
very rapidly. A fast transducer, such as a photomulti- changes in the monochromator efficiency with wave-
plier tub is required. The mechanical rapid-scanning length.
spectromeier can achieve complete spectral scans in In some molecular and all atomic spectrometers,
seconds or even milliseconds in some cases. These spec- the reference beam does not pass through the reference
trometers suffer from mechanical complexity and the container, but is monitored directly by the reference
difficulty in maintaining optical alignment. Mechanical detector, often without wavelength selection. Here the
rapid-scannirng system have practically been replaced double-beam system only provides compensation for
by multichannel systems. source intensity fluctuations.
Another approach to sequential, multiwavelength
spectrometry is known as the slew-scan approach. Here
the information at certain fixed wavelengths across a
broad spectral range mighi ue of interest. The slew-scan Multichannel spectrometers increase sample through-
snochromator is programmed such that it rapidly scans put and reduce sample consumption where many spe-
(slews) through the undesired wavelengths and then cies per sample must be measured. The multichannel
stops at or slowly scans across the desired wavelength spectrometer measures the spectral information at mul-
region. The major advantage is that a monochromator tiple wavelengths simultaneously. It is still a dispersive
is used, for simplicity and low cost, and little time is system in that the spectral information is dispersed spa-
wasted in spectral regions that are uninteresting. tially by a grating or prism, owever, the multichannel
The image dissector tube described in Section 4- spectrometer is either a polychromator with multiple
4 is another approach to rapidly scanning a spectrum. exit slits and detectars or a spectrograph with a mul-
The IDT has the advantage of speed in that scanning tichannel detector (see Chapter 2).
is accomplished electromagnetically rather than me-
chanically. The IDT can also scan in two dimensions, Direct Readers. The direct-reading spectrome-
which makes it suitable for use with echelle spectrom- ter has been commercially available for emission spec-
eters. Like any PMT, the IDT has high sensitivity and trometry for many years. Typically, an array of slits and
a wide linear dynamic range. Unlike solid-state array photomultipliers are used, one for each wavelength to
detectors, the IDT does not integrate the incident in- be detected. A typical system using a Rowland circle
tensity, which can be a disadvantage in some situations. configuration is discussed in Chapter 3, Figure 3-54.
However, the major drawback of the IDT at present is Usually, high-dispersion poiychromators with focal
its high initial cost (= $10,000), which has severely lim- lengths of 0.75 m or greater used in these systems to
ited its use in spectrochemical instrumentations. make it easier to position the slits and detectors. In-
struments with as many as 57 exit slits and detectors
Double-Beam Spectrometers. Double-beam have been described.
systems are used in absorption and photoluminescence The direct-reading spectrometer is widely used
measurements. Here the excitation beam is split into where the same group of wavelengths must be moni-
Chap. 4 1 Optical Sources, Transducers, and Measurement Systems
Sample Sample
cell detector
8-
Source
Monochromator kc,L
nce
detector U
Signal
processing
Chopper-beam
splitter Motor Sample
I / cell
w
Source Monochromator
cell
I I
(b)
FIGURE 4-41 Double-beam spectrometers. In the double-beam in-space
configuration (a), the excitation beam is split and two separate detectors
are used. In the dmble-beam in-time configuration (b), the two beams
are recombined and alternately strike a single detector. In either case the
signal processor calculates the ratio of the sample signal magnitude to that
of the reference signal (T) for absorption measurements or the difference
between the sample signal strength and the blank signal strength for pho-
toluminescence measurements. Wavelength selection can occur before or
after the beams pass through the cells.
tored routinely, as in multielement emission spectros- olution systems. They allow spectra to be obtained in
copy. This type of system is expensive and is not very situations where the concentrations of the monitored
flexible because alignment of the exit slits is a tedious species are changing in time. One commercial emission
and time-consuming task. As a result, wavelengths are spectroscopy system uses an echelle grating and a two-
changed only infrequently. Correction for background dimensional vidicon detector to give high-resolution,
is also difficult with a direct reader since fixed wave- multiple-wavelength coverage. Several commercial mo-
lengths are used. Oscillating slit arrangements and vi- lecular absorption spectrophotometers use diode ar-
brating quartz plates in front of the exit slits as discussed rays; most can provide a complete UV-visible absorp-
in the preceding section have been used to scan a small tion spectrum in seconds. Such spectrophotometers are
spectral region around the fixed wavelength and thus now becoming quite popular and could make conven-
allow background correction. tional scanning UV-visible instruments obsolete in the
near future.
Spectrographs w i t h Multichannel Detec-
tors. An alternative approach to the direct reader is Dual-Channel Spectrometers. A dual-channel
to remove the exit slits and to obtain all the spectral spectrometer is a special case of a multichannel spec-
information simultaneously at the focal plane. A true trometer where two wavelengths are monitored. It can
multichannel detector is then required. The photo- be implemented with a direct-reader approach, but is
graphic plate has been used with spectrographs for many often constructed by using two filters and detectors or
years. Recently, however, it is being replaced by the two monochromators and detectors. Such spectrome-
video-type multichannel detectors discussed in Section ters are used primarily for background or blank cor-
4-4. Because of their small sizes and rapid response rection. One channel is used to monitor the analytical
speeds, diode arrays and vidicons are used in small, wavelength (analyte plus background signal) and the
medium-resolution spectrographs as well as in high-res- other channel monitors a wavelength or wavelength range
Sec. 4-6 1 Optical Spectrometers 131
where only the background signal is present (or signif- into account the instrumental parameters discussed in
icant). If the background signal at this second wave- this chapter. We consider here a fixed-wavelength spec-
length is equal to the background signal at the analytical trometer of throughput factor Y ( X ) and analog signal
wavelength, the difference in the signals from the two processing with voltage readout. A portion of the ra-
channels represents the corrected analytical signal. Note diation emitted or transmitted by the chemical system
that channels in direct readers can be dedicated to back- and wavelength selector is assumed to be incident on a
ground correction and that with multichannel detectors PMT transducer of radiant cathodic responsivity R(h)
signals from appropriate wavelengths can be selected and gain m . For simplicity, here and in later chapters
to obtain the required information. Scanning a dual- we will assume that the PMT collection efficiency is 1.
channel spectrometer can be used to obtain derivative The gain of the signal modifier in V AP1 is G . Note
information. that if a current-to-voltage converter is the only signal
processing element used, G = Rf,the feedback resist-
Multiplexing Systems ance.
Multiplexing systems can be classified in general as time-
division multiplexing or frequency-division multiplex- Emission and Chemiluminescence Spectrom-
ing systems. Time-division multiplexing allows the eters. The analytical emission signal EE is obtained
timesharing of a single optical path or single detector by subtraction of a blank signal E,, from the total meas-
by multiple optical signals. The double-beam-in-time ured signal EtE (EE = E,, - E,,). The signal EE is
spectrometer is an example of time-division multiplexed related to instrumental parameters by equations 4-33
spectrometer for two optical beams. Time-division mul- (where i, is the appropriate PMT anodic current or
tiplexing has also been used in atomic absorption and product of the photocathodic current and the PMT gain),
fluorescence spectrometry. 4-30, 3-69, and 3-71:
In the frequency-division multiplex approach all
wavelengths strike a single-channel detector simulta-
neously; the spectral information at each wavelength is
encoded in such a manner that it can be recovered using
mathematical transform techniques. In Fourier trans- where B,, is the spectral radiance due to analyte emis-
form spectroscopy the spectral information is encoded sion and i, is the analyte emission photocathodic cur-
with a Michelson interferometer. Since the radiation is rent. Here and elsewhere, for simplicity, we drop the
not dispersed, the throughput can be very high. Also, subscript c for cathodic unless there is possible confu-
the detection of all wavelengths simultaneously leads sion between cathodic and anodic current. The blank
to a SIN advantage in cases where detector noise is signal is composed of a dark signal Ed and a background
limiting. Fourier transform methods are most widely emission signal E,,. The dark signal is given by
used in the IR spectral region because detectors are
often detector noise limited; in the UV and visible re-
gions the SIN is almost always limited by other sources
of noise.
In Hadamard transform spectroscopy a conven- while the background emission signal is
tional dispersive spectrometer is used and a movable
mask that contains a series of slits is placed in the focal
E,, = mGi,, = mG BAbEY(h)R(h)
dh (4-36)
plane. The positions of the transparent and opaque parts
of the mask correspond to the elements of a Hadamard
matrix. Computer solution to the linear equations that where BAbEis the spectral radiance of the background
result from intensity measurements for different mask emission, ,i is the background emission photocathodic
positions produces a conventional spectrum. Some also current, and id is the cathodic dark current. For quan-
have a slit mask for the entrance slit. Like Fourier trans- titative analysis B,, can be related to the analyte con-
form methods, Hadamard transform techniques have centration as is shown in later chapters.
been found most useful in the IR region of the spec-
trum, where detector noise determines the SIN.
Photoluminescence Spectrometers. In pho-
Quantitative Readout Expressions toluminescence the analyte luminescence signal EL is
obtained after blank subtraction from the total lumi-
We are now prepared to expand the general readout nescence signal E, (EL = EtL - E,,) if analyte emis-
expressions presented in Chapter 2 for emission, pho- sion is negligible. The photoluminescence signal is re-
toluminescence, and absorption spectrometry to take lated to the spectral radiance of analyte photo-
132 Chap. 4 I Optical Sources, Transducers, and Measurement Systems
luminescence BALby from the total sample signal E,,. and E, is obtained by
subtracting E,,, from the 100% T signal El,. With the
foregoing assumptions. E,,, is due to dark current and
background emission given by equations 4-35 and 4-36,
respectively. The signals E, and E, are given by
where i, is the photocathodic current due to analyte
luminescence. The blank signal E,, is composed of a
dark signal. given by equation 4-35, a background emis-
sion signal, a scattering signal, and a background lu-
minescence signal. These latter signals are all of the
and
form given by equation 4-37 with their respective spec-
tral radiances substituted for B,,. Again BhLis related
to the analyte concentration as described in later chap-
ters. Note that for emission and photoluminescence
measurements (see equations 4-34, 4-36, and 4-37), the where B, is the source spectral radiance, T,is the sam-
observed signal is the desired spectral radiance distorted ple transmission factor, TI is the reference transmission
by the spectrometer response function [e.g., Y (X)R(X)]. factor, and i, and i, are the reference and sample pho-
Therefore, in a spectral scan, the shape and intensities tocathodic currents. The value of T, is related to the
of the spectral bands or lines obtained are dependent analyte concentration as detailed in later chapters. Often
on the instrumental response function. experimental conditions are arranged such that the an-
alyte absorptivity does not vary over the wavelength
Absorption Spectrometers. In absorption range incident on the sample. In this case, the instru-
spectrometry the transmittance T is given by T = mental response function does not appear in a trans-
E,IEr, where Es is the signal with the sample in place mittance or absorbance measurement because a ratio
and E, is that with the reference in place. If we assume is measured.
that background luminescence, analyte emission, and For direct absorbance measurements the voltage
analyte luminescence are absent, the transmittance is proportional to the analyte absorbance E, is related to
the signals E, and E, by equations 2-33 and 2-34. In the
case of modulated systems, the radiances and cathodic
currents in the expressions above are the average val-
ues. For photon counting systems the signals are in
Here E, is obtained by subtracting the 0% T signal E,, terms of n . the number of counts in time t .
PROBLEMS
4-1. Usually when one buys a radiation source, the man- A1,0, doped with Cr. Assume that the Cr concentra-
ufacturer will supply spectral radiance data so that one tion is 1.58 x 101'cm-j and the transition cross section
can judge if the intensity is sufficient for a given ap- is 1.27 x cm2 at the lasing wavelength of 694
plication. For tungsten lamps, the spectral radiance nm.
can be estimated from blackbody parameters. (a) Calculate the transmittance of the ruby rod in the
(a) Calculate the spectral radiance of a tungsten lamp absence of the pump.
at 500 nm with a color temperature of 2700 K , (b) Calculate the threshold population inversion and
E~ = 0.40, and T , = 0.92 in W sr-' cm-' n m - ' the number density of the upper energy level of
and photons s-' cm-2 sr-I nm-'. the lasing transition at threshold.
(b) Apply propagation of error mathematics to the 4-3. The anode of a 1P28 photomultiplier is connected to
blackbody equation to determine how the uncer- a current-to-voltage converter with a 1.0 x 10'-Q feed-
tainty in temperature relates to an uncertainty in back resistor. The PMT voltage is adjusted to 800 V
radiance. Evaluate the standard deviation and RSD and the collection efficiency is 1.00. Assume that
in radiance at 500 nm caused by a 5•‹Ctemperature 1.0 x 10-I' W at 500 nrn is incident on the photo-
standard deviation under the conditions in part cathode. Calculate or indicate the following values (the
(a). specification sheet in Appendix I)is required).
4-2. Consider a ruby laser made with a 10-cm-long ruby (a) Current gain (m).
rod and mirrors of reflectivity 0.99 and 0.74. Ruby is (b) Photocathodic current (i,,).
Chap. 4 I References
(c) Photoanodic current (i,). 4-12. How do the total radiance and wavelength of maxi-
) Photocathodic pulse rate (r,). mum emission of blackbodies at 2000 K and 3000 K
(e) Anodic dark current (i,,). compare?
(f) Cathodic dark current (i,,). -13. For an atomic transition from the first excited state of
(g) Photoanodic voltage (E,). degeneracy 2 to a nondegenerate ground state, the
(h) Anodic dark current voltage (E,). Einstein coefficient for spontaneous emission is 1.0 x
How does one decide between a lock-in amplifier and 10' s-I and the wavelength is 300 nm. Calculate the
a boxcar integrator as a signal processor for modulated following values.
signals? (a) The Einstein B coefficients for stimulated absorp-
When does one choose a multichannel spectrometer tion and emission.
instead of a single-channel spectrometer? (b) The absorption oscillator strength (refer to Ap-
Contrast the characteristics and uses of photon detec- pendix F).
tors versus thermal detectors. -14. Calculate the longitudual mode separation for a
A vacuum phototube has a radiant cathodic respon- 0.25-m laser in Hz and nm for a lasing wavelength of
sivity of 0.08 A W-' at 400 nm. 488 nm.
(a) Find the quantum efficiency at 400 nm. 4-15. The D" value for a detector with an area of 0.10 cm2
(b) If the incident photon flux at 400 nm is 2.75 x is 1.0 x 1OY cm Hz1I2W- '. Calculate the noise equiv-
lo5 photons s-', find the anodic pulse rate and the alent power with a 1-Hz noise equivalent bandpass.
photoanodic current for a collection efficiency of 4-16. Consider a linear diode array with diodes that have a
0.90. saturation charge of a 3.6 pC and a responsivity at 500
A photomultiplier tube has a radiant cathodic respon- nm of 12 pA pW- ' cm'. Calculate the exposure time
sivity of 8.0 mA W-I at 365 nm. It is operated under with an incident irradiance of 1.0 pW ~ m that - pro-
~
conditions that give a gain m of 1.5 x 10' and a col- duces saturation. How many electron-holes pairs are
lection efficiency y of 0.88. If 5.8 X 10.- l 1 A of anodic produced at saturation?
current can just be detected, what is the minimum 4-17. In photon counting, nonlinearity occurs at high photon
detectable photon flux in photons s--'? If photon fluxes because of pulse overlap effects (i.e., the prob-
counting were used instead of dc current signal pro- ability of two photons generating photoelectrons within
cessing, what is the count rate corresponding to the the response time or dead time of the detector or signal
foregoing photon flux? Note that a lower light levcl is processor becomes significant). For some photon
normally detectable with photon counting. counting systems denoted paralyzable systems, the ob-
A PMT has a current gain of 4.5 x lo5 and a quantum served count rate r' is related to the true count rate r
efficiency of 0.36 at 450 nm. The PMT output is con- by r' = r(l - r ~ if) r7 < 0.13, where T is the system
nected to a current-to-voltage converter with a feed- deadtime in seconds. Calculate the true count rate for
back resistor of 10 MR. A n~onochromaticsource with which the observed count rate is 1 % low for a photon
a flux @, of 2.0 x 10"hotons s 1 is incident on the counting system with a dead time of 2 ns.
PMT. What is the output voltage of the I-Vconverter? 4-18. Nonlinear effects are only observed with laser radia-
Assume y = 1. tion. Calculate the electric field and irradiance nec-
Compare and contrast the microchannel plate inten- essary for the polarization due to second-order effects
sified photodiode array with a photomultiplier tube to be as large at that due to first-order effects. Assume
for the detection of low radiation levels. Give the ma- the second-order electric susceptibility for the material
jor advantages and disadvantages of both detectors. irradiated to be of the first-order susceptibility.
Calculate the spectral energy density for a 2500-K
blackbody at 600 nm in J c m 3n m l and photons c m i
nm-'.
The following are general references on optical sources and/ ductive, photoemissive, thermal, bolometers, pyroelec-
or detectors including optical measurements. tric, and charge-transfer devices (CCDs and CIDs).
1. F. Grum and R. J. Becherer, Optical Radiation Meas- 3. G . J. Zissis and A. J. Larocca, "Optical Radiators and
urements; vol. 1, Radiometry, Academic Press, New York, Sources," in Handbook of Optics, W. G. Driscoll and
1979. Has a good discussion of sources of radiation and W. Vaughan, eds., McGraw-Hill, New York, 1978. A
detection. practical description of various optical sources.
2. E. L. Dereniak and D . 6. Growe, Optical Radiation 4. S. F. Jacobs, "Nonimaging Detectors," in Handbook of
Detectors, Wiley, New York, 1984. Good discussion of Optics, W. G. Driscoll and W. Vaughan, eds., McGraw-
all optical detectors. Includes photovoltaic, photocon- Hill, New York, 1978.
134 Chap. 4 / Optical Sources, Transducers, and Measurement Systems
H . T. Betz and G. L. Johnson, "Spectroradiometric 15. J. M. Harris, "Pulsed Laser Systems," in Lasers in
Principles," in Analytical Emission Spectroscopy, vol. I , Chemical Analysis, G . M . Hieftje, J. C. Travis. and F.
pt. 11, E . L. Grove, ed., Marcel Dekker. New York. E . Lytle, eds., Humana Press, Clifton, N.J., 1981. A
1971, pp. 323-381. good discussion of pulsed lasers.
R . C. Elser, "Optical Instrumentation," in Trace Anal- 16. E. H. Piepmeier, ed., Analytical Applications of Lasers,
ysis; Spectroscopic Methods for Elements, chap. V. Wiley-Interscience, New York, 1986.
J. D . Winefordner, ed., Wiley-Interscience, New York, 17. A. Yariv, Optical Electronics, 3rd ed., Holt, Rinehart
1976. A good discussion of optical measurements, in- and Winston, New York, 1985. An excellent laser book.
cluding transducer, signal modifier? and readout sys- Also discusses beam propagation in fibers: modulation,
tems. atomic systems, detectors, and many. other topics.
R. W. Engstrom, R C A Photomultiplier Handbook, PMT-
62, RCA Corp., Solid State Division. Lancaster, Pa., Listed below are more specialized references providing ad-
1980. A classic reference book for users of photomul- ditional details on specific topics covered in this chapter.
tiplier tubes. Contains a wealth of useful information.
Electro-Optics Handbook, Technical Series EOH-11, 18. J. C. Wright, "Nonlinear Optics," in Lasers in Chemical
RCA Corp., Lancaster, Pa., 1974. Analysis, G . M . Hieftje, J. C. Travis, and F. E. Lytle,
G. F. Kirkbright and M. Sargent, Atomic Absorption eds., Humana Press, Clifton, N.J., 1981. Includes a good
and Fluorescence Spectroscopy, Academic Press, Lon- discussion of nonlinear polarization effects.
don, 1974. Has an excellent chapter on spectral light 19. H . V. Malmstadt, C. G. Enke, and S. R. Crouch, Elec-
sources, including hollow cathode lamps, electrodeless tronics and Instrumentation for Scientists, Benjamin-
discharge lamps, and others. Cummings, Menlo Park, Calif.. 1981. A modern text
that discusses instrumentation and measurement prin-
The following references are devoted to the principles and ciples.
application of lasers.
20. R. J . Higgins, Electronics with Digital and Analog In-
M. D . Levenson, Introduction to Nonlinear Laser Spec- tegrated Circuits, Prentice-Hall, Englewood Cliffs. N.J.,
troscopy, Academic Press, New York. 1982. 1983. A good discussion of modern electronics with em-
N. Omenetto. Analytical Laser Spectroscopy, Wiley, New phasis on integrated circuits.
York, 1979. An excellent book dealing with laser prin- 21. Y. Talmi, ed., Image Devices in Spectroscopy, ACS
ciples and the applications of lasers in analytical chem- Symposium Series 102, American Chemical Society,
istry. Washington. D.C., 1982. Papers were presented in a
G. M. Hieftje, ed., New Applications of Lasers to Chem- symposium on image detectors.
istry, American Chemical Society, Washington, D . C . , 22. Y. Talmi, ed., Multichannel Image Detectors, vol. 2,
1978. Contains excellent chapters on laser applications ACS Symposium Series 236, American Chemical Soci-
written by experts in the field. ety, Washington, D.C., 1983. A more recent book based
F. E . Lytle, "Laser Fundamentals," in Lasers in Chem- on a 1982 symposium by leaders in the field.
ical Analysis, G . M . Hieftje, J. C. Travis, and F. E . 23. D . G. Jones, "Photodiode Array Detectors in UV-VIS
Lytle, eds., Humana Press, Clifton, N.J., 1981. A good Spectroscopy: I and 11," Anal. Chem., 57,1057A, 1207A
introduction to laser principles. (1985). Recent reviews on array detectors.
M. J. Wirth, "Tunable Laser Systems," in Lasers in 24. G. Horlick and E. G. Codding, "Photodiode Arrays for
Chemical Analysis, G . M. Hieftje, J. C. Travis. and F. Spectrochemical Measurements," in Contemporary Topics
E . Lytle, eds., Humana Press, Clifton, N.J., 1981. A in Analytical and Clinical Chemistry, D . M. Hercules,
practical discussion of the state of the art as of 1981 in G. M. Hieftje, L. R. Snyder, and M. A . Evenson, eds.,
tunable lasers. vol. 1, chap. 4, Plenum Press, New York, 1977.
The precision of instrumental measurements, including dependencies on experimental variables are presented
spectrochemical measurements, is often limited by noise next. Complete SIN expressions are developed for emis-
in the measured signals. Noise is defined here as un- sion, luminescence, and absorption techniques. The use
wanted fluctuations in the desired signal which obscure of these expressions for optimizing precision is empha-
its measurement. As an analogy, consider the problem sized. Finally, SIN enhancement techniques are re-
of listening to one person in a crowded room with a viewed.
multitude of conversations. The noise in this case is the
audible noise from all the other conversations which if
too loud can obscure the signal, the voice of the person
you are trying to hear.
Because noise affects the quality and precision of
spectrochemical measurements, the optimization of ex-
perimental variables is often based on reducing the rel- A typical recorder tracing of a dc signal from a spec-
ative amount of noise or increasing the signal-to-noise trometer is shown in Figure 5-1. The resolution of the
ratio (SIN). Noise can be introduced by the sample recorder is sufficient to observe the fluctuations or noise
presentation system, the optical information encoder, in the signal. The signal is characterized by its mean
the radiation transducer, and the signal processing and value, which can be estimated by drawing a line through
readout system. Random errors in sample acquisition the center of the fluctuations as shown. Alternatively,
and sample preparation can also limit the precision of n independent measurements of Eican be made during
a technique. Irreproducibility due to these sources is the observation period, and the arithmetic mean cal-
normally not considered noise, although it is sometimes culated from
Time
FIGURE 5-1 Recorder tracing of dc voltage with noise. The mean value
of the signal is E. The magnitude of the noise is the root-mean-square
(rms) noise, ,s, the average power content of the noise. The rms noise is
approximately one-fifth of the peak-to-peak noise (E,.,). In (a) the R C
time constant T is 0.1 S , while in (b). T is 1.0 s. The larger time constant
is seen to reduce the higher-frequency noise components and thus to
reduce the total amount of noise observed.
to follow the changes of interest faithfully. High-fre- since the RSD equals 0.1%. Although the noise and
quency fluctuations due to undesirable noise can be signal in equation 5-3 are expressed in terms of voltages,
attenuated by integration or by a low-pass filter, as they can be expressed in any appropriate signal units
described in Section 5-2. or for any observation point within an instrument.
Frequency
I I
FIGURE 5-3 Some ac signals and their power spectra. Most of the signal
information in the chopped signal (a) is contained at the fundamental
frequency f,. Note that chopping produces odd harmonics, as in a square
wave. T o attenuate noise at other frequencies, a bandpass filter or a tuned
amplifier can be used to pass or amplify a narrow frequency interval
centered at f,. The peak-shaped signal (b) could result from a spectral
scan. Note that its power spectrum is similar to that of a dc signal, but
the signal power may extend to relatively high frequencies.
Sec. 5-2 I Frequency Characteristics of Signal Processing and Readout Systems
where e, and ei, are the instantaneous output and input IE,IElnI vs. f . Similarly, the phase shift between output
voltages, respectively. The transfer function depends and input can be measured as a function off.
on frequency (o = 27rf); the j operator (G) is a Three typical amplitude transfer functions are
reminder that there may be a phase difference between plotted in Figure 5-4. The amplitude transfer function
e, and el,. Note that we use f to represent the electrical illustrated by curve a is typical of instruments that use
frequency, usually in the audio range, while v denotes a single-stage R C low-pass filter for measuring dc sig-
the optical frequency. In our discussion, we will con- nals. If the signal is ac, or has been converted to ac by
sider only the am e transfer function lH(f)l and not modulation, a bandpass filter can be employed. The
concern ourselves with phase shifts. It must be kept in response of a bandpass filter with a central frequency
mind, however, that phase shifts are introduced by most of 100 Hz is shown in curve b of Figure 5-4. The fre-
reactive circuits used for filtering and narrow-band am- quency of maximum response is adjusted to the signal
plification. frequency while the width of IH(f)l is adjusted to min-
The amplitude transfer function is defined by imize the range of noise frequencies passed. Curve c in
Figure 5-4 shows the frequency response of an integra-
tor with a l-s integration time.
where E,, is the amplitude of the input signal (or noise) The amplitude transfer function determines the range
and E, is the amplitude of the output signal (or noise). of frequencies passed by the frequency-limiting com-
A plot of IN(f)l vs. frequency is the frequency response ponents; this range can be expressed as a signal fre-
of the system. The amplitude transfer functions for sev- quency bandpass or a noise equivalent bandpass. The
eral common signal processing circuits were defined in signal frequency bandpass is the range of frequencies
Figures 4-35 and 4-36. For simple passive filters (filters for which the attenuation is less than 3 decibels (dB)
without gain), the transfer function is unity at the fre- (IH(f)l > 0.707) [dB = 20 log (E,IEi,)]. The noise equiv-
quency of maximum response and falls off rapidly at alent bandpass is defined by
higher or lower frequencies. In a spectrochemical in-
strument, adjustments are made so that IH(f)l = 1 for
the frequency at which the analytical signal is encoded
and zero or near zero for frequencies removed from the
encoding frequency. Thus noise at frequencies removed and represents the frequency range of electrical power
from the signal frequency is significantly attenuated. passed by an ideal filter with a rectangular amplitude
The amplitude transfer function, as well as the transfer function [i.e., IH(f)l equals 1 or 01, as shown
phase shift, can be calculated as a function of frequency by rectangles d, e, and f in Figure 5-4. The signal fre-
for specific filtering networks. It can also be determined quency bandpass and the noise equivalent bandpass are
experimentally by using a sine-wave signal generator as compared in Table 5-1 for a low-pass filter of time con-
input to the frequency-limiting circuitry. The input and stant T (T = RC), an integrator of integration time t,
output voltages are measured sequentially for different and a bandpass filter with low and high cutoff frequen-
signal frequencies to generate a plot of IW(f)I or cies (3 dB points) off, and f,.
TABLE 5- 1
Band pass characteristics of frequency-limiting circuitsa
Signal Noise
Lower cutoff Upper cutoff frequency equivalent
frequency, f, frequency. f, bandpass, bandpass,
Circuit HCf)l (3 dB point) (3 dB point) f~ - fi (Hz) Af (Hz)
Low-pass [l + (21~f7)~]-'" 0 ( 2 ~ 7-) (27s)- I (47) - I
filter
Integrator sin ~ f t
- 0 -
0.433 0.433
- (24-
~ft t t
Bandpass - fi fi f- -
Tfm
filter Q 2Q
-- --
9, Time constant (RC) of low-pass filter (s); t, integration time of dc integrator (s): Q, quality factor for bandpass filter = fm#(f2 - f,):f,,.
central frequency of bandpass filter (Hz).
To illustrate the differences in the signal bandpass noise. However, there are practical limitations. For low-
and the noise equivalent bandpass, consider a low-pass pass filters, a small Af is obtained at the expense of a
filter with a T value of 1 s. The signal bandpass (see large time constant. The time constant affects the re-
Table 5-1) is lI(2m) = 0.16 Hz, while the noise equiv- sponse time of the instrument. The response time is the
alent bandpass is 11(47) = 0.25 Hz. At frequencies higher time it takes the output signal to reach some percentage
than the 3-dB point (0.16 Hz), the attenuation of the (e.g., 99.7 or 98%) of its equilibrium or final value after
low-pass filter increases with frequency until it asymp- a step signal is applied to the input. For a simple low-
totically approaches 20 dBIdecade. pass R C filter, the response time is often defined as 57
(5RC), the time for 99.7% of full response.
Noise in Readout Signals For dc signals, the time constant of a simple low-
pass filter is chosen as a compromise between noise
Limiting the bandpass of the signal processing system filtering and response time. Thus for a 7 of 1 s, one
can drastically attenuate noise, especially if it is white must wait 5 s after the analytical sample is introduced
noise [constant power density P(e) at all frequencies]. to the spectrochemical information encoder before tak-
The mean-square noise (variance) in the readout signal ing a signal reading. Time constants larger than 10 s
is given by are rarely used because it would take 50 s or more to
obtain one sample reading. Also, reduction of Af below
about 0.02 Hz rarely provides any SIN additional ad-
vantage, due to the dominance of llf noise and drift.
For single-event, transient optical signals (a peak-
where P(e) is the noise density in V2 H Z - l . If the NPS shaped, ramp, or exponential time dependence), 7 is
is flat or due only to white noise, usually chosen to be as large as possible to reduce Af,
but still small enough to prevent distortion or atten-
uation of the signal information. Generally, T should
be no greater than one-tenth of the time duration over
which the transient signal is measured. In the case of
Equations 5-6 and 5-7 indicate that the amount of recording spectra, the time constant should be about
noise observed decreases as the frequency bandpass of one-tenth of the time it takes to scan through the nar-
the instrument is reduced. In the case of white noise, rowest peak. Here one is making a compromise be-
s2, is proportional to Af, or the rms noise (s,) is pro- tween the time to scan a spectrum (e.g., the scan rate
portional to Af1I2. For 1If noise, decreasing Af by a in nm s-l) and Af.
factor of 2 reduces the rms noise by less than a factor With an R C filtering system, noise is effectively
of 2lI2 because P(e) is greater at lower frequencies. The averaged over a few time constants. Higher-frequency
dependence of the magnitude of the lif noise observed noise components are highly attenuated because the
on Af is controlled by the shape of its NPS. In some period of their fluctuations is much less than the re-
cases, there may be no significant improvement or even sponse time of the R C network (see Figure 5-1). Sim-
an increase in the noise observed when Af is decreased. ilarly, with an integration system, the contributions of
The equations above indicate that Af should be higher-frequency fluctuations with periods much less
made as small as possible to reduce the observed rms than the integration time tend to average to nearly zero
Sec. 5-3 I Noise Sources 1
because many such fluctuations occur during the inte- cations for photodiodes, phototubes, and other types
gration period. If the noise is at a fixed frequency, such of transducers are discussed later in this chapter. Two
as 60-Hz noise from the ac power lines, choosing the common signal processor-readout systems, analog and
integration time to be an integral multiple of the noise photon counting, are considered. In the analog signal
eriod can provide nearly perfect noise rejection since processing system, the signal processor contains circui-
an integral number of noise cycles are averaged. Many try for converting the transducer anodic current into a
integrating analog-to-digital converters use this princi- voltage (possibly with additional stages of amplifica-
rovide very high rejection of 60-Hz noise. tion) before display on a voltage readout device. The
hen integrators are used for noise reduction in electrical bandwidth is controlled by an analog filter (or
dc signals, the integration period is chosen to be large integrator). The photon counting signal processor sys-
to reduce Af, but small enough for time convenience. tem consists of a fast amplifier, discriminator, and pulse
Integration times much greater than 1min are not usu- counter; the integration or counting time can be varied
ally used for the same reasons that very large time con- (see Section 4-5).
stants are not employed. Let us establish at the outset several general prin-
Integration systems can also be used for transient ciples. First, in each type of signal (dark, background,
signals, but the integration time is often chosen to be and analytical), there can be a fundamental and a non-
small enough that the signal does not change signifi- fundamental noise component. Additionally, the signal
cantly over t. If the signal amplitude does change over processing system generates noise (both fundamental
t, the time integral of the signal is recorded; this can and nonfundamental) in the absence of any signals from
be used to find, for example, the area of a peak-shaped the optical transducer. Second, the progression of sig-
signal. nals through an instrunlent can be considered to be a
An integration system can provide higher through- series of sequential transformations where both the sig-
put since there is not an RC response time. An RC nal and noise experience the same transformation.
circuit with a .r value of 1.0 s and an integration system Moreover, additional noise or signals can be injected
with an integration time t of 0.5 s have identical noise during each transformation step. The transformation
bandwid.ths (Af = 0.5 Hz). However, signal data can steps are illustrated in Figure 5-5.
be obtained in 1 s with the integrator, but the RC filter
circuit requires 2.5 s to respond fully.
A simple RC filter is a first-order filter. Higher-
order low-pass filters with two or more RC networks
are available. Compared to first-order filters, higher- First, consider the flux of background and signal pho-
order filters, particularly active filters based on oper- tons (from the optical information encoder and selector
ational amplifiers, can provide steeper roll-off at higher system) that strikes the photocathode of the optical
frequencies and equivalent Af with a faster response transducer. If we could count the number of photons
time. This means that greater noise reduction can be impingent per unit time, we could establish a mean
achieved (i.e., smaller Af values) without waiting as value and a standard deviation in the radiant power
long for the filter response and with less distortion and expressed in photons per second or watts. In many cases,
attentuation of transient signals. the photons arrive randomly (the spacing between pho-
Finally, it should be noted that with computer data tons is random) because the emission of photons from
acquisition filtering can be carried out with software the excitation source or from the analyte is random.
rather than hardware, as discussed in Section 5-6. Under these conditions, the Poisson probability distri-
bution function applies,
/
Incident
photons -
' Current gain by dynodes Signal
processor
Readout
Photon
counting
signal
processing:
' P J ,' P , B
(photon s-')
KO)
____t
' C S , ' C B , ',d
(electrons s-')
: 'OS' ' O B > rod
(anodic pulses s-')
- tfd ns: n~ ad
(counts)
counted in integration time t and is given by square root of n and can be improved by increasing a,
or t. The equations above apply equally to signal and
background photons. However, the signal in equation
5-11 is normally taken to be only due to signal photons.
Here and in further discussions, we will drop the bar The quantum noise in the original photon flux is
and understand that the mean value of n or other signal seen in the signals at the photocathode, the anode, and
quantities is implied. The standard deviation or rms the readout device because all conversions are random
noise in the number of photons counted in time t (u,) or multiply the signal and noise by linear factors. Quan-
is found from tum noise is often called Schottky noise or shot noise.
Schottky noise is observed whenever a current passes
through or is generated at an interface. The signal and
quantum noise equations that apply at the transfor-
The subscript q denotes quantum noise, the fundamen- mation points for both analog and photon counting sig-
tal noise due to the random emission of photons from nal processing are summarized in Table 5-2. In this
a source. If the background photon flux is negligible, discussion n , i, and E are the symbols for signals ex-
the SIN is given by pressed in terms of the number of events or pulses
counted, the current, or the readout voltage, respec-
tively. These symbols are also used as subscripts with
u to denote the noise in a particular type of signal.
Subscripts c, a, and r are used to designate the signal
Equation 5-11 indicates that the SIN increases as the at the photocathode, the anode, or the readout device
See. 5-3 1 Noise Sources
(only for photon counting), respectively. For simplicity, s) to denote specifically noise in the anodic current or
these subscripts are not used with o ; the application of readout voltage signals that is due both to quantum
the symbol to the noise at the photocathode, the anode, noise (subscript q) and PMT multiplication noise (sub-
or the readout device is determined from the context. T multiplication or secon
Let us first consider the signals and noise at the noise accounts for random fluctuations in the gain due
photocathode. Equations 1 to 4 in Table 5-2 apply to to the random nature of secondary emission of electrons
the analytical signal and the background signal, where at the dynodes. This is a multiple Poisson process so
r, is either the signal or background photoelectron pulse that the expression for the PMT multiplication noise in
rate. The SIN due to quantum noise in the analytical the anodic current has the form of the quantum noise
signal is n,lo, = nfI2or iJo, = (i,tIe)1'2. Notice that this equation. Thus equation 8 of Table 5-2 is a summation
SIN is less than the SIN calculated in the original photon of quantum noise [(u,), = (mei,It)112]and multiplication
beam (n1l2)because some information has been lost at noise [((T,), = (amei,lt)112],where all2is just some frac-
the photocathode due to the quantum efficiency being tion of the quantum noise in the anodic current. The
less than 1. However, the characteristic dependence of factor a is often between 0.1 and 0.5 for a good-quality
the SIN on the square root of the signal and the square PMT and can be estimated to be (6 - 1 ) - I where 6 is
root of the integration time under quantum-noise-lim- the gain per dynode stage (see Section 4-4). This for-
ited conditions is still obvious. mula for a does not correctly account for the PMT
The photocathode is a transformation point. The multiplication noise if secondary emission noise at the
cathodic dark signal and dark signal quantum noise are dynodes is not uniform or if the statistics are not Poisson
injected at this point and can be calculated from equa- (e.g., exponential).
tions l to 4 if the cathodic dark pulse rate is known. Equations 9 to 12 of Table 5-2 are the readout
Equations 5 to 8 of Table 5-2 apply to the ana- equations for the mean signal and noise. For photon
lytical, background, and dark anodic signals or the noise counting, equations 9 and 10 can be used to calculate
in these signals. We will use the termshot noise (subscript the mean (n,, n,, and n,) and shot noise [(a,),, (o,),,
TABLE 5-2
Signal and noise expressions for quantum or shot noise at transformation pointsa
Vzn,,mean number of photocathodic electrons counted in time t (dimensionless); (a,,),,SD in 12,. r l , , or 11, due to quantum noisc, dimensionless; r,, photocathodic
electron pulse rate (s-I); i,, mean cathodic current (A); (u,),, SD in i, or i , due to quantum noise when integrated for time t (A); r r , , Incan number of photoanodic
pulses counted in time t (dimensionless); i,, mean anodic current (A); (u,),, SD in i , due to shot noisc when integrated for time r (A); 1 1 , . readout counts in time t
(dimensionless); f,, discriminator level (dimensionless); E , readout voltage (V); G, gain factor (V A-I); (u,),, SD in E due to shot noise when integrated for time
t (V); a , secondary emission factor (di.nensionless).
'
bAt the photocathode, the photon flux is converted with quantum efficiency K(X) into a photoelectron pulse rate in photoelectrons s or a photocathodic current
in A . Conversion of n or u,,to the analog current domain 1s seen to involve multiplication by the factor (elr). Division of r z or u,, by r converts counts to counts per
second and multiplication by e (1.6 x lo-" A) converts counts per second into amperes. Alternatively, i, can bc expressed as @R(X) or the product of the incident
radiant power (@) in W and the cathodic responsivity [R(X)] in A W-'.
"With a PMT, the photoelectrons receive an average gain of m , or the average number of electrons in an anod~cpulse packet is m . The gain process does not change
the pulse rate so the cathodic pulse rate (r,) and the anodic pulse rate (r,) are the same (assummg a collection efficiency of 1). as are n, and n,,. The conversion
factor from cathodic to anodic current for the signal is the PMT gain m , while for the noise is m ( l + a)", whcre a takes into account the PMT multiplication
noise.
dFor photon counting, a fraction of the anodic pulses reach the readout counter as determined by the discriminator level fd. With analog signal processing, thc anodic
currents are converted to voltages according to the gain factor C of the signal processing system in V A - ' .
144 Chap 5 1 Signal-lo-Noise Ratio Considerations
TABLE 5-3
Signal and noise expressions for quantum and shot noise in analytical, background, and dark
signais
and (ad),], in the analytical, background, and dark sig- gain (m = 1) and the lack of secondary emission noise
nals in counts, respectively, as shown in Table 5-3. The ( a = 0). Phototransistors and avalanche photodiodes
shot noise in this case is only due to quantum noise have gain and the equations for shot noise are similar
because variations in the gain from pulse to pulse change to those in Table 5-3. With multichannel array detectors
the pulse heights but not the pulse rate. (vidicons and photodiode arrays), rn = 1, cr = 0, and
For analog signal processing, it is convenient to the quantity related to photoelectron rate is the number
write equation 12 of Table 5-2 in terms of Af so that it of electron-hole pairs generated per unit time. Al-
can be employed with analog signal processing using though these devices integrate the flux over a certain
filters or integrators. Thus if Af = (2t)-l is used for an integration time, their outputs can be related to an av-
integration system, erage input flux, and equations similar to those in Table
5-3 apply. Intensified arrays (SIT tubes, intensified diode
arrays) have gain and thus multiplication noise. Al-
though the noise observed with thermal detectors has
where Af = (47)-l when a simple low-pass filter re- shot noise components, they are almost invariably lim-
stricts the noise equivalent bandpass. Equation 5-12 can ited by thermal noise, which is independent of illumi-
be further simplified by defining a bandwidth constant nation level, as discussed later in this section.
K equal to 2eAf(l + a ) ; thus
Flicker Noise
point in the instrument (e.g., photocathode, photo- sion signal is uncorrelated with the foregoing source
anode) is the product of the mean signal at that point flicker noises, so that a different flicker factor must be
and the flicker factor. used.
A flicker factor must be evaluated experimentally In all spectrometric measurements and, in partic-
for a particular situation since it depends on the noise ular, absorption measurements, the fluctuations in the
equivalent bandpass and the magnitude of other ex- transmission characteristics of the sample container (in-
perimental variables. Different symbols are used for the dependent of the analyte) can be significant. This flicker
analytical signal and background flicker factors since noise is denoted transmission or convection flicker noise
they can be independent of each other. However, in and is characterized by a transmission flicker factor,
some cases, they can be equivalent if the background denoted k,. Here one can see that the absolute fluc-
and signal flicker noise have the same origin. The flicker tuations in a light beam transmitted through a sample
noise in either the analytical or background signal is container with fluctuating transmission characteristics
often composed of two or more components (i.e., sep- are proportional to incident radiant power, but that the
arate flicker factors for each noise component) of in- relative fluctuations are independent of the incident
dependent origins. radiant power.
The direct proportionality between the magnitude Particularly in atomic spectrometry, the analyte
of the absolute flicker noise and the signal level is not population in the probed volume element in the sample
derived from fundamental principles as is the square- container can vary with time. This analyte flicker noise
root relationship between noise and signal level in the is often caused by fluctuations in the sample presen-
case of shot noise. Rather, it is empirically observed tation system. In emission and chemiluminescence, flicker
that in many cases, fluctuations in the variables which noise can also be caused by fluctuations in the excitation
affect the analytical or background signal radiance are conditions.
such that the relative fluctuation in the radiance is in- The flicker noise equations (Table 5-4) apply to
dependent of the absolute radiant power. Consider any radiation transducer if the appropriate output quan-
viewing a light source through a filter whose transmit- tity is used and multiplied by the flicker factor.
tance varies randomly in time with a 1% RSD. The
RSD in the transmitted photon flux will be 1% if the
RSD in the photon flux incident on the filter due to Other Noise Sources
quantum noise is much less than 1%. If the incident
photon flux is doubled, the absolute fluctuation in the The equations for dark current noise in Table 5-3 are
transmitted flux will double, but the relative fluctuation incomplete if there are significant contributions from
will still be 1% . sources other than thermal emission from the photo-
Specific sources of flicker noise are described in cathode. Some additional sources are thermal emission
some detail in discussion of specific techniques in later from the dynodes, cold field emission, ohmic leakage,
chapters. Here only a few general sources of flicker and radioactivity. Often the shot noise contribution from
noise will be identified. In absorption and photolumi- thermal emission from the dynodes is small because the
nescence spectrometry, fluctuations in the readout sig- dynode thermal dark current is small and receives a
nals are often caused by flicker noise in the excitation gain less than in. In analog signal processing systems,
source radiance. Source flicker noise is caused by var- the noise in the dark current voltage due to nonthermai
iations in experimental variables that control the source sources is denoted excess dark current noise with the
radiance, such as electrical power (not 60-Hz ripple) or symbol (o,),,. As the PMT bias voltage increases, the
temperature; it can also be caused by variations that relative contribution of ohmic leakage to the total dark
affect the source radiant power viewed, such as vibra- current decreases, but that due to cold field emission
tions. Thus a source can be characterized by a source increases.
flicker factor, denoted 5,. The noise observed in the absence of analytical,
Source flicker can cause both analytical and back- background, and dark current signals is amplifier-read-
ground flicker noise in a photoluminescence experiment out noise, a,,. It is typically measured with the bias
because the analyte fluorescence, background (non- voltage of the PMT or PT turned off. This noise is due
analyte) fluorescence, and scattering signals are all pro- to fundamental and nonfundamental noise generated
portional to the excitation source radiant power and in the signal processing and readout circuitry (a,) and
suffer an equivalent fluctuation due to source flicker. quantization noise (o,). Flicker noise in electronic cir-
In this case the flicker noise in the analytical and the cuitry can be caused by the random appearance of im-
background signals due to source flicker noise are to- purity centers in components. Quantization noise is due
tally correlated. Flicker noise in the background emis- to the finite resolution of any readout device and is
146 Chap. 5 I Signal-to-Noise Ratio Considerations
be subdivided into their individual components. The the equations in Table 5-3 and 5-4 to yield
blank noise is due to noise in the background signal
(shot and flicker), dark current noise (shot and excess),
and amplifier-readout noise or
where (u,), is the dark current rms shot noise and (u,),,
is the dark current excess rms noise.
The rms noise in the analytical signal (us) is due
to shot and flicker noise, so that
for the blank measurement). In this case the SIN is given proved by adjusting experimental conditions to increase
by the analytical signal or to reduce the dark current. The
latter can be accomplished by cooling the PMT or by
selecting a PMT with lower dark current but the same
cathodic responsivity.
If background signal shot noise is limiting, the
We will use the simpler definition of the SIN (ESIu,); SIN is proportional to i,iii2. The SIN can be improved
all the foregoing SIN expressions can be converted to by adjusting experimental variables to increase the fore-
this alternative form by multiplying all noise variances going ratio.
due to the blank by 2. The ramifications of equation
5-31 can be better understood by considering several Nonfundamental Noise-Limited Case. If dark
limiting cases. current and background signal shot noise are negligible,
the SIN is given by
lank-Noise-limite Expressions
The SIN can be improved by increasing is or by reducing ground flicker noise, excess dark current noise, or am-
the dark current signal or noise as noted previously. In plifier-readout noise becomes dominant.
a more general sense, any detec
ma1 detector) can be characteriz
IT,. One measures the total rms
output o,,when the detector is no e analytical signal flicker
and subtracts the noise contribution of the signal pro- han signal shot noise be-
cessing electronics [u, = (u;, - m&)112].In infrared
spectroscopy, the detector noise from thermal detectors signal flicker noise limit is r
is often dominant at all analytical signal levels. 5-31 reduces to
Detector noise characteristics can also be ex-
pressed as a noise equivalent power (NEP or @), or
detectivity ( D ) , as discussed in Section 4-4. The NEP
is a useful figure of merit because it indicates the min-
imum radiant power that can be detected. If the radiant Mere the SIN can be improved by reducing the noise
power incident on the detector equals the NEP, bandwidth or by adjusting experimental conditions to
SIN is unity. The detectivity D , the inverse of the N reduce the magnit e of the signal flicker factor. The
is higher for detectors with less noise and represents dependence of (u or 5 on Af must be determined
the theoretical SIN for a photon flux of 1 W on the experimentally; in general, the improvement is less than
detector, if only detector noise is significant. The D:" for shot noise (SIN improves as " where x < i).
value is just the detectivity normalized to a noise equiv- Often, 5 has only a minor dependence on experimental
alent bandpass of 1 Hz and an area of 1 cm2. Often variables and significant improvements can be achieved
detector noise is proportional to area, so that D* allows only through substantial redesign of the spectrometer.
different types of detectors with different areas to be
compared. Typical values of D" are found in Tables
4-5 and 4-6.
For NEP calculations, the detector and signal pro- The dependence of the SIN on the analytical signal
cessor gain factors must be taken into account since the (expressed as a photocathodic current) under different
NEP is usually defined as o,lR(h), where u, is the rms conditions is shown in Figure 5-6. The curves also rep-
dark noise before gain. For a P T limited by shot noise resent plots of SIN versus analyte concentration in the
in the thermal dark current, the detector (dark cur- linear portion of the calibration curve and are calculate
rent) noise in the readout voltage equals (mCKE,)1'2. from the equation
Thus 5, referenced to the photocathode equals (Ki,)lI2,
NEP = (Ki,)1'21R(X), D = R(X)/(Kid)'12, and D* =
R(X)A1'21[2e(l + a)i,ll". If excess dark current noise
is important, a plot of D vs. the P T bias voltage can
be used to choose the optimum bias voltage (maximum for different values of 5 and a,, (normalized to the
Dl. otocathode) with Af = 1 Hz and a = 0.3. Curve a
ashed line) represents t e dependence of SIN on is if
only signal shot noise is p sent. Thus curve a indicates
the highest SIN obtainable for a given photocathodic
current at the noise equivalent bandpass chosen; the
Eventually, as is is increased by changing experimental log-log slope is 9 2 because of the square root depend-
conditions or by increasing the analyte concentration ence of signal shot noise on signal. The range of pho-
until i, > i, and a, > a,,, signal shot noise or the Ki, tocathodic currents plotted (10-I' to 1W8A) represents
component in equation 5-31 dominates, and we have the typical range of signal levels expected in most emis-
sion and luminescence measurements.
In curves b to e, blank andlor signal flicker noise
is significant or dominant co ed to signal shot noise
at certain signal levels. Th ence of signal flicker
Here we see that the SIN is proportional to the square noise or blank noise reduces the SIN to a value below
root of the analytical signal and also proportional to that achievable if only signal shot noise is present. Curve
K-lI2 or Af-lf2. This holds true until the noise band- b illustrates that increasing the analyte concentration
width is reduced to the point that either signal or back- from the detection li it to higher concentrations gives
Chap. 5 1 Signal-to-Noise Ratio Considerations
ance is thus given by in measuring E,, and E,,, but the approximation is good
to within a factor of 2lI2.
a, -
0.430, - - The noise in Es,is due to signal flicker noise, signal
A TA TIn T shot noise, and 0% T noise, as shown by
A T UT uA WT UA UT 'JA
of signal shot noise to uA increases as T-lI2, and the In a real instrument a combination of the different
contribution of 0% T noise to UA increases as T-l. This limiting noise sources determines the dependence of
occurs because u, is proportional to o,,/T (equations u,/A on A , as shown in curve d of Figure 5-7. As T
5-44 and 5-49), due to the logarithmic relationship be- decreases (A increases), E, decreases, so that signal
tween A and T. flicker noise becomes insignificant compared to the sig-
The dependence of uAIA shown in Figure 5-7 (plots nal shot noise. Eventually, the signal shot noise is in-
of equation 5-50) is seen to be more complex because significant compared to the 0% T noise. Thus the
u, is constant or increasing and A is increasing as T u,/A curve is between curves a and c in Figure 5-7 (for
decreases. In all cases u,/A initially decreases from its a given value of uStat T = 1).
value at T = 1 (A = 0) because A increases faster than If we divide equation 5-50 by mG to express the
uA increases. In the case of the 0% T noise limit (curve reference signal in terms of the photocathodic current,
a), o, increases faster than A increases above A = 0.4.
For the signal-shot-noise-limiting case (curve b), the
absorbance at which a, increases faster than A (o,lA
increases) is larger (A = 0.9), the precision is better at
a given absorbance, and u,/A increases at high absorb-
ances at a slower rate than for the 0% T noise-limiting
case. Finally, for the signal-flicker-noise-limiting case Thus we see that aA/A for a given K is strongly de-
(curve c), uA/A continually improves because o, is con- pendent on the magnitudes of i,, t,, and a,,. We also
stant. note that m, G, or the radiant power incident on the
sample container is usually adjusted so that E, = Ef,,
where Efs is the full-scale setting of the readout device
(100% T voltage). Thus mG = E@,. Substitution of
this relationship into equation 5-51 yields
0% T Noise-Limited Expressions
Expansion of the noise terms yields in which case u,lA continually improves with increasing
A (if (, is independent of T ) until the point is reached
that signal shot or 0% T noise become limiting. The
sample flicker factor is given by
where (a,,), = (mGKEbE)la= mG(KibE)la,(u,,)~ =
xEbE = xmGibE,and (u,), = (mGKEd)lD= mG(KiJ1".
Thus, for a given T under 0% T noise-limiting condi- Here 5, is the sour r factor or the RSD of the
tions, the precision can be improved by reducing the source spectral rad er the measurement band-
relative 0% T noise or uOtIEf,through reduction of a,, width due to flicker noise. The flicker factor 5, is the
for a given i, or by increasing i, for a given IT,,.A higher Ee container transmission icker Bctor or the RSD
value of i, allows reduction of G, which reduces the of the transmission of the sample container due to fluc-
relative contribution of background emission shot and tuating transmission properties independent of the an-
flicker noise, detector or dark current noise, and pos- alyte. (It is sometimes called the convection or cell po-
sibly amplifier noise if it is dependent on G . Similarly, sitioning flicker factor.) Thus u,/A can be improved at
if i, is increased, m can be reduced, which reduces the <,
a given T by reducing through use of a more stable
contribution from background emission noise and dark light source or double-beam compensation, or by re-
current shot noise. When 0% T noise is predominantly ducing <, through better control of the factors influ-
readout or quantization noise, equation 5-53 can be encing it.
written as
Signal-Flicker-Noise-Limite
For measurements that are signal flicker noise limited, where IT;,is the 0%, T noise at the input of the loga-
equation 5-52 reduces to rithmic circuitry, and a:, is the rms noise due to the
logarithmic conversion circuitry or the readout device.
The shot and flicker noise and E, are also referenced
to the input of the logarithmic circuitry. If a:, is the
154 Chap. 5 I Signal-to-Noise Ratio Considerations
limiting noise source, equation 5-59 simplifies to If signal carried noise is limiting (a, >> o,,),one
must determine which component (signal shot or signal
a,- d, flicker noise) dominates by comparing the noise in the
A Ak' analytical signal (cr,) to the signal shot noise calculated
from the appropriate form of the Schottky equation.
Note that under these conditions, a,IA continues to Alternatively, the dependence of cr, on the analytical
improve as A is increased as for the signal-flicker-lim- signal in the analyte concentration range of interest
ited case since the standard deviation in absorbance is indicates if signal shot noise (a,x i:l2)or signal flicker
independent of A . When the logarithmic conversion is noise (as x is) dominates. Similar techniques can be
performed by computer software, the SIN expressions applied to absorption measurements where a,,is first
for transmittance readout still apply. If round-off errors compared to o,, (light source blocked) to determine if
in the software are negligible, the precision can be lim- lamp signal carried noise or noise independent of the
ited by the resolution of the analog-to-digital converter lamp signal (0% T noise) dominates.
used to measure E,, and a,,in equations 5-52 and 5-53 Once the limiting noise sources are identified, ex-
is determined by the resolution of the ADC. perimental conditions can be changed to optimize the
In Chapters 10, 13, and 14, the SIN expressions SIN. It must be remembered that one is often optimizing
for absorption are considered further as we discuss de- the SIN for a particular analyte concentration range;
tection limits, optimization methods, and the precision the SIN may be decreased for a different analyte con-
of specific absorption techniques. centration range. Also, conditions that optimize pre-
cision may have adverse effects on linearity or on in-
terferences.
5-6 SIGNAL-TO-NOISE RATIO
ENHANCEMENT TECHNIQUES
If precision is limited by electrical noise, the relative We have already discussed the techniques of low-pass
amount of noise can be decreased (SIN enhancement) filtering and integration to enhance the SIN for dc sig-
to the point that other factors limit the precision. It is nals. These techniques have two major limitations: the
crucial to identify the SIN expression applicable to a noise bandwidth extends to 0 Hz (dc), which is the
given type of spectrometric measurement and to the region of greatest llf noise, and measurements are lim-
specific instrument and experimental conditions used; ited to dc or slowly changing signals. Because filtering
this allows one to pinpoint the limiting noise sources. a signal can be considered as a process of weighting the
As discussed in Sections 5-4 and 5-5, the strategy data (convolution), filtering can be readily carried out
for SIN optimization depends on the limiting noise by digital techniques in hardware or in software. Two
source(s). Thus identification of these source(s) is the general schemes are employed: time-domain filtering
first step in carrying out a systematic SIN optimization. (see later in this section) and frequency-domain filter-
Measurement of the noise in the readout signal under ing.
a few sets of conditions can quickly identify its source. In the frequency-domain approach, the data are
For example, consider emission or luminescence meas- digitized at a high-enough rate to follow the signal
urements. Measurement of the rms noise or SD in the changes. The Fourier transform of the signal is taken
total signal (a,) for an analyte solution and then a blank to convert the information from the time to frequency
(a,,) indicates if o, is due to blank noise (a, = a,,) or domain, and then multiplied by the desired frequency
signal-carried shot or flicker noise (a, >> a,,). The response I H ( ~ ) of the filter. An important theorem of
blank-noise-limiting case occurs for low analyte con- Fourier transformation states that multiplication in the
centrations where the analytical signal is small. Note frequency domain is equivalent to convolution in the
that if the blank does not contain concomitants which time domain. The inverse Fourier transform of
contribute significantly to the blank component of the the product then yields the filtered signal in the time
total signal and noise, the blank noise measurement domain. Essentially, any frequency response desired
does not reflect the true blank noise in the total signal. can be programmed. Filters with no phase shift, higher-
If the blank signal noise is dominant, blank noise can order filters, sharp cutoff filters, and unique bandpass
be compared to the total dark current noise (o,,), meas- filters are all readily implemented. Digital filtering in
ured with all radiation to the detector blocked, to as- the frequency domain is analogous to smoothing in the
certain if background signal noise is dominant (a, = time domain (see later in this section). The frequency-
a,, >> udr)or if dark current or amplifier-readout noise domain approach is more versatile and allows visual-
is limiting (a,, = a,,). ization of the filtering operation.
See. 5-6 1 Signal-to-Noise Ratio Enhancement Techniques
above, but can be significant if noise from thermal emis- ing the potential SIN advantage provided by modulation
sion at the dynodes or excess dark current noise is sig- techniques. Additive noise is independent of the ana-
nificant. With photon counting, amplifier-readout noise lytical signal and includes detector noise, amplifier noise,
is essentiaily zero, and the signal and noise due to sources and background signal noise (0% Tin absorption meas-
of dark current other than photocathode thermal emis- urements). Multiplicative noise is carried by the ana-
sion are significantly reduced. The SIN advantage of lytical signal. Although signal shot noise is analytical
photon counting is particularly significant when long signal-carried noise, multiplicative noise usually refers
integration times are required to accumulate enough only to analytical signal flicker noise that is directly
signal to obtain a reasonable SIN. Photon counting is proportional to the signal.
less susceptible to llf noise and to slow drifts in dc offset Signals and noise are carried by the modulation
levels, in amplifier gains, and in photomultiplier gains. waveform if they are present only during the on-cycle
of the modulation waveform. Thus the noise and the
Modulation Techniques signal carried affect the amplitude or appear as side-
bands of the modulation waveform. Multiplicative noise
Amplitude modulation techniques translate the signal is always carried by the modulation waveform because
information to a higher-frequency region by encoding it is present only during the on-cycle of modulation
it as the amplitude of a carrier waveform. This process when the analyte is probed or viewed. Background sig-
can move dc and low-frequency signal information out nals and noise are not carried if they are present during
of the region of high background llf noise into a region the on and off cycles of the modulation waveform. They
of lower noise, as shown in Figure 5-8. In spectrometric contribute to the magnitude of the baseline but not to
systems, the techniques of source modulation. sample the amplitude of the ac waveform. Additive noise may
modulation, or wavelength modulation are used to or may not be carried by the modulation waveform.
translate the signal information to higher frequencies, Dark current and amplifier noise are always additive
as discussed in Section 4-5. noises that are not carried by the modulation waveform.
The type of modulation in a given spectrometer tech-
SIN Advantages. It is useful to define the terms nique determines which types of background signals and
additive and multiplicative noise as an aid to understand- additive background noise are carried by the modula-
tion waveform.
Modulation improves the SIN when low-frequency
additive noise that is not carried by the modulation
waveform is limiting, and the analytical signal is moved
to a frequency region where the additive noise in the
interval Af is less (i.e.. the additive noise has lif char-
acter). No SINimprovement results when multiplicative
noise or carried additive noise dominates because the
noise originally at 0 Hz is moved to f,.
In emission experiments with source modulation
(e.g., a mechanical chopper), both the background and
analyte emission signals are translated to the modula-
Frequency tion frequency; the background and analyte emission
FIGURE 5-8 Advantage of modulation. Mod- flicker noise are carried. No SIN advantage results in
ulation is used to encode the signal information this case unless l I f noise in the dark signal or amplifier
at the modulation frequency f,,.The background noise is limiting. As discussed in later chapters, sample
noise observed is less at frequency f,,, (region B) and wavelength modulation techniques can discriminate
than at dc frequencies (region A) for the same against lif background emission flicker noise. With these
noise equivalent bandpass (Af). The rms noise is modulation techniques, some types of background
proportional to the square root of the area under emission signals are present during the on and off cycles
the NPS defined by hf.To discriminate against of modulation, and hence the lif noise at 0 Hz in these
l i f noise, the signal processor is adjusted to re-
background emission signals is not moved to f,.
spond only to the signal encoded at f, and the
noise in the bandwidth centered around f,. It is In photoluminescence experiments with modula-
important to choose f, to be high enough that tion of the excitation source, neither the analyte nor
the lif noise has fallen off and to be at a frequency the background emission signal is carried by the mod-
where interference noise is negligible Cf, should ulation waveform. Thus lif noise in these signals is ad-
not be in region C). ditive and is discriminated against. This applies equally
Sec. 5-6 Signal-to-Noise Ratio Enhancement Techniques
eraged. These operations can be performed without the and noise are observed for 11100 of the time in the
lif noise and drift problems from additive noise which pulsed experiment. This means that 11100 of the back-
is not carried by the modulation waveform because a ground emission signal photons or thermal dark current
high-pass or a bandpass filter is used to remove the low- electrons are observed per unit time compared to the
frequency components before demodulation. The re- dc experiment. Thus the absolute background emission
sulting bandlimited waveform is demodulated and then noise and dark current shot noise are reduced by a
low-pass filtered to convert the amplitude of the ac factor of 10 (1001'2) and the background signal flicker
waveform into a proportional dc signal. Low-pass fil- noise (that is carried) by a factor of 100. Also, in some
tering or another SIN enhancement technique can now cases, the aperture time can be delayed to measure the
be used more effectively because llf noise components analytical signal after a background signal has decayed
have been removed to leave only white additive noise. to a smaller magnitude.
With square-wave or sinusoidal modulation, a lock- Although modulation techniques usually employ
in amplifier is usually employed, as discussed in Sec- a sinusoidal, square wave, or pulse waveform, any pe-
tion 4-5. The carrier waveform is multiplied by a riodic waveform or even nonperiodic waveforms can be
reference waveform of the same frequency. This con- used. The cross-correlation function [C,,,(T)] is defined
tinually reinforces the signal amplitude information; noise as
at frequencies or phases different from the reference
waveform produces an average output of zero. The syn-
chronous demodulation step produces an ac output at
twice the reference frequency with the ac amplitude
proportional to the signal information. A low-pass filter where El(t) and E,(t) are two functions describing the
removes the ac component and determines the overall dependence of voltage on time and 7 is the delay time.
noise equivalent bandpass. Even though the lock-in am- If El(t) = E,(t), then C1,2(~) is the autocorrelation func-
plifier output averages zero for noncarried background tion [C1,l(~)].Thus correlation techniques depend on
or dark signals, the actual dc signal level (i, or id) must the relationship between a signal and a delayed version
be used to calculate the background or dark current of itself (autocorrelation) or of another signal (cross
shot noise in SIN expressions. correlation). A plot of C1,,(7) VS.T is called a correlo-
For pulse modulation waveforms (duty cycle less gram. Correlation techniques are less susceptible to im-
than 50%), such as with a pulsed laser, a boxcar inte- pulse noise since the effect of noise spikes will show up
grator is used. As discussed in Section 4-5, a boxcar at only a certain T in a correlogram.
integrator is a gated integrator. The signal is switched Autocorrelation is useful when there is no refer-
to an integrator during an aperture time controlled by ence signal or means of synchronizing a sampling system
a reference signal. Thus the aperture window can be to a signal. Only at T = 0 will random noise be cor-
synchronized to pass signal information only during the related in an autocorrelogram. At T different from zero,
actual signal pulse. The signal need not be periodic, as random noise decays to zero, but the desired signal
with a lock-in amplifier, but must be repetitive and information is enhanced at certain values of T or scanned
synchronized to a reference waveform. Typically a high- across the waveform.
pass filter is used to remove noncarried lif additive A lock-in amplifier is a single point correlator where
noise and signals prior to the integrator. As with a lock- E l is the measured optical signal and E, is the reference
in amplifier, a low-pass filter at the output can be used modulation signal with T adjusted to zero. Similarly, a
to determine Af. Alternatively, the signal information boxcar integrator is a single-point correlator, but in this
from each pulse can be summed. If n pulses of inte- case, T can be adjusted to any desired value.
gration time t are summed, the total averaging time is
nt. Thus, as a first approximation Af = li(2nt). It can Dsuble-Beam and Dual-Channel Techniques
be shown that actually, Af is equal to 0.886!(nt).
An additional SIN advantage can result from using Flicker noise and drift in signals can be reduced by
a high-intensity pulsed source in absorption, and par- double-beam or dual-channel optical arrangements (see
ticularly, photoluminescence experiments. Consider the Section 4-6), which monitor both the analytical signal
difference between a dc measurement with a continuous and another signal correlated to it. For double-beam
light source and a measurement with a pulsed light source instrumentation in absorption and photoluminescence
of 100 times greater peak intensity and a 1% duty cycle. experiments, the source radiation is split into an ana-
The signal shot noise in both experi~nentsis the same lytical beam which probes the sample, and a reference
since the number of photons observed per unit time is beam, which is viewed directly or after passing through
the same. However, the dc background emission signal the blank. These two beams are detected simultane-
Sec. 5-6 1 Signal-to-Noise Ratio Enhancement Techniques
ously with two optical transducers (double beam in- filter in dc (low-pass filter) or ac (lock-in amplifier)
space) or alternatively with a single optical transducer systems. Alternatively, n measurements of the readout
(houble beam in-time). Ratioing the sample and ref- signal from an integrating system (box-car integrator,
erence signals compensates for drift and low-frequency photon counting system) can be summed. In a broader
flicker noise due to the source since both signals track sense, the SIN or precision can be improved by making
the source intensity. This compensates for source flicker repetitive measurements of spectra, of total and blank
noise in absorption measurements and analyte lumi- signal pairs, or of sample and standard pairs or even
nescence flicker noise, background luminescence flicker by repetitions of the complete measurement scheme
noise, and scattering flicker noise due to source flicker (samples, standards, and establishment of the calibra-
in luminescence measurements. tion function).
Oftcn in double-beam in-time spectrometers, the The SIN does not always improve with n1I2.The
amplitudes of both the analytical and reference signals improvement depends on the limiting types of noise
are measured relative to a baseline established by block- and their nature, the SIN enhancement techniques used
ing the source radiation. This source modulation pro- prior to ensemble averaging, and the way measure-
vides discrimination against nonwhite noise that is in- ments are conducted. If white noise (e.g., shot noise in
dependent of the source. The chopping frequency is the analytical, background, or dark signals) is limiting,
chosen to be in a frequency region where llf additive the SIN improves with n1I2.If nonwhite noise such as
noise is insignificant. llf noise is limiting, the SIN may improve as n is in-
Particularly in emission and photoluminescence creased with the maximum improvement limited by the
measurements, the dual-channel approach can be used nl" rule. In some cases, the SIN can even decrease with
to compensate for llf noise in background signals. Here repetitive measurements.
the reference channel is adjusted to a wavelength where A related question is whether it is better to make
the analytical signal is zero but the background signal one measurement (or spectral scan) for time T or to
is the same as for the analytical channel. The signal average n measurements (or spectral scans) of time t
processor continuously calculates the difference be- where T = nt. The SIN is equivalent in these two cases
tween the analytical and reference channel signals. This if white noise is limiting. If nonwhite noise such as llf
compensates for flicker noise if the background flicker noise (drift) is limiting, the use of n repetitive measure-
noise in both channels is correlated. Note that wave- ments in time T generally provides higher SIN up to a
length modulation performs this operation in time. factor of n1I2compared to one measurement over time
None of the double-beam or dual-channel ap- T.
proaches compensate for any type of shot noise since Again, the SINenhancement observed if nonwhite
different photons are viewed by two detectors or by one noise is limiting depends on the nature of the noise and
detector during two different time periods. Thus when the procedure used. For example, consider a case where
shot noise limits measurement precision, the dual-chan- there is low-frequency noise or drift only in the blank
nel or double-beam approach can result in a reduced signal. For this situation, it is better to make 10 paired
SIN since the shot noise in the reference signal adds to 1-s measurements (difference between sequential 1-s
the total noise. integrations of total and blank signal) than one paired
10-s measurement. Note that this is effectively manual
sample modulation and that pairing blank and sample
readings discriminates against any drift or low-fre-
Time-domain filtering, signal averaging, or ense quency noise in the blank signal with a period much
averaging is accomplished by summing n repetitive greater than the time between blank and sample meas-
measurements of a signal. It can be employed for re- urements. Similarly, making many sample and standard
petitive signals and is essentially an instrumental su- measurements in time T can provide higher precision
perposition of a number of signal traces accomplished than one sample and standard measurement in time T
by recording and storing n signal traces in exactly the if multiplicative noise or flicker noise or drift in the
same way (i.e., same period of time in the signal wav- analytical signal is limiting.
eform). Thus, if a repetitive waveform is coherent, the Digital time-domain filtering, as digital frequency-
signal will increase in direct proportion to n, but the domain filtering discussed previously, can also be used
random noise will sum as n1I2to provide an increase in to smooth a signal spectrum or transient signal. Here
the SIN proportional to n1I2.This is the principle used the signal data are digitized at a high enough rate to
with boxcar integrators and signal averagers. follow a time-dependent signal faithfully with a rela-
Ensemble averaging is often used to process the tively large Af and then stored in computer memory.
signal after frequency-domain filtering with an electrical Software is then used to take a weighted average of a
160 Chap. 5 1 Signal-to-Noise Ratio Considerations
selected number of signal data points over a chosen somewhat different because many or all wavelengths
time period, which provides software control of Af. Such are simultaneously striking a single detector. For sim-
a filter window can be sequentially moved over the data plicity consider the comparison of a spectrum taken with
points selected to smooth out the data, such as in the a conventional single-channel scanning spectrometer to
case of a spectrum. The algorithm developed by Savit- the spectrum taken with a multiplex spectrometer with
sky and Golay is now widely used in digital filtering. equivalent wavelength coverage, number of resolution
Other digital filtering algorithms are gaining popularity elements n, and measurement time T. As a first ap-
in certain applications. The availability of low-cost mi- proximation, the multiplex spectrometer provides a
crocomputers has resulted in a rapid increase in the use nl/* SIN advantage if detector or amplifier noise is lim-
of digital filtering techniques. iting because the number of photons observed per spec-
tral element in time T is n times greater and the noise
Multichannel and Multiplex Systems is only n112 times greater. This is why the multiplex
approach is commonly used in infrared spectrometers
Multichannel spectrometers [direct readers or systems limited by detector noise.
based on multichannel detectors (diode arrays, vidi- If shot or flicker noise is limiting, the SIN com-
cons)] and multiplex [interferometers and Hadamard parison depends on the nature and complexity of the
spectrometers (see Section 4-6)] can provide a multiplex spectrum. Note that shot or flicker noise in the multi-
advantage by allowing many analytical wavelengths to plex spectrometer can arise from signal or background
be monitored simultaneously. Contrast a spectrum taken photons in the resolution element of interest or from
by scanning with a conventional monochromator-PMT photons in other resolution elements. If the spectrum
configuration to a spectrum obtained with a spectro- consists of only one strong line in one resolution ele-
graph and multichannel detector where the wavelength ment, a n1I2SIN advantage is realized with the multiplex
coverage and resolution are equivalent. If the spectrum spectrometer if it is signal or background shot noise
is taken over time T for both cases, the number of limited; no SIN advantage is realized if it is signal or
photons observed per spectral resolution element is n background flicker noise limited.
times greater for the multichannel detector, where n is When a significant number of photons is observed
the number of spectral resolution elements. If meas- from other resolution elements (e.g., a complex or con-
urements with both systems are limited by amplifier tinuum spectrum), the SIN advantage decreases or a
white noise, dark signal white noise, signal shot noise, SIN disadvantage (the multiplex disadvantage) results.
or background shot noise of equivalent magnitude, the For example, consider a spectrum with 10 equally in-
multichannel detector provides a n112 SIN advantage. tense lines each occupying one of 100 possible spectral
No advantage results if both detectors are limited by resolution elements. Compared to the scanning spec-
signal or background flicker noise. trometer case, the number of signal photons observed
A precise comparison is more complex since in in time T in the spectral element of interest increases
the discussion above it is assumed that the same amount by 100, but the total number of photons observed in-
of light from the sample container strikes the PMT pho- creases by 1000. This yields a SIN advantage of
tocathode or a given array element; that the detector 1001(1000)1'2 or loll2 rather than 10 under shot-noise-
responsivities, gain, and multiplication noise are equiv- limiting conditions. For the case of 100 equally intense
alent; and that the amplifier-readout and dark signal lines (a continuum) or a weak line of interest and 10
noise are equivalent. The SIN advantage provided by strong lines (10 times as intense), no SIN advantage
multichannel detectors for spectrum acquisition is usu- results with the multiplex spectrometer. If flicker noise
ally less than predicted above because of the small de- is limiting in the foregoing situation of 10 equally intense
tector element area and larger relative amplifier-read- lines, the signal increases by 100, but the noise increases
out and dark signal noise of multichannel detectors. by 10 x 100 = 1000, yielding a factor of 10 decrease
Readout noise from multichannel detectors is often in SIN. For a continuum spread across all resolution
independent of the integration time (it arises only dur- elements (or a weak line of interest plus 10 other lines
ing the readout process). Under these readout-noise- of 10 times the intensity), the SIN disadvantage is a
limiting conditions, the SIN is directly proportional to factor of 100. For the flicker noise case above, it is
the integration time until other noise sources become assumed the flicker noise for all resolution elements is
significant. Here it is better to allow signal levels to totally dependent or correlated. When the flicker noise
approach detector saturation levels than to perform n in each channel is totally independent, the SIN disad-
readouts in the same time (i.e., a n1I2 SIN advantage vantages above are reduced to 1O1I2 and 10.
results). Again a precise comparison is complex and must
The SIN considerations for multiplex systems are take into account the throughput of the device and dif-
Chap. 5 1 Problems 1
ferences in detector responsivity vs. wavelength and in The discussion above indicates the multichannel
detector noise. With a Michelson interferometer the and multiplex spectrometers provide a significant SIN
solid angle of collection, and hence the throughput, are advantage in certain situations, particularly when meas-
much larger than for a monochromator with an entrance urements are limited by detector noise. In the case of
slit. However, the SIN advantage is slightly reduced a line spectrum consisting of a few lines of interest, the
(factor of 21J2to 81iz) due to such factors as light loss at SIN with a single-channel spectrometer can be improved
the beam splitter. For a Hadamard transform spec- by slew scanning between the analytical lines so that
trometer, only a fraction of all spectral elements are most of the measurement time is spent at the analytical
viewed simultaneously due to the design of the slit wavelengths of interest.
5-1. A PMT, op amp I-Vconverter combination was used (d) Determine the effect of i, and E,, on the results in
to monitor the emission of 10 ng mL-' Na in a flame. parts (b) and (6).
The PMT current gain was 1.0 x loh, the feedback
resistor was 1.0 x 10' 0,the secondary emission factor In this problem, the SIN characteristics of a PMT, I-
was 0.30, the analyte emission flicker factor (5,) was V converter configuration are compared to a PT, I-
0.50 x the background emission flicker factor V converter configuration. Assume that the back-
(x)was 2.0 x the time constant was 0.50 s, the ground signal noise is negligible, as well as amplifier
amplifier-readout noise was 1.0 x V, the analyte noise, readout noise, and excess dark current noise.
emission signal was 1.0 V, the background emission Thus the only noise sources to consider are signal and
signal was 0.10 V, and the dark current voltage was dark current shot noise, signal flicker noise, and John-
0.0010 V. Assume that the excess dark current noise son noise. Assume that the Johnson noise is due solely
is negligible. to the feedback resistor of the op amp I-V converter
(a) Calculate the following. Here S is always taken as and that the resistance is 1.0 x lo6 0 (25•‹C).Assume
the analytical signal and all noises are rms noises. a 0.25s time constant, a PMT gain of 1.0 x lo6, a
(1) Af, K, and mGK. cathodic dark current of 1.0 x 10-Is A, a collection
(2) Dark current shot noise. efficiency of 1, a signal flicker factor of 1.0 x
(3) Analytical signal shot noise. and a = 0.30.
(4) Background signal shot noise. (a) Write a SIN expression for each configuration ref-
(5) Analytical signal flicker noise. erenced to the output of the op amp.
(6) Background signal flicker noise. (b) Calculate the photocathodic current at which the
(7) Total noise and SIN for measurement of the SIN = 1 for each configuration. What is the lim-
analytical signal. iting noise at this point?
(b) What are the dominant noise sources for this meas- (c) At what photocathodic current do measurements
urement? become signal flicker noise limited (i.e., two-thirds
(c) Determine the effect of Af, m , a , <,, and x on the of the total noise) for both configurations?
results in part (a). (d) Under what conditions would you use each con-
5-2. In molecular absorption, the relative precision (a,iA) figuration?
depends on the absorbance, the photocathodic current Calculate the rms Johnson noise for a lo8-0 resistor
employed, the signal flicker factors, and the 0% T at 25•‹Cwith a 1-Hz noise equivalent bandwidth.
noise. The reference photocathodic current can vary What specific types of noise becoming limiting with
from about 1.0 x 10' l2 to 1.0 x A depending large time constants or long integration times?
on the wavelength of analysis, the radiance of the lamp, Show that the noise equivalent bandpass for an inte-
the slit width employed, and the Fin and optical ef- grator equals (2t) - I .
ficiency of the monochromator. For this problem as-
Why is signal flicker noise often limiting at high con-
sume that E, = 2.0 x a,, = 4.0 x V, 100%
centrations in emission and luminescence measure-
T or full scale on the readout device (E,,) is 1.00 V,
and a 0.50-s integration time, a = 0.30, and i, = ments but at low concentrations in absorption meas-
urements?
1.0 x lo-" A.
(a) Calculate Af, K, and mGK. If the standard deviations in the blank and sample
(b) Calculate uA/A at A = 1 and 2 and a, at A = 0. measurements are quite different, what are the pos-
(c) Indicate the limiting noise sources at each absorb- sible limiting noise sources?
ance. When are modulation techniques advantageous?
162 Chap. 5 I Signal-to-Noise Ratio Considerations
In an emission experiment the analyte concentration each of the following types of noise is limiting: signal
is increased by a factor of 10. Identify the possible shot noise, background shot noise, signal flicker noise.
limiting noise sources for the following conditions. 5-18. In an atomic emission experiment, the analyte emis-
(a) The noise in the total signal increases by a factor sion signal, background emission signal, and SIN are
of 10. measured before and after the slit width is doubled.
(b) The noise in the total signal increases by a factor The increase in slit width causes the analytical signal
of 10"'. to double and the background signal to quadruple. For
(c) The noise in the total signal remains constant. each of the cases below, indicate what types of noise
Why is it imprecise to say that measurements are lim- are limiting.
ited by shot noise? (a) The SIN does not change.
A photomultiplier tube is operated under illumina- (b) The SIN improves by a factor of 2 l I 2 .
tion conditions that produce a photoanodic current of (c) The SIN improves by a factor of 2.
6.5 x A. Repeated measurements of the pho- (d) The SIN decreases by a factor of 2.
toanodic current give a standard deviation of 1.61 x 5-19. A series of noise measurements are made in a pho-
10-l3 A. When the illumination level is reduced by a toluminescence experiment. From the information given
factor of 10, the standard deviation in the photoanodic in each step, indicate the types of noise that are not
current is 5.1 x lo-'". Is the limiting source of limiting in the measurement of the sample signal.
noise in the measurement source flicker noise. pho- (a) The noise in the blank signal and total sample
tocurrent shot noise, or dark current shot noise? signal are approximately equal.
What types of noise must be limiting in a photolu- (b) The PMT shutter is closed and the noise decreases
minescence measurement for source modulation to im- by a factor of 10.
prove the SIN? (c) The noise in the blank signal increases by a factor
The rms noise in a signal is measured to be 2.0 mV. of 2 when the excitation radiant power is doubled.
The resolution of the readout device is 1 mV. Does 5-20. For a photon counting measurement system and a ther-
quantization noise make a significant contribution to mal detector radiant power monitor, determine the
the total noise? detector dark noise, the noise equivalent power, and
the minimum detectable radiant power (SIN = 2) in
In an absorption measurement, the RSD in the ab-
watts and photons per second at 600 nm with a noise
sorbance continually improves as the absorbance is
increased from I to 2. What type of noise is limiting? equivalent bandpass of 1 Hz. For the photon counting
system, assume that the photocathode quantum effi-
If signal shot noise is limiting, what instrumental var- ciency is 10% and that the dark current pulse rate is
iable can be changed to improve the SIN? 100 s-'. For the thermal detector, assume a typical
A spectrum is obtained with a multichannel spectrom- detectivity of 1.0 x lo9 W-I limited by detector ther-
eter with 500 pixels and with a scanning spectrometer mal noise and a responsivity of 10 V W-I. What in-
with equivalent wavelength resolution. With the for- cident radiant power would be necessary for the signal
mer, an integration time of 10 s is employed, while shot noise to be equal to the thermal detector noise
the scan time with the latter is 100 s. Determine the with the thermal detector?
SIN advantage of the multichannel spectrometer when
REFERENCES
1. W. J. McCarthy, "The Signal-to-Noise Ratio in Spec- 5. K. D. C. MacDonald, Noise and Fluctuations, A n In-
trochemical Analysis," appendix in Spectrochemical troduction, Wiley, New York, 1962.
Methods of Analysis, J . D . Winefordner, ed., Advances 6. A. Van der Ziel, Noise in Measurements, Wiley, New
in Analytical Chemistry and Instrumentation Series, vol. York, 1976.
9, Wiley-Interscience, New York, 1971.
7 . W. R. Bennett, Electrical Noise, McGraw-Hill, New York,
2. C. Th. J. Alkemade, G. D . Boutilier, et al., "A Review
1960.
and Tutorial Discussion of Noise and Signal-to-Noise
Ratios in Analytical Spectrometry," Spectrochim. Acta, 8. M. S. Epstein and J. D . Winefordner, "Summary of the
33B, 383 (1978); 35B, 261 (1980). Usefulness of Signal-to-Noise Ratio Treatment in An-
3. G. M. Hieftje, "Signal-to-Noise Enhancement through alytical Spectrometry, Prog. Anal. At. Spectrosc., 7(1)
Instrumental Techniques," Anal. Chem., 44, 81A (May (1984). This comprehensive article also contains a very
1972); 69A (June 1972). complete reference list.
4. R. King, Electrical Noise, Chapman & Hall. London, 9. J . D . Ingle, Jr., and S. R. Crouch, "A Critical Com-
1966. parison of Photon Counting and Direct Current Meas-
Chap. 5 1 References 3
case of solutions, the same solvent is used for analytical solute methods can be in error for the same reasons as
samples and external standards (e.g., aqueous external relative methods based on calibration standards except
standards are used for analysis of water samples). there are no errors involving preparation and measure-
In some cases one can use a real sample as an ment of standards. However, the error due to the dif-
external standard for other samples if the analyte con- ference between the true calibration function and the
centration in the calibration sample has previously been precalibrated calibration function must be considered.
determined. The National Bureau of Standards (NBS)
and other organizations provide many types of real sam-
ples (e.g., geological, biological) in which the concen-
tration of many species is certified. Thus one can choose
as a standard a calibration sample with a matrix similar
to the analytical sample. The calibration sample can be A systematic error (see Appendix A) in a determination
used to check the suitability of the technique and any is an error in which the analyte concentration deter-
external standards prepared from purified chemicals. mined in a given sample is either too low or too high
To determine the analytical signal, a blank meas- compared to the true analyte concentration regardless
urement is required. The ideal blank is identical to the of the number of repetitive measurements made on the
sample except that the analyte concentration is zero. sample. Systematic errors can be classified as matrix
Real blanks are usually solvent or reagent blanks. The errors, calibration errors, sample acquisition and prep-
same solvent in which the sample is dissolved is used aration errors, or measurement errors. Any classifica-
. A reagent blank contains the solvent tion is somewhat arbitrary since certain types of errors
plus all reagents used in the sample preparation pro- have characteristics of two or more classes. A systematic
cedure at the same concentrations as in the analytical error results in an error in determination only if the
sample. magnitude of the error is different for samples and
The basic assumption of the external standard pro- standards.
cedure is that sample and standard of the same analyte
concentration (c, = c, = c,) will yield the same ana-
lytical signal. Thus, for an accurate determination, the
calibration function established from the standards must
apply equally to the sample. There will be an error in Matrix errors arise because of differences between the
the determination (c, determined is incorrect) if this matrix of the analytical sample and the matrix of
basic assumption is not true. the external standards or blanks used for calibration.
Several factors can be responsible for the break- The analytical sample matrix can be different because
down of the basic assumption. Matrix effects, due to of the matrix of the original bulk sample or because of
the presence of species in the analytical sample but not species added or removed during sample preparation.
in the standards or blank, can alter the sample analytical If a technique has perfect specificity, the difference be-
signal relative to that from a standard of the same an- tween ideal and real standards or blanks does not cause
alyte concentration. Thus significant differences be- an error.
tween an ideal and real standard or an ideal and real Often matrix errors are called interference errors
blank manifest themselves in the sample presentation since they are due to the presence of concomitants or
or measurement steps. Differences in the magnitudes interferents in the analytical sample. An interferent is
of experimental variables (instrumental, physical, or a species or material (e.g., particle) in the sample,
chemical) at the times of sample, standard, and blank standard, or blank which affects the mean value of the
measurements, random errors, or noise can alter the analytical signal determined. Interference effects are
analytical signals determined and the validity of the often quantitatively described by indicating the error in
calibration function established. Even if the basic as- determination caused by the specified concentration of
sumption is correct, there can be error in the deter- the concomitant at a particular analyte concentration
mination if there is loss or gain of analyte (contami- under defined experimental conditions. For example,
nation) in the sample acquisition or preparation steps. the interference effect of vitamin B, on the determi-
In a few cases, calibration with standards is not nation of vitamin B, might be stated as follows: The
employed and absolute measurements are made. This presence of l o W 6M vitamin B, causes the determined
implies that the instrument is precalibrated and that the concentration of l o W 8M vitamin B, to be high by 1 %
calibration function, and hence the calibration curve, (a 1% positive error). atrix errors or interferences
are known so that the analytical signal determined for can be subdivided into blank and analyte interferences
a sample is converted directly to concentration. Ab- as discussed below.
166 Chap. 6 / Methodology in Spectrochemical Analysis
Blank Interferences. A blank interference comitant in the sample that is not present in the stand-
(sometimes called an additive interference) is due to a ards. although it could be caused by improper prepa-
species or material which produces an uncompensated ration of the standards such that they contain species
signal independent of the analyte concentration. A blank not in the analytical sample. If the blank interferent is
interference causes an error in determination if the blank absent from the blank, but equivalent in the standard
component of the total signal measured for the sample and sample, the analytical signal extracted from the
is different from the blank component of the total signal total signal for the standard and sample is incorrect,
measured for the standards. This is shown in Figure 6- but no systematic error in determination results. If the
1. species responsible for a blank interference is present
Usually, a blank interference is caused by a con- in all the standards at the same concentration, the cal-
ibration curve exhibits a nonzero intercept.
Many blank interferences in spectrochemical
methods are spectral interferences. A spectral interfer-
ence is caused by an overlap within the monitored wave-
length range (e.g.. the spectral bandpass) of the emis-
sion, luminescence, scattering, or absorption spectral
profile of the analyte with one or more such profiles
due to concomitants in the sample. An interference
effect can also result if stray radiation effects cause the
background radiation from concomitants to appear in
the monitored wavelength range. A blank spectral in-
terference causes the analytical signal determined to be
too high or too low if the blank measurement does not
FIGURE 6-1 Systematic errors caused by blank compensate for background spectral signals or for ab-
and analyte interference. The total, blank, and sorption by the concomitants in the sample. Specific
analytical signals are shown for a given analyte types of spectral interferences are summarized in Table
concentration with (a) no interferences, (b) a blank 6-1.
interference, and (c) an analyte interference. In Some blank interferences are not spectral inter-
(a) subtraction of the blank signal EL, from the ferences. A nonspectral interference is caused by a spe-
total signal E, yields the proper analytical signal
cies or substance which itself does not necessarily emit
E,. For (b) the total signal Ei is larger due to a
larger blank signal component E,,. If the blank or absorb radiation over the wavelength range moni-
in (a) is used, the analytical signal E: determined tored, but which affects the spectral signals from other
is too high and a blank interference results. In species. A nonspectral blank interference effect occurs
(c), E; is too low and hence the magnitude of if a concomitant depresses or enhances the background
E: is too low because an analyte interference has spectral signal from the sample container or another
suppressed the analytical signal. concomitant in the sample.
TABLE 6- 7
Types of blank spectral interferences
fitting the calibration curve. The determination of the centration in the analytical sample is not representative
analyte in the sample can be no more accurate than of the analyte concentration in the bulk sample. This
the accuracy with which the analyte concentrations in can occur if the original sample acquired is not repre-
the standards are known. The accuracy of the standards sentative of the bulk sample or if some of the analyte
is determined by the accuracy of the gravimetric and is lost during sample acquisition, storage, or prepara-
volumetric techniques and equipment employed. For tion. Also. the analyte concentration determined can
the highest accuracy work, temperature and pressure be too high, due to contamination in the steps noted
effects on mass and volume measurements must be con- above. The accuracy of volumetric and gravimetric op-
sidered. The purity and stability of materials used to erations must be considered carefully as mentioned for
prepare the standards must be confirmed. standards.
The chemical form of the standards should be Systematic errors can arise during sample pres-
identical to that of the analyte since the state of oxi- entation and measurement of samples, standards, and
dation, complexation, or isomerization of the analyte blanks due to numerous factors, including "cockpit er-
could influence the analytical signal. Often, the chem- ror" (i.e., blunders). Drifts in critical experiment var-
ical speciation of the analyte must be considered. Thus iables (e.g., T, pH) that affect the analytical or blank
one must be careful to designate exactly what is meant signals cause errors if the magnitude of these variables
by the "analyte." Consider the determination of chro- is significantly different between sample, standard, and
mium. In one case, total chromium might be of interest, blank measurements. If drifts occur in the blank signal,
while in another determination. the amount present as standard and blank measurements or sample and blank
Cr(II1) or Cr(V1) might be important. If Cr(II1) is to measurements should be paired and made close to one
be determined, is it the total Cr(II1) or just the Cr(II1) another in time. If drifts in the analytical signal are
that is complexed with a specific ligand that is of in- significant, sample measurements should be bracketed
terest? Such chemical speciation information is very by standard measurements.
important because the chemical form of an element
affects its biological activity, transport, and environ-
mental impact. 6-3 RANDOM ERRORS IN
Once the standards are prepared, care must be SPECTROCHEMICAL MEASUREMENTS
taken, especially for trace analysis, to ensure that the
analyte concentration in the standard is the same at the It is important to consider the precision of measure-
time of measurement as when it was prepared. During ments since random errors also affect accuracy. Ran-
storage, the analyte concentration can decrease due to dom errors are caused by fluctuations in the magnitudes
decomposition, volatilization, or adsorption onto con- of experimental variables or by the particle nature of
tainer walls. The analyte concentration can be higher light and matter; they result in the measured values
than expected due to contamination. Contamination being distributed around the mean value. The relative
arises from desorption of the analyte from surfaces of uncertainty due to random error decreases with the
the apparatus used to prepare or transfer the standards number of measurements averaged since positive and
to the sample container or of the sample container itself. negative detiations tend to cancel. Some knowledge of
Dust particles and reagent impurities are additional statistics is required to assess the influence of random
sources of contamination. error, and Appendix A should be consulted for review
Systematic errors can also result from bias or im- purposes.
proper fitting of the calibration curve to the data as can
occur when using a straight-line least-squares program etermination of Standard Deviation
on nonlinear data. Enough standards must be measured in Concentration
to ensure that the calibration curve is well established
since usually the analytical signal for the sample is found The magnitude of the random error is obtained by mak-
by interpolation between the analytical signals from the ing n repetitive determinations of the analytical signal
standards. for a given sample or standard. From these measure-
ments, the mean $ and standard deviation ss of the
Sample Acquisition, Preparation, analytical signal are calculated. The magnitude of s, in
and Measurement Errors a given technique can be related, with propagation of
uncertainty mathematics, to the magnitude of the stand-
The accuracy of a determination is limited by the in- ard deviation (SD) for each of the measurements used
tegrity of the analytical sample presented for measure- to extract the analytical signal as shown in Table 6-2.
ment. Systematic errors can result if the analyte con- With these equations, one can estimate the SD for de-
Sec. 6-3 / Random Errors in Spectrochemicat Measurements 169
TABLE 6-2
Relationships with measured component signals and the determined analytical signal
Means and standard Calculation of Calculation of standard
Component signals deviation of mean analytical deviation for determining
Technique measured component signals signal analytical signal
- - -
Emission or E,, Ebk E,, Ebk Es = E, - Ehk ss = (s: + stk)'I2
luminescence
Absorption with
transmittance
readout
termining the analytical signal from repetitive meas- uncertainty of the results (see Appendix A). For nor-
urements of each of the signal components (e.g., n mally distributed data, the interval defined by the equa-
measurements of the blank signal followed by n meas- tion
urements of the total signal) to evaluate if measurement
of a particular signal component limits the precision.
However, this procedure can yield too high an estimate
of s, if significant drift occurs during repetitive meas- encloses the true concentration pc with a confidence
urements of a given signal component. Thus one usually level of (1 - 2a).Where sc is the SD for an individual
makes n repetitive determinations of the analytical sig- measurement of the analyte concentration calculated
nal (all component signals measured in each repetition), from the SD of the measured analytical signal for the
since this reflects the manner in which measurements sample (equation 6-4) and t, is the Student t statistic
are normally conducted and minimizes drifts in the val- (see appendix A). If more than 30 measurements are
ues of variables which affect all signal component meas- made to determine the SD, the z statistic can be sub-
urements. stituted for the t statistic. The confidence level chosen
The SD for determining the analytical signal can depends on the situation, but is typically in the range
be used to determine the uncertainty or SD in the an- 90 to 99%. Often data are reported as the mean
alytical concentration. Application of propagation of ? SDIn112,which corresponds to a confidence level of
uncertainty mathematics to the calibration function only 68% for n 2 30 or less if n < 30 (50% for n = 5).
[S = f(c,); see Chapter 11 yields ss = (dfldc)sc or It must be stressed that the interval defined by
equation 6-5 encloses the true value with only the con-
fidence level chosen and only if random errors described
by the normal distribution are dominant. A systematic
error could cause an error much larger than t,sclnl".
where (dfldc), is the slope of the calibration curve at In some cases, one may question whether the con-
the analyte concentration of interest and c is understood centration determined is significantly different from an
to be the analyte concentration c,. accepted value (pc) (such as for analysis of an NBS
certified material). Here the experimental value o f t (or
Statistical Statements z if n r 30) is calculated from the equation
If the statistical distribution of the data is known, one
can make quantitative statements about the random
170 Chap. 6 1 Methodology in Spectrochemical Analysis
assuming normally distributed data. The largest value sample acquisition or preparation are more difficult to
oft, that is still less than t is found in a probability table assess; they result from inhomogeneity of the bulk Sam-
(see Table A-2). From the value of a corresponding to ple and random variations in analyte concentration due
this t,, one can say that the difference between the to contamination or losses. Random errors due to vol-
experimental mean and the accepted mean is not totally umetric and gravimetric operations in these steps can
due to random error with a confidence level greater be predicted more quantitatively.
than (1 - a ) . If (1 - a ) is large (typically e 90%), Equation 6-4 does not account for random error
one suspects a significant systematic error, although in the calibration function due to imprecision in the
there is still a probability a that the difference is due preparation, presentation. and measurement of the
to random error, and in any case, the observed differ- standards and the blank. The uncertainty in the cali-
ence can be due to both random and systematic error. bration function can be evaluated from the measured
More complex tests are available to account for the SDs of the analytical signals for the standards. For ex-
uncertainty in the "accepted" value. ample, with a linear calibration curve, the SD in the
slope and intercept can be evaluated. From this infor-
mation, the SD of the analytical concentration in the
Other Considerations sample can be calculated; this reflects both the SD in
determining the sample analytical signal and the SD in
The SD in the analytical signal can arise from impre- the calibration function. Calibration curve uncertainty
cision in all steps that affect its magnitude as follows: is illustrated pictorially in Figure 6-2.
Experimentally, the SD of the complete analytical
procedure can be determined from taking n samples
through the entire analytical scheme, including for each
where (s,), is the sample acquisition SD due to inho- sample the preparation of standards and a blank and
mogeneity of the starting materials, (s,), is the sample the establishment of a calibration curve. This SD can
treatment SD, (s,) is the sample placement SD, and be compared to the SD obtained with only one cali-
(s,), is the sample measurement SD. Often, the SD in bration curve determination to evaluate if calibration
the analytical signal, used to calculate the SD in the imprecision is critical.
concentration or to make statistical statements, is the
SD calculated from repetitive paired measurements on
a given analytical sample and blank. This procedure
only takes into account imprecision in the sample place-
ment and measurement steps and thus often underes-
timates the SD of the total analysis procedure. Com-
parison of t h e S D calculated f r o m repetitive
measurements under different conditions allows one to
determine what step or steps limit the precision of a
total analytical scheme. For example, if discrete sam-
pling is employed. the SD from repetitive measure-
ments on one portion of an analytical sample can be
compared to the SD from repetitive measurements of
equivalent samples (e.g., different 1-mL aliquots from
100 mL of a given analytical sample solution) to deter-
mine if sample placement imprecision is critical. These
SDs can be compared to the SD obtained from meas- FIGURE 6-2 Effect of calibration curve uncer-
urements of n separate samples from the same bulk tainty. The uncertainty in the analyte concentra-
sample each carried through the complete sample treat- tion [(s,),] is usually evaluated from the SD in
the determined analytical signal (s,) for the sam-
ment, placement, and measurement scheme to deter-
ple and the calibration function (light dashed lines).
mine if sample acquisition or treatment imprecision is The heavy dashed curved lines are uncertainty
critical. limits for a certain l e ~ e lof confidence for the
The factors affecting measurement precision and calibration curve. When the additional uncer-
sometimes sample placement precision can be calcu- tainty in the calibration curve is considered, the
lated and understood with SIN theory and measure- actual uncertainty in the determined analyte con-
ments as discussed in Chapter 5 . Random errors in centration (s,), is seen to be greater.
See. 6-4 1 Sensitivity and Detection Limit 1
TAB6E 6-3
Basic definitions of sensitivity
one-fifth of the peak-to-peak noise in the blank signal prove that there is analyte in the sample, the null hy-
(see Section 5-1). It is prudent to measure a series of pothesis that the analyte concentration is zero is tested.
standards of decreasing concentration down to concen- Thus p, = 0, s, = s,,, and z = Sh,, or S = zs,,. Since
trations below the D L to confirm that concentrations operationally the D L is defined as the concentration
near the D L can indeed be detected and that the slope where S = ks,,, the confidence factor k corresponds
of the calibration curve near the D L is linear and known. to the z statistic. If an analytical signal S equal to ks,,
Often, contamination problems can limit the ability to is obtained, the analyte is detected (S is significantly
prepare solutions below a certain concentration and different from zero) with a (1 - a ) significance level,
cause the calibration slope to approach zero at low con- where a is the probability associated with z = k.
centrations. The most common values of k are 1 , 2 , and 3; these
The D L is a useful figure of merit because it de- correspond to levels of confidence (see Table A-2) of
fines the lower end of the dynamic range of an analytical 84.1, 97.7, and 99.9%. Note that a one-tailed test is used
technique and instrument. If several analytical tech- here since negative concentrations are physically impos-
niques are available, one would not normally choose sible. Thus the alternative hypothesis (S > 0) is the basis
the technique which had a D L higher than the suspected for the common statement that defining the DL as the
analyte concentration in the sample, although sample concentration yielding an analytical signal equal to twice
preconcentration could be employed. One would prefer the SD of the blank corresponds to a confidence level of
not to use a technique where the analyte concentration 97.7%. Unfortunately, this is not correct because it does
in the sample is at or slightly above the DL. By defi- not take into account that two measurements (i.e., total
nition the RSD at the D L is 50% (k = 2). Thus meas- and blank) are required to obtain the analytical signal.
urements are normally made at concentrations at least Thus the SD for determining the analytical signal is really
a factor of 10 above the DL. S, = (s: + s;,)~'~,not st. So for zero analyte concentra-
The DL reported by an instrument manufacturer tion, s, = s,, and s, = a s,. By definition, S = ks,,
or a researcher may not apply to real samples. Usually, at the D L and z = Sls, = SIV? s,,. Hence k = @ z
such DLs are obtained with a completely optimized and a confidence factor of k corresponds to the level of
instrument and pure standards. The presence of con- confidence associated with the value of z = ki*. For
comitants in a real sample can affect the slope of the k = 1, 2, and 3, the corresponding confidence levels are
calibration curve or the blank SD. In real samples, it 76.1, 92.1, or 98.3%, respectively. A 97.7% confidence
is often the sample-to-sample variation of the uncom- level applies only if the DL is defined as the concentration
pensated blank component of the total signal (or of E, where S = 2 a s,,.
in absorption measurements) that limits detectability If s,, is determined with less than 30 measurements,
rather than the SD obtained from repetitive measure- the confidence level for k must be determined from the
ments of one reagent blank solution. t statistic [t = (S, - ps)ls, = S/@ sbk for pS = 0,
How the blank SD is evaluated is also important. s, = s,,, and n = 11. In an analogous fashion it can
Blank measurements should be made in a way that be shown that a confidence factor of k corresponds to the
accounts for sample placement imprecision. It is more level of confidence associated with t = k/@ rather than
realistic to make repetitive measurements on n sepa- t = k. Thus for k = 2, t = 2*! = 1.41, and for n =
rately prepared blank solutions than n measurements 8, the confidence level is 90% rather than the 92.1%
on one blank to account for variability in blank prep- predicted by the z statistic.
aration and contamination. Even though many blank measurements are com-
monly used to estimate s,,, the DL is usually calculated
Statistical Significance of Detection Limit. The as if only one paired measurement (n = 1) of the sample
D L should be defined as the concentration that yields and blank is made to determine if the analyte is detected.
a mean analytical signal which we are confident is not If n paired measurements of a sample are made, the D L
solely due to fluctuations in the blank measurement. is reduced by n1l2 assuming that random error limits the
The magnitude of the confidence factor k is chosen to blank measurement precision.
correspond to the desired confidence level. In some cases drift can be significant during the time
If the blank measurements are normally distrib- interval required to make a number of blank measure-
uted, the t or z statistic can be used. The z statistic ments; as a result the estimate of s,, is too high. Here it
requires sufficient blank measurements (n r 30) that is preferable to obtain the SD of the difference of n paired
the SD measured is a good estimate of the population blank measurements where the time separation between
SD (s,, = u b k )For one measurement of the mean paired blank measurements is the same as between nor-
(n = I), z can be expressed as z = (S - ps)!ss. To mal paired sampleblank measurements. In this case, s,,
Sec. 6-4 1 Sensitivity and Detection Limit
centration which produces an analytical signal equal to analyte loss or gain (contamination). Still, in many sit-
a factor of 10 to 20 times the blank standard deviation. uations, the only way to eliminate an interferent is to
More complex methods have been proposed to remove it, and such procedures are actually gaining
calculate the detection limit. These methods take into popularity with the development of automated instru-
account the uncertainty in the slope and intercept of ments which incorporate separation technology.
the calibration curve. Many separation techniques also preconcentrate
the analyte by transferring the analyte in the original
sample to a smaller volume. This is necessary if the
6-5 TECHNIQUES FOR MINIMIZATION OF analyte concentration is below the D L of a technique
SYSTEMATIC AND RANDOM ERRORS or is only slightly above the D L where measurement
precision is poor.
The ideal spectrometric technique is totally selective in Spectrometric detectors are now widely used in
that the presence of concomitants does not affect the liquid chromatography, in particular with high-perform-
measured analytical signal (i.e., no blank or analyte ance liquid chromatography, and to some extent in gas
interferences), and simple external standards can be chromatography. The most common spectrometric HPLC
employed for calibration. This is, of course, rarely the detectors are based on refraction, UV-visible molecular
case, especially for complex samples in which the an- absorption, and molecular fluorescence.
alyte concentration is orders of magnitude lower than There is growing need for detectors with greater
the concentration of many species in the analytical sam- selectivity that compensate for imperfect chromatog-
ple matrix. In this section, many of the sample treat- raphy. Often, it is difficult to separate completely all
ment and instrumental techniques used to minimize or components in a complex sample; two or more com-
correct for systematic errors caused by the presence of ponents can coelute or have approximately the same
interferents are reviewed. Some of these techniques can retention time. In such cases it is advantageous to use
also reduce random error and improve the SIN if they a detector that responds to only one of the unresolved
reduce the magnitude of background signals (i.e., blank components in a chromatographic band. Refractive in-
interferences) and thus background noise. dex detectors are good universal detectors but are quite
In an ideal instrument, the experimental variables unselective. Molecular absorption detectors provide
which affect the magnitude of the analytical and blank moderate selectivity and are discussed in more detail
signals are controlled so that they do not cause signif- in Section 13-5. Molecular fluorescence detection yields
icant errors. However, fluctuations and drifts in the even greater selectivity, as detailed in Section 15-3.
magnitude of experimental variables do occur and cause Other spectrometric phenomena on which HPLC
random and systematic errors. detection can be based include IR absorption (see
The methods discussed in this section primarily Chapter 14), circular dichroism, optical activity, pho-
minimize or correct for errors involved in the sample toionization, the photoacoustic effect, and thermal len-
measurement step. It must be stressed that errors in sing. (See Chapter 17 for a discussion of the last three
calibration, sample acquisition, and sample preparation techniques.) In addition to detectors based on molec-
can limit the overall accuracy and precision of an an- ular properties, it is becoming increasingly common to
alytical procedure. It is assumed here that relevant ex- use atomic emission, absorption, or fluorescence ele-
perimental variables (wavelength, slit width, reagent ment-specific detection. With this type of detection, the
concentrations) have been adjusted to minimize the ef- detector responds only to compounds that contain the
fect of interferences and to optimize the SIB or SIN. element of interest. This makes it possible to separate
and detect different chemical forms of the same metal
Separations (e.g., methyl mercury and ethyl mercury).
Various separation techniques, including filtration, di- Saturation, Buffer, and Masking Methods
alysis, solvent extraction, volatilization, ion exchange,
precipitation, and chromatography, can be used for Certain analyte interferences depress or enhance the
sample cleanup (see Appendix E). The cleanup can analytical signal by a fixed fraction once the interferent
involve separation of the analyte from the major con- concentration is above a certain level. In the saturation
stituents in the sample matrix or removal of the primary method, the interferent is added to all samples, stand-
interferences from the sample matrix. leaving the an- ards, and blanks to the same high concentration level
alyte behind in the analytical sample. Although sepa- such that the interference effect is independent of the
rations are extremely valuable, they increase the time original concentration of interferent in the sample (i.e.,
required for sample preparation and the probability of the analyte in the sample and standards experiences the
Sec. 6-5 1 Techniques for Minimization of Systematic and Random Errors
same interference effect). The fractional change in the concentration level. The degree of dilution is limited
analytical signal due to the presence of the interferent by the ratio of the original analyte concentration to the
must be independent of the analyte concentration. If analyte detection limit.
the analytical signal is significantly depressed, the de- ation, or synthetic du-
tection limit can be degraded. plication method, an attempt is made to duplicate the
In some cases, the presence of another species can matrix of the analytical sample in the standard and blank
suppress the effect of a blank or analyte interference. solutions. This can compensate for certain blank or an-
In the buffer method, this third species, often called the alyte interferences if the interference effect is the same
buffer or matrix modifier, is added to all samples, stand- for samples, standards, and blank. For example, in the
ards, and blanks in sufficient amounts that the analytical analysis of seawater for trace species, standards can be
signal is independent of the interferent concentration, prepared in synthetic seawater which contains all the
as shown in Figure 6-6. A pH buffer is used in many major components of seawater (e.g., Na+, K t , C1-,
wet-chemical procedures to keep the pH of a reaction Ca2+, Mg2+, COZ-). The matrix match method can
mixture constant (within limits) regardless of the initial also be used for industrial products where the concen-
sample pH. Sometimes the buffer is a masking agent trations of major components are known and con-
which selectively reacts with the interferent to form a trolled. Sometimes the matrix solution can be prepared
complex which does not contribute a blank signal or from a real sample where the analyte is removed from
alter the analyte signal. Some buffers are added to en- the original sample matrix; known amounts of the an-
sure that bulk properties such as ionic strength are con- alyte are then added to make standards.
stant. The parametric method is a special case of the
In many solution techniques, the buffer is part of matrix match method; it is usually employed if there is
the reagent solution that is mixed with the analytical only one major blank or analyte interference. The in-
sample. Care must be taken in buffer methods that the terferent is added to all standards and blanks at the
buffer is not contaminated with a significant amount of same concentration level as it is in the analytical sample.
analyte or other interferents and that the buffer reagent A series of calibration curves is prepared with different
does not itself cause an interference effect. concentrations of the interferent in the standards; the
appropriate calibration curve that matches the inter-
ferent concentration in the sample is then used for the
determination. Of course, this means that the interfer-
ent concentration must be known or accurately deter-
Simply diluting the sample can sometimes minimize blank mined.
or analyte matrix interference if the interferent pro- In either the matrix match or parametric methods,
duces no significant interference effect below a certain care must be taken that the substances added are not
contaminated with analyte. Also, the added interferents
may reduce precision or increase the detection limit,
due to a decrease in the calibration sensitivity or an
increase in the background signal and noise.
signal levels are not reduced, but their average contri- ted radiation polarize
bution to the measured total signal is zero. The noise a magnetic field surro
from the background signal at the modulation fre- discrimination against background absorption in Zee-
quency is still present, although llf additive noise that man atomic absorption spectrophotometry. Differences
is not carried by the modulation waveform is discrim- in the polarization of fluorescence and scattering signals
inated against as discussed in Section 5-6. Wavelength can sometimes be used to attenuate scattering signals
modulation is often used in atomic spectrometry to dis- in fluorescence measurements (see Section 15-6)
criminate against continuum background signals (see ecules of different molecular weight can be
Figure 4-34) and in molecular fluorescence spectrom- guished based on the degree to which their fluorescence
etry to minimize spectral interference from concomi- radiation is depolarized (see Section 15-6).
tants (see Section 15-1).
There are many other examples in which time
dependence is or could be exploited to increase selec-
tivity, and we will certainly see more use being made Primarily in molecular absorption and luminescence
of time-discrimination techniques in the future. spectrometry, the selectivity for the analyte is often
enhanced by using chemical reactions of the analyte
Spatial Discrimination. Spatial position is an with selective reagents. Often, these reactions convert
important selection criterion, particularly in atomic the analyte into a form more suitable for spectrochem-
emission, absorption, and fluorescence measurements. ical measurement. For example, in molecular absorp-
In these techniques, the number density of the atomic tion spectrophotometry many analyte species do not
or molecular species of interest varies substantially with significantly absorb radiation in the UV or visible re-
the volume element viewed or probed. Analytical flames, gions of the spectrum; these must be converted into
inductively coupled plasmas, dc arcs, high-voltage sparks, absorbing species by chemical means.
and electrothermal cells are examples of atomization based on a selective analytical or deriv
sources that produce position-dependent information. tions are equilibrium-based methods since spectrometric
If the analyte signal can be monitored in a region of measurements are made after the analytical reaction
the sample container that is relatively free from back- has come to equilibrium. The magnitude of the meas-
ground, significant improvements in SIB'S can be ured steady-state signal (e.g., absorbance) from the re-
achieved, Many commercial atomic spectrometer sys- action product is related to the analyte concentration.
tems allow the user to change the viewing region as well Chemical selectivity is determined by the availa-
as the region irradiated by the external source in atomic bility of selective analytical reagents. Other species can
absorption and fluorescence measurements so as to op- cause a blank interference by reacting with the analy-
timize SIB'S. The complete spatial profiles of analyte tical reagent to form reaction products that
and background signals are of great assistance in op- optical signal at the analytical wavelength. Also, analyte
timization. In atomic absorption and atomic fluores- interferences can occur if the magnitude of the analy-
cence spectrometry, optimization of the region that is tical signal from the new chemical form of the analyte
irradiated by the external source and detected can also is affected or the efficiency or rate of production of the
be of benefit. reaction product is altered by the presence of concom-
itants. A very high degree of chemical selectivity is
Polarization Discrimination. The interaction of possible with enzymatic reactions (see Section 13-5) or
polarized radiation with optically active substances is antibody-antigen reactions as used in imunoassays (see
the basis of polarimetry, optical rotatory dispersion,
and circular dichroism (see Section 3-2). The direction hen analytical reactions are used, the blank
of polarization of Raman scattering can give informa- measurement is usually based on a mixture of the an-
tion about the symmetry of molecular vibrations (see alytical reagent and the solvent. Sometimes an internal
Section 16-2). Depolarization of polarized fluorescence blank measurement is made on the sample mixture mi-
radiation by molecular rotation is used to study the nus one of the necessary analytical reagents. This in-
characteristics of macromolecules (see Section 12-6). ternal blank does not compensate for concomitants that
Differences in the interaction of polarized radia- react with the analytical reagent to produce interfering
tion with the analyte and concomitants provide the means spectral signals.
to enhance selectivity in absorption and photolumi- In spectrophotometric reaction-rate (kinetics-based)
nescence techniques. For example, as discussed in Sec- methods, the rate of change of absorbance with tune is
tion 10-3, the differences in the intensities of transmit- the analytical signal measured. This represents a case
182 Chap. 6 I Methodology in Spectrochemical Analysis
where both chemical and time selectivity are employed. ters. The reference signal is provided by an optical
Concomitants that contribute only a steady-state back- transducer that monitors the light source intensity. If
ground absorbance or that react with the analytical re- the light source intensity suddenly decreases 1%, the
agent at a significantly different rate than the analyte optical feedback circuit increases the lamp power supply
do not interfere. current or voltage to increase the intensity to its initial
In spectrophotometric titrations, the equivalence value.
point in a titration is indicated by a rapid change in the With mathematical correction schemes, the ref-
solution absorbance. Concomitants which absorb ra- erence signal is not used to maintain the variable mag-
diation at the analytical wavelength, but which do not nitude at a constant level. Instead, the reference signal
react with the titrant, do not interfere with the detection provides a means to track changes in the magnitude of
of the endpoint. the variable. The measured analytical signal is corrected
Derivatization reactions, reaction-rate methods, for the changes in the variable magnitude that occur in
and spectrometric titrations are covered in detail in Sec- accordance with the previously established mathemat-
tion 13-5 because these techniques are most commonly ical relationship between the analytical signal and the
used with molecular absorption spectrophotometry. variable magnitude. As for feedback correction, the
These same techniques are also used to increase selec- most common example of mathematical correction in-
tivity in molecular fluorescence spectrometry (see Sec- volves compensating for changes in the intensity of the
tion 15-3). In chemiluminescence spectrometry, a excitation source in absorption and luminescence spec-
chemical reaction is mandatory to produce the lumi- trometers. As discussed in Sections 4-6 and 5-6, the
nescent species (see Section 15-5). reference beam in double-beam spectrometers provides
Selective chemical reactions are normally not used a signal proportional to the light source intensity. The
with atomic spectrometric techniques because the atom- correction in this case involves electronic or software
ization process converts all chemical forms of the ele- ratioing of the analytical signal to the reference signal
ment of interest into an atomic vapor. However, selec- and provides compensation for source flicker noise and
tive chemical reactions are sometimes used for sample drift. Note that we use the terms correction and com-
introduction. For example, some elements can be chem- pensation interchangeably even though the term cor-
ically converted to volatile products which are swept rection sometimes refers only to feedback correction
out of the sample solution and directed into the at- schemes.
omizer (see Section 10-1). Instrumental correction techniques can be ex-
tended to any variable that has a significant effect on
Instrumental Correction Methods the magnitude of the analytical signal (e.g., p H or tem-
perature in solution measurements). Note that the in-
Fluctuations and drifts in the magnitude of experimen- ternal standard method is a type of mathematical cor-
tal variables that affect the analytical signal can cause rection technique. The signal from the reference species
random and systematic errors. These types of errors is used to correct for changes in experimental condi-
can be minimized by better controlling the critical var- tions, or even analyte interferences, that affect the an-
iables or by correcting the measured analytical signal alyte and reference analytical signals in an equivalent
for changes in the magnitude of the variable that occur manner. Microcomputer-based instrumentation ena-
during the measurement process. Instrumental correc- bles mathematical correction schemes to be more widely
tion techniques for changes in experimental conditions used because acquisition and correction can be imple-
can be based on either feedback systems (e.g., servo mented on-line.
systems) or mathematical schemes. In either case, it is Instrumental correction techniques are also used
necessary to obtain a reference signal that is related to to compensate for spectral interference from concom-
the magnitude of the variable of interest. itants. In cases where the analytical signal contains op-
In feedback correction schemes, the reference sig- tical signal contributions from concomitants uncom-
nal is used to control the magnitude of the experimental pensated by the blank measurement (i.e., a blank
variable. The reference signal is continually compared interference), a measurement under different condi-
to a preset signal level and any error signal detected tions (e.g., at a different wavelength, with a different
generates a response that minimizes the error signal and source, or with a different polarization state) can yield
thus maintains the variable value at its preset value. the background signal, which, in turn, is subtracted
The most common example is optical feedback correc- from the measured analytical signal. For example, in
tion in absorption and photoluminescence spectrome- atomic spectrometry it is common to correct for con-
Sec. 6-6 1 Automated Spectrochemical Measurements
tinuum background signals. the analytical signal containers on the sampler. By rotation or other means,
contains optical contribution both the analyte line each container is sequentially positioned under a sam-
and from the continuum produced by concomitants. pling tube, which is then lowered into the test solution.
The background signal at the analytical wavelength can Each test solution is pumped through tubing into the
be estimated by measuring the signal at a wavelength spectrochemical encoder in a sequential manner. Often,
slightly removed from the analytical line and then sub- the sampling tube dips into a container containing rinse
tracted from the analytical signal. This can be accom- solution between samples to minimize contamination
plished with dual-channel (dual-wavelength) spectrom- between samples. Many automated spectrometric sys-
eters or spectrometers fitted with refractor plates (see tems include additional hardware for adding reagents,
Figure 4-34). Several other techniques are used to es- mixing, or performing sample cleanup.
timate the continuum background absorption signal in Automated systems are widely used in clinical and
atomic absorption spectrophotometry (see Section 10- water analysis laboratories where a few species must be
3). Measurements at two or more wavelengths are also determined in a large number of similar samples. In
used in molecular absorption and photoluminescence such applications, the high sample throughput rate of
spectrometry to compensate for absorption or lumi- automated systems is critical. Such systems require less
nescence by concomitants (see Sections 13-1 and 15-1). skilled operators and often can be operated unattended.
Note also that the modulation techniques discussed ear- Operator blunders are minimized and precision can be
lier in this section often are used as instrumental tech- improved because all operations are strictly timed and
niques for background signal correction. performed by mechanical or electrical devices.
In some mathematical correction techniques, sig- In this section we emphasize automated spectro-
nal information at different wavelengths is used to es- metric systems based on molecular absorption or mo-
timate a background signal correction term. For ex- lecular fluorescence detection as these molecular de-
ample, a multichannel atomic spectrometer can measure tection schemes are most commonly used. Usually, it
analytical signals (atomic line intensities) from many is necessary to convert the analyte to an appropriate
elements. If 10 pg m L p l of element X is known to chemical form for measurement. Thus the sample must
produce (due to spectral overlap) an apparent signal in be mixed with reagents before the reaction mixture is
the channel for element Y equivalent to the signal from sent to a sample cell. Automated systems using atomic
0.1 kg m L p l of Y, the concentration of X can be meas- spectrometric detection are briefly covered at the end
ured and used to subtract an appropriate signal from of this section. The basic characteristics of batch and
the measured signal in channel Y. continuous automated analyzers are now reviewed.
Different types of spectral signals can also be used allyaers, the sample is main-
ematical correction. For example, it is possible tained as a separate entity in a separate vessel throughout
to correct for the attenuation of photoluminescence sig- the analysis sequence. Most batch analyzers are sequential
nals by concomitants that absorb a significant fraction analyzers. Pumps, valves, or automatic syringes are con-
of the excitation or emission radiation if the absorbance figured and controlled to dispense one sample plus ap-
at the excitation and emission wavelengths is measured propriate reagents and diluents into a sample cell. After
(see Sections 15-1 and 15-2). In this case the correction mixing, an appropriate delay time, and possibly heating
scheme compensates for analyte rather than blank spec- to speed slow analytical reactions, the molecular spectro-
tral interferences. metric measurement is made. If one sample cell is em-
ployed, it is evacuated and rinsed between samples. A1-
ternatively, different sample cells for each sample are
transported sequentially to the spectrometer sample com-
partment. In this case it is often possible to obtain dis-
posable sample cells with prepackaged reagents; it is nec-
In automated instruments, often called automated ana- essary only to add the sample.
lyzers, a spectrometer or another detection system is A unique version of a batch analyzer is the cen-
one component in a larger system which has provision trifugal analyzer; it is a parallel analyzer which allows
for automating sample placement or delivery steps, and many samples to be analyzed simultaneously. The op-
often sample treatment steps. At a minimum, an au- eration of the instrument is based on a plastic rotor
tomated system usually includes a pump and an auto- which typically has 17 dual compartments arranged con-
mated sampler (e.g., a sample turntable). The samples, centrically (see Figure 6-10a). Each sample, standard,
standards, and blanks are first loaded into the separate or blank is dispensed into one of the compartments
184 Chap. 6 1 Methodology in Spectrochemical Analysis
<'
\
samples and enhance mixing between the sample and case, the amount of sample dispersion is controlled
reagents. by the flow rate, the sample size, and the length and
Other modules can be added to the flow stream diameter of the tubing. It is possible to stop the
to cleanup the sample by dialysis, solvent extraction, flow when the sample plug reaches the flow cell to
or filtration. Normally, a debubbler removes the bub- make reaction-rate measurements. Solvent extraction
bles before the flow stream is sent to the flow cell. or dialysis modules can be incorporated. Typical
Alternatively, a bubble gate is used to trigger the ac- sample throughput rates range from 60 to 300 samples
quisition of spectrometric data only between bubbles. per hour.
In unsegmented continuous flow analyzers (Figure Automated atomic spectrometric systems are also
6-12a), a sample loop valve is often used to inject a available. Because sample treatment is not often re-
precise volume of sample (typically 10 to 100 pL) into quired, simple automated systems consist of an auto-
the flow stream. Peristaltic or positive-displacement matic sampler and pump that allow sequential delivery
pumps force the carrier stream through the valve and of samples and standards to the atomizer. In more so-
to the flow cell. The sample plug disperses into the phisticated systems, there is provision for automated
carrier stream as it flows through tubing to the flow cell standard addition or addition of matrix modifiers to
as illustrated in Figure 6-12b. If the carrier stream is a reduce interference effects. Flow injection analysis sys-
reagent mixture, the analyte and reagents react to yield tems are well suited for high-throughput, automated
a peak-shaped reaction product profile. In some cases, atomic spectrometric measurements.
the sample and reagent are injected with separate sam- Microcomputers have greatly enhanced the power
ple loop valves into two carrier streams. With proper and ease of use of all automated analyzers. They allow
timing, the two plugs merge at a tee and are mixed as convenient and precise control of all operations and
the combined plug is carried to the flow cell. In either automated data manipulation and reporting.
186 Chap. 6 I Methodology in Spectrochemical Analysis
Waste
71I
Sample initially
injected as
Diffusion
PROBLEMS
6-1. Copper was determined in a river water sample by ards or the unknown with an appropriate reagent blank
flame atomic absorption spectroscopy with the method (c, = 0) and all sample measurements are paired with
of standard additions. For the standard addition, 1.00 a blank measurement.
mL of a 1000-kg mL-I Cu standard was added to
1.00 L of sample. The following data were obtained: c, E,E Ebk
(ng mL-') (v) (V)
Reagent blank: A = 0.050
Sample: A = 0.550 0.0
Sample +
std. addition: A = 1.050 0.0
0.0
(a) Calculate the concentration of Cu in the sample. 0.0
(b) Further studies revealed that the reagent blank 0.0
10.0
was not adequate and that the actual blank ab-
10.0
sorbance in the sample was 0.200. What is the 10.0
magnitude of the error caused by using an im- Unknown
proper blank? Unknown
6-2. The following atomic emission measurements were Unknown
made for Na. All measurements were made on stand-
Chap. 6 1 Problems 1
Calculate the following values. (e) The detection limit for 97.5% confidence based
(a) The mean analyte emission signal for 0.0 and 10.0 on the z statistic and the t statistic. Note that s,
ng-' mL Na and the unknown. for c, = 0 equals fl s,. For the z-statistic cal-
(b) The standard deviation (s,) for the measurement culation, assume that s, = u,.
of E, for 0 and 10 ng mL-I Na and the unknown. ( f ) The characteristic concentration (atomic absorp-
Notice that there is an apparent drift in the blank tion sensitivity).
signal, so be sure to use the paired measurement (g) The range of Z n concentration in which the true
scheme for all calculations. concentration of Z n in the unknown is contained
(c) The calibration sensitivity (m) for 0 - 10 ng mL ' with a 95% level of confidence. Assume no error
(assume linearity). in the calibration curve.
(d) The analytical sensitivity (y) at 10 ng mL-'. In general, watch the significant figures and the differ-
(e) The detection limit based on a confidence factor cnce between a one- and a two-tailed test. Be sure to
(k) of 3. T o what level of confidence (within 1 % ) use the paired measurement scheme for all calculations.
does this correspond? 6-4. Can a single concomitant cause both spectral and non-
( f ) The range of voltages in which the population mean spectral interference?
for 10 ng mL-I Na is contained with a 98% level 6-5. Will a spectral interferent in the standards or the blank
of confidence. result in a positive or negative intercept for a calibra-
(g) The concentration of Na in the unknown has been tion curve? Discuss the possibilities.
determined accurately by another method to be 6-6. Strontium was determined in seawater by flame emission
5.20 ng mL-I. D o the data indicate that there spectrometry. The standard addition method was used.
might be some systematic error or interfercncc in The following table indicates the volume of a 3.00 g L-'
this determination by flame emission? Assume that Sr solution added to 1000 mL of the sample solution and
there is no uncertainty in the calibration curve. the blank-corrected emission signal recorded.
In general, watch the significant figures and the
difference between a one- and a two-tailed test. Be
Volume of Sr added Analytical signal
sure to use the paired measurement scheme for all
calculations. Note that s, for c, = 0 equals fls,,. (mL) (mv)
6-3. The following atomic absorption measurements were
made for Zn. All measurements were made on stand-
ards with an appropriate reagent blank (c, = 0) and
all sample measurements are paired with a reference
measurement. The signals due to dark current, back-
ground and analyte emission, and background and an-
alyte fluorescence are negligible.
Calculate the concentration of Sr in the unknown by
t h e graphical method a n d by t h e single-addition
technique for 2 mL.
6-7. In absorption measurements, what methods can be
0 used to correct for the effect of analyte luminescence
0 on the measured absorbance signal?
0
0 6-8. When do noise calculations or measurements give a
10.0 poor estimate of the detection limit?
10.0 6-9. A photoluminescence measurement is limited by back-
10.0 ground luminescence noise. Calculate the detection
Unknown limit for a confidence factor of 2 based on the following
Unknown information: signal processor gain, 1.0 x 10hV A-';
Unknown
PMT gain, 1.0 x lo6; noise equivalent bandpass, 1.0
Hz; secondary emission factor, 0.30; calibration curve
slope, 1.0 mV nM-I; E,, 0.10 V.
Calculate the following values.
(a) The mean absorbance for 0 and 10 ng mL-' Z n 6-10. What advantages d o automated instruments provide?
and the unknown. 6-11. What techniques can be used to compensate for non-
(b) The standard deviation for the measurement of A fundamental variations in the excitation source inten-
(s,) for 0 and 10 ng mL-' and the unknown. sity that occur during a series of measurements?
(c) The calibration sensitivity (m) for 0 - 10 ng mL-' 6-12. What is indicated when the slope of a calibration curve
(assume linearity). prepared from external standards and the slope of a
(d) The analytical sensitivity (y) at 10 ng mL-'. standard addition plot differ significantly?
1 Chap. 6 I Methodology in Spectrochernical Analysis
Figures of merit such as the detection limit and sensitivity are M. Zief and J. W. Mitchell, Contamination Control in
discussed in the following references. Trace Element Analysis, Wiley-Interscience, New York,
1976.
G . L. Long and J . D. Winefordner, "Limit of Detec-
tion," Anal. Chem., 55; 712A (1983). L. Larsen, N. A. Hartmann and J. J. Wagner, '.Esti-
mating Precision for the Method of Standard Addi-
A. L. Wilson, "Performance-Characteristics of Analyt-
ical Methods: I-IV," Talanta, 17.21 (1970); I 7 , 3 1 (1970); tions,'' Anal. Chem., 45, 1511 (1973).
20, 725 (1973); 21, 1109 (1974). E. B. Sandell, "Errors in Chemical Analysis," chap. 2
H . Kaiser, "Quantitation in Elemental Analysis," Anal. in Treatise on Analytical Chemistry, pt. I , vol. 1, p. 19,
Chem., 42, 24A (1970); 42, 26A (1970). Wiley-Interscience, New York, 1959.
B. Kratochvil and J. K. Taylor, "Sampling for Chemical
P. W. J. M. Boumans, "A Tutorial Review of Some
Analysis," Anal. Chem., 53, 925A (1981).
Elementary Concepts in the Statistical Evaluation of Trace
Element Measurements," Spectrochim. Acta, 33B, 625 I. M. Kolthoff and P. J. Elving, eds., Treatise on An-
(1978). alytical Chemistry, 2nd ed., pt. I, vol. 1, sec. B, Wiley-
Interscience, New York, 1978. Chapters 3, 4, 5. and 6
R . K. Skogerboe, "Concepts of the Definitions of the
are excellent reviews of analytical methodology, sources
Terms Sensitivity and Detection Limit," Spectrosc. Lett..
3, 215 (1970). of error, and sampling.
G. H . Morrison, "General Aspects of Trace Analytical
Methods: I. Methods of Calibration in Trace Analysis," Automated analysis is considered in the following references.
Pure Appl. Chem., 41, 397 (1975). M. Margoshes and D. A. Burns, "Automation: Instru-
J . D . Ingle, Jr., "Sensitivity and Limit of Detection in mentation for Analysis Systems," in Treatise on Ana-
Quantitative Spectrometric Methods," J. Chem. Educ., lytical Chemistry, 2nd ed., pt. I, vol. 4. P. J. Elving and
51: 100 (1974). I. M. Kolthoff, eds., Wiley. New York, 1984.
J. D. Winefordner, "Quantitation in Analysis." appen- W. B. Furman, Continuous Flow Analysis-Theory and
dix A in Trace Analysis: Spectroscopic Methods for Ele- Practice, Marcel Dekker, New York, 1976.
ments, J . D . Winefordner, ed., Wiley-Interscience, New D. Betteridge, "Flow Injection Analysis," Anal. Chem.,
York, 1976. 50, 832A (1978).
K. Eckschlager and V. Stepanek, Information Theory L. Snyder; J. Levine, R . Stoy and A. Conetta, "Au-
as Applied to Chemical Analysis, Wiley-Interscience, New tomated Chemical Analysis: Update on Continuous-Flow
York, 1979. Analysis," Anal. Chem., 48, 942A (1976).
Analytical methodology, errors, and data analysis are con- C. B. Ranger, "Flow Injection Analysis," Anal. Chem.,
sidered in the following references. 53, 20A (1981).
K. K. Stewart, "Flow Injection Analysis." Anal. Chem.,
R. Klein, Jr., and C. Hach, "Standard Additions-Uses
55, 931A (1983).
and Limitations in Spectrophotometric Analysis," Am.
Lab.. 9 (7), 21 (1977). J. Rizitka, "Flow Injection Analysis," Anal. Chem.,
55, 3040A (1983).
M. A. Sharaf, D . L. Illman, and B. R. Kowalski, Che-
mometrics, Wiley-Interscience, New York, 1986. C. D . Scott and C. A . Burtis, "A Miniature Fast Ana-
H . A. Laitinen and W. E . Harris, Chemical Analysis, lyzer System," Anal. Chem. : 45, 327A (1973).
2nd ed., chaps. 26 and 27; McGraw-Hill, New York, J. RiziEka and E. H . H. Hansen. Flow Injection Anal-
1975. ysis, Wiley, New York, 1981.
T. C. O'Haver, "Analytical Considerations," chap. I1 Anal. Chim. Acta. 179 (1986). A special issue devoted
in Trace Analysis: Spectroscopic Methods for Elements, to flow analysis.
J . D. Winefordner, ed., Wiley-Interscience, New York, Flow Injection Analysis Bibliography, Tecator, Ho-
1976. ganas, Sweden, 1985.
In previous chapters we have considered the aspects of any of the interferences obscrved in emission,
spectrochemical analyses that are generally applicable on, and fluorescence have similar origins that
to all methods. In the remaining chapters of this book, can be traced to inefficiences in the sample introduction
specific spectrochemical methods are described. We fol- or atomization steps. e of these similarities, it is
low here the conventional classification of such methods useful to consider th on interferences together.
into atomic spectrochemical methods and molecular Similarly, all atomic spectrochemical methods deal with
spectrochemical methods. The specific atomic tech- similar electronic transitions of the analyte atoms. The
niques to be discussed are flame and plasma atomic electronic states of atoms and the term symbols that are
emission (Chapter 8), arc and spark atomic emission used to describe these states are reviewed here.
(Chapter 9), atomic absorption (Chapter lo), and atomic spectral line profiles of analyte atoms play an important
fluorescence (Chapter 11) spectrometry. Although these role in determining the shapes of calibration curves, the
methods involve different excitation and radiative proc- selectivities sf the techniques, and the spectral line in-
esses, they have much in common. For example, all ities are a major factor in
depend on having the analyte present in the form of a tection of the methods. T
free vapor of neutral atoms (or occasionally ions). A this chapter concludes with a discussion of spectral line
sample presentation system, consisting of a sample in- profiles and line intensities. The general principles and
troduction system and the atomization device, converts concepts presented here are intended to lay an impor-
the analyte into the appropriate form for spectrochem- tant foundation for the specific atomic spectrochemical
ical encoding (recall Figure 2-1). The sample introduc- methods that follow.
tion step is extremely important in atomic spectrochem-
ical methods and is often the step that limits the amount
of analyte converted into the appropriate form for sub-
sequent observation. The most widely employed sample
introduction methods are treated here along with the
processes that lead to the formation of atomic vapors e consider first an overview of the sample introduc-
since these are common to atomic spectrochemical tion and atomization steps that must occur in all atomic
methods. spectrochemical methods. Later in this section, specific
190 Chap. 7 1 Introduction to Atomic Spectroscopy
Overview
duced is a transient because of the limited amount of into flame or plasma excitation sources by means of
sample. In flow injection techniques, a plug of the an- electrothermal atomizers. Here the first atomizer con-
alytical sample solution in a carrier stream is introduced verts the analyte into an atomic vapor, while the flame
into the atomizer as a mist with the aid of a nebulizer. or plasma must now only supply the energy needed to
Dry aerosols and vapors can also be introduced as plugs excite the sample.
of material. With vapor introduction (e.g., hydride niques, a laser beam is
techniques), the volatile analyte species is often stripped directed onto a small portion of a solid sample. The
from the analytical solution and carried by a gas to the sample is vaporized and atomized by radiative heating.
atomizer. This stripping step can be preceded by a spe- Either the plume of sample formed can be directly probed
cific chemical reaction that converts the analyte into a by the encoding system or the vapor produced can be
volatile form. It is now common to use gas and liquid swept into a second atomization cell for observation of
chromatography to introduce samples into atomizers. emission, absorption, or fluorescence.
Nebulizers are often used for liquid chromatographic Detailed descriptions of many atomizers and sam-
eluates. Chromatographic introduction is a type of dis- ple introduction systems are presented in Chapters 8 to
continuous sample introduction since the atomizer re- 11, which deal with specific methods. To provide an
ceives a time-varying concentration of analyte as com- understanding of the atomization process, general char-
ponents elute from the column. acteristics of continuous and noncontinuous sample in-
With arcs and sparks, the sample introduction and troduction/atomization systems are discussed below. In
atomization processes are more difficult to separate. In both systems, the analytical sample is assumed to be a
many arc and spark determinations, solid samples are solution, which is the most common sample form.
employed. These are often shaped into the form of an
electrode, and the discharge struck between the sample Processes Occurring during Atomization. Let
electrode and a second electrode. Alternatively, the us consider the processes that must take place in order
sample electrode can be in the form of a cup into which to transform a solution sample into an atomic vapor
powdered samples are packed, a porous cup, or a ro- with continuous sample introduction into a continuous
tating disk for solutions. In any case the atomic signals atomizer. As shown in Figure 7-2, the sample intro-
produced are transient because the discharge conditions duction system disperses the sample into the high-tem-
vary as the discrete amount of sample is atomized. perature environment of a flame or plasma, usually as
In some cases one atomizer can be used as a sam- a fine spray or mist. This process is called nebulization.
ple introduction system for a second atomizer. This can Heat from the flame or plasma evaporates the solvent
be particularly advantageous in atomic emission, where and volatilizes the dry aerosol that remains as Figure
the atomization device is usually called upon to excite 7-2 illustrates. Once free atoms are formed, they can
the analyte as well as atomize it. Separating the sam- be excited by collisions to produce characteristic spec-
plinglatomization step from the excitation step can al- tral lines or an external radiation source can be used to
low optimization of the energy input for each step. As obtain atomic absorption or fluorescence information.
an example of this approach, samples can be introduced The processes that occur with discrete sample in-
do'o,
a
Nebulization -?;go
n~;ol:W
o'A
-">
0 .
O.
" "0 0
0
Desolvation
A n d y tical Spray Vapor
sample
solution
C
Molecular
Analytical
sample
*
Transport Dlscrete
droplet
-i-,
Desolvation
e
Solid
particle
Volatilization
Vapor
solution
FIGURE 7-3 Formation of atomic vapor with discrete sample introduc-
tion. A discrete sample is transported to the atomization device. The
atomizer heats the sample to the boiling point of the solvent and a solid
particle remains. The solid particle is then heated to a much higher tem-
perature to produce vapor-phase species. With electrothermal atomizers
the heating stages are separated in time. The volatilization step is often
carried out in two stages, one of moderate temperature to drive off organic
material (ashing or charing), and one of high temperature to vaporize the
analyte material (atomization).
troduction into a continuous or noncontinuous atomizer opening that concentrically surrounds the capillary tube,
are illustrated in Figure 7-3. With continuous sample causing a reduced pressure at the tip and thus suction
introduction, a spray is created, while with discrete sam- of the sample solution from the container (Bernoulli
ple introduction, a fixed amount of sample is trans- effect). In the angular or cross-flow nebulizer (Figure
ported to the atomizer. With electrothermal atomizers, 7-4b), a flow of compressed gas over the sample cap-
the various heating stages (desolvation and vaporiza- illary at right angles produces the same Bernoulli effect
tion) are often separated in time by a temperature pro- and aspirates the sample. In most cases the flow of
gram, while with continuous atomizers these steps occur solution is laminar, and the aspiration rate is propor-
sequentially under the influence of heat from the atom- tional to the pressure drop along the capillary and to
ization device. With arc and spark discharges and so- the fourth power of the capillary diameter; it is inversely
lution samples, these same processes occur, although it proportional to the viscosity of the solution. In some
is difficult to separate the stages. Solid samples intro- cases a pump (peristaltic or syringe pump) can be used
duce additional complexities as discussed in Chapter 9. to transport the solution at a rate that is independent
of the compressed gas pressure.
Nebulizers The solution drawn up the capillary tube encoun-
ters the high velocity of the nebulizing gas, which causes
The formation of free atoms in flames and plasmas the formation of droplets of various sizes. Typically,
depends critically on the properties of the sample trans- pneumatic nebulizers produce droplet diameters in the
port-nebulizer-atomizer system. The nebulization step range 1 to 50 km. In most modern nebulizers, the mist
and the remaining steps shown in Figure 7-2 are all produced enters a nebulization chamber, where most
interactive, and the details of the processes that occur of the droplets with diameters larger than a certain cut-
are very complex. The type of nebulizer used influences off diameter are removed from the stream and drained
directly the efficiency of the nebulization, desolvation, away. Devices that remove large droplets from the stream
and volatilization steps. We consider here the most are often called aerosol modifiers. These can be impact
common types of nebulizers employed in atomic spec- beads, as shown in Figure 7-4, spray chamber surfaces,
trometry and leave it for later chapters to discuss more or impaction surfaces. Impact beads may also produce
specialized devices. small droplets from larger drops by causing them to
shatter. Only the smaller droplets are transported to
Pneumatic Nebulizers. The most commonly the atomization source. For plasmas, the nebulization
used nebulization devices are pneumatic devices in which gas (e.g., Ar) containing the fine sample mist is directed
a jet of compressed gas (the nebulization gas) aspirates into the plasma (usually formed from the same gas).
and nebulizes the solution. Two common types are shown For flame systems, the nebulization gas is usually the
in Figure 7-4. The transport of solution to the nebulizer oxidant, and the fuel is brought into the nebulization
tip is known as aspiration. With the concentric nebulizer chamber through a separate port. The mixed oxidant,
(Figure 7-4a) the nebulization gas flows through an fuel, and sample mist are then carried to the burner
Sec. 7- 7 I Sample Introduction and Atomization
Sample
apiary
,-4 Spray
TO plasma or
flame
4
the nebulizer tip. These pneumatic, concentric nebu-
lizer-burners are called total consumption nebulizer-
burners because all the aspirated sample is directed into
the flame without droplet size selection. They are dis-
cussed in more detail in Chapter 8.
-
To plasma or
flame
effects may also be related to the process of droplet
size discrimination within the spray chamber. The glass
frit nebulizer shown in Figure 7-5 produces a much finer
aerosol than the conventional pneumatic nebulizer. It
is not as susceptible to clogging as a conventional ne-
. . bulizer and has thus found some use with samples of a
Nebulizing
gas high salt or particulate content. A mean droplet di-
I'J)
ameter of less than 1 pm has been found for this ne-
bulizer; transport efficiencies on the order of 90% have
FIGURE 7-4 Pneumatic nebulizers. In (a) a
concentric tube nebulizer is shown that is used
been obtained. To achieve this efficiency, the total sam-
with a spray chamber. The nebulizing gas flows ple flow rate must be quite low (0.1 mL min-l) so that
through an orifice that surrounds the sample-con- the overall mass transport rate is not large. However,
taining capillary concentrically. In the angular or this nebulizer may be advantageous where sample vol-
crossed-flow nebulizer (b), the nebulizing gas flows umes are limited and where long nebulization times
over the sample capillary at right angles and causes must be used (e.g., for some multielement determi-
aspiration and nebulization of the sample solu- nations).
tion. Both nebulizers are examples of pneumatic
nebulizers in which a jet of compressed gas as- Ultrasonic Nebulizers. The ultrasonic nebu-
pirates and nebulizes the solution. A spray cham- lizer has often been suggested as a replacement for the
ber separates large droplets from small droplets.
The latter are carried into the plasma or flame,
1 Argon
while the former are drained away. The nebulizer
can be made of glass or metal.
Water
OU t
Aerosol
RF in
Water
in
Argon
(a) ('J)
FIGURE 7-6 Ultrasonic nebulizer. Solution is FIGURE 7-7 Babington-type nebulizers. In the
fed onto the surface of the piezoelectric crystal normal Babington nebulizer (a), the sample is fed
by gravity flow or by a pump. Vibrations of the by gravity or a pump and forms a film over the
crystal cause the solution to break into small outside surface of a sphere or rounded tip with a
droplets, which are transported by the carrier gas small orifice for the nebulizing gas. The gas blows
to the flame or plasma. a hole through the film to produce the aerosol.
In the V-groove nebulizer (b) the sample solution
conventional pneumatic nebulizer. With these devices passes down a V-groove with a small hole in the
a piezoelectric crystal (a ceramic crystal transducer) is center for the nebulizing gas. Both nebulizers can
be made of glass, metal or plastic materials such
vibrated at ultrasonic frequencies (20 kHz to 5 MHz). as Teflon.
Ultrasonic vibrations are coupled to the sample solution
by a coupling liquid, by a velocity transformer, or by
directly flowing the solution onto the vibrating surface. in pneumatic nebulizers have typical inside diameters
The vibrations cause the solution to break up into small of 0.1 mm and are thus subject to clogging when samples
droplets which are transported by the carrier gas through with a high particulate content are aspirated. The ne-
the nebulization chamber to the flame or plasma. In bulizer designs shown in Figure 7-7 have been found to
the vertical, direct-coupled design shown in Figure be quite tolerant to such solutions. Both nebulizers are
7-6, the sample is fed onto the surface of the vibrating variations on the Babington design, which was origi-
crystal, where nebulization occurs. Such nebulizers can nally developed for spraying paint. In both designs the
produce dense aerosols that are more homogeneous in solution is delivered through a tube of much larger in-
droplet size than those produced by pneumatic nebu- side diameter than those used with pneumatic nebu-
lizers. Ultrasonic nebulizers have the advantage that lizers. Thus these nebulizers are essentially free from
the nebulizer parameters (frequency of vibration, power clogging, and they are excellent nebulizers when high
applied to the transducer) are independent of any flame particulates cannot be avoided. The Babington-type de-
or plasma gas flow rates so that separate optimizations vices are, however, less efficient than pneumatic ne-
can be made. The solution flow rates must, however, bulizers in producing small droplets with normal sam-
be fairly low with these nebulizers (0.1 to 1.0 mL min - l). ples. Because of this the processes of desolvation,
The droplet diameters produced depend on the fre- volatilization, and atomization may be less efficient than
quency of the ultrasonic energy as well as the surface with conventional nebulizers and some interference ef-
tension and viscosity of the liquid. High frequencies fects may be enhanced.
favor the formation of small droplets. With a frequency
of 1 MHz and a solution flow rate of 0.3 mL min-l, Isolated Droplet Generator, In order to study
droplets with diameters of 1.5 to 2.5 pm have been closely the processes of desolvation, vaporization, and
produced with up to 30% efficiency. The major limi- atomization, solution samples have been introduced into
tation of ultrasonic nebulizers is their poor efficiency flames as individual, isolated droplets. Figure 7-8 shows
with viscous solutions and with solutions that have high a system for producing isolated droplets. Droplet di-
particulate content. ameters depend on the size of the capillary used and
are typically in the range 10 to 200 pm. By charging
High Solids Nebulizers. The capillary tubes used some of the droplets produced and electrostatically de-
Sec. 7- 1 I Sample Introduction and Atomization 1
fleeting them, the introduction rate can be varied from relatively low gas flow rates, which increase residence
1 Hz to 2 x lo5 Hz. Observation of the size of the times. Organic solvents evaporate more rapidly than
droplets injected into a flame allows the desolvation does water, and with flame atomizers, the desolvation
and volatilization processes to be studied, while meas- can be accelerated by the heat of combustion of the
urement of the emission in a spatially isolated region vapor. Desolvation efficiencies can be quite high for
of the atomizer allows investigation of the atomization pneumatic nebulizers with spray chambers that reject
process. larger droplets. With the total consumption nebulizers,
once popular for flame emission, all of the droplets may
not reside in the high-temperature environment long
enough for complete solvent evaporation, particularly
Free-Atom Formation after Nebulization with watzr as the solvent. When flames are used, water
also tends to lower the flame temperature.
Once the sample solution has been nebulized and trans-
ported to the atomization device, a variety of complex
processes begins. We consider here the desolvation of
the droplets, the vaporization of the solid or molten
particles, the dissociation of molecular species, the ion-
ization of analyte atoms, and the influence of these Volatilization. The solid or molten particle re-
processes on the shapes of calibration curves. maining after desolvation must be vaporized to obtain
free atoms. Incomplete volatilization leads to a loss of
analyte free atoms and thus a reduction in the analytical
signal. It may also cause nonlinearity in analytical curves,
Desolvation. Desolvation of the droplets is the continuous background emission from incandescent
first step that must occur after nebulization. Desolva- particles, and light scattering in the case of fluorescence
tion leaves a dry aerosol of molten or solid particles measurements. The volatilization efficiency depends on
and often begins in the nebulization chamber. The ef- a number of factors, including the atomizer tempera-
ficiency of the desolvation step is determined by several ture, the composition of the analytical sample (nature
experimental variables, such as the atomizer tempera- and concentration of the analyte, solvent, and concom-
ture, the trajectories, diameters and residence times of itants), the size distribution of the dry aerosol, the tra-
the droplets in the atomization cell, the nature of the jectories and residence times of the particles, and the
solvent, and the design of the nebulizer. High-temper- type of nebulizer. For many metals, oxides are less
ature atomizers increase desolvation efficiency, as do volatile than the metal itself or salts of the metal. The
I96 Chap. 7 1 Introduction to Atomic Spectroscopy
bond dissociation energies of the compounds that are The ionization of metal atoms can also be consid-
formed with the analyte are important in determining ered an equilibrium process in many flames and plas-
volatilization efficiencies. With flames, high tempera- mas, at least in localized regions. Such ionization
tures and a reducing environment tend to increase the (M S M + + e-) can be described by the Saha equa-
volatilization efficiency and reduce the formation of tion for the ionization constant K,:
refractory oxides. The volatilization rate increases as
the size of the droplets introduced into the atomizer
decreases.
Dissociation and Ionization. In the vapor phase where n, is the number density of free electrons. Again
the analyte can exist as free atoms, molecules, or ions. ionization constants can be calculated from statistical
The formation of molecular species and ions reduces mechanics and are of the form
the concentration of free atoms and thus degrades the
detection limit. Molecular species, such as CaO, CaOH,
KC1, and LiOH, can be formed by reactions of analyte log K, = 15.684 + log,- 4-
+ 2 (log T) -
atoms with gaseous flame constituents or with volatil-
ized species from the analytical sample. Ionic species,
such as K + , Na+ , CaOHt , and Ca' , are formed by
the loss of an electron by an atom or molecule. where the 2,'s are partition functions for the ion and
In many cases we consider molecules to be in equi- metal (see Section 7-3) and E,,, is the ionization energy
librium with free atoms, at least in localized regions of in eV. Small values of E,,, and high temperatures favor
the atomizer (flame or plasma). If chemical equilibrium the formation of ions. For example, if all electrons come
exists, the law of mass action describes the degree of from the ionization reaction, at moderate concentra-
dissociation of molecular species, and we can write a tions, K (E,,, = 4.34 eV) is about 50% ionized at 2500
dissociation constant for a molecular species (MX) into K, whereas Na (E,,, = 5.14 eV) is only 7% ionized at
its components (MX $ M + X) as the same temperature. Usually, only the first ionization
needs to be considered in most flames and in the ob-
servation region sf most plasmas.
The fraction atomized, also called the free atom
fraction, pa, is a measurable quantity that describes the
where n is the number density (number per cm3) and loss of analyte atoms due to compound formation and
the subscript indicates the species. From statistical me- ionization. It can be expressed as
chanics, the dissociation constant for a diatomic mol-
ecule is a function of temperature and the type of reac-
tants according to
ization efficiency P, and the local free atom fraction that the ideal gas law holds. From the latter, plRT is
p,. Thus the number of moles per unit volume. The number
density n, is then n,, = pNlRT = 7.34 x lo2' PIT.
concentrations and to c at high concentrations. Thus a continuous sample introduction, there are many factors
nonlinearity is predicted at low values of c. that can lead to nonlinearities and many substances that
In most atomizers, n, is determined by the ioni- cause interference in the conversion of analyte to ground-
zation of several species. not just the analyte. If cur- state free atoms.
vature due to ionization is a problem for a particular
analyte, a large concentration of a more easily ionized
element can be added to all samples and standards to 7-2 INTERFERENCES IN ATOMIC
suppress ionization of the analyte. We see from equa- SPECTROSCOPY
tion 7-3 that if n, >> n,, the ratio n,+/n, and the
fraction ionized a, are constant and n, n, .x c. Fur- Many of the interferences caused by concomitants are
thermore, if n, >> K,. a, = 0. quite similar in all atomic spectrometric techniques.
Similar nonlinearities arise due to dissociation However, certain techniques may show freedom from
equilibria (equation 7-2) for small values of K,. As for some types of interference or exhibit increased suscep-
ionization, the fraction dissociated is larger at small tibility to others. In this section some of the interfer-
concentrations compared to large concentrations. Since, ences that were introduced in Chapter 6 are discussed
in this case, the dissociated species M is measured, a in the context of atomic emission, absorption, and flu-
negative deviation in the calibration curve can result. orescence spectroscopy. Interference effects that are
Many other factors affect linearity in specific tech- specific to the medium used (flame, plasma, electro-
niques, as we shall see in later chapters, but the causes thermal atomizer, arc, etc.) are discussed in later chap-
noted above are common for all atomic spectrochemical ters dealing with specific techniques.
methods. We will see in the next section that many
interferences in atomic spectrochemical methods occur Blank lnterferences
because the conversion of the analyte into free atoms
is affected by a concomitant that changes the overall As discussed in Chapter 6, a blank or additive inter-
nebulization-atomization efficiency for a given amount ference produces an uncompensated signal independent
of analyte. of the analyte concentration. In atomic spectroscopic
methods these are usually spectral in nature, although
Free-Atom Formation with Discrete Sample nonspectral blank interferences are possible.
lntroduction
Atomic Emission lnterferences. Spectral in-
With discrete sample introduction into a furnace, a terferences, sometimes called cross-spectral interfer-
droplet of solution is deposited directly in the atomizer ences, are most troublesome in emission spectroscopy
(usually a furnace). Hence there is no nebulization term because the selectivity is normally determined by a
in the expression for the overall atomization efficiency wavelength selection device (filter, monochromator,
(e, = P,P,P,). These factors are usually time dependent polychromator, etc.). Here any concomitant that emits
because of the discrete sample and the fact that the radiation within the bandpass of the wavelength selec-
furnace is usually operated in a noncontinuous mode. tion device or radiation that appears in that bandpass
In most cases the furnace is heated to the boiling point due to stray radiation causes such a spectral interfer-
of the solvent for a time long enough that complete ence. A quick look at any fairly complete set of wave-
desolvation occurs (P, = 1). With these differences in length tables, such as the MIT tables, will convince the
mind, the expression for the instantaneous ground-state reader that almost every sensitive emission line has sev-
free atom density n,(t) is similar to that given in equa- eral potential interfering lines very near it. Whether a
tion 7-9 except that F is replaced by V, the volume of spectral interference is observed or not depends on the
solution delivered to the furnace. Note that e,, ep Z ( T ) , composition of the sample and the technique used to
and thus n,(t) are all time dependent because the at- excite emission. Because arcs, sparks, and plasmas have
omizer temperature varies with time and the amount higher excitation energies than flames, spectral inter-
of sample is limited. ferences are potentially more troublesome with these
In many cases we expect n,(t) to be linearly related excitation sources.
to the analyte concentration [n,(t) = kc]. For this to Atomic line interferences in emission methods can,
be true, F,, Q, ef, and Z ( T ) must be constant and in- in principle, be reduced by improving the spectral res-
dependent of analyte concentration. This implies that olution of the wavelength selection device since true
measurements must be made at constant temperature, spectral line overlaps are rare. Thus high excitation
time, flow rate of support gas, observation height, energy emission sources normally use (and need) much
atomization efficiency, and so on. Just as was true for higher resolution wavelength selection devices than flame
Sec. 7-2 1 Interferences in Atomic Spectroscopy
sources, which often use moderate resolution mono- metallic impurities and the filler gas are also emitted.
chromators or even filters. Spectral interferences are, Such lines can cause spectral interferences if they are
however, common in flame emission methods. Since absorbed and not separated from the analyte line by
the practicing analyst rarely has the option of switching the wavelength selection device. Multielement line
to a higher-resolution spectrometer when spectral in- sources are also employed in A A and these can increase
terferences occur, the choices usually involve decreas- the risk of spectral interferences. Continuum source
ing the spectral bandpass (if possible), changing analyte AA, like flame emission, depends highly on the spectral
spectral lines, changing source excitation conditions, or resolution of the wavelength selection device for selec-
trying to compensate or correct for the effect (see Sec- tivity. If the spectral bandpass is large, the probability
tion 6-5). of absorption by other atomic species is enhanced.
Because of their dependence on excitation con- Background emission is normally not a problem
ditions and spectral resolution, spectral interferences in in A A because modulation and ac detection are used
emission methods are highly instrument dependent. For to discriminate against it. However, absorption by mo-
example, in flame emission with filter photometers, an lecular species and scattering of the source radiation by
interference of the n triplet (403.1, 403.3, and 403.4 nonvolatile salt particles or oxides can occur and can
found on the K doublet (404.4, 404.7 nm). Since give rise to a blank interference (see Section 10-3).
n interfering lines are 1 nm or more from the Absorption by molecular species is particularly a prob-
t K line, a moderate-resolution instrument can lem with electrothermal atomizers and in relatively cool
eliminate this interference. On the other hand, the in- flames. Scattering is also said to cause an interference
terference of Ga (403.298 nm) on Mn (403.307 nm) is in cool flames. At wavelengths shorter than 220 nm,
much more difficult to overcome. This latter interfer- the flame itself becomes absorbing. While this is nor-
ence has been eliminated by using an echelle grating mally taken care of by the blank measurement, any
spectrometer operating in the twelfth order. Another change in the background absorption of the flame by a
troublesome interference is that of Na (285.28 nm) on concomitant in the sample causes a blank interference.
Mg (285.21 nm) in flame emission. This interference is
rarely noted, however, in plasma or arc emission meth- A tomic Fluorescence Interferences. Flame
ods with higher-resolution spectrometers. atomic fluorescence (AF) with line source excitation is
Molecular band emission can occur with flame potentially as free from spectral interferences as line
excitation when the sample contains combustible ma- source AA. Again continuum sources enhance the risk
terials or constituents that give rise to molecular species. that concomitants will produce fluorescence within the
gh concentrations of Ca in the sample bandpass of the wavelength selection device.
band emission, which can be a blank In AF, normal background emission from the
interference if it appears at the analyte wavelength. In atomization system is usually compensated by modu-
contrast to atomic line interferences, the narrow atomic lation techniques. As in AA, alteration of the back-
analyte line is superimposed on the broad emission band ground by concomitants in the sample can cause a non-
from the molecular species, and thus band emission spectral blank interference. Resonance AF is particularly
interferences cannot be eliminated by improving the susceptible to interference from scattering of the source
spectral resolution of the spectrometer. In flame emis- radiation by droplets and nonvolatilized salt particles.
sion the normal background emission from the flame Since resonance A F is measured at the source wave-
itself is not usually an interference since the blank meas- length, it is often difficult to distinguish such scattering
urement can compensate for unchanging background. from true fluorescence. Nonresonance A F methods do
owever, if a concomitant alters the background emis- not suffer from this interference since the fluorescence
sion, it can cause a nonspectral blank interference. signal is measured at a wavelength different from the
source excitation wavelength.
Atomic Absorption lnletferences. Atomic line
interferences are rarely observed in atomic absorption
spectroscopy with narrow line sources since the source
width effectively determines the spectral resolution. As Analyte or multiplicative interferences alter the mag-
discussed in Chapter 4, hollow cathode lamps produce nitude of the analytical signal itself. In atomic spec-
atomic lines with FWHM values on the order of 0.001 troscopy they are invariably nonspectral interferences
nm. In flame A A , for example, there is no interference because of the narrowness of the lines involved.
of Na (285.28 nm) on the Mg 285.21-nm line because
of this high selectivity. Line sources in A A , however, Nonspecific lntetferences. Nonspecific inter-
emit more than just the desired analyte line; lines from ferences, often called physical interferences, are fairly
200 Chap. 7 I Introduction to Atomic Spectroscopy
alters the atomizer temperature and thus the fraction gorize spectral transitions according to spectroscopic
of analyte atoms excited. Since the temperature de- term symbols, shorthand descriptions of electronic states.
pendence of emission is a function of excitation energy, The statistical weights of states are discussed, and the
these interferences can be considered to be analyte spe- concept of the partition function is introduced.
cific. Although excitation interferences occur only in
emission, it should not be implied that changes in at- Quantum Numbers
omizer temperature do not also influence absorption
and fluorescence. Atomizer temperature also affects The electronic configuration of the hydrogen atom can
desolvation, solute volatilization, dissociation of com- be specified by four quantum numbers: n , the principal
pounds, and analyte ionization, and thus changes in quantum number; 1, the orbital angular momentum
temperature influence all three techniques. quantum number; m,, the orbital magnetic quantum
In atomic fluorescence spectroscopy, concomi- number; and m,, the spin magnetic quantum number.
tants in the sample can alter atomizer composition and The description of each of these quantum numbers and
thus change the fluorescence quantum yield. Fluores- their allowed values and symbols are given in Table
cence efficiencies are usually higher in inert-gas at- 7-1. We are all familiar with the simple atomic orbital
mospheres. The presence of oxygen or nitrogen in the description of the electronic configurations of poly-
atomization gas can decrease the fluorescence yield electronic atoms. Here the electrons are assigned to
through quenching. If such an effect is produced by hydrogen-like atomic orbitals using the quantum num-
sample constituents, a quenching interference results. bers of Table 7-1. For many purposes the state of an
It is unlikely that direct quenching by collisions with atom can be classified by the quantum numbers n and
concomitant atoms or molecules occurs with most at- 1 for each electron in the atom, because these are the
omizers because of the low concentrations compared to two quantum numbers that largely determine the en-
the atomizer gases. ergy of the atom. The value of 1 determines the shape
of the orbital. An orbital with 1 = 0 is spherically sym-
metric, has no orbital angular momentum, and for his-
7-3 ELECTRONIC STATES OF ATOMS torical reasons. is designated as an s orbital. Orbitals
with 1 = 1 are called p orbitals, those with 1 = 2 are d
The general characteristics of atomic spectra were in- orbitals, and so on. There are three p orbitals corre-
troduced briefly in Chapter 1. Here we examine in more sponding to the allowed values of m,, five d orbitals,
detail the electronic configurations of atoms and cate- and so on.
TABLE 7-1
Summary of quantum numbers for individual electrons
A given assignment of n and 1 values to all the vector, is called the multiplicity of the state. The
electrons in an atom produces an electronic configu- exclusion principle requires that S = 0 for a completely
ration. The ground-state configuration of an atom can filled subshell.
often be deduced by knowing the number of electrons For atoms with weak spin-orbit interactions, the
in the atom (equal to its atomic number Z), the max- resultant orbital and spin angular momenta couple to
imum number of electrons in a given orbital type (2 for produce a total angular momentum denoted by quan-
s orbitals, 6 for p orbitals, 10 for d orbitals), and the tum number 7, sometimes called the inner quantum
order of filling orbitals. This Au number,'for the entire atom. This Russell-Saunders or
3s, 3p, 4s, 3d, 4p, 5s, 4d, 5p, 6s, 4f, 5d, 6p, 7s, and so LS coupling scheme leads to the allowed values of J in
on. Thus, for carbon (Z = 6), the ground-state config- Table 7-2.
uration is designated ls2 2s2 2p2, where the superscript When spin-orbit interactions are large, LS cou-
indicates the number of electrons in the orbital. For pling no longer describes the electron interaction and
sodium ( Z = 11), the ground-state configuration is ls2 the jj coupling scheme is employed. Here a resultant j
for each electron is computed according to the allowed
auli exclusion principle states that no two values j = 1 + s, 1 + s - 1, ... I1 - sl. The individual
electrons in the same atom can have the same four j's then combine to give a J value for the entire atom.
quantum numbers. Thus, for He ( Z = 2 with config- These vector models provide a convenient phys-
uration ls2), the two electrons must have m, values of ical picture and are useful in calculations. However,
- 4 and -I respectively (opposite spins). neither the LS nor the jj coupling scheme is a completely
adequate description. Instead these are two limits with
intermediate cases possible. The two limiting cases blend
smoothly into each other as a function of atomic num-
For many-electron atoms, the hydrogen quantum num- ber. In fact, even a heavy atom, such as lead, does not
bers can be thought of as describing the individual elec- exhibit pure jj coupling. We shall assume for the re-
trons, but they are not "good" quantum numbers for mainder of this discussion that we can use the L S cou-
the entire atom. In the language of quantum mechanics, pling scheme.
good quantum numbers are associated with operators No matter which coupling scheme applies, the in-
(squared angular momentum and z-component of an- ternal energy, the Jvalue and the parity are well-defined
gular momentum) that commute with the total atom quantities for a given state. According to quantum me-
Hamiltonian. For polyelectronic atoms, three resultant chanics, the parity is odd if the atomic wave function
quantum numbers, L , S, and J, are good quantum num- changes sign upon inversion of the electron coordinates
bers for the entire atom. For light atoms ( Z < 30), with and even if it does not. Configurations with 21, an even
weak spin-orbit interactions, a resultant orbital angular number lead to states with even parity, and those with
momentum specified by quantum number L is produced this sum an odd number lead to states of odd parity.
by coupling the orbital angular momenta of each elec- The electrons in closed subshells contribute to the sum
tron, a process of vector addition. The allowed values by an even number; therefore, the parity depends only
of L and the symbols used to abbreviate them are given on the valence electrons.
in Table 7-2. For a single electron the resultant L is
equal to 1 for that electron.
For more than two electrons, the overall coupling
can be broken down into a chain of pairwise couplings A multiplet of closely spaced states with the same L
of the individual I,. Closed shells possess no net orbital and S values, but different J values, is called a s
motion, and thus the electrons in these shells do not scopic term. A complete term symbol is a designation
need to be considered when the resultant is computed. for a particular electronic state and is written n2S+1{L},,
The individual spin moments couple to give a re- where n is the principal quantum number for the valence
sultant spin quantum number S as shown in Table 7-2. electron(s), {L) is the symbol (S, P , D, F, etc.) for the
Two electrons thus give S = 1 and S = 0 since s, = quantum number L , 2S + 1 is the multiplicity, and J
s2 = i. States with S = 0 are called singlets since there is the total angular momentum quantum number. Thus
is only a single value for Ms, the resultant spin magnetic the complete term symbol for the ground state of so-
quantum number. States with S = 1 are doublets, those dium is 32S,,2. Sometimes the entire electronic config-
with S = 2 are triplets, and so on. The value of uration is used instead of n in the term symbol and
2S + 1, which corresponds to the number of possible often the value of n is omitted. Thus the ground state
orientations of the resultant spin angular momentum of sodium can be written ls2 2s2 2p6 3s2 Sl,2or just
204 Chap. 7 1 Introduction to Atomic Spectroscopy
TABLE 7-2
Quantum numbers for many-electron atoms with LS coupling
L Resultant orbital L = l ~ + 1 2 , 1 1 - 1 2 - l. . . . , I l 1 - l 2 l O = S
angular momentum for two electrons with orbital angular l = P
quantum number; momentum quantum numbers I , and l2 2 = D
determines 3 = F
magnitude of orbital etc.
angular momentum
Resultant orbital ML = C (m,),
magnetic quantum 2L + 1 possible values
number; describes
orientation of
angular momentum
vector
Resultant spin S = s , + s 2 . s , + s 2 - 1 , . . . , S , -szl For 2S + 1:
quantum number; for two electrons with spin quantum 0 = singlet
determines numbers s, and s2 1 = doublet
magnitude of spin 2 = triplet
angular momentum etc.
Ms Resultant spin MS = V m , ) ,
magnetic quantum 2s + 1 possible values
number; describes
orientation of spin
angular momentum
vector
Total angulai
momentum quantum
number; determines
magnitude of total
angular momentum
of entire atom
MJ Resultant total M, = J , J - 1 , . . . . - J
magnetic quantum 25 + 1 possible values
number; describes
orientation of total
angular momentum
vector
2S,,,. Here for the single 3s valence electron, L = 1 = the table. We can see that there must be a D term
0 S = s = $ J = L + S = i . because there are states with M L = 1 2 . For this term,
For atoms with several valence electrons, a given the spins must be paired by the exclusion principle
configuration can give rise to many states. With carbon ( S = 0); thus the D term must be a singlet term. The
( l s 2 2s2 2p2), for example, there are two open-shell p D term is composed of five states with M L = + 2 , 1,
electrons to consider. One 2p electron has n = 2, 1 = 0, - 1 , - 2 and M s = 0. Similarly, there must be a
1 , three possible values of m,, and two possible values triplet term because of the pre,ence of the values
of m,, a total of six possible sets of quantum numbers. M s = 11. The latter values appear only in association
For the second 2p electron (same n and I ) there are five with M L = 11. Hence this is a tripiet P term ( L = 1,
possible sets because of the exclusion principle. For S = 1 ) composed of nine states with M L = + 1 , 0 ,
both electrons, there are (6 x 5)12 = 15 possible as- - 1 and M s = -t1, 0. - 1. The remaining term has
signments since the electrons are indistinguishable. Since L = 0, S = 0 and is thus a IS term. Note that all L
I, = 1, = 1 , L can be 2 , 1, or 0 , and there are D , P, values up to the maximum are represented. Because
and S terms. the electrons are indistinguishable, it is impossible in
Table 7-3 gives the possible arrangements and the some cases to assign a specific microstate to a particular
resulting terms for the carbon ground electronic con- term. Hence states that differ only in the spins of the
figuration. Finding the terms is a matter of picking out electrons are associated with more than one term as
those multiplets that can give rise to all the states in can be seen in Table 7-3. The J values associated with
Sec. 7-3 1 Electronic States of Atoms
the terms for carbon are J = 2 for the D term, J = 2, orbital. In this case n = 3, S = 1, 2 s + 1 = 2,
1, 0 for the 3P term, and J = 0 for the IS term. The L= 1 ( P term), J = %, t. There are two closely spaced
complete set of term symbols for the ground electronic levels in the excited term of sodium with term symbols
configuration of carbon is thus ID,, 3P2, 3P1, 3P0, and ,PI,, and 2P312.The state with J = 4 is of slightly lower
IS,. Term symbols for almost all practical configurations energy than the state with J = t (Hund's rule 3). The
have been tabulated in E. U . Condon and G. H . Shor- well-known yellow sodium D lines result from transi-
tley, The Theory of Atomic Spectra (Cambridge Uni- tions from this pair of excited levels to the ground level
versity Press, Cambridge, 1963). as shown below.
States with unlike term symbols differ in energy
because of different degrees of electronic repulsion and 2P,12 + ,SII2 5896 (Dl line)
correlation. The lowest-energy state for an atom with
several possibilities can be determined with the aid of 2P,12 2S3,2 5890 A (D2 line)
sitions. Transitions that are dipole forbidden may occur allowed. For L S coupling, the change in J is restricted
because they are electric quadrupole or magnetic dipole to jAJ1 5 2; the change in L is limited to lALl 5 2.
allowed. Furthermore, the approximation is made that For alkali metal atoms the two-photon selection rules
the transitions involve only single electrons; that is, allow only 4 L = 0 and I4LI = 2.
there are no changes in the orbitals of all the other The selection ruies noted above can also account
electrons or mixing of states. This is not strictly correct, for the existence of long-lived atomic states with ener-
but the selection rule model works fairly well for de- gies above the ground state. Such states are often called
scribing spectra and for calculating the intensities of metastable states. These cannot lose their energy by
atomic lines. emitting a photon because the transition required is
For single-photon transitions the general selection forbidden. For example, the 3Postate of mercury cannot
rules that must be strictly obeyed for an allowed tran- decay to the ISo ground state because the transition is
sition are: forbidden by general selection rule 3 (J = 0 to J = 0
is forbidden). Metastable atoms are often quite useful
1. The parities of the upper and lower level must carriers of energy in sparks, plasmas, and flames.
be different. Some of the allowed single-photon transitions in
2. A1 = i. 1. Na are shown in Figure 7-lla. Note that the allowed
3. AJ = 0 or ? 1, but J = 0 to J = 0 is for- transitions are determined by the selection rule A1 =
bidden. t 1. The emission spectrum of sodium (and other alkali
metals) consists of three major series of spectral lines.
In addition, there are special selection rules that depend Transitions between P states and the 3S ground state
on which coupling scheme (LS or jj) prevails. For LS form the principal series. Transitions from excited S
coupling and single-photon transitions, these special se- states to one of the 3P states form the sharp series,
lection rules are: while those from D states to one of the 3P states form
the diffuse series. For hydrogen and alkali metals the
1. 4s = 0. spectral series converge at the ionization limit. For more
2. AL = 0 , o r i 1 , b u t L = OtoL = Ois complex atoms different series may converge to differ-
forbidden. ent limits. The alkaline earth elements give spectra quite
different from the alkali metals. From the energy-level
For two-photon processes (see Chapter l l ) , only diagram for Ca in Figure 7-llb, it can be seen that the
transitions between levels that have the same parity are resonance line at 422.7 nm is a single line and not a
Sec. 7-3 1 Electronic States of Atoms
oublet as was the case with Na. Note also that the with respect to the field. Figure 7-12a illustrates the
triplet terms do not give resonance lines at all, as the energy splittings for several different values of J . It can
transition to the ground state is spin forbidden. The 4p be shown theoretically that the selection rule for M, is
triplet terms are, in fact, metastable. AM, = 0, i- 1. Singlet terms give rise to the nor
If an electron other than the outermost is excited Zeemae effect, in which the splitting between compo-
in an atom or if two electrons are excited simultane- nents is the same in different states of the atom, as
ously, the energy of the resulting state may be high illustrated in Figure 7-12b for Ca. For the normal Zee-
enough that it can pass into a ground-state ion and a man effect, the magnitude of the splitting is directly
free electron with excess kinetic energy. Such states are proportional to the magnetic field strength.
said to be autoisnizing states. They result from spectral For terms with S > 0, the Zeeman splitting be-
lines with upper terms above the normal ionization limit. tween components is no longer the same in different
These lines are often observed to be broadened due to states, and this anomalous Zeeman effect can give rise
the very short lifetime of the upper state (see the next to a more complicated splitting pattern, as illustrated
section). Spectral lines from such states may also have in Figure 7-12c for the Na D l lines. The magnitude of
anomalous intensities. The existence of a level above the splitting in this case is a complex function of the
the normal ionization limit implies that the recombi- field strength. The Zeeman effect is used for back-
nation of a free electron of the appropriate energy with ground correction in atomic absorption spectrophotom-
the ion has an enhanced probability. High current den- etry (see Section 10-3).
sities in electrical discharges are, in fact, often found In the presence of an electric field spectral lines
to enhance lines which show autoionization because of can also be split into closely spaced components by the
recombination followed by emission. Stark effect. For hydrogen the Stark effect removes the
degeneracy between states of the same n , but different
1, producing an effect in which the splitting is propor-
tional to the field strength (first-order Stark effect). In
An additional splitting of terms can occur because of more complicated atoms there is no degeneracy be-
magnetic coupling of the spin and orbital motion of the tween states of different I, and the splitting is propor-
electrons in atoms with the nuclear spin. Such splitting tional to the square of the electric field (quadratic Stark
results in hyperfine structure. For Na the 2S,,2 ground effect). Line broadening due to the Stark effect is im-
level is split into two close levels, which leads to ad- portant in arcs, sparks, and plasmas, as discussed in the
ditional splitting of each D line. For Na, the hyperfine next section. The Stark effect is widely used in analyzing
splitting is on the order of 0.02 A and not readily de- microwave spectra of molecules.
tected. Other atoms, such as Cu and In, exhibit larger
hyperfine splittings that are important in analytical ap-
plications.
When several isotopes of an element are present,
a splitting of atomic lines can result from the issts We have seen in the preceding section that atomic states
shift. The resonance lines of 'jLi and 7Li are separated differing only in their M, values are degenerate. The
by about 0.16 A, enough to determine their concentra- t of a level is the number of degenerate
quantum states making up that level. It is important in
tion ratios by atomic absorption.
In external fields atoms undergo additional term calculations of population distributions and spectral line
radiances because we must account for the degeneracy
splitting. The splitting of spectral lines in a magnetic
of a particular level by weighting it according to the
field is known as the Zeeman effect. The quantum num-
numbers of quantum states it contains. An electronic
ber M, (see Table 7-2) describes the orientations (pro-
state with total angular momentum quantum number J
jections) of the J vector; it assumes values from +J to
has a statistical weight g given by
-J that differ by unity. In the absence of a magnetic
field, states with the same values of L, S, and J but with
different M, values are all of equivalent energy and are g = 2 J + 1 (7-12)
said to be degenerate states. For carbon, for example,
the five states with the term symbol ID, are degenerate. The 2P,,2excited state of Na has a statistical weight of
Similarly, for calcium (see Figure 7-llb), the three states 2 and the 2P3,2 state has a statistical weight of 4. In some
with the term symbol IP, are degenerate. In a magnetic cases fine structure splitting is ignored and the statistical
field, however, there are slight energy differences cor- weight refers to the multiplet or term as a whole. In
responding to the different orientations of the J vector this case g = (2L + 1)(2S + 1). For the Na 2Pterm,
208 Chap. 7 I Introduction to Atomic Spectroscopy
FIGURE 7-12 Zeeman splitting of spectral lines. In (a) the energy levels
corresponding to different MJ values are shown for 4 values of J in the
absence (to the left of the dashed lines) and in the presence of a magnetic
field (to the right). The normal Zeeman splitting pattern for the Ca 7326-
A line is shown in (b). The upper and lower levels are split by equal
amounts in the normal Zeeman effect giving rise to three lines corre-
sponding to the three allowed values of AM,. Components with AM, =
6are called n components, while those with A M , = k 1 are called cr
components. The u transition occurs at lower energy than the a+ tran-
sition. In (c) the anomalous Zeeman effect is illustrated for the Na D,
line. Here the upper and lower levels are split by different amounts so
t s the same AM, are no longer of equivalent
that the c o m p o ~ ~ nwith
energy. This gives rise to four lines. The same effect is observed for the
Na D, line ('P,,,-t ,S,,,).
ignoring the fine structure splitting gives rise to a sta- of the number of states available (g, weighted by the
tistical weight of 6 for the doublet as a whole. Boltzmann factor e-"IkT). It is the function needed to
The internal partition function for an atom is de- obtain the statistical mechanical properties of a system,
fined as as we have seen earlier in this chapter with dissociation
and ionization constants. As shown in Chapter 2 (equa-
tion 2-14), the partition function is also needed in the
normalization of the Boltzmann distribution to allow
calculation of the fractional population of an energy
where g, is the statistical weight and E, is the energy of level. As equation 7-13 shows, partition functions de-
the ith level. For the ground state E, = 0. The partition pend on temperature.
function or "sum-over-states" is thus a weighted sum One problem in calculating the partition functions
Sec. 7-4 I Spectral Line Profiles 9
for atoms is that the number of energy levels is infinite croscopic process must equal the reverse rate of that
since the principal quantum number n is unlimited. Since same process, which is known as the principle of
the Boltzmann factors in equation 7-13 are limited to balancing. This principle allows us to state that the pho-
values below the ionization limit, the partition function tons emitted and absorbed from a continuous radiation
would also be infinite. In practice this dilemma is avoided field in equilibrium have the same spectral distribution,
by assuming an upper cutoff of the summation; the sum namely S,. Thus we shall consider here the factors that
is bounded because with a very large orbital radius the contribute to the distribution and keep in mind that the
electron begins to interact with other atoms or the walls results apply equally well to absorption and emission
of the container. Fortunately, if the atom has only a as long as equilibrium conditions prevail.
single ground level and a lowest excited level at least 2
eV above the ground level, the contributions of higher
go is the statistical weight of the ground state. In cases Consider a two-level atomic system undergoing the var-
ious radiational and nonradiational processes shown in
where the ground state is a multiplet with fine-structure
splitting, however, the assumption above is not valid Figure 7-13. Because of emission and absorption from
the radiation field and collisional processes, states j and
and other terms must be included. Partition functions
i have finite lifetimes, and this gives rise to uncertainties
have been calculated for several atoms at temperatures
existing in common media [see, e.g., L. deGalan, R.
Smith, and J. D. Winefordner, Spectrochim. Acta, 23B, -
in the energies of both states according to the Heisen-
berg relationship AEAt h12n. Because photons emit-
ted or absorbed by the atomic system have frequencies
521 (1968)l.
determined by v = (E, - E,(lh, uncertainties in the
energies of the states give rise to a frequency distri-
bution of photons.
The upper-state lifetime 7, is determined by ra-
diative processes (spontaneous and stimulated emis-
When atomic emission, absorption, or fluorescence sion) and collisional deactivation. In fact, the excited-
spectra are recorded, narrow spectral lines are ob- state lifetime is the reciprocal of the sum of the rate
tained. With ordinary spectrometers, the widths of the constants for deactivation as given below.
lines obtained are determined not by the atomic system,
but by the properties of the spectrometer employed (slit
function and spectral bandpass). With very high reso- 7, = (A,, + B,,U, + k,) ' -
Iline
S, dv = 1 or
I
line
S,, dX = 1
Absorption Collisional
deexcitation
Stimulated
emission
We shall deal most often with the spectral distribution
FIGURE 7-13 Excitation/deexcitation processes
in terms of frequency, because line-broadening expres-
in a two-level atom. Radiational proc&ses are
sions are simpler and easier to interpret than in wave- absorption, spontaneous emission, and stimu-
length units. The distribution S, has the units of time lated emission. Nonradiational processes are col-
or H z r l ; it can be converted to S, in (length)-I units lisional excitation and deexcitation. The rate con-
(e.g., nm-') by S, = S,(cih2,), where A, is the peak stants for these processes are given in the diagram.
wavelength. The radiation field has energy density U, at fre-
In thermal equilibrium, the forward rate of a mi- quency v,.
210 Chap. 7 1 Introduction to Atomic Spectroscopy
uncertainty in its energy AE, given by It results from the natural or radiative lifetime T, given
by
atom and partners that are a different species (atoms, partners (cm s-l), a, is the optical cross section for
molecules, or ions), the broadening is called foreign gas adiabatic collision broadening (cm2), and nx is the den-
broadening or Lorentz broadening. When collisions are sity of perturbers. Here we define u, as ~ ( r +, r,)2,
between like atoms, the broadening is called Holtzmark where r, and r2 are the radii of the collision partners.
roadening or resonance broadening. From the kinetic theory of gases, the average relative
Diabatic collisions reduce the lifetime of the initial velocity can be expressed as & = (8kTlnp)112,where
state and give rise to an exponential decay of the ex- p is the reduced mass of the collision partners in grams.
cited-state population with time. The diabatic collision If we use this value in equation 7-22 and substitute 7,
lifetime 7, is given by into equation 7-21, the adiabatic collision half-width is
expressed as
value of Ah, is -
For Na in a 2500-K flame diluted with N,, the
35 mA. This value greatly exceeds
the natural half-width and is greater than the diabatic
Note that the collisional deactivation rate constant k,
collision half-width. In addition to broadening the lines,
(s-l) may be replaced by kln,, where k; is the second-
adiabatic collisions can also cause a small shift in the
order rate constant (cm3 s p l ) and nx is the number
line center (usually toward the red) and an asymmetry
density of collision partners x ( ~ m - ~The
) . spectral line
in the far line wings.
profile for a line broadened by diabatic collisions is
Lorentzian and of the form of equation 7-18 with Av,
replaced by Av,. The values of Av, are highly media Total Lorentzian Profile. If natural and colli-
dependent. For Na in flames containing Ar as a diluent, sional broadening are assumed to be mutually inde-
AX, is as small as 0.3 mA. For flames diluted with N,, pendent, the resulting half-intensity width of the Lor-
a much more effective collision partner, AX, can be as entzian profile Av, is Av, = Av, + Av,, where Av, is
high as 10 mA (0.01 A). In flames diabatic collision the half-intensity width from both adiabatic and diabatic
broadening is often, but not always, small compared to collisions. The latter is not the simple sum of Av, and
adiabatic collision broadening. Av,, because the two types of collisions are not mutually
Adiabatic collisions are more difficult to treat the- independent. The total spectral profile S,, is then a
oretically, although extensive classical, semiclassical, and Lorentzian of the form
quantum mechanical treatments have been made. In
the classical sense an adiabatic collision results in a
change in phase of the atomic oscillator. Such phase-
changing collisions can be characterized by a correlation
time T,, which can be interpreted as the average time Here we have neglected any shifts in the line center or
between two phase-changing collisions. The more fre- asymmetry caused by adiabatic collisions. In flames and
quent the collisions, the shorter the correlation time.
The half-width of the spectral distribution that results
from adiabatic collisions Av, is then
Av, -
plasmas, it is usually found that Av, >> Av,, so
Av,.
1
Av, = -
"i7,
The Doppler effect is an additional source of broad-
and the spectral profile is Lorentzian as in equation ening in most atomic spectroscopic experiments. It re-
7-18 with Av, replacing Av,. The reciprocal of the cor- sults because of a statistical distribution of velocities of
relation time is the average number of effective colli- the emitting or absorbing atoms along the observation
sions per unit time. For conditions of thermal equilib- path. Because atoms are in motion with respect to the
rium 7;' is observation line, the Doppler effect causes a statistical
distribution in the frequencies observed that is directly
related to the velocity distribution. In thermal equilib-
rium the distribution of velocities is given by Maxwell's
where G,is the average relative velocity of the collision law, which expresses the fraction, f(v,) dv,, of particles
212 Chap, 7 I Introduction to Atomic Spectroscopy
moving along the x axis that have velocities in the in- specified time interval have an equal probability of ab-
terval v, to v, + dv,. Maxwell's law predicts a Gaussian sorbing or emitting any frequency within the absorption
distribution of velocities given by line. Inhomogeneous broadening occurs when different
atoms are responsible for different parts of the line
( v )v = ) 312
e -"":~dv,
profile. Natural broadening is always homogeneous, but
collision broadening can be inhomogeneous if the time
interval for absorption of a photon is short compared
where m is the mass of the particle, T is its temperature, to the duration of the disturbance of the atomic energy
and k is Boltzmann's constant. If the velocity of an atom levels. The distinction between these two types of
does not change while radiating, the profile of a Dop- broadening is important when intense, monochromatic
pler-broadened line S,, is also Gaussian with a half- laser beams interact with atomic systems. In this case
width Av,: only those atoms having a velocity component in the
beam direction that satisfies the Doppler relationship
will contribute to the absorption and induced emission.
Such inhomogeneous broadening can be used to gen-
erate Doppler-free spectra (see Section 17-4).
The maximum value of the Doppler-broadened spectral
profile (S,,), occurs when v = urn and is given by Other Causes of Line Broadening
The quantity 6(a, v,) is the Voigt integral given by becomes a dominant part of the Voigt function in the
line wings. For values of a < I , as shown. the Voigt
a -= -P function resembles a Gaussian profile near the line cen-
6(a, v,) =- dy (7-30)
- a +( - y)2 ter. Figure 7-14b shows Voigt profiles for various values
of the a-parameter.
where y is an integration variable. The a-parameter or The convolution process implicit in the Voigt pro-
damping constant a is related to the ratio of the Lorentz file is just one way to account for the simultaneous
half-width to the Doppler half-width by action of various broadening processes. It assumes that
the natural, diabatic, and adiabatic profiles are Lor-
entzian functions and that Doppler broadening is a
Gaussian function. The major assumption is that the
Gaussian and Lorentzian components are mutually in-
The parameter v, is the relathe frequency compared to dependent. The mutual independence of the various
AvD and is given by broadening processes has been questioned by several
authors. Quantum mechanical theories of line broad-
ening have also been developed. These do not require
the foregoing assumption, and are formally at least more
satisfactory. However, they do require an interaction
potential to account for the broadening effects before
In the limit as a -+ 0, equation 7-29 simplifies to a pure numerical calculations become feasible. The form of
Gaussian profile (equation 7-25), and as a -+ m , to a the interaction potential is not known and thus its in-
pure Lorentzian profile (equation 7-24). clusion also involves several assumptions. Experimental
The Voigt distribution function (equation 7-29) is results of line broadening studies in flames have shown
fully determined by the a-parameter, v,, and AvD. We that the line core and the near wings are adequately
shall see in the next section that the a-parameter plays described by the Voigt profile. However, the decrease
an important role in describing the intensity of a spectral in intensity in the far line wings has been found to
line as a function of the atom density. deviate from the v ; ~dependence predicted by the Voigt
Numerical values of the Voigt integral, the ratio profile, Similar results have been obtained in gas cells
of the normalized Voigt profile to the normalized Dop- at pressures between l o p 2 and 1 atm.
pler profile, can be found as a function of the a-param-
eter and v, in many tabulations. Within lo%, the fol-
lowing approximate expressions hold for the Voigt 7-5 SPECTRAL LINE INTENSITIES
integral at the line center (v, = 0):
The intensities of atomic emission, absorption, and flu-
orescence lines are functions of many variables. We
and shall consider here general expressions for line intens-
ities as a function of atomic population densities. In
later chapters that deal with specific atomic spectro-
chemical methods, these expressions are expanded to
include specific experimental and instrumental param-
Note that the Voigt integral and the a-parameter are eters.
identical if profiles are expressed in wavelength or in
frequency units. Thermal Emission
One cannot find the half-intensity width Av, of
the Voigt profile by simple addition of hv, and AvD. We shall find here the radiance of a spectral line emitted
Within about 1 % the following empirical expression by a collection of atoms in thermal equilibrium. In
holds for all a-parameter values. Chapter 2 (equation 2-15), the radiant power of emis-
sion was shown to be a function of the population den-
sity of excited atoms, the number of photons emitted
per second by each excited atom (the Einstein A coef-
ficient), the energy of each photon hv, and the observed
Figure 7-14a shows an example of normalized Lor- volume element V of the emitting system. If we consider
entzian, Gaussian, and Voigt profiles for Av, = AvD emission from atoms with upper level j and lower level
(a = 0.84). It can be seen that the Lorentzian profile i, equation 2-15 becomes, after substitution of the Boltz-
Sec. 7-5 1 Spectral Line Intensities
mann distribution (equation 2-14), (m, cm, nm). Thus the line radiance can be calculated
if A, can be evaluated, since Bfmis known from Planck's
law.
The absorption factor a@) is related to the atomic
absorption coefficient k(X), in cm-l or mP1, by Beer's
where n, is the total number of free atoms per cm3. law:
This simple equation holds at low population densities.
At high population densities, a significant fraction
of the photons emitted by excited atoms is absorbed by where I is the pathlength (cm or m). The atomic ab-
atoms in the lower level. This phenomenon of self-ab- sorption coefficient can be written in terms of the pop-
sorption is particularly a problem when the lower level ulation density ni (atoms ~ m - of ~ the
) lower level and
is the ground level (resonance transition) because the Einstein coefficients or oscillator strengths as discussed
majority of atoms are present as ground-state atoms in Appendix F. The most useful relationships for this
even in very hot environments. Self-absorption could discussion are equations F-26 and F-28, which are re-
be included in equation 7-31 by multiplying @, by a peated here:
factor that describes the fraction of emitted photons
that escape the emission source without being absorbed
by lower-level atoms. However, we will take here an
alternative approach that includes the self-absorption
phenomenon in a natural way.
The spectral radiance BAEof any thermal emitter
at temperature T is related to that of a blackbody ra-
diator Bf at the same wavelength and temperature by
its emissivity &(A)(see Section 4-1). Thus
where A,, is the Einstein coefficient for spontaneous
emission, S, is the spectral profile function (we will omit
the subscript V for Voigt profile for simplicity), h is
For a radiator in thermal equilibrium, the absorptance Planck's constant, the g values are the statistical weights,
a(X) equals the emissivity at the same wavelength so is the permittivity of free space, me is the mass of
[a@) = &(A)].Hence the electron, c is the speed of light, and f , is the ab-
sorption oscillator strength. The constant in equation
7-36 has the units of crn with S, in cm-I and A in cm.
The spectral profile can also be expressed in terms
The spectral absorptance a(A), often called simply the of (SAD),, the peak value of the Doppler profile, and
absorption factor, is related to the spectral transmit- &(a,vr), the Voigt integral, or
tance of the medium by a(X) = 1 - T(X).
The total line radiance BE emitted by the thermal
radiator is found by integrating BAEover the entire line:
where (S,,), is given by 2 Z/ji;;-Z/(Ah,fi) (equation
7-26). Using this equation in equation 7-35 yields
BE = 1line
B,, dA = 1
line
Bfa(A) dA (7-32)
An approximation can be made to evaluate the sectional area of the cell (area x I = V) and the total
line radiance if the product of k(A) and 1 in equation solid angle over which emission occurs (4n sr).
7-34 is much smaller than unity, which usually occurs In the case of high optical thickness [k(h)l >> 11,
at low values of n,. In this case the system is said to an asymptotic expression can be derived for the integral
be optically thin and we can approximate 1 - eck(",' absorption A,. The result is
as k(A)l. The line radiance is then
(Bdk(,,,~ = Btm J
line k(h)l dh (7-37)
- e2h$nihy1
a, = 1 - exp
The maximum absorption coefficient can readily For optically thin conditions (nil + 0), series ex-
be evaluated. It is worthwhile to define an effective pansion of the exponential term in equation 7-46 gives,
width Aheff as the width of an equivalent rectangular after evaluation of the constants,
profile with the same peak value and same area as S,.
Thus
After evaluating the constants equation 7-48 becomes gion over which ((3,)" exists is determined by the spec-
tral bandpass s of the monochromator. We will assume
that (@,), is constant over s and equal to (@,), the
spectral radiant power at the absorption line center.
Since the bandpass s is much larger than the absorption
Note here that A, is predicted to be strictly a linear line width, we can express a, as
function of n,. In practice nonlinearity occurs because
of the variation of k(X) over the source width (poly-
chromatic radiation), because of stray radiation, and
because not all light rays travel the same path 1 through
the absorbing gas. Unresolved hyperfine structure of where all integrals are taken over A, t s. Since k(X) is
the atomic line can also cause deviations from linearity, finite only over the absorption line width, the integral
because of different k(X) values for the hyperfine com- in the numerator is just A,, the integral absorption. The
ponents. integral in the denominator is just s, the spectral band-
In the case of a continuum source (e.g., xenon arc pass. Hence
lamp), the incident spectral profile is much broader than
the absorption profile, as shown in Figure 7-17a. In fact,
the incident spectral profile detected is usually deter-
mined by the spectral bandpass s of the monochromator
Note that A, has the limiting values given by equation
that follows the atomic vapor cell, since the mono-
7-38 for optically thin conditions and by equation 7-41
chromator bandpass is much larger than the width of
for optically thick conditions. A plot of log a, vs. log
the absorption profile.
n,, for a given bandpass s, is shown in Figure 7-17b and
To find the absorption factor a, for a continuum
has the same shape as the growth curves of Figure
source, we evaluate equation 7-42, where now the re-
7-15. For optically thin conditions, evaluating the con-
stants in A, in equation 7-51 gives
Atomic Fluorescence
log r z ,
Many of the concepts and expressions described earlier
(b)
in this section apply as well to atomic fluorescence (AF)
FIGURE 7-17 Spectral profiles (a) and depend- because fluorescence involves absorption of incident
ency of absorption factor a, on atomic population radiation as in A A , emission of fluorescence radiation,
(b) for a continuum source. The source spectral
and possible self-absorption of the emitted radiation as
profile in (a) is determined by the monochro-
mator slit function t ( X ) and the half-width is the
in AE. Although there are many types of A F transi-
monochromator bandpasss. Note in (a) that (QA),, tions, we will restrict the discussion here primarily to
is essentially constant over the absorption profile. resonance fluorescence, which is the most widely used
The hatched area in (a) represents the wave- type. Other A F excitation/deexcitation schemes are
lengths where absorption can occur. considered in Chapter 11.
220 Chap. 7 / Introduction to Atomic Spectroscopy
At large atomic concentrations (i.e., high n,l), A, We will consider only the qualitative effects of
in equation 7-58 varies as the square root of n, (see self-absorption on resonance fluorescence growth curves.
equation 7-41). Thus under these conditions, @bc is Exact expressions have been derived, but they are com-
proportional to fi. plex and highly dependent on cell geometry. As a first
If the source is a line source that emits at A, from approximation, the self-absorption factorf, exhibits the
equation 7-43 we have proportionality
prime symbol distinguishes the fluorescence radiant FIGURE 7-18 Growth curves for atomic fluo-
power produced in volume element V from that ob- rescence for continuum source and line source
served. excitation.
222 Chap. 7 I Introduction to Atomic Spectroscopy
TABLE 7-4
Dependencies of line strengths o n atomic population densitiesa
- - - - -- -
PROBLEMS
7-1. Find the ground-state term symbols for the following while in atomic absorption systems the monochro-
atoms and show how you arrived at them. mator often has a spectral bandpass in the range
0.2 to 1.0 nm.
Ne, B, Li, A r ; Ca, Al For the Ca 422.7-nm resonance line in a flame at 3000
K, calculate the Doppler half-width in nanometers.
7-2. The total pressure of sodium (atoms plus ions) in a For Na atoms in a 2500 K flame diluted with N, at
2800-K flame is 1.0 x atm. The ionization energy atmospheric pressure, the value of the adiabatic col-
of Na is 5.14 eV, and the equilibrium constant for lision half-width for the 589.0 nm D line is approxi-
ionization is K, = 7.4 x atm. Find the fraction mately 35 mA. Find the optical cross section for adi-
of Na present as nonionized atoms for the following abatic collision broadening cr, in A2.
conditions. Oscillator strengths for absorption transitions are often
(a) All electrons in the flame arise from Na ionization. tabulated as gf values, where g is the statistical weight
(b) The partial pressure of electrons is buffered at l o 7 of the lower level and f is the emission oscillator strength.
atm by adding potassium. For the Na 2Sl,,-+ ,P3 transition at 5890 A, gf = 1.3.
7-3. Two solutions used in flame emission spectrometry Find the value of the Einstein transition probability
with an air-C,H, flame contain identical amounts of (see Appendix F for the relationship between A and
calcium. Solution 1 is a standard, and solution 2 is the f), and the radiative lifetime of the upper level.
sample. For each of the measurement conditions be- Consider flame atomic spectrometric measurements of
low, compare the atomic ground-state number densi- Cu at 324.7 nm in a typical stoichiometric N,0/C2H,
ties with solutions 1 and 2 in a qualitative manner flame. (See Table 8-3 for temperature and stoichi-
[(n,), = (n,),, (n,), > (n,),, relative populations in- ometry information.) W e will b e concerned with tran-
determinant, etc.]. Justify your answers. sitions between the first excited state and the ground
(a) The flow rate of air, the nebulizing gas, is higher state at A = 324.7 nm. Assume that the solution trans-
when solution 2 is sprayed than when solution 1 port rate is 5.0 cm3 min-', E, = 0.030, E, = 0.050, Q
is sprayed. = 10 L min-I, Z ( T ) = go, A,, = 0.95 x lo8 s-', go
(b) The viscosity of solution 2 is greater than that of = 2, and g, = 4. The test solution is a 2.0 pg mL-'
solution 1. aqueous solution of Cu. Calculate the following val-
(c) Solution 2 contains 1000 pg mL-' KC1 in addition ues.
to calcium. (a) The values of no and n,.
(d) Both solutions contain a large excess of phosphate. (b) The total radiant power emitted in 1.0 cm3, a,.
However, solution 2 was made 0.01 M in E D T A (c) The radiance due to Cu emission from a 1.0 cm3
prior to introducing it into the flame. cube of atomic vapor. B E .
7-4. Give a short (one or two sentence) scientific reason For the same system and conditions as in problem
why the statements below are true. 7-8, calculate the following values.
(a) Spectral lines in flames often have half-widths (full (a) The natural, Doppler, collisional, total Lorent-
widths at half maximum intensity) o n the order of zian, and total (Voigt) half-widths. (Calculate in
0.005 to 0.01 nm, whereas natural broadening is Hz and in nm, and assume that adiabatic collisions
predicted to give a line width on the order of are dominant and that the average atomic mass of
2 x nm. the species colliding with Cu is 23 g, the number
(b) A monochromator with a spectral bandpass < 0.1 density of perturbers is 2.3 x 1018~ m - and ~ , the
nm is usually found in atomic emission systems, collision cross section is 300 A,.)
Chap. 7 / References 223
(b) The a parameter. (a) List the allowed transitions and justify according
(c) The value of the Voigt integral at the maximum. to the selection rules.
(d) The maximum value of the Doppler-broadened (b) If an external continuum source is used to observe
profile in Hz-' and nm-'. the absorption of Hg atoms in a room-temperature
(e) The maximum value of the Voigt profile in Hz-' vapor, list the transitions that will be seen if the
and nm-'. selection rules are strictly obeyed.
(0 The effective half-width in nm. If the 3P2,,,0states of Hg are thermally populated in
(g) The peak absorption coefficient for pure Doppler a flame at 2700 K, calculate the ratio of the populations
broadening and for the Voigt profile. of the 3P2and 3P0levels. The splitting is 6398 cm-'.
(h) The fraction absorbed a, and the absorbance with Compare continuous and noncontinuous atomizers.
a line source, A,, with a burner of 5 cm pathlength.
Discuss why the "optimum" temperature might be dif-
(i) The total fluorescence radiant power emitted from
ferent for AA, AE, or A F measurements of a given
a 1.0-cm3 cube of atomic vapor if the incident ra-
element.
diant power is 1.0 mW and the power yield is 0.2.
Why do the shapes of A A and A F calibration curves
7-10. The accompanying figure is a partial term diagram for
differ between line and continuum source excitation?
Hg. Use the information in the diagram to answer the
following questions. For CaOH, the dissociation energy Ed = 4.3 eV. Find
the dissociation constant K, at 1000, 1500, and 2000
Singlets Triplets K. You can assume that the ratio Z,,Z,,/Z ,,,, = 1.
IS, IP, 3 ~ 1 3p2,l,o For LiOH, the dissociation energy Ed = 103 kcal
I mol-l. If the partition function ratio is unity, find
K, at 2000 K.
REFERENCES
The following are reference books and reviews dealing with The following are references having a great deal of data on
atomic structure, atomic spectra, and atomic energy levels. energy levels, transition probabilities, and wavelengths of
1. H. G. Kuhn, Atomic Spectra, Academic Press, New York, spectral lines.
1962. A very readable book on atomic spectra. 6. S. Bashkin and J. 0. Stoner, Jr., Atomic Energy Levels
2. E. U. Condon and G. H. Shortley, The Theory of Atomic and Grotrian Diagrams, vols. 1-4, North-Holland, Am-
Spectra, Cambridge University Press, London, 1967. One sterdam, 1975-1982.
of the classics on the quantum mechanical treatment of 7. C. E. Moore, Atomic Energy Levels, vols. 1-111, NBS
atomic spectra. Particularly good on term symbols and Circular 467 (vol. I), NSDRS-NBS-35 (vols. 11, 111),
coupling modes. National Bureau of Standards, Washington, D.C., 1949
3. G. Herzberg, Atomic Spectra and Atomic Structure, (vol. I), 1970 (vols. 11, 111).
Dover, New York, 1944. A classic. 8. C. H. Corliss and W. R. Bozman, Experimental Tran-
4. J. J. Devlin, "Origins of Atomic Spectra," in Analytical sition Probabilities for Spectral Lines of Seventy Ele-
Emission Spectroscopy, vol. 1, pt. 11, E . L. Grove, ed., ments, NBS Monograph 53, National Bureau of Stand-
Marcel Dekker, New York, 1971, pp. 73-130. ards, Washington, D.C., 1962.
5. A. C. G. Mitchell and M. W. Zemansky, Resonance 9. W. L. Wiese, M. W. Smith, and B. M. Glennon, Atomic
Radiation and Excited Atoms, Cambridge University Press Transition Probabilities, Hydrogen through Neon,
(New York and London), 1961. The classic work on NSDRS-NBS-4, vol. I, National Bureau of Standards,
atomic spectroscopy. Washington, D.C., 1969.
Chap. 7 1 Introduction to Atomic Spectroscopy
W. L. Wiese, M. W. Smith, and B. M. Miles, Atomic The following deal with more specialized topics of concern in
Transition Probabilities, Sodium through Calcium, this chapter.
NSDRS-NBS-22, vol. 11, National Bureau of Standards,
P. J. Th. Zeegers, R. Smith, and J. D. Winefordner,
Washington, D.C., 1969.
"Shapes of Analytical Curves in Flame Spectroscopy,"
J. Reader, C. H. Corliss, W. L. Wiese, and G. A. Mar- Anal. Chem., 40, 13 (1968).
tin, Wavelengths and Transition Probabilities for Atoms
J. D . Winefordner, V. Svoboda, and L. J. Cline, "Crit-
and Atomic Ions, NSDRS-NBS-68, National Bureau of
ical Comparison of Flame Spectrometric Methods," CRC
Standards, Washington, D.C., 1980.
Crit. Rev. Anal. Chem., 1, 233 (1970).
W. F. Meggers, C. H. Corliss, and B. F. Scribner, Tables
J. D. Winefordner, "Spectroscopic Methods," in Trace
of Spectral Line Intensities, pt. I, Arranged by Element;
Analysis: Spectroscopic Methods for Elements, chap. IV,
pt. 11, Arranged by Wavelength, U.S. Government Print-
J . D. Winefordner, ed., Wiley-Interscience, New York,
ing Office, Washington, D.C., 1961.
1976.
G. R. Harrison, Wavelength Tables, MIT Press, Cam-
A. Syty, "Developments in Methods of Sample Injection
bridge, Mass., 1969.
and Atomization," CRC Crit. Rev. Anal. Chem., 4, 155
A. N. Zaidel, V. K. Prokof'ev, S. M. Raiskii, V. A. (1974).
Slavnyi, and E. Ya. Shreider, Tables of Spectral Lines,
E . J. Meehan, "Optical Methods: Emission and Ab-
Plenum Press, New York, 1970.
sorption of Radiant Energy," in Treatise on Analytical
Chemistry, 2nd ed., pt. 1, vol. 7, chap. 1, I. M. Kolthoff
The references below deal with the principles of analytical and P. J. Elving, eds., Wiley, New York, 1981.
atomic spectroscopy. R. F. Browner and A. W. Boorn, "Sample Introduction:
C. Th. J. Alkemade, Tj. Hollander, W. Snelleman, and The Achilles Heel of Atomic Spectroscopy," Anal. Chem.,
P. J. Th. Zeegers, Metal Vapors in Flames, Pergamon 56, 787A (1984). This article and the following one are
Press, Elmsford, N.Y., 1982. Everything you wanted to excellent discussions of the problems and techniques of
know about line broadening, intensities of spectral lines, sample introduction in atomic spectrometry.
and anything fundamental in flames. R. F. Browner and A. W. Boorn, "Sample Introduction
C. Th. J. Alkemade and R. Herrmann, Fundamentals Techniques for Atomic Spectroscopy," Anal. Chem., 56,
of Analytical Flame Spectroscopy, Halsted Press, New 875A (1984).
York, 1979. An excellent discussion of flame spectrom- R. K. Skogerboe and S. J. Freeland, "Experimental
etry from a fundamental, yet qualitative viewpoint. Par- Characterization of Aerosol Production, Transport, Va-
ticularly good discussion of interferences in atomic spec- porization, and Atomization Systems: Part I. Factors
trometry. Controlling Aspiration Rates; Part 2. Factors Control-
P. W. J. M. Boumans, Theory of Spectrochemical Ex- ling Aerosol Size Distributions Produced," Appl. Spec-
citation, Adam Hilger, Bristol, England, 1966. Another trosc., 39, 916 (pt. I), 920 (pt. 11) (1985).
classic reference on atomic spectrometric theory. R. K. Skogerboe and S. J. Freeland, "Effect of Solution
G. F. Kirkbright and M. Sargent, Atomic Absorption Composition on the Physical Characteristics of Aerosols
and Fluorescence Spectroscopy, Academic Press, Lon- Produced by Nebulization," Appl. Spectrosc., 39, 925
don, 1974. Includes a chapter on spectroscopic theory (1985).
and chapters on the theory of AA and AF measure- L. deGalan, "New Directions in Optical Atomic Spec-
ments. Also includes a chapter on introduction of liquids trometry," Anal. Chem., 58, 697A (1986).
into flames. M. L. Parsons, B. W. Smith, and G. E. Bentley, Hand-
C. Th. J. Alkemade and P. J. Th. Zeegers, "Excitation book of Flame Spectroscopy, Plenum Press, New York,
and De-excitation Processes in Flames," in Spectro- 1975.
chemical Methods of Analysis, J . D . Winefordner, ed., R. W. B. Pearse and A. G. Gaydon, The Identification
Wiley-Interscience, New York, 1971, pp. 3-125. An of Molecular Spectra, 3rd ed., Chapman & Hall, Lon-
excellent treatment of the processes occurring in flames don, 1963.
from a fundamental point of view. "Atomic Absorption, Atomic Fluorescence, and Flame
T. J. Vickers and J. D. Winefordner, "Flame Spectrom- Emission Spectrometry." This review appears every two
etry ," in Analytical Emission Spectroscopy, vol. 1, pt. years in the Fundamental Reviews issue of Analytical
11, E . L. Grove, ed., Marcel Dekker, New York, 1972. Chemistry.
Emission spectrometry has great potential as a quali- rope. Flame emission rapidly grew to become an
tative and quantitative tool since all elements can be important analytical technique, particularly after the
made to emit characteristic spectra under the appro- introduction of Gilbert's total consumption burner by
priate circumstances. Unfortunately, no single excita- Beckman Instruments in 1951.
tion source exists that can excite all the elements in an In the 1940s interest developed in materials that
optimum manner. This chapter considers emission spec- were difficult to analyze by flame emission methods.
troscopy with flame and plasma excitation. These two The war years provided great impetus for the devel-
techniques are considered together because instrumen- opment of more energetic excitation sources than com-
tal and sample introduction considerations are similar. mon flames. Hence arc and spark discharges became
Flame emission is the first of the atomic spectro- important sources particularly for the analysis of solids
chemical techniques that we will consider. It is not only and refractory materials. We discuss arc and spark tech-
the simplest atomic spectrochemical method, but also niques in detail in Chapter 9.
the oldest; the first observations of the colors produced The atomic absorption (AA) technique, discussed
by metallic salts in flames date back to the eighteenth in Chapter 10, was introduced as a new flame spectro-
century. The pioneering work of Kirchhoff and Bunsen metric method in 1955, and commercial instruments for
in the mid-1800s led to a general recognition of the AA became widely available in the early 1960s. The
power of flame emission for the qualitative identifica- growth of the AA method, and particularly the growth
tion of many elements. In fact, these workers discov- in commercial AA instruments, soon relegated flame
ered the elements cesium, thallium, indium, and gal- emission to a lesser role. At present flame emission is
lium by spectrochemical methods. Quantitative flame little used except for easily excited elements such as
emission was not fully established until the 1920s, when alkali metals.
the Swedish plant physiologist Lundegardh developed Interest in new excitation sources increased dra-
quantitative methods for studying plant metabolism. matically in the 1960s with the development of stable
Commercial interest soon followed, and the first com- high-frequency and dc plasma devices. Like the com-
mercially available flame emission instrument was in- bustion flame, plasma devices are relatively easy and
troduced in the mid-1930s by Siemens and Zeiss in Eu- convenient to operate and capable of high precision.
226 Chap. 8 / Flame and Plasma Atomic Emission Spectrometry
Flame emission remains useful in determining alkali jor, minor, and trace constituents of a sample. Because
metals and other easy-to-excite elements. However, all elements emit characteristic spectral lines whose ra-
common flames are not very energetic and this limits diances are proportional to concentration, at least over
the detectability of elements with high excitation ener- a limited range, emission spectroscopy is potentially a
gies. Because plasma sources are much more energetic very powerful elemental analysis technique. In this sec-
than common flames, they can provide higher atomi- tion we investigate the properties and characteristics of
zation efficiencies, high degrees of excitation. and lower the ideal atomic emission spectrometric system in order
detection limits for many elements. Following the in- to place the actual excitation sources and spectrometers
troduction of commercial inductively coupled and dc discussed later into proper perspective.
plasmas in the mid-1970s, these sources became rapidly
accepted as analytical tools. Plasma sources now dom- Information Desired from Emission Spectra
inate the sales of emission spectrometers; most of the
research on emission sources deals with plasma devices. What type of information would the ideal emission sys-
This chapter begins with a consideration of the tem provide? Since we are dealing with techniques ca-
ideal source for emission spectrochemical analysis. The pable of atomizing samples, we will obtain information
characteristics of the ideal source provide a basis for concerning mainly the atomic composition of the sam-
determining the strengths and weaknesses of the real ple; we will leave it to molecular spectrometric tech-
sources that are used. Flames are next considered as niques to give us information concerning the molecular
excitation sources. Their structures. combustion proc- makeup of the sample. Information that would allow
esses, temperatures, and spectral background emission identification of the elements in the sample is certainly
are important not only in flame emission, but also in desirable. Thus the ideal emission system should be
flame A A and flame atomic fluorescence (AF) . Plasma capable of rapid, qualitative elemental analysis so that
atomic emission sources, including the inductively cou- samples with unknown constituents can be identified
pled plasma, the dc plasma, and the microwave plasma, and rapid comparisons made of different samples. With
are then described and their characteristics compared the prevailing interest in trace constituents, the tech-
to those of the ideal source. Complete atomic emission nique should cover a wide concentration range (pg mL-I
spectrometers useful in single- and multielement de- to mg ml-l, or nine orders of magnitude, would be
terminations are introduced and described. To gain some desirable).
feeling for how the readout signal in emission spec- For quantitative determinations of a single ele-
trometry depends on instrumental parameters and an- ment, we now demand that trace methods be capable
alyte characteristics, mathematical expressions are de- of accuracies and relative standard deviations of a few
veloped for the signals obtained, and signal-to-noise percent. Beyond this, however, an ideal system would
considerations in emission spectrometry are discussed. be capable of true, simultaneous multielement deter-
The SIN expressions are useful in predicting conditions minations with similar precision and accuracy. We can
for achieving high precision or low detection limits. The imagine for elemental profiling purposes (moon rocks
factors that influence linearity, accuracy, precision, and or pollution studies, for example) that it would be de-
detection limits are developed next. This chapter con- sirable to determine as many as 50 elements simulta-
cludes by describing how the practical methodology, neously. Although not all situations are this demanding,
discussed in Chapter 6, is adapted to meet the specific the ability to determine one element at a time or many
requirements of plasma and flame emission methods. elements simultaneously would give the technique great
The determination of Na and K in blood serum is used flexibility. These lofty ideals are not satisfied at present
to illustrate several practical aspects of flame emission by any existing emission system. However, modern
spectrometry. Similarly, the application of plasma emis- emission sources and spectrometers are quite versatile
sion methods to water testing and marine sediment anal- tools that can give a wealth of information.
ysis illustrates some of the considerations with these
sources. Characteristics of the Ideal Emission Source
TABLE 8-7
Characteristics of the ideal atomic emission source
-~ -
Atomizer temperature not only influences the popu-
Complete atomization of all elements
lation of ground-state atoms through the atomization
Controllable excitation energy
Sufficient excitation energy to excite all elements efficiency, but also the fraction of excited atoms through
Inert chemical environment the Boltzrnann factor e E I I k TTable
. 8-2 gives the Boltz-
No background mann factor for several different atomizer temperatures
Accept solutions, gases, or solids and excitation energies. If the upper and lower levels
Tolerant to various solution conditions and solvents
are of equal statistical weight, the factors shown are
Simultaneous multielement analysis
Reproducible atomization and excitation conditions equal to the fraction of atoms excited. It should be
Accurate and precise analytical results noted that in highly ionized plasmas, even though the
Inexpensive to purchase and maintain Boltzmann factors are relatively high, the absolute neu-
Ease of operation tral atom populations can be low because of significant
ionization.
source should be an ideal atomizer as well as an ideal
excitation source. Table 8-1 lists a few of the charac-
teristics of such an ideal emission source. In addition E ATOMIC EMlSSlO
to the primary features discussed above, we want the
source to have an excitation energy that can be con- The flames used in spectrochemical analysis are hot,
trolled by the user. This would allow optimization of chemical flames in which a highly exoergic, autocata-
source conditions for the determination of easy-to-ex- lytic chemical reaction takes place in the gas phase be-
cite elements as well as those with high excitation ener- tween a fuel (hydrogen, acetylene, propane) and an
gies. Atomization and excitation should occur in an oxidant (oxygen, air, nitrous oxide). After the flame is
inert chemical environment to minimize compound for- ignited, it propagates rapidly and spontaneously through
mation and background emission. With sufficient en- the combustible mixture. Flames are made stationary
ergy and an inert environment, the chemical, physical, by means of a burner which supplies the reacting gases
and spectral interferences that plague many techniques and supports the combustion reaction. The fuel and
should be minimal. We would also like the source to oxidant are continuously supplied to the burner and
be able to accept samples as solids, liquids, and gases ignited on top of the burner head. The flames are usu-
with minimal (ideally no) sample pretreatment. It should ally premixed flames, in which the fuel and oxidant are
be tolerant of samples that contain organic solvents, are mixed prior to the region where combustion takes place,
viscous, or contain particulates. The source should but they can be unpremixed flames, in which the fuel
maintain reproducible atomization and excitation con- gas and oxidant are first mixed in the combustion region
ditions in order to give precise and accurate single ele- itself. It is extremely important to control gas flow rates
ment and multielement results. Finally, the source should carefully. Normally, the fuel and oxidant pressures from
be inexpensive to purchase and maintain, and quite compressed gas cylinders are regulated by a two-stage
simple to operate. Needless to say, the real sources that regulator. Instruments that use flames also have pres-
will be considered in this chapter do not meet all the sure gauges or flow meters preceeding the nebulizer1
criteria listed in Table 8-1. However, as we shall see, burner for fine-adjustment purposes.
some sources approach these ideals and, indeed, this is In flame emission, the sample to be analyzed is
a major reason for their popularity. usually nebulized into the high-temperature environment
TABLE 8-2
Boltzmann factors for several excitation energies and atomizer temperatures
e-E~/kI
Resonance line
wavelength,
Am (nm) E,, J (eV) 2000 K 3000 K 4000 K 5000 K 6000 K
228 Chap. 8 1 Flame and Plasma Atomic Emission Spectrometry
Properties of c o m m o n flames
..- p~
Theoretical
stoichiometric Maximum
temperature burning velocity
Fuel- oxidant Combustion reactiona (K) (cm s-')
stantially lower than theoretical values. For example, This is the region of a stable flame. At yet higher flow
a temperature = 2950 K has been obtained for a fuel- rates the flame continues to rise and blows off the burner.
rich N,O-C,H, flame, which is lower than the theo- Flames that burn with air as the oxidant can be
retical stoichiometric value of 3150 K. The oxycyanogen premixed because of the low burning velocities (see
flame (not listed in Table 8-3) has occasionally been Table 8-3). The N,O-C,H, flame can likewise be pre-
employed for analytical purposes. Its temperature has mixed without problems. However, mixtures of oxygen
been experimentally determined to be in excess of 4500 and acetylene require a great deal of care to burn safely
K, one of the hottest flames known. With aspiration of after premixing. Oxygen and hydrogen, if diluted with
aqueous solutions, the temperature decreases due to argon or helium, can be premixed.
the energy expended for solvent evaporation.
Note from Table 8-3 that the use of oxygen in Flame Background. Background emission can
place of air increases the theoretical flame temperature be the limiting factor in determining the precision of
by 700 to 800 K. This is because the inert N, from the analytical measurements or the limits of detection. The
air absorbs heat. The theoretical maximum tempera- blank measurement in flame emission spectrometry can
tures of the flames listed in Table 8-3 are often slightly compensate for the background emission only if it does
higher (20 to 100 K) than the stoichiometric tempera- not change between sample and blank determinations.
tures and, for acetylene flames, are often achieved under This implies that the background emission does not
fuel-rich conditions because the combustion products fluctuate with time or with slight variations in condi-
(CO or CO,) are partly dissociated, which withdraws tions, such as fuel and oxidant flow rates, and that the
heat from the flame gases. For hydrogen flames, the background emission is not influenced by concomitants
maximum temperature occurs very near the stoichio- in the sample that are not present in the blank. If analyte
metric fuel-to-oxidant ratio. emission occurs in a spectral region of high background
The burning velocities listed in Table 8-3 are crit- emission, spectral overlap can occur, and any fluctua-
ical flame parameters. Flames achieve stability only within tions in background emission will degrade precision and
a certain region of gas flow rates. If gas rise velocities the limits of detection. For these reasons it is important
do not exceed the burning velocity, the flame propa- to examine flame background emission in some detail.
gates inside the burner, resulting in a flashback con- We shall consider here only background emission pro-
dition. As the gas flow is increased, the rise velocity duced by the flame itself; concomitants in the sample
soon exceeds the burning velocity at all points, and the can, of course, also give rise to background emission.
flame rises until it reaches a point above the burner Most often analyte emission is measured in the
where the rise and burning velocities are just equal. region above the primary combustion zone. The back-
230 Chap. 8 1 Flame and Plasma Atomic Emission Spectrometrv
ground emission observed is from both the secondary in hydrogen flames. but also bands from such hydro-
combustion zone and the interzonal region itself, unless carbon radicals as CH, which has bandheads at 431.5,
a separated flame is used in which case only the latter 390.0, and 314.3 nm. The primary combustion zone also
region need be considered. Background emission spec- contains emission from C,, the Swan bands at 474, 516,
tra for three common analytical flames are shown in and 563 nm. Oxygen-acetylene flames also produce
Figure 8-2. There are two types of flame background some C O bands in the region 205 to 245 nm. In fuel-
that are commonly observed: spectral continua and band rich nitrous oxide-acetylene flames, emission from CN,
spectra. Band spectra are emitted by molecular species. C,, CH, and NH can be seen in the region 300 to 700
Hydrogen flames show intense emission from O H rad- nm.
icals with bandheads at 281.1. 306.4 (strongest), and The continuum can be caused by blackbody emis-
342.8 nm. These radicals are formed from reactions sion from hot solids (e.g., incandescent carbon parti-
such as cles) or from unquantized recombination reactions. In
0,-H, flames the reaction
and
NO + 0 -+ NO, + hv
Burner
also produce continuum emission. When flames contain head
a high concentration of an easily ionized metal, the
radiative recombination of ions and electrons produces Sample
an increase in background emission according to solution
'
Aspirating
gas
II
Drain
From Figure 8-2 it can readily be seen that the FIGURE 8-3 Premixed, spray chamber nebu-
background emission is lower and simpler in flames lizeriburner. The sample is aspirated and nebu-
fueled with hydrogen. The exact intensity of the back- lized into the spray chamber by a concentric ne-
ground emission is a strong function of the fuel-to-ox- bulizer with oxidant as the nebulizing gas. Often
aerosol modifiers (spoilers) are used to break up
idant ratio, gas purity, the type of burner, the type of
larger droplets. All but the finest spray particles
solvent being sprayed, and the observation region in condense and are drained away (see Chapter 7).
the flame. When the spectrometer resolution is low, the The sample spray is mixed in the spray chamber
molecular bands shown in Figure 8-2 may appear diffuse with the fuel and additional oxidant gas to control
and more like continua than bands. In addition, con- the fuel-to-oxidant ratio. Only the small (5-20
comitants in the sample that form hard to dissociate bm)-diameter particles enter the flame. Typical
molecular species can give rise to additional emission solution flow rates are 2 to 5 mL min-'.
bands, and atomized concomitants can produce atomic
lines. while the sheath reduces air entrainment, and renders
the flame stable against room drafts. Inert-gas sheathing
can also reduce background emission and minimize flame
Sample introduction into Flames
flicker due to external air movement. The Alkemade
burner is usually employed with a relatively low tem-
Solution samples are usually introduced into flames by
perature flame (0,-Ar-H, or air-C,H,); it does not
means of a nebulizerlburner system. We will consider
have the high atomization efficiency typical of most
here several nebulizerlburner systems. Nebulizers and
analytical flame systems.
the principles of atom formation with this type of sample
introduction were described in Section 7-1.
Total Consumption Burners. Many older flame and lower temperatures than theoretically achievable.
emission spectrometers use the total consumption burner. Along with the shortness of the flame, these charac-
often called a direct injection burner. shown in Figure teristics are responsible for the short analyte residence
8-5. With this burner, all the sample mist is injected times, which can increase nonspectral interferences.
into the flame (E, = I), including the large droplets
that would be removed with spray chamber-premixed Atomization and Excitation Characteristics
burner designs. The residence time for the large drop-
lets is often not long enough to ensure complete de- The choice of what flame to use in a given application
solvation and vaporization. Hence desolvation efficien- depends on many factors. For flame emission, it is de-
cies are often significantly less than unity. The larger sirable to choose a flame consistent with high atomi-
amount of water entering the flame (1 to 2 mL min-') zation efficiency, freedom from interferences, and suf-
compared to chamber burners cools the flame upon ficient excitation energy for the analyte. The hotter
evaporation. The total consumption burner is also sub- flames generally produce higher background emission,
ject to salt encrustation at the burner tip which can more ionization, and richer sample spectra with sub-
distort the flame. sequent spectral overlap problems. Although hydro-
The unpremixed flames produced by total con- carbon flames produce more background emission than
sumption burners are turbulent flames or diffusion flames. hydrogen flames, they also have lower burning veloc-
Since fuel and oxidant are mixed above the burner tip, ities and can provide longer residence times for analyte
highly explosive gas mixtures with high burning veloc- species. Hydrocarbon flames also produce, under fuel-
ities, such as 0,-H, and 8,-C,H,, can safely be burned, rich conditions, reducing carbon-containing radicals (C,
and combustible samples (i.e., gasoline) can safely be C,, CN, CH), which can minimize the formation of
aspirated. refractory oxides with metals. The N20-C,H, flame is
Unpremixed, turbulent flames have much less dis- particularly useful in this respect. Table 8-4 shows ex-
tinct structure than do premixed flames. The primary perimentally measured free atom fractions (Pa values)
combustion zone can still be distinguished, but it is for several elements in air-C,H, and N20-C,H, flames.
usually much more diffuse because of turbulent mixing. Many of the elements that form stable oxides (Al, Ba,
These flames are acoustically noisier than premixed Mn, Sn) and hydroxides (Ca, Ba, Mg) are more highly
flames because of the irregularity of the flame front and dissociated (higher Pa values) when nitrous oxide is em-
its erratic, changing velocity and direction. There is an ployed.
interzonal region which attains some degree of ther- The thermal energy ( k T ) in typical flames, 2000
modynamic equilibrium, but the turbulent flame is not to 4000 K, ranges from 2.8 x J. which is 0.17
at all homogeneous in its properties; the secondary com- eV, to 5.5 x J , which is 0.35 eV. As can be seen
bustion zone is often not easily recognized. Turbulent in Table 8-2, the Boltzmann factors and hence the frac-
flames are usually characterized by higher rise velocities tion of atoms excited is quite low for UV transitions
Canillarv Burner
TABLE 8-4
Atomization efficiencies (pavalues) in acetylene flames
Element C2H2-air flame C2H2-N20flame
A1 <lO-j 0.42-0.59
Ba 0.0011-0.009 0.15"-0.30"
Ca 0.066-0.14 0.69"-1.4"
CU 0.87-l.OOb l.OOb
Fe 0.38-0.66 -
Li 0.21-0.26" 0.44"
ME 0.59-0.84 1.5-2.3
Mn 0.45-0.93 0.76-0.80
Na 0.50-1.00" 0.33"-0.65"
Capillary
Sn <lo-"0.078 0.71-0.76
Zn 0.45-1.10 0.91-1.00
FIGURE 8-5 Total consumption nebulizer1
burner. The pneumatic concentric nebulizer as- SOURCE:J. D. Winefordner. S. G. Schulman, and T. C. O'Haver. Lurnines-
cence Spectrorneirj in Analytical Chernisiry. Wiley-Interscience, New York,
pirates and nebulizes the solution at the nebulizer 1972.
tip. The fuel flows through the outer concentric "onization suppressor added.
ring around the burner tip. "ome methods were relative :a Cu for which P, = 1.00 was assigned.
Sec. 8-3 I Plasma Atomic Emission Sources
with flames. Even for visible transitions in the range The plasmas are energize -frequency elec-
500 to 700 nm, the Boltzmann factors are 0.1% or less tromagnetic fields (RF or energy) or with
for normal flame temperatures. hen combined with a high-quality
For A A , the considerations in choosing a flame la sources provide many of the char-
are similar to those for flame emission, except that ex- al source. The WF, inductively cou-
citation energy is not a concern. For AF, quenching by pled plasma is the most prevalent of the plasma devices
flame constituents must also be considered. Many work- and is discussed first. Then variou licrowave plasmas
ers have used relatively cool flames for fluorescence and the dc plasma are described. enever appropri-
measurements. Flames such as H,--0,-Ar produce high ate, the characteristics of these sources are compared
fluorescence yields for alkali metals (see Chapter 11). to those of combustion flames and to our ideal atomic
emission source.
K
Plasma
~nductidn
coil
bQuartz
tube
Two different argon flows are used. A relatively gas expansion is the highest. Such a shape thus results
low flow velocity, about 1 L min-l, is used to transport in inefficient heating of the sample. The annular plasma,
the sample to the plasma. A much higher flow velocity, obtained at higher frequencies (Figure 8-7b), gives rise
typically 10 L min-l, is introduced tangentially as shown to efficient desolvation, volatilization, and excitation of
in Figure 8-6c. The latter flow of Ar thermally isolates the sample. This annular-shaped plasma looks much
the plasma from the outer quartz confinement tube and like a doughnut when viewed from below. The shape
prevents overheating. The plasma itself is located near results from the skin-depth effect of induction heating.
the exit end of the concentric tubes. As the frequency of the R F generator is increased, the
eddy current paths move closer to the outer or skin
Sample Introduction. One of the early prob- region of the plasma. The greatest heating occurs in the
lems with the ICP was introduction of the sample. If region of high eddy currents. The central region of the
the plasma is operated at frequencies of about 5 MHz, plasma (the doughnut hole) is thus somewhat cooler,
a plasma shape much like a teardrop is obtained, as which reduces the resistance to sample introduction.
shown in Figure 8-7a. The sample tends to avoid the Because of efficient desolvation and volatilization
high-temperature region, where the resistance due to in the ICP, the most widely used mode of sample in-
Sec. 8-3 1 Plasma Atomic Emission Sources 2
0 6,
FIGURE 8-8 Background emission spectra from the argon ICP with
deionized water being aspirated into the plasma. The upper recording was
taken under conditions favoring molecular band emission, while the lower
recording was taken under conditions minimizing molecular band emis-
sion. The flow rates listed are those for the outer, tangential argon flow.
All other conditions were the same for the two recordings. Note the
presence of Ar lines, some OH band emission, and some other molecular
bands, particularly in the upper trace. In the lower recording most of the
molecular bands have disappeared and there are mainly Ar lines. (Re-
printed with permission from R . K. Winge, V. A. Fassel, V. J. Peterson,
and M. A. Floyd, Inductively Coupled Plasma-Atomic Emission Spec-
troscopy: A n Atlas of Spectral Information, Elsevier, Amsterdam, 1985.)
several measures of plasma temperature (gas kinetic load coil. To date the most comprehensive model is a
temperature, excitation temperature, ionization tem- collisional-radiative model that includes radiation trap-
perature, etc.). Electron number densities are quite high ping and transport processes.
in a pure Ar ICP; measurements based on Stark broad- If we compare the ICP to the ideal emission source
ening indicate values in the range 1014 to 1016 cmp3. characterized in Table 8-1, we can see that it meets
These high values explain the lack of ionization inter- many of the criteria listed. While the ICP energy is not
ferences. The electron, and hence argon ion, number readily controlled, it is capable of exciting a wide range
densities are, however, higher than predicted by the of elements within the observation zone. This signifi-
spectroscopically measured excitation temperatures. The cantly enhances the simultaneous, multielement capa-
values found for excitation temperatures increase as bilities of the source. The ICP does have sufficient
the upper level of the transition used increases in en- energy to excite all common elements of interest. Atom-
ergy. In other words, the overpopulation of an energy ization and excitation occur in an inert argon atmos-
level increases with energy. It has also been found that phere, which lessens the probabilities for chemical in-
ion line-to-atom line intensity ratios for a given element terferences. Solutions and gases can be injected directly
are significantly higher than expected from the LTE into the plasma, but solids are analyzed only with some
model. difficulty. Direct insertion probes made from carbon or
Several excitation models have been presented for graphite have been used to introduce solids. A spark
the argon ICP. In one model a high population of argon discharge is used in one commercial system to vaporize
metastable levels is used to explain the observed ion1 solids for introduction into the ICP (see Chapter 9).
atom intensity ratios. Another model explains the over- The ICP is a reproducible source and capable of pro-
population of high energy levels, particularly argon ducing accurate and precise results (see later). ICP
metastable levels, by a radiation-trapping model (see sources are less than ideal, however, in several cate-
Section 7-5). Yet a third model uses ambipolar diffusion gories. First, there are spectral overlap interferences,
to explain observations made in the analytical region even though these are minimal in many cases. Second,
based on conditions present in the region inside the ICP emission systems are not inexpensive either to pur-
Sec. 8-3 1 Plasma Atomic Emission Sources
Tuning
stub
Although they have not been studied as extensively as Tuning
ICPs, microwave plasmas have been used for many stub
emission spectrometric applications. These sources are
generally operated at lower powers than ICP sources
and, of course, at microwave frequencies.
Capilla~y
Nebulizer DC Plasmas
-<-1---
- --- - -
- -. +Sample
Direct-current plasmas (DCPs), also called plasma jets,
are also extensively used in emission spectroscopy.
Heated spray t
Nebul~zer
Commercial instrumentation based on DCPs is avail-
gas
able and this has undoubtedly stimulated the use of
these devices. In a DCP, a flow of partially ionized gas
t
T o drain
is forced out of a small orifice at high velocity. The
(underwater)
plasma is produced by a dc discharge, similar to a dc
arc (see Section 9-I), struck between two or more elec-
FIGURE 8-10 Nebulizer with desolvation sys- trodes. To obtain high current densities, the discharge
tem. Solutions are sprayed with a pneumatic ne- is made to constrict by cooling the outer regions of the
bulizer into a heated chamber where solvent is
plasma with a high-velocity gas stream. The latter makes
vaporized. A condenser cooled to about 10•‹C
causes the solvent to condense. Solvent flows out the outer regions of the arc less conductive and de-
the drain and desolvated sample is carried away creases the current channel area.
to the source by the nebulizer gas. Nearly com- Early dc plasmas were made from ring-shaped car-
plete desolvation can be achieved with overall bon electrodes and an upper cathode. They had the
efficiencies of 35%. disadvantage that observations were made in a region
Sec. 8-4 I Flame and Plasma Emission Spectrometers
where the background continuum due to electron-ion and its acceptance of solutions with relatively high solids
recombination was relatively intense. In the early 1970s content. The small region where optimum line-to-back-
the background problem was partially overcome by a ground ratios occur is the principal disadvantage of this
new electrode geometry which positioned the anode DCP. With a conventional spectrometer, a much larger
and cathode at an angle of 30" to one another. The region is ordinarily viewed (5 to 10 mm slit height)
plasma then took the shape of an upside-down V. Aer- unless optics are used to magnify the source image. For
osol was sprayed from between the two electrodes, and this reason echelle spectrometers are most often used
emission was observed above the apex of the arc. This with this DCP. Different elements are found to have
observation zone avoided the strongly emitting plasma their optimum line-to-background ratios in different
core and resulted in higher signal-to-background ratios. spatial regions, which makes the selection of compro-
The third major development in DCPs occurred mise conditions for multielement analysis, particularly
with the introduction of a commercial three-electrode near the limits of detection, somewhat difficult. It has
system by Spectrametrics, Inc. This three-electrode DCP also been found that plasma properties change when
is shown in Figure 8-11. Here two independent plasma high concentrations of easily ionized materials (e.g.,
jets have a single common cathode, and the overall alkali metals) are introduced. Like the ICP, a DCP
plasma burns in the form of an upside down Y. A flow spectrometer is neither inexpensive to purchase or
of argon gas is directed over each electrode at a velocity maintain. For an operator to use DCP systems effec-
high enough to cool the ceramic sleeves and prevent tively requires considerable training. Despite these
their melting. The sample is also nebulized with Ar gas, problems, the three-electrode DCP is a versatile and
and the total Ar consumption is about 8 L min-I. useful source especially when a broad range of analyt-
Experiments have shown that although excitation ical matrices is encountered.
temperatures can reach 6000 K in this DCP, sample
volatilization is not complete because residence times
in the plasma are relatively short. This can be trouble-
some with samples that contain difficult-to-volatilize
materials. Electron number densities in the three-elec-
trode DCP are quite similar to those in the ICP. The In addition to the excitation source and sample intro-
major advantages of the three-electrode DCP are its duction system, an emission spectrometer must contain
good stability, its low power requirements (<1 kW), its a wavelength selection device with entrance optics, one
ability to handle organic as well as aqueous solutions, or more transducers, an electronic processing system,
and a readout system, as shown in the block diagram
Tungsten of Figure 8-12. Although the trend in recent years has
been toward multichannel spectrometers, single-chan-
Cathode nel systems are still widely used, and, for flame emis-
block
sion, simple filter photometers can sometimes be suf-
ficient. We consider first the optical components of the
/I Excitation emission spectrometer system; later in this section the
electronic components and readout system are de-
scribed along with computer-controlled emission sys-
tems.
Flame or
plasma
<--
-.
Or:: Monochromator, Readout
polychromator,
or spectrograph
x Sample
near the analyte line. We will look first at the entrance emission spectrometry. For routine flame emission de-
optics, which play an important role in the first goal terminations (e.g., Na or K in blood serum), interfer-
above. ence filter photometers are often employed. Because
of their low resolution, filter photometers are used only
Entrance Optics. Typically, a lens or mirror is where spectral interferences are unlikely. The major
used to focus an image of the source (flame or plasma) advantages of filter instruments are their low cost, small
onto the wavelength selection device. With dispersive size, and high light throughput. Several commercial flame
devices, the Fin of the optical element and its position photometers (see Section 8-7) designed specifically for
are chosen to match the monochromator (or polychro- clinical applications are based on interference filters.
rnator) solid angle (Q) of collection. For 1:l imaging, The grating monochromators discussed in Chapter
the Fin is one-half the Fin of the monochromator, and 3 are most often used in single-channel instruments.
the excitation source and entrance slit are placed 2f (f The most versatile monochromators are capable of
is the focal length of the lens or mirror) from the en- moderate to high resolution (< 0.1 nm bandpass) and
trance optic. With lenses, chromatic aberrations can have variable slit widths to allow optimization of the
cause the throughput to vary with wavelength. The en- SIN (see the next section). Automatic scanning is highly
trance slit dimensions determine the portion of the source desirable to allow the background emission to be ob-
that is observed. With flames the viewing window is tained on either or both sides of the analytical line.
readily adjusted by mounting the burner on a transla- Alternatively, wavelength modulation (see Section 4-
tional stage and moving the flame vertically with respect 5) can be employed for automatic background subtrac-
to the optical element. With plasmas it is usually easier tion. With single-channel systems, the response time of
to adjust the optics or the spectrometer than to move the measurement electronics can limit the maximum
the plasma. In some systems a concave mirror is placed scan rates achievable. If more than a few elements are
behind the excitation source with its center of curvature to be determined, or the analysis wavelengths are widely
located within the source to increase the solid angle of separated, linear scanning can lead to intolerable anal-
collection. ysis times and excessive sample consumption.
If ac or lock-in detection is to be employed, a
chopper must be placed in the optical train unless wave-
Sequential Multielement Instruments. Several
length or sample modulation is employed. In most cases,
a mechanical chopper is positioned between the source different types of sequential spectrometers, are em-
ployed for emission spectrometry. Slew-scan mono-
and the spectrometer entrance slit (see Section 4-5).
chromators can provide a significant savings in analysis
Single-Channel Systems. Single-channel in- time and sample consumption over conventional linear
struments are very common in both flame and plasma scanning monochromators. Slew-scan monochromators
Sec. 8-4 1 Flame and Plasma Emission Spectrometers
can be programmed to scan slowly over or to momen- Multichannel Spectrometers. For true simul-
tarily stop at wavelength regions of interest, but to move taneous multielement determinations, instruments ca-
rapidly (slew) through uninteresting regions. Both con- pable of monitoring at multiple wavelengths in parallel
ventional grating monochromators and echelle grating are required. In the past spectrographs with photo-
monochromators can be made to slew-scan. Of course, graphic detection were commonly used. Photographic
for developmental work in choosing appropriate ana- emulsions, however, have limited linearity, are difficult
lytical lines or in investigating spectral characteristics, to calibrate, must be developed, and must be subjected
it is desirable to have both linear scanning and slew- to densitometry to obtain intensity information. For
scanning capabilities. For example, one commercial these reasons, photographic emulsions are used now
echelle spectrometer used with a dc plasma allows linear primarily for qualitative spectral observations. Modern
scanning over long- or short-wavelength regions as well multichannel emission systems use either polychroma-
as slew scanning for multielement determinations. The tors with photomultiplier detectors or spectrographs with
spectrometer can slew from 190 to 900 nm in less than array detectors.
3 s. Typical slewing times between lines are 1.5 s or A modern direct-reading spectrometer used with
less. Slew-scan systems also allow simple background an ICP source is shown in Figure 8-13. A commercial
correction methods to be used. The spectrometer can instrument of this design has provision for more than
be programmed to measure the background intensity 60 exit slits and photomultiplier tubes; smaller 48- and
on either or both sides of the analytical line. 36-channel versions are available. All channels can be
Photomultiplier
-- .- L.
tubes
'oncave
-
.
'
.
Stepper
motor
dedicated to analytes or some can be used for the meas- small adjustments to be made in the wavelengths pass-
urement of background. In some cases multiple lines ing through the slit. Because of the difficulties in align-
of the same e1emen.t can be used for analysis. Alignment ing the exit slits, direct-reading spectrometers are not
of the individual exit slits in a direct reader is extremely very flexible systems. Many commercial instruments,
tedious and time consuming. Hence analysis lines are however, provide an extra observation port to which a
added or changed only very rarely. Unfortunately, scanning monochromator can be added for additional
alignment also depends on temperature and even the versatility (the n + 1 channel).
refractive index of the air. Some spectrometers use one Background correction is also difficult with direct-
channel to monitor an intense spectral line from a hol- reading spectrometers. Quartz refractor plates which
low cathode or mercury lamp. A servo system contin- scan a small wavelength interval around the analytical
uously adjusts the position of the grating or exit-slit line are often used in many background correction
assembly to keep this reference line centered. In other schemes (see Figure 4-34). One refractor plate can be
systems quartz refractor plates in front of each slit allow used behind the entrance slit to scan all channels or
plates in front of each exit slit can be used to spectroscopic applications, particularly with crossed-
scan each channel individually. The entrance slit can dispersion, echelle grating spectrographs, but at present
also be oscillated which effectively scans all the slit im- they are quite expensive and difficult to use. With two-
ages dispersed in the focal plane. However, these can dimensional arrays, it is possible to intercept the entire
add complexity to a system alrcady difficult to use. The two-dimensional spectrum generated by the echelle
high cost and low optical speed of these spectrometers spectrograph.
are additional considerations. Signal-to-noise ratio cal- Multiplex spectrometers (see Chapter 4), such as
culations have shown that for a fixed total measurement Hadamard and Fourier transform spectrometers, have
time the multichannel techniques provide the highest also been proposed for multielement emission spec-
SIN'S. Slew-scan techniques approach multichannel trometry. These techniques have been widely used in
techniques only when few lines must be measured. the IR spectral region, where detector noise is the lim-
Echelle spectrometers are also commonly used as iting source. They have not been very successful in the
direct readers. Recall that the echelle grating with a UV-visible region, where quantum noise or flicker noise
prism order sorter disperses the spectrum in two di- is often the limiting noise source.
mensions. Hence multiple exit slits and photomulti-
pliers can be placed in the two-dimensional focal plane
as illustrated in Figure 8-14. Excellent resolution can
be obtained in a compact spectrometer by using an Transducers and Electronic Components
echelle grating. One commercial instrument based on
the design of Figure 8-14 allows operation in the se- Photomultiplier tubes are used as transducers in the
quential mode (Figure 8-14b) as well as the true si- majority of emission spectrometers, although some sim-
multaneous mode (Figure 8-14a). In either mode the ple filter photometers for flame emission have used
exit slit plate can be moved off the analysis line to enable vacuum phototubes or photodiodes. Since the radiant
background correction or to allow the selected line to flux emitted by atoms (ions) can vary over several or-
be scanned. One advantage of the fixed optics approach ders of magnitude, it is important that the transducer,
shown in Figure 8-14 is that the grating is never moved the signal processing electronics, and the readout sys-
and the optical components are mounted in a thermally tem have a wide dynamic range. Most emission sources
and vibrationally isolated housing for maximum stabil- are also somewhat noisy. Because photomultipliers are
ity. The throughput of echelle polychromators o r rapid response detectors, some signal averaging is usu-
monochromators can be less than for conventional grat- ally needed for SIN improvement. Instruments that are
ing systems because smaller slit heights are used and dc-based have used current-to-voltage converters with
only one order is monitored. dc amplifiers (if needed) to obtain voltages in the range
Another approach to multichannel systems is to 0.1 to 10 V. Typically, these instruments use low-pass
use a spectrograph with an array detector. At present filtering with time constants of a few seconds. Another
there is no perfectly suitable solid-state replacement for approach that is often used with dc-based plasma emis-
the photographic plate in emission applications. De- sion systems is to use a linear integrator with an inte-
vices such as diode arrays are too small (1 in. long or gration time of 1to 30 s. For an n-channel direct reader,
less) to achieve both the desired resolution and the n integrators are therefore required for averaging. In
desired wavelength coverage for multielement deter- one commercial 60-channel system, 60 operational am-
minations. They are also not as sensitive as photomul- plifier integrators are used. To increase the dynamic
tiplier tubes unless operated with an intensifier. One range, the integration capacitor is usually charged to
recent spectrometer combines the direct reader ap- full scale and discharged several times during the meas-
proach with a diode array detector. A grating coarsely urement. The processing system then counts the num-
disperses the radiation, and several wavelength regions ber of full-scale integration cycles and adds this to the
are allowed to pass through a mask to an echelle grating. final integrator output. The integrator sensitivity (value
The latter further disperses the radiation from each of of integration capacitor) is different for weak lines than
the selected wavelength windows onto the array. As a for strong lines. Hence the electronics for each channel
result, several wavelength regions (a few nm each) ap- are somewhat customized. The output of each integra-
pear side by side on the array. A different mask is tor is sent to a multiplexed analog-to-digital conversion
required for a different selection of elements, but since system for conversion to the digital domain. A com-
the masks are coarse openings, they are not expensive. puter system stores the acquired data for each channel.
Some of the newer solid-state devices, such as the In another approach, the PMT outputs are converted
charge-coupled device and the charge-injection device, to voltages by operational amplifier circuits. The volt-
appear to have considerable potential in high-resolution age output of each channel is then acquired with an
244 Chap. 8 I Flame and Plasma Atomic Emission Spectrometry
integrating analog-to-digital converter, such as a volt- 8-5 SIGNAL AND NOISE CONSIDERATIONS
age-to-frequency-to-digital converter. In either case
computers are required to store and analyze the large Expressions for the readout signal, the noise, and the
amounts of data obtained. signal-to-noise ratio (SIN) can give insights into the fac-
With modulation (source, sample, or wavelength), tors that influence these quantities. Such expressions
current-to-voltage conversion followed by either ac am- can be used as guidelines in optimizing measurement
plification and demodulation or by lock-in amplification precision. By combining several equations considered
is employed. Except for specific clinical analyzers, it is previously, an expression is developed here for the
difficult to find new commercial instruments that are ded- readout voltage in flame and plasma emission spec-
icated to flame emission determinations. Many flame A A trometry. The theoretical equation for the SIN that was
instruments (see Chapter 10) have provision for flame developed in Chapter 5 is then modified to a form ap-
AE measurements. Often a mechanical chopper is acti- propriate for emission measurements, and limiting noise
vated to modulate the flame emission signal at the mod- sources are identified. Optimization of SIN'S and the
ulation frequency of the A A hollow cathode lamp (not influence of experimental variables on detection limits
employed for emission) so that the same ac electronics are then discussed.
can be used. AA instruments are not always well suited
for AE measurements because automatic scanning is usu-
ally not provided and their resolution may not be suffi- Readout Signals
cient.
Let us consider the flame or plasma emission spectrom-
eter of Figure 8-12 and expand the equation for the
Computer Control readout voltage EE given in equation 4-34 and repeated
here:
Nearly all recently manufactured emission spectrometer
systems include a microprocessor or a microcomputer
system. The degree of computer involvement in the
experiment varies from model to model. In some cases
the computer merely acquires the emission data, com- where G is the gain of the signal modifier (V A-i), m
putes calibration data from standards, calculates the is the transducer gain, BkEis the spectral radiance due
concentrations of unknowns from stored calibration to analyte emission, Y(A) is the monochromator (or
equations, and outputs the data to a printer or video polychromator) throughput factor, and R(A) is the ra-
display. Some programs allow emission data to be cor- diant cathode responsivity of the transducer. We con-
rected for matrix effects by including various empirical sider here the normal emission case where a narrow
correction factors. Others allow background correction spectral line is monitored (Ah,,, << s). The transition
to be implemented under program control. Many in- monitored is assumed to be a resonance transition from
struments also provide an automatic sampler and in the first excited level (level 1) to the ground level (level
some cases the computer can control the sampler. 0) and self-absorption is presumed negligible. The spec-
With some instruments the computer takes a more trometer throughput factor, given by equation 3-70, and
active role in the experiment. For example, many se- R(h) are constant over the narrow emission line width.
quential or slew-scanned monochromators are placed Thus
under computer control so that all scanning functions
(e.g., scan range, scan rate, analysis wavelengths, and
integration time for each element) can be programmed.
This permits routines to be developed for various de-
terminations and stored in a library for later reuse. One-to-one imaging is assumed and To, includes
Ideally, one would also like to control the excitation the entrance optics transmission factor. The integral in
source via the computer. Parameters such as gas flow equation 8-2 has been shown in equation 7-32 to be the
rates, nebulizer flow rates, observation windows, and total line radiance, BE. Thus we can write
many others are amenable to control by the computer.
Placing as many parameters as possible under computer
control allows self-optimization routines to be used to
develop optimum analysis conditions as discussed If we assume that self-absorption is negligible (op-
Section 8-5. tically thin limit), we can use equation 7-39 for the total
Sec. 8-5 1 Signal and Noise Considerations
line radiance, which gives Let us roughly compare flames and plasmas with
the aid of equation 8-5. The gas flow rates (Q values)
are roughly the same for the two media, but the solution
flow rate F is usually lower in plasmas because of the
low flow rate nebulizers used. However, with plasmas,
Here we have made use of the relationship E l = the values of the expansion factor ef and the Boltzmann
hclX, for a resonance transition in order to express the factor ecE1lkTare significantly higher because of the
exponential in equation 8-4 in terms of the upper-state higher gas kinetic and excitation temperatures. The larger
energy. For sources that are not in thermal equilibrium, Boltzmann factors along with the larger atomization
T refers specifically to the excitation temperature. The efficiencies are usually more than enough to yield higher
ground-state number density n, can be expressed in readout signals in plasma emission.
aEE(iE),analyte emission voltage (cathodic current) [V(A)]; E,,(i,,), background emission voltage (cathodic current) [V(A)];
Ed(id),dark voltage (cathodic current) [V(A)]; m, transducer gain (dimensionless); G, signal processor gain (V A-I); K,
bandwidth constant (C s r l ) ; cE,
analyte emission flicker factor (dimensionless); x,. background emission flicker factor (di-
mensionless); (ad),,, rms dark current excess noise (V); a,,, rms amplifier-readout noise (V).
246 Chap. 8 1 Flame and Plasma Atomic Emission Spectrometry
the noise is limited by such factors as background emis- For example, changing the fuel-to-oxidant ratio affects
sion flicker or shot noise and dark current noise which not only Q, but also T, which in turn affects s,, ef,BbE,
are independent of EE. Thus at low analyte concentra- and x,. In practice, the SIN is usually measured ex-
tions, the SIN is directly proportional to concentration perimentally as a function of the variable of interest
(SIN sc EE or i,). Usually, background emission levels with theoretical expressions as a guide. Some variables
are high enough in emission spectrometry that dark show optimum values while others do not. The most
current noise and amplifier-readout noise are negligible important instrumental variables are monochromator
(EbE >> Ed). At intermediate analyte concentrations, slit width, W, noise bandwidth Af (included in K), and
the first term in the denominator of the equations in certain atomizer factors (gas flow rates, nebulizer var-
Table 8-6 (shot noise term) can become dominant with iables, height of observation, etc.).
EE >> (EbE + Ed). When analyte emission shot noise Increasing the monochromator slit width increases
dominates, the SIN is proportional to the square root EE and E,,. Since E, W while EbE W2, from equa-
of concentration [SIN a (E,)lI2 or (i,)1'2]. At high con- tion 8-10 SIN r WI[kl(W2 + k,) + k,W71'2. where the
centrations, the analyte emission flicker noise term k's are constants and the background emission is as-
dominates and the SIN becomes independent of con- sumed to be a continuum. At very narrow slit widths,
centration [SIN # f (E, or i,)]. dark current shot noise predominates and the SIN is
proportional to W because the noise is independent of
W. As the slits become wider. background emission shot
Signal-to-Noise Optimization noise becomes appreciable and the SIN becomes in-
dependent of W. Eventually, background emission flicker
The equations of Table 8-6 can give us insight into how noise dominates, and the SIN is proportional to W-l.
experimental variables influence the SIN and how the A typical plot of SIN vs. W shows a maximum at a
SIN can be maximized. If measurement precision is lim- particular optimum value of slit width W,,. If the back-
ited by random noise, maximizing the SIN minimizes ground emission includes lines from concomitants, the
the relative standard deviation (RSD) of the measure- SIN can improve dramatically if the slit width is de-
ments. Let us first consider low analyte concentrations creased enough to eliminate an intense background line.
where the contribution of analyte emission shot and At low analyte concentrations and small slit widths,
flicker noise is negligible. For the rest of this discussion when background emission flicker is low, equation 8-
we will assume that excess dark current noise and am- 10 predicts that decreasing the noise equivalent band-
plifier-readout noise can also be neglected, and that the pass Af and thus K increases the SIN. Since the back-
gains (mG) are adjusted so that measurements are not ground emission flicker noise often shows a llf behavior
readout resolution limited. In this case, equation 8-8 at low frequencies, decreasing Af will eventually cause
reduces for low concentrations to background emission flicker noise to predominate and
the SIN becomes independent of Af or may even de-
crease. With flame emission, an optimum Af on the
order of 0.1 to 1 Hz is normally found experimentally.
With a mechanical chopper and source modula-
To improve the SIN at low concentrations, the tion, the background emission signal and noise are car-
signal-to-background ratio EEIEbE can be increased, ried along with the emission signal so that synchronous
the bandwidth constant K can be reduced, and the back- ac detection provides no advantage if background emis-
ground emission flicker factor xE can be reduced. Equa- sion flicker noise is the limiting noise source. Improve-
tion 8-5 gives the dependence of EE on experimental ments in the SIN at low concentrations and the detection
variables. Unfortunately, many of the factors that in- limit have been achieved with wavelength and sample
crease the analyte emission signal EE also increase the modulation since in this case the background emission
background emission signal EbE. signal and low-frequency background emission noise
We could substitute the detailed expressions for are not carried.
EE and EbE(equations 8-5 and 8-7) into equation 8-10, Gas flow rates, nebulizer parameters, slit width,
to provide a SIN expression that indicates the depend- slit height, and observation height are adjusted for max-
ence of the SIN on all variables. Use of such an expres- imum signal or SIN, although the effects of these var-
sion for optimization is difficult since the dependence iables are very difficult to predict theoretically. With
of the magnitude of some of the variables on other flames, the type of fuel and oxidant and their flow rates
variables is not known or the variables are interrelated. can influence both EE and EbEby changing flame tem-
Sec. 8-6 1 Performance Characteristics
perature, solution flow rate, atomization efficiency, and we will assume that determinations are carried out with
other factors. Optimum values are often found exper- solutions. Experimental methods for evaluating and im-
imentally. With plasmas, gas flow rates, the solution proving performance are introduced.
flow rate, and the applied RF power influence both EE
and EbE. Similarly, in flames and plasmas the optimum
observation height must be found experimentally since inearity
EE, EbE, and x vary with position.
Background emission flicker noise can result from Equation 8-4 gives the general relationship between
variations in atomizer conditions or in the sample in- readout signal EE and ground-state number density no
troduction system. The flicker factor depends on noise in all emission techniques. In addition, equation 8-5
equivalent bandpass and many other variables. For a gives the relationship between EE and solution concen-
given situation, xE is determined experimentally. For tration c for techniques that introduce samples by con-
flames and plasmas, typical values are 0.3 to 3% for Af tinuous nebulization. The proportionality between
in the range 0.1 to 1 Hz. readout signal and concentration is invariably estab-
At high analyte concentrations, analyte emission lished by means of a calibration curve. Here EE values
flicker often becomes the dominant noise source. Under are obtained for several standards of known concen-
these conditions, equation 8-8 reduces to trations, and EE is plotted vs. c or a log-log plot is made.
For many reasons it is desirable that such calibration
curves be linear or at least follow predicted relation-
ships. We expect to obtain linear calibration curves if
the emission intensity is proportional to no, if no is pro-
The only way to improve the SIN is to reduce the portional to c, and if instrumental and source factors
magnitude of the analyte emission flicker factor 5,. The are constant from one standard to the next. Verification
fluctuations in analyte emission are caused mainly by of linearity is then experimental evidence that the in-
variations in the free-atom population in the viewed strumental system is performing as expected.
region of the flame or plasma and variations in exci- A major cause of nonlinearity when resonance
tation conditions. Sometimes the analyte emission noise lines are used is self-absorption at high concentrations.
is called nebulizer noise because it is believed to be due Self-absorption causes the slope of a log-log plot of EE
to fluctuations in the sample introduction rate. As a vs. c to deviate from its low concentration value of 1
first approximation tEis assumed independent of an- and approach its high concentration limiting value of
alyte concentration so that the SIN becomes independ- % (see Figure 7-15). In flame emission, the onset of
ent of c at high concentrations. At concentrations where nonlinearity due to self-absorption typically occurs in
self-absorption is significant, fluctuations in self-ab- the concentration range 10 to 100 kg mLP1. The onset
sorption may cause the relative flicker noise and hence of self-absorption in ICPs often occurs at higher con-
EE to increase. Again 6, is determined experimentally centrations than in combustion flames. Although high
and is typically 0.3 to 3% in flames or plasmas for Af concentrations can give lower calibration curve slopes,
values in the range 0.1 to 1 Hz. and thus lower sensitivity, useful analytical results can
still be obtained in the region where self-absorption
occurs. Thus nonlinearity at high concentrations should
be expected and not taken as an instrumental failure
MANCE CHARACTE unless the theoretical relationship is not obeyed.
At low concentrations, ionization of the analyte
There are several ways to evaluate the performance of can cause nonlinearity (see Section 7-2). With flames
an emission spectrometer or a particular analytical it is common to use an ionization buffer with easily
method. Among the factors that are important in char- ionized elements to overcome this effect. With ICP and
acterizing performance are the linearity of calibration DCP sources the high electron densities tend to buffer
curves, the accuracy of determinations, the precision of the plasma against changes in the extent of ionization
repetitive determinations, and the experimentally with concentration. Also, with ICP sources ionic emis-
achievable limits of detection. In this section these char- sion lines are often used for easily ionized elements and
acteristics are discussed, and the factors leading to non- these are often less susceptible than neutral atom lines
ideal behavior are described. Because solution samples to additional ionization. Changes in solution flow rate,
are most often employed with flame and plasma sources, atomization efficiency, and atomizer temperature with
248 Chap. 8 1 Flame and Plasma Atomic Emission Spectrometry
/O
/ Cr Precision
measured signal E, depends on temperature as For new analytical methods, samples should also be
EE = k'ecEJkTand that k' is independent of temper- analyzed by independent methods when they are avail-
ature (not really true), then able.
An optimized emission spectrometer and analyt-
ical procedure should be capable of precision (random
error)-limited accuracy. Since RSDs are typically in the
Since E,IkT is on the order of 10 in flames, the range 1 to 5% for many procedures, accuracies in this
RSD in temperature crJT should be less than 0.1% for range are often the goal. Precision-limited accuracy can
the RSD in EE to be less than 1%. Hence in a 2500-K be achieved, of course, only if systematic errors are
flame, cr, would need to be ? 2.5 K for the RSD in E, minimized or eliminated.
to be 1 % . In reality, the influence of flame temperature With external standards, the accuracy depends di-
is much more complex since it also affects desolvation, rectly on standard preparations and how well the stand-
solute volatilization, compound dissociation, ioniza- ards approach the ideal standard (see Chapter 6). In-
tion, and flame background. Because of the many var- correctly prepared standards lead to calibration errors
iables affected, flame temperature fluctuations can give which are usually seen as a deviation from expected
rise to larger or smaller values of the RSD than pre- calibration curve behavior or as an error when samples
dicted by excitation considerations alone. of known concentration are analyzed. If there is doubt
Signal-to-noise ratio considerations with plasma as to the reliability of the standards, they should be
emission spectrometers are very similar to those for analyzed by independent methods.
flame emission systems, although the expressions can Less easily detected and corrected are the blank
be considerably more complex when multielement de- and analyte interferences discussed in Section 7-2. Blank
terminations are included. It is not uncommon with ICP or additive spectral interferences are generally detected
and DCP sources to achieve relative standard deviations by using blank solutions that contain the suspected in-
on standards in the range 1 to 5% and on real samples terferent(s) at varying concentration levels and by
in the range 1 to 10%. With many of the sources con- changing the monochromator spectral bandpass. The
sidered here, the stability of the source itself can limit high excitation energies of the plasma sources can give
precision through its effect on temperature and indi- rise to very rich emission spectra and make spectral
rectly on atomization and excitation efficiencies. overlap problems more severe than with flames. Stray
Of course, the ultimate limit of precision of the radiation can result from the scattering of intense spec-
measurement step is imposed by random noise, as de- tral lines by grating imperfections or by other compo-
scribed in Section 8-5. The goal of an experimentalist nents in the spectrometer, or it can result from over-
should always be to improve the precision of all the lapping orders. I n any of these cases a spectral
noninstrumental steps until electrical noise dominates. interference can result. For these reasons, moderate-
Then the SIN optimization methods mentioned previ- to high-resolution spectrometers are preferred in plasma
ously can be used to advantage and reflected in the emission methods. However, the 0.75-m spectrometers
precision of the entire procedure. used with many ICP sources may not possess sufficient
resolution for trace-element determinations in complex
Accuracy samples. The echelle grating spectrometers possess higher
resolution in a moderate-sized instrument. With many
The accuracy of determinations must necessarily be plasma sources, background emission from the plasma
evaluated by inference since the analyte concentration continuum and from molecular bands is also dependent
is unknown in an actual sample. Usually, periodic de- on the composition of the sample. Because of the dif-
terminations of the concentrations of standard refer- ficulty of applying background correction methods with
ence materials or samples of known composition with many multichannel spectrometers, such changes in the
matrices similar to the analytical sample are used to background can lead to inaccurate results. These effects
estimate accuracy and uncover errors in procedure. In often seem more severe in multielement determinations
certain areas, such as clinical chemistry, preanalyzed because so many elements are being determined si-
blood serum samples with normal and abnormal analyte multaneously under compromise conditions. Elaborate
concentrations are available for evaluation. With other correction procedures have been established to com-
sample types, standards certified by the National Bu- pensate for additive interference effects with multi-
reau of Standards are used. If the errors are outside channel spectrometers. These typically involve multi-
the expected or desired range, a systematic search can channel measurements on a series of single-element
be made for their source and for ways to minimize them. standards. All responses are then used to construct an
250 Chap. 8 1 Flame and Plasma Atomic Emission Spectrometry
interference matrix that accounts for the additive ef- ionization interference by a concomitant can be rec-
fects. Computerized systems are, of course, required to ognized by simultaneous emission measurements of an
acquire and process the large amount of data. For the atomic and an ion line. Because of the high tempera-
ICP, extensive tabulations of interelement effects and tures, many of the analyte specific interferences found
interelement corrections are available from manufac- so commonly in flames are negligible or reduced to
turers of ICP instruments. Most require a fairly good tolerable proportions in plasma sources. For example,
knowledge of the major sample constituents. the Ca neutral atom line was found to undergo only a
Nonspecific analyte or multiplicative interferences 2 to 3% enhancement when the Na concentration was
can be distinguished from specific analyte interferences changed from 0 to 0.7 wt 5% in an ICP source. Similarly,
by determining a different analyte in the same sample. the depressant effect of on Ca was found to be
A nonspecific effect of HC1 on solution viscosity, sur- negligibly small in the ICP. Enhancements of atomic
face tension, density, or vapor pressure, for example, and ionic emission have been found, however, in the
alters the flow rate of solution into the atomizer and DCP when high concentrations of alkali metals were
influences different analytes in the same manner. A present. These have been attributed to changes in the
specific interference of HC1 on dissociation or ioniza- plasma properties and not to ionization suppression.
tion should not affect two different analytes in exactly Some interferences can be distinguished by plot-
the same way. Excitation interferences can be discov- ting the measured analyte signal vs, interferent con-
ered by using several analyte lines of different excitation centration and noting the shape of the plot. In other
energies. Since temperature changes influence the frac- cases, measuring the analyte emission as a function of
tion excited through the Boltzmann factor ( e - E 1 l k fthe
), the height of observation in the flame or plasma can
percentage change in signal should be different for lines give insight into the type of interference. Once the type
of differing ELvalues. An excitation interference can of interference has been discovered, procedures can be
also be distinguished from a nonspecific interference by designed to compensate or correct for the resulting er-
observing the same line in absorption and emission. An rors as discussed in Chapter 6.
TABLE 8-7
Some flame emission detection limits
TABLE 8-8
ICP and DCP detection limits
The detection limit (DL) for an element on a given
Element ICP (ng mL-') DCP (ng mL-')
instrument is quite useful as a figure of merit since it
defines the lower limit of the concentration range that
can be used. Detection limits are evaluated as discussed
in Chapter 6 by measuring the calibration curve slope
m and the blank standard deviation (DL = ks,,Im). It
is frequently found that DLs calculated from standards
prepared in a matrix that duplicates that of the analyt-
ical samplc as closely as feasible are significantly higher
than those obtained on standards prepared in a pure
solvent. Since the latter are usually reported by instru-
ment manufacturers, they should be used with some
caution. In any case, DLs obtained experimentally or
reported by others can often determine whether or not
a particular determination is even feasible. Small dif-
ferences in DLs between workers or instruments are
usually insignificant, while differences of one or more
orders of magnitude are usually significant.
The best detection limits reported to date for flame
emission methods are listed in Table 8-7. These were
obtained with several different instruments, nebulizer-
burner types, flame types, and measurement electron-
ics. The DLs reported do give an excellent indication
of the elements for which flame emission is highly sen-
sitive. As would be expected, the alkali metals and
alkaline earths have quite low DLs by flame emission
spectrometry, whereas elements with UV resonance lines
(e.g., Zn) and small Boltzmann factors have poor DLs.
Detection limits are reported in Table 8-8 for ICP
and DCP sources. These are the best detection limits
obtained to date with pneumatic nebulization and re-
search-grade instrumentation. In general it can be seen
that the ICP achieves superior powers of detection com-
pared to the DCP, but often by less than one order of
magnitude. For many elements a comparison with Table
8-7 reveals that ICP detection limits are significantly
lower than those reported by flame emission. The alkali precision near the DL, SIN theory can be used to predict
metals are the major exceptions. For some elements DLs and their variation with experimental variables.
with the ICP source, detection limits have been re- Thus the discussion in Section 8-5 on optimizing the
ported in matrices other than distilled water. These SIN at low analyte concentrations applies to optim-
have revealed that powers of detection do not signifi- izing the DL.
cantly deteriorate in some "real-life" matrices. Detec-
tion limits can vary significantly, however, with the
spectrometer quality, throughput, and the type of 8-7 METHODOLOGY AND APPLlCATlO
nebulizer. In many cases, particularly for the ICP, DLs
obtained in practice with commercial instruments may The analytical sample presented to the flame or plasma
be significantly worse (e.g., by a factor of 10 or more) source is most often in the form of a solution. Hence
than the values shown in Table 8-8. With ultrasonic the bulk sample, if it is not initially in solution form,
nebulization and research-grade instrumentation, the must first be dissolved andlor decomposed. Next, sam-
best DLs obtained can be a factor of 10 lower than ple cleanup methods are used, if necessary, for sepa-
those given in Table 8-8. ration of the analyte, removal of primary interferences,
If random noise in the blank limits measurement or preconcentration of the analyte. Finally, the analyt-
252 Chap. 8 I Flame and Plasma Atomic Emission Spectrometry
ical sample itself is prepared along with the appropriate cathode lamps or other standard spectral sources (e.g.,
blank and standard solutions. The exact treatment of low-pressure Hg arc lamps). The presence of spectral
the samples and standards depends on the specific method multiplets often greatly assists assignment of lines to
to be utilized. The most common sample dissolution elements because of their fingerprint nature and the
and separation methods employed in atomic spectro- stability of their intensity ratios under various condi-
chemical methods are reviewed in Appendix E. tions. Once a particular wavelength has been assigned,
A major advantage of the emission spectrometric it is useful to determine whether the assignment is cor-
methods is their ability to accept samples without a great rect by using standards of different concentrations. With
deal of sample pretreatment. Because of the low de- multichannel spectrometers, the positions of the various
tection limits of the plasma emission methods, precon- exit slits are often preset at the factory and not readily
centration methods are not often needed. Although some changed by the user. The exact wavelength regions passed
of the procedures described in Appendix E may be by each slit can often be varied by refractor plate ad-
needed for sample dissolution, in general, extensive justments. These are usually made while radiation from
separation and sample treatment steps are not required a stable, narrow line source, such as a hollow cathode
with these sources. Flame emission methods more often lamp, is incident on the spectrometer. The refractor
require sample preconcentration and the addition of plate position is adjusted to give a maximum photo-
chemicals such as ionization buffers and complexing or multiplier tube output signal.
releasing agents.
In addition to the sample treatment discussed Selection of Analysis Lines. With flame emis-
above, emission spectrochemical methods usually re- sion it is often an easy task to pick the exact emission
quire a choice of instrument and conditions as well as line to be used for the analysis. For most elements, the
the specific methodology to be employed (external resonance lines are the most sensitive in flames, and
standard, internal standard, standard addition method, they are thus used in the majority of flame methods.
etc.). The particular solvent to be used may also be an With plasma sources, the choice is more difficult be-
important factor because of sample insolubility in aqueous cause several emission lines from neutral atoms or ions
media. Samples in organic matrices, for example, may of the same element may appear useful in quantitative
dictate that a DCP or flame technique be used, since determinations. These often vary widely in relative in-
ICPs are less tolerant of the solvent. This section begins tensities and in the abundance of nearby, potentially
by considering the identification and selection of the interfering lines from other elements. With some direct
emission lines that will be employed in quantitative readers, of course, the selection may have been sub-
methods. Then we consider briefly some of the most stantially narrowed by the initial setup of the exit slits
common procedures that are used in emission spec- and detectors. Reference books, atlases of spectral lines,
troscopy and the trade-offs involved in choosing a spe- journal articles, and application notes from instrument
cific method. Finally, a few specific applications are manufacturers can be extremely helpful in choosing a
presented to illustrate how the techniques are used in good plasma analysis line. These often give linear ranges
practical situations. and potential interferences for a given emission line
with a particular sourcelspectrometer combination. Often
the expected concentration range will dictate whether
ldentification and Selection of Analytical to use a neutral atom resonance line, an ion line. or a
Lines line arising from transitions between excited atomic states.
Resonance lines are, of course, most useful for trace
In order to use emission spectroscopy the analysis line(s) constituents, but they are also susceptible to self-ab-
must be identified and selected. For qualitative analysis, sorption at high concentrations. Lines of lower relative
wavelength identification is the primary task, while with intensities are often used for minor and major constit-
quantitative methods selection of the appropriate line uents. If an internal standard is being used (see later),
is of paramount importance. the availability of a suitable internal standard line also
influences the choice of analysis line. In many cases two
Wavelength Identification. With flame emis- or three lines are monitored during the initial method
sion the scarcity of lines makes wavelength identifica- development stages, and the most suitable of these is
tion quite straightforward. Since plasma sources pro- chosen for final use. With some computerized direct
duce much richer spectra, identification can be more readers, it is possible to use more than one line in the
difficult. Scanning spectrometers can simplify the task final analysis procedure and obtain results that are the
of wavelength identification. However, errors in the average of several lines. Also, more than one line can
grating drive system and the wavelength readout should be used to extend the concentration range if samples
be assessed frequently by recording spectra from hollow vary widely in analyte concentration.
Sec. 8-7 / Methodology and Applications
If multiple elements are to be determined with a In order to make effective use of the internal
scanning or slew-scan monochromator, it is usually de- standard method, the two line intensities should be
sirable that the lines chosen be near each other in wave- measured simultaneously. With a direct-reading spec-
length. This minimizes the time required to scan or slew trometer or a spectrograph with multichannel detection,
between lines and allows the same detector to be used simultaneous acquisition of intensities is readily achieved.
for all the lines. The specific source and spectrometer The internal standard method is, however, difficult to
often plays a major role in choosing an appropriate line. implement with a slew-scan or linear scan monochro-
mator since measurements are necessarily taken at dif-
Analytical Procedures ferent times.
There are several requirements for a good internal
Most of the analytical methodology discussed in Chap- standard element. First, the internal standard element
ter 6 has been applied to emission spectrochemical anal- should not be present in the samples or standards so
yses. With flame emission, external standard methods that a known concentration can be added. This require-
are most common. In the usual analytical procedure, ment is often difficult to meet with complex samples.
the flame emission spectrometer is first optimized for In some cases a constituent already present in the sam-
maximum signal or SIN as described in Section 8-5. ples at a fixed level can be used as an internal standard.
Then the analytical signal EE (total signal minus blank) Second, as can be seen from equation 8-12, the exci-
is obtained for a series of standards of known analyte tation energies of the two transitions should be similar
concentration c, preferably with added constituents to to compensate for changes in source conditions (tem-
simulate the matrix of the analytical sample. The blank perature, electron density, etc.). Third, factors that re-
should be a reagent blank that contains all reagents used late gas-phase concentrations to solution or solid con-
in the procedure at identical concentrations as in the centrations should be affected similarly by changes in
sample. A calibration curve is obtained by plotting EE conditions. For solution samples and standards, for ex-
vs. c, or log E,vs. log c,. The net signal is then measured ample, this requirement means that the two elements
for the analytical sample, and the analyte concentration should have similar ionization potentials, similar chem-
obtained from the calibration curve or its mathematical ical compound formation tendencies, and similar vol-
equivalent. atilities so that changes in atomization efficiencies are
With plasma emission, external standard methods compensated. Changes in solution introduction rates
are also used, but they require careful control over should be well compensated by the internal standard
source conditions if high accuracy and precision are to method.
be achieved. Because line intensities are highly de- It is rare to find an internal standard element that
pendent on source parameters, small changes between meets all the requirements given. Instead compromises
standards and samples can cause large errors. must be made, particularly with multielement deter-
minations, where a single internal standard often serves
Internal Standard Methods. The internal for several analytes. The internal standard method can,
standard method (see Section 6-5) is widely used in however, be quite successful in reducing variations due
emission spectrometry, particularly in flame emission to changes in source temperature, source position, and
determinations of Na and K, as discussed later in this sample introduction rate.
section. In internal standard methods, a known con-
centration of the reference element is added to all sam-
ples, standards, and blanks, and the ratio of the inten- Standard Additions. The method of standard
sity of the analyte to that of the reference element is additions is also frequently used in emission spectrom-
obtained. If we consider the equation relating readout etry for solution samples (see Section 6-5). Usually, the
signal to ground-state number density (equation 8-4), method of multiple additions is preferred because of its
we can see that a number of factors cancel or are in built-in check of linearity. Standard additions have also
constant proportion when the ratio of two line inten- been applied to multielement determinations. Here a
sities is obtained. Equation 8-12 expresses the measured multielement standard is used to make the additions.
voltage signal ratio as a function of the analyte number Concentrated standards are added in small volumes to
density, (n,),, the reference element number density, avoid changing the matrix of the analytical sample.
(n,),, and the difference in upper state energies
( E , - E,) for both transitions being resonance transi-
tions.
Emission spectrochernical analysis has been used for a
wide variety of sample types and almost every stable
element. Here we consider one practical application of
254 Chap. 8 I Flame and Plasma Atomic Emission Spectrometry
flame emission and two applications that use plasma L-l of NaC and 2 to 8 rneq L-' of K'. Many clinical
excitation. analyzers provide automatic dilutions so that standards
and serum samples are identically diluted with the in-
Determination of Na and K in Blood Serum. ternal standard. A blank containing only the internal
Although the use of flame emission has been decreasing standard is used for the zero adjust. Often only a one-
in recent years, it is still widely used in clinical methods. or two-point calibration is used. Control serum samples
Even here, however, many laboratories have switched with known amounts of Na and K are frequently ana-
to the use of ion selective electrodes, particularly for lyzed for accuracy assessment. Typical reproducibilities
determining Ca in serum. Flame emission results give are .= 1 rneq L p l for Na and 0.1 rneq L-l for K.
the total calcium present, which is not as clinically useful
as the free, ionized calcium. For Na and K, flame emis-
sion is still widely used. Water Testing with an ICP Spectrometer.
The normal ranges in serum are 135 to 142 rneq Several water resource laboratories use ICP emission
L-I (3105 to 3266 pg mL-l) for Na and 3.5 to 5.0 rneq spectrometry for testing the quality of various ground-
L-I (137 to 195 pg mL-I) for K. Typically, samples and surface-water samples. In one method [J. R. Gar-
(usually, -
are diluted 1:1000 with a Li internal standard solution
15 rneq L p l ) before introduction to the
flame spectrometer. Simple dilution produces a solution
barino and H. E. Taylor, Spectrochim. Acta, 38B, 323
(1983)], 17 elements (Ba, Be, Ca, Cd, Co, Cu, Fe, Li,
Mg, Mn, Mo, Na, Pb, Si, Sr, V, and Zn) were deter-
that can be readily nebulized without additional treat- mined simultaneously with an automated spectrometer.
ment (deproteinization or ashing). A computer provided acquisition, processing, and stor-
To take full advantage of the internal standard age of data generated by the spectrometer system. In
method, a three-channel photometer, such as that shown addition, the computer controlled an automatic sampler
in Figure 8-16, is used for simultaneous ratio recording. and an automatic standardization system for introduc-
This ratio recording system outputs directly the ratio of ing the external standards. A rotating refractor plate
the Na signal to the Li internal standard signal and the (spectrum shifter) was used for dynamic background
ratio of the K signal to the Li signal. Still other com- corrections. Interelement corrections were applied to
mercial systems allow Na and K to be measured relative analytes that suffered spectral interferences. One par-
to a Li internal standard and Li to be measured relative ticular standard was analyzed frequently for quality con-
to a K internal standard. Because of the low excitation trol of the analytical results. Approximately 100 sam-
energy needed for alkali metals and to avoid ionization, ples could be analyzed daily for the elements listed
air-propane flames are commonly used. above; the element profiles were automatically entered
Standard solutions containing both Na and K are into a data-base management system on a laboratory
prepared. Usually, these range from 100 to 160 rneq computer.
/
/ Balance
/
Marine Sediment Analysis with a DCP Spec- ards were matrix matched as closely as possible. T h e
trometer. Marine sediments taken near offshore drill- high concentrations of Ca were found t o result in stray
ing installations often show heavy metal accumulations. radiation errors for all elements except Cd. T h e errors
These can b e monitored with emission spectrometry. were reduced t o tolerable levels by adding Ca t o all
In o n e study [W. C. Grogan, Spectrochirn. Acta, 38B, standards. I n a related study, shellfish extracts were
357 (1983)], a DCP system was used because of its analyzed by t h e same method. Stray radiation errors
ability to handle solutions with a high content of dis- were found t o arise from both C a and Mg. Correction
solved solids. Sediments were digested with HNO, and equations were used t o subtract t h e contribution of stray
diluted. A high concentration set was analyzed and t h e radiation. Excellent agreement between DCP results
trace elements, C d , C r , Cu, Ni, P b , V, and Z n deter- and certified values were found when standard mate-
mined. Further dilution was necessary in order t o de- rials of similar matrices were analyzed.
termine the major elements B a , Ca, Fe, and Sr. Stand-
Give a brief scientific reason why the following state- 10, F = 5.0mLmin-l, E, = 0.03, Q = 1 0 L m i n - I ,
ments are true. A,, = 0.95 x lo8 s-l, go = 2, and g, = 4. The
(a) Ionization interferences are usually not as severe pathlength 1 = 1 cm. A monochromator is used with
in the inductively coupled plasma (ICP) as in flames. a slit width W of 50 ym, a slit height H of 5 mm, a
(b) Echelle-grating monochromators or polychroma- solid angle SZ of 0.02 sr, and an optics transmission
tors are increasingly being found in commercial factor To, = 0.6. A PMT is used with a gain m of
ICP atomic emission systems. 5 x lo5 and a cathodic responsivity R(h) of 15 mA
The flame emission determination of Na in a biolog- W-I. The current-to-voltage converter has a transfer
ically derived fluid is being considered. Burners and function of lo7 V A-I. Calculate the analyte emission
gases for all common flames listed in Table 8-3 are signal voltage E, under these conditions.
available. Discuss the most important factors that will What types of data suggest that local thermodynamic
influence the choice of which flame to use. Include the equilibrium does not exist in an ICP?
advantages and disadvantages of low and high tem-
peratures for this determination. Compare and contrast the thermal energy ( k T ) in a
flame of 3500 K to that in the axial channel of an ICP
Compare and contrast free-atom formation from a so- at 7500 K.
lution sample for a N,O-C2H, flame and an ICP. List
all of the steps involved and draw conclusions as to Discuss the atomizer and excitation parameters that
which excitation source is most efficient for each step. would be influenced by a sudden increase in fuel gas
Justify. flow rate in a flame emission determination. How would
Compare and contrast the DCP and the ICP for the these parameters change, and what would be the effect
on the emission signal for Na in an air-H, flame? If
following sample types.
the oxidant is also the nebulizing gas, what parameters
(a) An aqueous solution sample.
would be affected by a sudden increase in oxidant flow
(b) A sample in an organic solvent.
rate? Will the Na emission signal increase or decrease
(c) A sample with a high particulate content.
or is the result unpredictable?
Consider the list of ICP characteristics given in Table
8-5. Do these apply to the MIP and DCP as well? For In an atomic emission spectrometry determination, the
each item in the table state whether it is applicable to concentration of the analyte in the sample solution is
these sources and why or why not. in the region of the calibration curve where self-ab-
sorption is noted with standards. Discuss the various
Contrast H, and C2H2flames in terms of background
options that are available to obtain accurate analytical
and atomization characteristics.
results.
Calculate the SIN and SIB in an emission measurement
limited by background emission shot and flicker noise What would be the effect on the SIN of working in
with a slit width of 100 ym if E, = 0.010 V, EbE = the self-absorption region? If dilution by a factor of
0.050 V , mGK = 4.0 x V, and X, = 0.010. at least 20 is needed to put the sample emission signal
Calculate the SIN and SIB at slit widths of 200 and 400 into the linear region, is it better from an SIN stand-
ym assuming that the background emission is a con- point to work in the nonlinear region or to dilute the
tinuum. Justify the observed dependence on slit width. sample? Assume that signal shot noise is limiting for
Consider the flame emission measurement of a 50-yg both cases.
mL-' Cu solution at 324.7 nm in a 2000-K flame. The Discuss how the electron number density might be
atomizer and analyte spectroscopic factors are ef = measured in a particular spatial region of an ICP.
256 Chap. 8 I Flame and Plasma Atomic Emission Spectrometry
REFERENCES
The following references deal with flames and flame emission 1-196 (1986). This entire issue of Spectrochimica Acta
spectroscopy. is devoted to proceedings of a conference on plasma
spectrochemistry. Articles from earlier conferences may
C. Th. J. Alkemade, Tj. Hollander, W. Snelleman, and be found in "Plasma Spectrochemistry 11," Spectrochim.
P. J. Th. Zeegers, Metal Vapors in Flames, Pergamon Acta, 40B, 1-412 (1984) and "Plasma Spectrochemistry
Press, Elmsford, N.Y., 1982. I," Spectrochim. Acta, 38B, 1-445 (1983).
C. Th. J. Alkemade, and R. Herrmann, Fundamentals 14. H. R. Griem, Plasma Spectroscopy, McGraw-Hill, New
of Analytical Flame Spectroscopy, Halsted Press, New York, 1964. A leading book on plasma diagnostics and
York, 1979. physics from a spectroscopic perspective.
C. Th. J. Alkemade and P. J. Th. Zeegers, "Excitation 15. R. K. Winge, V. A. Fassel, V. J. Peterson, and M. A.
and De-excitation Processes in Flames," in Spectro- Floyd, Inductively Coupled Plasma-Atomic Emission
chemical Methods of Analysis, J . D . Winefordner, ed., Spectroscopy: A n Atlas of Spectral Information, Elsev-
Wiley-Interscience, New York, 1971, pp. 3-125. An ier, New York, 1985.
excellent treatment of the processes occurring in flames
16. M. L. Parsons, A. Forster, and D. Anderson, A n Atlas
from a fundamental point of view.
of Spectral Interferences in ICP Spectroscopy, Plenum
R. Mavrodineanu, Analytical Flame Spectroscopy-Se- Press, New York, 1980.
lected Topics, Springer-Verlag, New York, 1970.
17. R. M. Barnes, "Sample Preparation and Presentation
A. Syty, "Flame Emission Spectroscopy," in Treatise o n in Inductively Coupled Plasma Spectrometry," Spec-
Analytical Chemistry, 2nd ed., pt. 1, vol. 7, chap. 7, troscopy, 1(5), 24 (1986).
I. M. Kolthoff and P. J. Elving, eds., Wiley-Interscience,
18. R. S. Houk, "Mass Spectrometry of Inductively Coupled
New York, 1981.
Plasmas," Anal. Chem., 58, 97A (1986).
A. G. Gaydon, The Spectroscopy of Flames, 2nd ed.,
19. P. W. J. M. Boumans, ed., Inductively Coupled Plasma
Halsted Press, New York, 1974. An excellent introduc-
Emission Spectroscopy, pts. I and 11, Wiley-Interscience,
tion to flame phenomena from a spectroscopic view-
New York, 1987. Covers the fundamentals, instrumen-
point.
tation, methodology, and applications of ICP emission
A. G. Gaydon and H . G. Wolfhard, Flames, Their Struc- spectroscopy.
ture, Radiation and Temperature, 3rd ed., Chapman &
20. A. Montaser and D . W. Golightly, eds., Inductively
Hall, London, 1971. A widely used book on the fun-
Coupled Plasmas in Analy tical Atomic Spectrometry, VCH
damental properties of flames.
Publishers, Inc., New York, 1987. Sixteen chapters writ-
J. A . Dean and T. C. Rains, eds., Flame Emission and ten by experts on ICP devices and their applications.
Atomic Absorption Spectrometry, Vols. 1-111, Marcel
Dekker, New York, 1969, 1971, 1975. A multiple vol- The following are general references on emission spectros-
ume work written by experts in the field. copy and spectrochemical excitation (see also Chapter 7 ref-
R. Mavrodineanu and H. Boiteux, Flame Spectroscopy, erences).
Wiley, New York, 1965. 21. P. W. J. M. Boumans, Theory of Spectrochemical Ex-
The following references deal with plasmas and plasma spec- citation, Adam Hilger, Bristol, England, 1966.
troscopy. 22. C. Veillon, "Optical Atomic Spectroscopic Methods,"
in Trace Analysis: Spectroscopic Methods for Elements,
10. V. A. Fassel and R. N. Kniseley, "Inductively Coupled
Plasma Optical Emission Spectroscopy," Anal. Chem., chap. VI, J . D. Winefordner, ed., Wiley-Interscience,
New York, 1976.
46, lllOA, 1115A (1974). Among the early ICP papers.
23. R. M. Barnes, ed., Emission Spectroscopy, Dowden,
11. V. A. Fassel, "Simultaneous or Sequential Determi-
nation of the Elements at All Concentration Levels- Hutchinson & Ross, Stroudsburg, Pa., 1976. This book
the Renaissance of an Old Approach," Anal. Chem., is a collection of the "classic" papers in the field of
51, 1290A (1979). A review stressing the unique aspects emission spectrochemical analysis.
of the ICP in multielement analysis. 24. "Emission Spectrometry." A review of progress in the
field that appears once every two years in the Funda-
12. R. K. Skogerboe and G. N. Coleman, "Microwave Plasma
Emission Spectrometry ," Anal. Chem., 48, 611A (1976). mental Reviews issue of Analytical Chemistry.
13. "Plasma Spectrochemistry 111," Spectrochim. Acta, 41B,
The story of emission spectroscopy would certainly be semiquantitative analysis, because the precision of
incomplete without a consideration of arc and spark quantitative determinations is not as high as desirable.
excitation. These sources were primarily responsible for The high-voltage spark is even more energetic than the
the "coming of age" of emission spectroscopy. Before arc; even the rare gases and the halogens can be excited
arcs and sparks became available, only flames were in a spark discharge. The spark is capable of higher
widely used analytically to excite atomic emission. With precision than the dc arc, and is applied primarily in
arcs and sparks it became possible to excite nearly all quantitative determinations. Although plasmas have re-
the stable elements in the periodic table. Many of the cently become the most widely used excitation sources
instrumentation developments that were made with arc for emission spectrometry, arcs and sparks are still used,
and spark sources were carried over directly for use particularly for the analysis of solid samples. In fact, as
with the plasma sources that became popular in the we shall see, one commercial instrument uses a spark
1970s. discharge to vaporize solids prior to ICP excitation.
Arc and spark discharges have been used as ex- We saw in Chapter 8 that the ICP and DCP ap-
citation sources for qualitative and quantitative emis- proach our ideal atomic emission source. However, there
sion spectrometry since the 1920s. Many new devel- are many problems to be solved with plasmas. For ex-
opments in arc and spark excitation occurred during the ample, the plasma sources are less than ideal when it
war years of the 1940s, particularly during the Man- comes to dealing with solid samples or samples in or-
hattan Project. Commercial instrumentation became ganic matrices. Thus much research is currently being
available during this period, and use of the photomul- devoted to the development of new excitation sources
tiplier tube instead of the photographic plate increased for atomic emission spectrometry.
dramatically. Photoelectric detection greatly increased This chapter begins by describing the arc and spark
the throughput of quantitative determinations and led discharges that have proven useful in analytical emis-
to the development of many new quantitative proce- sion spectrometry. These sources are compared to the
dures. ideal atomic emission source developed in Chapter 8.
With the dc arc some 70 to 80 elements have been Several "miscellaneous" excitation sources are then
excited. The major use of the arc is in qualitative and considered. These include laser devices and reduced
258 Chap. 9 1 Arc and Spark Emission Spectrometry
likely to occur late in the burn when refractory materials provide substantial improvements in detection limits.
with rich spectra are volatilized. When operated in pure Ar atmospheres, the arc con-
Selective volatilization can also be used to advan- sumes electrodes and samples at extremely slow rates,
tage to enhance line-to-background ratios and minimize which can result in long analysis times. For this reason
interferences if the exposure period (integration time) a mixture of 80% Ar and 20% 0, is often used. The
is selected correctly. A low-boiling element such as gal- added 0, increases the rate of sample consumption
lium is sometimes intentionally added to the sample in without increasing CN emission.
the carrier distillation method. Although the exact The Stallwood jet shown in Figure 9-5 is com-
mechanism is not understood, when the low-boiling ele- monly used for controlled-atmosphere arc excitation.
ment volatilizes it is thought to carry with it other easily The particular gas mixture used establishes the exci-
volatilized elements, which can separate them from re- tation conditions. The gas flow helps to reduce arc wan-
fractory materials. Selective volatilization is often re- der, which improves analysis precision. The gas stream
duced by adding a spectrochemical buffer to the sample. also removes the absorbing vapor in the outer fringes
These are usually volatile, low-ionization-potential ma- of the arc, which reduces self-absorption. The cooling
terials such as alkali and alkaline earth carbonates or of the sample electrode helps to reduce selective vol-
halides. In many cases, graphite powder is added to the atilization.
buffer to increase conductivity. An excess of buffer tends
to keep the arc temperature constant and nearly in- AC and Intermittent Arcs. Two types of ac arcs
dependent of sample composition. Dilution with the have been employed. The high-voltage ac arc is oper-
buffer, of course, degrades detection limits. ated at 2000 to 4000 V, while low-voltage ac arcs employ
The dc arc emits band and continuum radiation, 100 to 400 V. In contrast to the continuous dc arc, the
which contribute to background. Band emission occurs ac arc extinguishes at the end of each half-cycle. High-
from molecules and radicals that are stable in the cooler voltage arcs reignite automatically when the applied
fringes of the arc. For example, in air, emission from voltage exceeds the breakdown voltage of the gas. Low-
the cyanogen radical (CN), formed from burning car- voltage arcs are reignited on each half-cycle by a low-
bon electrodes in the presence of nitroken, is quite in- current spark discharge. An intermittent arc is similar
tense. The CN band system can render the entire spec- to an ac arc except that the electrode polarity does not
tral region between 360.0 and 420.0 nm completely
unusable for analytical work. Continuum emission can
occur from the hot electrodes (blackbody radiation),
from incandescent particles in the arc, and from radia-
tive recombination and bremsstrahlung. One final prob-
lem with the free-burning dc arc is arc wander, which
occurs because the arc column contacts the electrodes
only at very small spots. Most of the volatilization of
sample and electrode material occurs at these cathode
and anode spots, which tend to move erratically over
the electrode surface, causing different portions of the
U
electrode to be sampled. The arc is also positionally
unstable because of thermal currents that are generated
during the burn. Arc wander is a major contributor to
the poor reproducibility usually associated with the dc Gas inlet
arc.
change on each half-cycle. In all cases the extinguishing Since the arc suffers from many nonidealities in
and reignition of the arc causes a different portion of its vaporization, atomization, and excitation roles, some
the electrode to be sampled on each cycle; this provides research has been aimed at separating the various func-
greater precision than the erratic, slow-moving dc arc. tions. For example, arcs have been used to vaporize
The ac arc is, however, less sensitive than the dc arc samples prior to their introduction into plasma excita-
because of the periodic changes in electrode polarity. tion sources. The microarc discharge shown in Figure
The arc behaves like a cathode layer arc on one half- 9-6 has been used for introducing small volumes of so-
cycle and like an arc with anodic sampling on the other. lution samples into MIPS and ICPs. The microarclplasma
Sample volatilization is more efficient when the sample combination separates in time the processes of desol-
electrode is anodic. On the cathodic half-cycle, signal- vation and volatilization from those of atomization and
to-background ratios arc reduced because there is less excitation so that these can be separately optimized.
sample vaporization. With the same current magnitude Introduction of samples with the microarc does not seem
and arc dimensions, there is less electrode heating in to degrade the precision and sensitivity of the plasma
an ac arc than in a dc arc. This can be a disadvantage source. Matrix interferences are not significantly dif-
for difficult-to-volatilizematerials, but an advantage with ferent from those normally experienced with the plasma.
metallic electrodes that might melt under continuous As can be inferred from the discussion above, arcs
arcing. The unidirectional, intermittent arc maintains are far from meeting the criteria established for our
the sample electrode as the anode on both half-cycles. ideal emission source. Arcs do not provide the con-
It thus has the sensitivity of the dc arc, while maintain- trollable excitation energy desired. In fact, the energy
ing the high precision of the ac arc. dissipated in the arc depends highly on the sample con-
stituents and their ionization potentials. The arc dis-
charge does not occur in an inert chemical environment,
and arcs are subject to severe matrix effects. Free-burn-
ing arcs often suffer from severe self-absorption and
Much of the recent work with arc excitation has in- even self-reversal of analytically useful lines. Back-
volved fundamental studies of the processes leading to ground emission can be severe unless controlled-at-
atomic emission. For example, studies have been made mosphere excitation is used. Although an arc is capable
of the effects of various inert and reactive gases in arc of exciting most elements, the reproducibility of the
formation, reproducibility and excitation. Various new atomization and excitation conditions is not high. For
geometries such as U-shaped arcs and V-shaped arcs these reasons the use of arc discharges in emission spec-
have been investigated. Several workers have also stud- troscopy has decreased dramatically in recent years,
ied the influence of a variety of external parameters, although we may find more uses being made of arc
such as magnetic fields and rotating electric fields on discharges in systems that separate sampling and exci-
arc behavior. tation.
To plasma
111
FIGURE 9-6 Diagram of a micro-
arc vaporizer for plasma atomic emis-
sion spectrometry. The arc is formed
between a tungsten cathode loop (A)
and a stainless steel anode (B). A
small volume of sample (1 to 10 yL)
is deposited on the cathode loop with 'Ace-tlued'
a syringe. A high-voltage, low-cur-
rent pulsating dc discharge (1500 V,
20 mA) desolvates the sample and
vaporizes it into the nebulizing gas
stream of the ICP or MIP. [From J. P.
Keilsohn, R. D. Deutsch, and C . M.
Hieftje, Appl. Spectrosc., 37, 101
(1983). Reprinted by permission of
Applied Spectroscopy.]
T
Nebulizer-gas
262 Chap. 9 1 Arc and Spark Emission Spectrometry
9-2 HIGH-VOLTAGE SPARKS AND OTHER sparking. Doubly ionized elements can produce quite
EMISSION SOURCES intense spectra in a high-voltage spark discharge. In
fact, ionization is so common in spark excitation that
High-voltage spark discharges have long been used as ion lines are often simply called spark lines. In spec-
excitation sources in emission spectrometry, especially troscopic terminology ion lines are designated as type
in the ferrous metal industry. These sources are char- (11), or type (111) lines, where the Roman numeral in-
acterized by the high precision with which they can dicates the state of ionization plus one. Thus a singly
vaporize and excite many solid samples. The high-volt- ionized element produces a type (11) line. a doubly
age spark discharge is an intermittent discharge rather ionized element a type (111) line, and so on. The larger
than a continuous discharge like the dc arc. A single number of ionized and excited states produced in sparks
spark normally lasts a few microseconds. Electrode ma- results in spectra that are more complex and less intense
terial is sampled many times during sparking in order than those produced by dc arcs.
to improve reproducibility. Because of the intermittent
nature of the spark and the resulting time-dependent Spark Sources. Simplified diagrams of two high-
atomic emission, the spark discharge is an extremely voltage spark source designs are shown in Figure 9-7.
complicated excitation source. The combination of the To regulate the breakdown of the analytical spark gap,
complex excitation processes and the inhomogeneities a control gap (Figure 9-7a) or an electronic switch (thyr-
associated with many solid samples often leads to "ma- atron tube in Figure 9-7b) is used. A fixed control gap
trix effects" with spark discharges. We have only re- allows the spark to fire only after a particular capacitor
cently begun to understand how sampling and excitation voltage is reached (voltage-thresholded spark), while a
occur with high-voltage spark discharges. Fundamental rotary gap can be used for time-thresholded operation.
mechanistic studies of spark excitation have led to sev- The thyraton-triggered source of Figure 9-7b can be
eral recent developments. including stabilized sparks operated in either a voltage- or a time-thresholded mode.
and controlled-waveform discharges. In addition to Once the analytical gap breaks down. a high current-
sparks, many other sources have been proposed and density discharge channel is formed between the two
studied for use in emission spectrometry. We consider electrodes. The duration of a single spark is generally
here two sources that use lasers to vaporize samples, a few microseconds. Conditions are arranged so that
several low-pressure discharges. and two types of sources several hundred breakdowns occur per second, and the
under development. radiation emitted by several thousand individual sparks
is integrated to improve precision. In spark excitation,
The High-Voltage Spark Discharge the sample electrode is arranged to be initially the cath-
ode because vaporization of cathodic electrodes is more
The sampling process in a high-voltage spark discharge rapid than that of anodic electrodes.
is more favorable than that in a dc arc discharge. An The sources shown in Figure 9-7 both operate by
arc tends to burn for some time at one or a few spots, overvolting the analytical gap. Walters and coworkers
which leads to selective and erratic volatilization. With have described an entirely different approach to spark
a spark, electrode material is sampled by many succes- source design in which gap breakdown occurs at the
sive discharges, striking different spots on the surface. open circuit end of a transmission line connected to a
This random, multiple sampling helps to improve the radio-frequency power supply. Once gap breakdown
precision of spark emission measurements because of occurs, a separate current source is used to inject cur-
averaging over multiple spots. However, the spark is rent into the conducting gap. An arbitrary current wave-
also not an ideal sampling device. The surface of the form can be injected into this quarter-wave source be-
material sampled tends to change with sparking time, cause the gap ignition circuitry is completely separated
which leads to changes in spark spectra with time. This from the current supply circuitry,
so-called "sparking-off" effect is often overcome by Samples for spark excitation are most often ma-
postponing measurements until a reproducible point in chined into the proper shape to serve as electrodes. In
the sampling process has been reached. Such a prespark some cases they can be solutions or solution residues.
or preburn time is typically 1 or 2 minutes. Conducting samples are often ground flat and used as
Although the spark discharge is an extremely en- one electrode with the other being a pointed graphite
ergetic source with high peak currents and high power electrode (point-to-plane configuration). Powders can
densities, limits of detection are usually higher than with be mixed with graphite and pressed into pellets, which
dc arc discharges because states of high internal energy then serve as the planar electrode. Solutions have been
are produced, and the extent of ionization is higher. introduced directly in the form of an aerosol from a
Also, very little sample is ordinarily consumed during pneumatic nebulizer both with and without prior de-
Set. 9-2 I High-Voltage Sparks and Other Emission Sources
in the discharge current, excitation conditions in such any single temperature to a spark plasma.) Ionic lines
a spark were highly time dependent. Today, however, tend to show intensity-time variations that follow roughly
most spark sources are unidirectional. Such sources give the current waveform. Neutral atom lines tend to reach
higher precision because of lower drift. Some of the their maximum intensities well after the current peaks.
newer spark source designs allow quite elegant control When the current decreases, ion-electron recombina-
over the discharge waveform. The quarter-wave source tion can occur along with various decay processes to
mentioned earlier allows arbitrary waveforms to be used. produce lower-energy states. Energy transfer from met-
Further investigations of tailored waveforms for high- astable species has also been proposed as a means of
voltage spark discharges are warranted. producing excited analyte atoms. Neutral atom emis-
sion is most pronounced in the radial wings of the dis-
Processes i n Spark F o r m a t i o n a n d De- charge as the current is decreasing.
cay. The processes that occur during the buildup and After the current ceases, emission disappears and
decay of a high-voltage spark are extremely complex. a postdischarge toroidal structure is formed. This torus
It is only in recent years that we have begun to under- has been observed by Schlieren photography. The exact
stand some of these processes and to take advantage of mechanism of torus formation and decay are not well
this knowledge. We will present here a qualitative de- understood, but the torus has been shown to contain
scription of the stages of spark formation and decay. ground-state analyte atoms as well as ions. Reexcitation
For more extensive treatments, the reader should con- of the torus material has been suggested as a way of
sult the references for this chapter. separating spark sampling frorn analyte excitation. The
We will assume here that a triggered, non-quarter- postdischarge vapor cloud can exist several hundred
wave spark source is used. After the source has been microseconds after the current ceases. Typically, there
triggered, the capacitor and gap voltage begin to in- are 100 ~s to 10 ms between discharges. The processes
crease. Typically this voltage buildup stage lasts from above are then repeated for the total number of sparks
100 to 500 ns. Initial spark formation begins with the used. It has been found that the breakdown voltage for
formation of a plasma in the gap. This plasma consists later discharges in a train of sparks is lower than that
of a thin streamer of charge carriers which cross the for single discharges or low-repetition-rate discharges.
gap and define the current channel between the elec- It is reasonable to presume that ions and metastable
trodes. Emission from ionized atmospheric species and species from previous discharges are present at the be-
continuum background are observed in the early stages ginning stages of later discharges and responsible for
of a spark. During these stages the channel is anchored this reduced breakdown voltage.
to the cathode, and a cathode spot forms. Current is
then conducted through the spark channel. This stage Time-Resolved Spectroscopy. As discussed
usually lasts about 100 ks. above. ionic and atomic analyte emission can vary dra-
The mechanism of energy transfer from the cur- matically with time during a spark discharge. In some
rent channel to the electrode surface is not entirely cases, the background emission and the analyte emis-
clear, but positive-ion bombardment (sputtering), pos- sion show significantly different intensity-time behav-
itive-ion neutralization, neutral-atom condensation, and ior. In such cases, time-resolved measurements of the
the return of previously ejected electrons to the elec- analyte emission signal, rather than integration
trode have been proposed. Bulk heating also occurs, throughout a complete train of sparks, have been shown
which leads to some surface melting. Charge transfer to improve the signal-to-background ratio substantially.
(neutralization) appears to be the dominant mechanism Several different approaches to time-resolved
in some sparks. Collisional excitation and ionization of measurements have been proposed. Usually, it is de-
electrode material may occur in the space-charge region sirable to obtain nlicrosecond or better resolution. Time
near the electrode surface or material may be vaporized resolution can be accomplished optically by rotating a
directly in excited or ionic states. During the sampling mirror so that the spark image is swept across the spec-
stage, intense continuum emission is observed near the trometer slit at the desired time. Alternatively, with
cathode as the current increases. Electrode vapor then electronically triggered spark sources, gated-integration
begins to propagate into the channel, where it interacts techniques (e.g., boxcar integration) can be used for
with plasma material. single wavelength measurements over a particular time
At the current peak, the temperature in the dis- window. It is also possible to gate an image intensifier
charge channel may reach 40,000 K, and the intensities tube so that multiwavelength measurements can be made.
of ion lines are quite high with respect to neutral atom Much of the fundamental information regarding
lines. (Because of the temporal and spatial inhomo- behavior of electrode material with time has been gath-
geneity it is probably improper to attempt to associate ered using time-resolved spectroscopy. Several workers
Sec. 9-2 / High- Voltage Sparks and Other Emission Sources
have also demonstrated the enhancement in signal-to- feasible. The application of modern computer technol-
noise ratios that can be achieved with such techniques. ark instrumentation should also lead to many
Unfortunately, only a very small portion of the elegant improvements. Although developments in plasma sources
work done by various researchers in the past 15 years have preoccupied manufacturers of emission spectros-
has found its way into commercial develo copy instrumentation, these sources have not solved the
problems associated with analyzing solid materials. It
Other Uses of Sparks. Although the most com- is here that the spark discharge still has an important
mon use of spark discharges is in the analysis of solid role to play.
samples, recent studies have shown that linear calibra-
tion curves over several decades in concentration can
be obtained with solution samples introduced after de-
solvation. A positionally stabilized, electronic-triggered Because no emission source truly meets all the criteria
spark source was used. Analyte emission was optimized established for the ideal source, much current research
relative to the background emission by gated-integra- is devoted to the development of new sources and the
tion techniques. study of their characteristics. We will consider here only
Spark discharges have been used as detectors for a few of the most useful or promising sources.
gas and liquid chromatography. The spark discharge is
easy to adapt for G C detection since the sam Laser Microprobe, A high-powered, pulsed laser
ready in the gas phase. Since it totally atomizes the G C beam (ruby, Nd:glass, Nd:UAG, flashlamp-pumped dye)
effluent, the spark is free from any structural effects. when focused to a small-diameter spot ( 5 to 50 pm) is
The analyte emission observed can be directly corre- capable of vaporizing solid materials, even if the sample
lated with concentration. Detection of nonmetals, such is not electrically conductive. When the laser radiation
as C, B, N, 0, and S, was demonstrated. In fact, it was strikes the surface, it produces a high-temperature va-
possible to calculate the empirical formulas of several por plume of atoms, ions, and molecules. The most
organic materials as they eluted from the G C column. successful applications of this principle have used the
Spark discharges have not been employed as success- laser for vaporization and a spark discharge for exci-
fully for detection of L C eluates. tation, as shown in Figure 9-9. The laser and spark are
Like arcs, sparks have also been used for intro- combined with a microscope objective for viewing the
ducing samples into other excitation sources. Because target, selecting the desired sampling area, and focusing
solid samples are not easily introduced into plasmas,
the spark discharge is used in one commercial instru- Prism
ment to vaporize solids into the support gas stream of
an ICP. The spark carries out the sampling and vapor-
ization of the solid material and the ICP finishes off the
\ Switch
ri
combination utilizes the high precision of spark sam-
pling and the high efficiency of the ICP for atomization
and excitation. Microscope
the laser beam. Indeed, a commercial laser microprobe inert-gas (e.g., Ar) atmosphere prior to laser spark for-
which used photographic detection was developed for mation. Solution samples have been introduced as either
analysis by this technique. With cross excitation by a dry aerosols, formed by desolvating the mist from a
spark, the spectrum obtained is determined primarily pneumatic nebulizer, or isolated droplets without de-
by the parameters of the spark source and not by the solvation. Although the LIBS technique is quite new,
characteristics of the laser. Such an instrument has it shows considerable promise for spectrochemical anal-
achieved quite spectacular results in determining trace ysis.
elements in selected portions of single blood cells, for
example. Unfortunately, laser microprobes are no longer Reduced Pressure Discharges. Gas discharges
being manufactured in the United States. With modern operated at reduced pressures have been well studied
array detector and computer technology, such an ap- by physicists. Some are beginning to be used in spec-
proach would seem quite attractive. trochemical analysis. The glow discharge is generated
between two planar electrodes in a cylindrical glass tube,
Laser-Induced Breakdown. A newer, related which is filled with gas to a pressure of a few torr. A
technique, known as laser-induced breakdown spec- dc voltage source and a current-controlling series resist-
troscopy (LIBS), uses a laser to cause dielectric break- ance are connected to the electrodes. Typically, a nor-
down of a gas located at the laser focal point. The mal glow discharge occurs with currents of to
fundamental frequency (1.06 p,m) from a Nd:YAG laser A and voltage drops of a few hundred volts across the
can produce power densities of several MW ~ r n - enough
~, electrodes. The presence of metallic electrodes in the
power to break down most gases, including ambient air. tube produces the spectrum of the metals present. In
The small spark that results produces atoms and ions the late 1960s Grimm took advantage of this and intro-
in excited states that radiate for a few microseconds duced a new type of glow discharge in which conductive
following the nanosecond laser pulse. Time-resolved solid samples could be readily inserted for analysis, as
techniques have been used to discriminate against the illustrated in Figure 9-10. In the Grimm discharge, Ar
intense continuum that is produced during and imme- is used as the filler gas at pressures of 10 to 12 torr.
diately following the laser pulse. The technique has This discharge has achieved detection limits in the
been used to analyze dust particles in the ambient air. pg g-I range for many elements. Grimm discharge
In addition, sample cells have been designed so that sources are commercially available.
gaseous samples or aerosols can be introduced into an The hollow cathode discharge has also been em-
Quartz
window
Anode
Hollow
cathode cup
Ar gas
inlet
Vacuum
pump 2
Vacuum
pump 1
End Gate
Enlarged view of
sample holder
\
Sample
FIGURE 9-10 Grimm discharge for emission spectrochemical analysis.
Solid samples are placed in the hollow cathodc: cup. The solid sample is
clamped to the cathode block and serves as the active cathode surface.
A r is used as the filter gas at pressures of 10 to 12 torr, and currents are
0.1 to 0.2 A . Positive A r ions formed by collisions with energetic electrons
sputter the sample from the surface. Sample vapor then enters the dis-
charge region where excitation and emission occur.
lnsrrumenraaon ana ren-ormance cnaracrerlstlcs 60/
ployed in atomic emission spectrometry in addition to studies of this promising method of excitation are under
its widespread use as a spectral source in atomic ab- way.
sorption and fluorescence. As discussed in Section 4-2,
the low gas temperatures in a hollow cathode tube result Theta Pinch Discharge. A theta pinch is cre-
in very narrow spectral lines, which are, of course, ad- ated by pulsing a high magnetic field around a diffuse
vantageous in emission techniques. Demountable hol- plasma. The field compresses and heats the plasma. The
low cathodes are required for emission spectrometry so plasma can be created by any of several convenient
that the cathode material can be changed. Unfortu- methods such as a microwave discharge or a spark dis-
nately, the hollow cathode tube must be brought to charge. Such pinched discharges are used in nuclear
atmospheric pressure each time the cathode is changed. fusion experiments. They can achieve temperatures an
The time-consuming evacuation and gas-filling opera- order of magnitude higher than those in sparks or ex-
tions make this emission source less than ideal for rapid, ploding thin films. Although research on the analytical
high-throughput determinations. Hollow cathode dis- potential of such discharges has only begun, they are
charges are advantageous, however, for solid samples promising discharges for sampling nonconducting solid
and for the analysis of refractory materials. Like the materials.
glow discharge, the primary mechanism for sample in-
troduction is sputtering. Since the cathode remains cool,
sputtering can be accomplished without significant se-
lective volatilization. Detection limits for many metals
are better than those obtained with atmospheric pres-
sure discharges. Extremely good detection limits have The optical and electronic instrumentation used with
also been achieved for nonmetallic elements, such as arc and spark emission is nearly identical to that dis-
sulfur, phosphorus, and the halogens. cussed in Chapter 8 in conjunction with flame and plasma
Low-pressure discharges have also been studied emission. Thus direct-reading spectrometers are used
as atomization sources for solid samples with atomic for simultaneous, multielement determinations, while
absorption or fluorescence detection. One company slew-scanned monochromators can be used for sequen-
markets a low-pressure discharge attachment for atomic tial determinations. Integration techniques are invari-
absorption spectrophotometers. The rate of sample va- ably used to increase signal-to-noise ratios and decrease
porization and atomic number density above the solid the effects of source instability. Many of the older emis-
sample surface are enhanced significantly by directing sion instruments, however, use photographic detection,
gas jets at the surface during the discharge. and a brief discussion of such systems is included here.
We also consider the performance characteristics of arc
and spark emission systems.
Exploding Thin Films. Pulsed electrical dis-
charges, such as sparks, are not convenient to use for etection for Arc an
nonconducting samples. Exploding conductors, how-
ever, have been shown by Sacks and coworkers to be
quite useful for analyzing such samples. While explod- The most common of the older instruments for arc and/
ing wires and foils have also been used, thin films have or spark emission used spectrographs with photographic
been shown to be more reproducible and convenient. emulsions to detect the radiant energy emitted. Even
The film is formed by vacuum depositing a metal with photoelectric detection, the photographic emul-
vapor on a flat, nonconducting substrate such as poly- sion remains an excellent qualitative detector to view
ethylene. The film, which may be as thin as 25 nm, is an entire region of the spectrum at once. The use of
exploded by discharging several hundred joules of photographic emulsions for qualitative identification is
energy through the high film resistance under a re- considered in Section 9-4. Here we focus on the use of
duced-pressure inert-gas atmosphere. Apparently, the photographic emulsions for quantitative measurements.
atmosphere near the film ionizes to initiate breakdown. When a photographic emulsion is exposed to ra-
Analytical applications are usually carried out by totally diant energy and later subjected to development (see
vaporizing the film. Section 4-4), we obtain a photographic response, which
The exploding thin-film technique has been shown is normally measured as the density of silver (weight of
to be capable of vaporizing materials with extremely silver produced per unit area). The response is related
high boiling points. For example, ZrC particulates have to the radiant exposure (irradiance integrated over the
been completely vaporized even though the material exposure time t ) . After development of the plate in an
boils at over 5000 K. Further mechanistic and analytical appropriate darkroom, the density D (similar to ab-
268 Chap. 9 1 Arc and Spark Emission Spectrometry
sorbance) is measured with a microphotometer called used so that the calibration can be made over a large
a densitometer. To calculate D , the densitometer meas- wavelength range. The sector, containing four to seven
ures the transmittance of a narrow beam of light through steps, is rotated rapidly in front of the entrance slit of
the exposed region of interest on the plate. Photoelec- the spectrograph. It is cut so that a series of exposures
tric detectors (phototubes or photomultiplier tubes) are reaches the plate stepwise upon rotation. Many other
commonly used. The 100% T signal (E, = E,, - Ed) techniques have been proposed to vary the exposure in
is obtained by moving the light beam to a clear area on a known manner.
the plate. The signal representing the transmitted flux The procedure for quantitative photographic work
(E, = E,, - Ed) is obtained by passing the beam through can be seen to be quite involved. First, the photographic
the exposed region of interest. The density is then re- emulsion is calibrated. Next, the spectra of interest are
lated to the measured signals by the equation obtained and the plate is removed to a darkroom for
development. After this, the plate is taken to a densi-
E tometer for measurement of the densities of the lines
D = -log T = log-, = log E,, - Ed (9-1) of interest. The densities obtained are then converted
Es E,, - Ed
to relative intensities via the emulsion calibration curve.
The relationship between density and relative ex- Of course in analytical spectroscopy, the relative in-
posure is established by means of a calibration curve tensities must be related to concentrations through an
known as a Hurter and Driffield or H and D curve, as additional calibration curve, the working curve.
shown in Figure 9-11. The slope of the linear region,
known as the gamma of the emulsion, is a measure of
the contrast. Unfortunately, gamma is also wavelength Performance Characteristics
dependent, so that the calibration should be carried out
at several wavelengths over the region of interest. The most commonly employed sample types with arc
A widely used method to obtain a known series and spark sources are solid samples, which makes it
of relative exposures is the rotating-step sector method. difficult to compare results obtained with these sources
Here a source with many lines, such as an Fe arc, is directly to those obtained with plasmas and flames on
solution samples. These sources must expend part of
their energy in sampling and vaporizing the solid ma-
terials to be excited.
Equation 8-4 gives the general relationship be-
tween readout signal E, and ground-state number den-
sity no in all emission techniques. For solid samples
introduced into arcs or sparks, we must modify equation
8-4 by introducing the relationship between ground-
state number density and solid-phase concentration.
Unfortunately, the latter relationship is extremely com-
plex because it depends on the homogeneity of the sam-
ple, the area sampled by the arc or spark, the efficiency
of vaporization, which may be time and matrix de-
pendent because of selective volatilization, and many
other factors. Linear ranges obtained thus depend highly
on the type of sample and matrix. It is not uncommon
to obtain working curves with arc discharges that bend
back over at high concentrations because of self-ab-
sorption, self-reversal, photographic saturation, or other
1 2 3 4 5 10 20 30 40 SO
Relative intensity causes. For solid samples, just obtaining appropriate
standard materials for calibration curves and testing can
FIGURE 9-11 H and D characteristic curve for
be a problem in itself. For powders that are to be mixed
photographic emulsion. A t moderate exposures
a linear relationship is obtained. The linear region
with spectrochemical buffers, pure standards can be
of the curve is called the latitude, while the slope added in different amounts. For metallic samples and
of the linear region is the gamma of the emulsion. alloys several standards are available in certified form
At low exposures the emulsion shows inertia, while from the NBS.
at high exposures photographic saturation be- Signal-to-noise considerations with arc and spark
comes apparent. emission spectrometers are very similar to those dis-
Sec. 9-4 1 Methodology and Applications
cussed in Section 8-5 for flame and plasma emission BLE 9-1
systems, although the expressions can be considerably Approximate detection limits for dc arc
more complex when solid sampling is included. In prac-
Approx. DL
tice, precision is almost always evaluated experimen- (wt %) Elements
tally by measuring replicate samples. With arcs and -
sparks, the stability of the source itself can limit pre- Li, Na, Cu, Ag
cision through its effect on temperature and indirectly K, Rb, Cs, Be, Mg, Ca, Y, La,
on atomization and excitation efficiencies. Similarly, Ti, Zr, V, Cr, Mo, Mn, Fe,
Ru, Co, Rh, Ni, Au, Z n ,
precision can be limited by sample homogeneity. High- Cd, B, Al, Ga, In, T1, Ge,
voltage spark excitation can under ideal circumstances Sn, Pb, Nd, Eu, Th, Dy, Pd,
give precision similar to that of an ICP (1 to 5% RSDs Ho, Er, Tm, Yb, Lu
with standards and 1 to 10% with real samples). By l~-~-lo-~ Hf, Nb, Ta, W, Re, Os, Ir, Pt,
contrast with free-burning dc arcs and solid samples that Hg, Si, P, As, Sb, Bi, F,
Th, U
show selective volatilization, RSDs of better than 10% 10-2-10-1 Se, Te, Ce, Sm, Gd
are rare.
Assessment of accuracy is usually a more difficult
experimental task, since accuracy depends highly on the ments can be detected in the range to wt %,
quality of the standards used and on the absence of with the majority being in the upper half of that range.
interferences. For new analytical methods, samples
should be analyzed by independent methods when they
are available. Again, standard reference materials
available from NBS should be used to assess accuracy
if needed. In some cases only semiquantitative results Dc arc emission spectroscopy is useful in qualitative and
are necessary. An entire semiquantitative methodology semiquantitative analysis as well as in quantitative de-
has been developed for use with dc arc excitation and terminations. Hence this section begins by discussing
photographic detection, as described in the next sec- this methodology. The general quantitative schemes that
tion. have proven useful for arcs andlor sparks are then de-
Interferences can be a major problem in trace scribed. The applications presented include the use of
analysis procedures, particularly with solid samples. Solid dc arc emission for samples of geological origin and the
samples are often extremely complex matrices with widely use of the high-voltage spark for the analysis of ferrous
varying compositions. This can greatly complicate the metals.
preparation of standards and lead to severe matrix ef-
fects. Sample inhomogeneity is also a major problem
with solid materials. Solids can, of course, be dissolved
by the methods described in Appendix E, and the re- The dc arc is particularly useful in qualitative analysis
sulting solution analyzed by ICP or DCP methods. because of its excellent detectability and its ability to
However, dissolution steps may introduce contaminants excite so many elements. To establish the presence or
and be excessively time consuming when results are absence of an element, two or three sensitive lines are
needed rapidly. Solids are most often determined by usually employed. More than one line is usually nec-
arc or spark excitation methods, which are well suited essary for positive identification because of the possi-
for direct analysis. bility of spectral interferences, stray radiation, and
Detection limits in arc and spark emission also background emission at the wavelength chosen. The
depend on a number of variables, such as sample ma- most sensitive lines of the element(s) of interest are
trix, source conditions, optical factors, and detector fac- normally found in standard wavelength tables, such as
tors (photographic plate vs. photoelectric detection). the MIT tables. Most tables give relative intensities of
Thus detection limits are not often meaningful because lines in the dc arc and high-voltage spark and the wave-
of the difficulty in reproducing them on other instru- lengths of these lines; some tables are arranged by ele-
ments or in other laboratories. Instead, Table 9-1 gives ment, while others are arranged by wavelength.
detection limits as ranges for several elements with a When spectra are recorded photographically, a
dc arc. Because solid samples are usually employed reference spectrum is usually recorded on each pho-
these ranges are given in wt %. tographic plate to facilitate in wavelength identification.
With spark discharge excitation, detection limits Typically, the spectrum from a low-current iron arc is
are usually somewhat poorer than those shown in Table used for this purpose. The Fe spectrum contains a mul-
9-1, as discussed previously. However, some 40 ele- titude of intense, narrow lines throughout the UV-vis-
270 Chap. 9 1 Arc and Spark Emission Spectrometry
ible region. Each photographic instrument should have ditions. By combining such sources with time- and spa-
available a standard plate that contains the Fe spectrum tial-resolution methods, it should be possible to achieve
and wavelength indications for the most intense lines. much higher precision and accuracy than obtainable
Identification of unknown samples is then made by com- previously. Indeed, one commercial system (Shimadzu,
parison of the sample and reference spectra to the Inc.) now uses time-resolved techniques for emission-
standard spectrum. intensity measurements.
Because of the time involved in using photo- In addition to the criteria given in Section 8-7 for
graphic detection for quantitative measurements, sev- choosing an appropriate internal standard, certain other
eral semiquantitative methods have been developed to factors must be considered with arc and spark excita-
provide more rapid results. In the simplest case, the tion. For solid samples, the analyte and the internal
method of quantitative estimates, the arc spectrum of standard should be similar in chemical properties so that
the sample is recorded photographically, and the plate their rates of vaporization are similar. If detection is
developed. The wavelengths to be used are then iden- photographic, the line pair should be close in wave-
tified and a visual estimate of film blackness on a scale length and of similar intensity to minimize emulsion
of 1 to 10 is made at each of the analytical wavelengths. calibration errors. If a good internal standard is used,
Published sensitivity factors are then used with the the internal standard method can be quite successful in
blackness estimate to give a rough ( k 50 to 100%) reducing variations due to changes in source tempera-
estimate of the weight percent of the element(s). ture and position (e.g., arc wander).
In the semiquantitative method of Harvey, a The method of standard additions is not often used
weighed amount of sample is mixed with graphite and in arc and spark spectroscopy, although it has been
loaded into the cavity of an electrode. The sample is applied to solid samples. The major disadvantage, when
completely burned in the dc arc under prescribed con- applied to solids, is the time required to weigh and mix
ditions, and the photographic plate is developed. Line- the standards and samples.
to-background ratios are then measured on a densitom-
eter and used along with published sensitivity factors Applications
to calculate the percent analyte in the sample. Since the
method uses the background emission as an internal Arc and spark spectrochemical analyses have been ap-
standard, careful control of conditions is necessary. When plied to many different analytes and sample types. We
used correctly, it is capable of accuracies on the order will consider here one common application of the dc
of i 30 to 50%. Neither emulsion calibration curves arc and one widely used high-voltage spark method.
nor working curves are needed.
Analysis of Silicate Minerals and Rocks. The
Quantitative Methods dc arc has found major application in the analysis of
minerals, soils, rocks, and meteorites. We will discuss
The considerations for true quantitative determinations here a general scheme for these materials, the scheme
are quite similar to those discussed in Chapter 8 for of Ahrens, which provides a good compromise between
plasma and flame emission except that sample disso- analysis speed and accuracy. The method takes advan-
lution is usually not required. First the analysis lines tage of selective volatilization to reduce matrix effects.
must be selected and tested for interferences during the Nearly all silicate rocks and minerals contain alkali
methods development stage. Then the specific meth- metals at concentration levels exceeding 0.5%. The al-
odology to be employed must be chosen. kali elements distill rapidly in the dc arc, and because
With traditional arc and spark sources, it is often of their low ionization potentials, they exert a strong
quite difficult to maintain reproducible source condi- influence on arc characteristics (e.g., temperature and
tions. Emission intensities are also highly sample de- electron density). The method of Ahrens divides the
pendent, particularly with solid samples. This often means elements to be determined into three groups: the alkali
that matrix matching of the standards to the sample is elements themselves, a group of volatile elements, and
required if external standards are to be used. Unfor- an involatile group. The alkali elements and the volatile
tunately, this means a prior knowledge of the sample group (Pb, Ga, Ag, Cu, T1, Zn, Sn, Ge, In, Bi, Sb,
composition. Hence quantitative arc and spark deter- and As) are determined during the alkali distillation
minations most often involve the use of internal stand- phase. The sodium present in the rock or mineral is
ards. used as the internal standard for the other alkali ele-
With the new generation spark sources (quarter- ments. Added lithium is used as the internal standard
wave, positionally stabilized, etc.) discussed previously, for sodium. For the volatile group, indium added as the
it is possible to achieve highly reproducible source con- oxide is used as the internal standard because its vol-
Chap. 9 1 References
atility is in the middle of the group and indium is nor- metals industry. Spark spectrometers are often found
mally present at undetectable levels in such samples. on the floors of large foundrys, where they can be used
The emission signals from the involatile group (V, to give rapid results. Direct-reading spectrometers are
Ni, Co, Zr, Cr, Y, Sc, La, Nd, S r , Ba, Mo, B, Be, Ti, often used for multielement determinations. It is im-
Ca, Fe, Mg, Si, and Mn) reach their maximum intens- portant in the steel industry to determine the compo-
ities in the arc well after the alkali metals have distilled. sition of a melt rapidly enough that it can be adjusted
For determinations of these elements, samples are mixed before the melt is poured. Hence samples of the melt
with graphite, and the arc is burned until complete con- are taken, cooled, and sparked in the point-to-plane
sumption has occurred. Photographic measurements are method. Major constituents can be used as internal
begun after the alkali metals have distilled as indicated standards or an added element can be employed. Rou-
by a change in the color and visual appearance of the tinely, more than 20 elements are monitored. Although
arc. Relative standard deviations on the order of 2 to some companies have switched to ICP excitation, spark
5% have been achieved with careful attention to details. excitation is likely to remain important in the steel in-
dustry because of the need for minimal sample prepa-
Determination of Metals in Steels. The high- ration, the speed with which results can be obtained,
voltage spark has been extensively used in the ferrous and the high precision of the spark.
A grating spectrograph has an angular dispersion of as well as the arc and spark methods of this chapter.
1.33 x rad nm-' in the first order and a focal Explain and justify your choices. Consider sample
length of 1.5 m. preparation time, accuracy, and precision in your an-
(a) What is the reciprocal linear dispersion in the first swer.
order? (a) Determination of Mo in a steel sample.
(b) Find the focal plane separation in mm between the (b) Determination of Cu, V, Fe, W, and Si in a deposit
3099.9- and 3100.7- A components of the iron tri- from a boiler.
plet. (c) Determination of Ca in a hard-water sample.
(c) Repeat parts (a) and (b) for a spectrograph with (d) Determination of 12 elements in a river water sam-
an angular dispersion of 1.25 x rad nm-I ple.
and a focal length of 2.5 m. (e) Determination of Cr, Fe, Ni, and Co in a crude
What must the resolution of a spectrograph be to sep- oil sample.
arate the iron triplet components at 3099.9 and 3100.3
A? 9-6. Describe the advantages and disadvantages caused by
It is desired to analyze a very inhomogeneous solid selective volatilization in a dc arc discharge.
disk sample. Which excitation source (dc arc or high- 9-70 What advantages are provided by controlling the at-
voltage spark) would be better suited for this analysis? mosphere around dc arcs?
Explain and justify. 9-8. Compare high-voltage spark sources to dc arc sources
A dc arc method for the determination of Cu in a in terms of spectra produced and performance char-
powder sample is being developed. The 3247-A line acteristics.
has been chosen. Photographic detection is all that is 9-9. What are the advantages of a unidirectional high-volt-
available. An internal standard must be chosen. List age spark compared to a damped oscillatory discharge
the characteristics that a good internal standard for waveform?
this determination would have. 9-10. Discuss how one might measure the spectroscopic tem-
Choose an appropriate emission spectrochemicai anal- perature present in an observed region of a dc arc
ysis technique for the following determinations. Con- discharge. What additional considerations are neces-
sider all the flame and plasma methods of Chapter 8 sary with a high-voltage spark discharge?
The following are general references on the theory and prin- 1. P. J. M. Boumans, " h i t a t i o n of Spectra," in An-
ciples of emission spectroscopy (see also Chapters 7 and 8 alytical Emission Spectroscopy, vol. I . , pt. 11,E. L. Grove,
references). ed., Marcel Dekker, New York, 1972. An excellent dis-
272 Chap. 9 I Arc a n d Spark Emission Spectrometry
cussion of arc and spark sources from the mechanistic Resolved Radiation Processes in a High Voltage Spark
point of view. Discharge," Anal. Chem., 42, 61 (1970).
2. P. W. J. M. Boumans, Theory of Spectrochemical Ex- 10. J. P. Walters, "Source Parameters and Excitation in a
citation, Adam Hilger, Bristol, England, 1966. Spark Discharge," Appl. Spectrosc., 26, 17 (1972).
3. R . M. Barnes, ed., Emission Spectroscopy, Dowden, 11. C. E . Harvey, Spectrochemical Procedures, Applied Re-
Hutchinson & Ross, Stroudsburg, Pa., 1976. This book search Laboratories, 1950. An older book on dc arc
is a collection of the "classic" papers in the field of procedures, including semiquantitative methods.
emission spectrochemical analysis. 12. L. H . Ahrens and S. R . Taylor, Spectrochemical Anal-
4. R. D . Sacks, "Emission Spectroscopy," in Treatise on ysis, Addison-Wesley, Reading, Mass., 1961. A classic
Analytical Chemistry, 2nd ed., pt. 1, vol. 7, chap. 6, book on the dc arc for emission spectrochemical analysis.
I. M. Kolthoff and P. J. Elving, eds., Wiley-Interscience,
New York, 1981. An excellent treatment of the princi- The following deal with more specialized topics discussed in
ples and instrumentation for arc, spark, and plasma this chapter.
emission methods. 13. W. F. Meggers, C. H . Corliss, and B. F. Scribner, Tables
5. M. Slavin, Emission Spectrochemical Analysis, Wiley- of Spectral Line Intensities, pt. I , Arranged by Element;
Interscience, New York, 1971. pt. 11, Arranged by Wavelength, U.S. Government Print-
ing Office, Washington, D.C., 1961.
The following deal specifically with arc andlor spark excitation
for emission spectrochemical analysis. 14. J. Reader, C. H . Corliss, W. L. Wiese, and G . A. Mar-
tin, Wavelengths and Transition Probabilities for Atoms
6. A . Scheeline, "High Voltage Discharges: Diagnostics and Atomic Ions, NSDRS-NBS-68, National Bureau of
and Opportunities," Prog. Analy. At. Spectrosc., 7 , 21 Standards, Washington, D.C., 1980.
(1984). 15. D. A. Cremers, L. J. Radziemski, and T . R. Loree,
7. J. P. Walters, "The Formation and Growth of a Stabi- "Spectrochemical Analysis of Liquids Using the Laser
lized Spark Discharge," Appl. Spectrosc., 26, 323 (1972). Spark," Appl. Spectrosc., 38, 721 (1984).
An excellent summary. 16. Yu. P. Razier, Laser-Induced Discharge Phenomena,
8. J. P. Walters and H . V. Malmstadt, "Emission Char- Consultants Bureau, New York, 1977.
acteristics and Sensitivity in a High-Voltage Spark Dis- 17. R. W. B. Pearse and A. G . Gaydon, The Identification
charge," Anal. Chem., 37, 1477 (1965). A classic. of Molecular Spectra, 3rd ed., Chapman & Hall, Lon-
9. R. D. Sacks and J. P. Walters, "Short-Time, Spatially- don, 1963.
Atomic absorption spectro otometry (AAS) has be- of elements determined by flame AAS. In the 1960s,
come the most widely us single-element technique L9vov and later Woodriff, Massman, and West pi-
for the determination of metals. It is based on the ab- oneered the development of electrothermal atomizers
sorption of radiation by neutral, ground-state atoms for AAS. By the 1970s, commercial electrothermal at-
produced by an atomizer. omizers became available and accepted as alternative
The phenomenon of atomic absorption was noted atomization sources to flames in AAS. Electrothermal
by Wollaston and Fraunhofer and explained by Kirch- atomizers provide superior detection limits for many
hoff and Bunsen in the nineteenth century. They ob- elements. Since the early 1970s, A A instrumentation
served dark lines in the solar spectrum. In 1912, Mal- has been further refined by the development of more
inowski measured the absorbance of mercury vapor as intense line sources, by the introduction of instrumen-
a function of mercury concentration. Paschen in 1914 tation for background absorption correction, and by the
developed the hollow cathode lamp ( incorporation of microcomputers.
primary source for AAS, but did not use this source The typical instrumental configuration for an A A
with flames. Despite these early studies, the use of AAS spectrophotometer is shown in Figure 10-1. The source
was restricted to astrophysics applications until the 1950s, is usually an HCL (Figure 4-71, although an electrode-
ermination of mercury in laboratory less discharge lamp (EDL) is often used for some ele-
sh in Australia and Alkemade and ments (As, Se, and Te) where the radiant power output
Milatz in Holland proposed and demonstrated analyt- of HCLs is low. The lamps are usually modulated at a
AS, in which radiation from a line source, 50% duty cycle by operation from a pulsed power sup-
CL, is absorbed by atomic vapor in a flame. ply. (Mechanical choppers were used in early commer-
The technique of flame AAS became accepted in the cial instruments.) Normally, 1:1 imaging is used where
1960s after the introduction of the first commercial flame lens L1 focuses the radiation in the center of the at-
A A spectrophotometer in 1959. omizer and L2 refocuses the radiation on the mono-
New developments soon contributed to the en- chromator entrance slit. Thus the Fin of the lenses is
hanced growth of AAS. The nitrous oxide-acetylene usually one-half that of the monochromator, which has
flame, introduced by illis in 1965, extended the range an Fin in the range F16 to FIlO. Mirrors are also used
274 Chap. 10 I Atomic Absorption Spectrophotometry
Radiatton Slgnal
Readout
source processor
v I
=1 Monochromator
FIGURE 10-1 Single-beam atomic absorption spectrophotometer. Ra-
diation from a line source is focused on the atomic vapor produced by
the atomizer and then directed to the monochromator, where the atomic
line of interest is isolated. The attenuation of the source radiation by the
analyte atomic vapor is detected by the PMT. The signal is appropriately
processed to yield an absorbance readout signal.
for focusing in some commercial instruments. The at- ceptance as reliable atomizers for AAS in many situ-
omizer is typically a flame or an electrothermal device. ations.
A monochromator with a spectral bandpass of 0.1
to 2 nm is required to isolate the resonance line of the Flame Atomizers
element to be determined from lamp impurity and filler
gas lines, and from the atomizer background emission. Combustion flames are still the most popular atomi-
The signal processor converts the photoanodic current zation sources for AAS. In most commercial A A spec-
from the PMT into a voltage. Amplification and de- trometers, a premixed, chamber-type nebulizer burner
modulation circuitry extract the amplitude information system is employed. The nebulizer is usually based on
from the carrier waveform. Hardware or software log- the concentric, pneumatic design (Figure 7-4a). Neb-
arithmic conversion is used to provide direct absorbance ulizer parts are fabricated with robust metals (e.g., Pt-
readout on a digital meter, video display, video ter- Ir alloy, Ta, Pt) to enhance the chemical resistance to
minal, or printer. Normally, the noise equivalent band- acidic solutions and other corrosive mixtures. Variable-
pass of the electronics is adjustable by the user, who flow-rate nebulizers allow solution flow rates to be re-
selects a time constant or integration time. A continuum duced for solutions that would normally yield high ab-
source with a high-resolution echelle monochromator sorbance~.Invariably, a slot burner head (Figure 8-4)
can also be used in place of a line source and a con- is used to provide a long pathlength (e.g., 5 to 10 cm)
ventional monochromator. Continuum source A A and for absorption. Special burner heads with wider slots
several other options and refinements are discussed in or three slots are available to minimize clogging with
Section 10-3. solutions of high salt content.
This chapter begins by considering the flame and The most popular flames are air-C,H, and N,O-
electrothermal atomizers that are commonly used in C,H,. The nitrous oxide-acetylene flame provides higher
AAS. Specialized sample introduction and atomization atomization efficiencies (see Table 8-4 for a comparison
schemes are also reviewed. Next, signal and SIN expres- of pa values) and thus better detection limits for re-
sions are presented and used to illustrate how critical fractory elements, such as Si, Al. Sc, Ti, V, Zr, and
experimental variables affect performance character- the rare earths. In addition, the hotter and more re-
istics. A discussion of various instrumental options and ducing N,O-C,H, flame minimizes interference effects
configurations follows. The chapter concludes with a (e.g., solute vaporization interferences) for many ele-
description of performance characteristics, methodol- ments such as Mg and Ca. An air-H, flame or nitro-
ogy, and several common applications. gen-H, entrained air flame is sometimes used for easy-
to-atomize elements such as As and Se with absorption
lines in the far-UV region.
10-1 ATOMIZERS The burner control unit is designed for convenient
and safe burner operation. Flow controllers with flow
For AAS, the ideal atomizer would provide complete meters allow adjustment of the fuelloxidant ratio, which
atomization (pa = 1) of the element of interest irre- is critical for some elements. With Cr, for example, a
spective of the sample matrix. For the lowest dete'ction' fuel-rich air-C,H, flame yields the best atomization
limits. the atomic vapor should not be highly diluted efficiency and detection limit.
by the atomizer gas so that a large ground-state neutral Due to potential flashback problems, a N,O-C,H,
atom population is produced. Excitation of the analyte flame cannot be ignited directly. First an air-C,H, flame
and other species should be minimal so that analyte and is lighted and then the N,O flow is turned on and in-
background emission noise is small. Although not ideal, creased as the air flow is simultaneously decreased to
flame and electrothermal atomizers have gained ac- zero. The procedure is reversed before extinguishing
Sec. 70-1 1 Atomizers 2
the flame. Automatic gas flow modules implement this interference effects can be more severe in a furnace
procedure without operator assistance. The slot length compared to a flame. As we shall see, much of the
and width for a N20-C,H, burner head (typically, a 5- development of electrothermal devices, methodology,
cm slot) is different than for an air-C2H2 burner head and ancillary equipment has been aimed at eliminating
(typically, a 10-cm slot) to prevent flashback with the or minimizing these interferences.
higher-burning-velocity flame (see Section 8-2). Burner Commercial electrothermal atomizers are essen-
interlocks are often provided to prevent use of the in- tially small, electrically heated tubular furnaces, as shown
correct burner head. Many instruments provide push- in Figure 10-2a. The furnace is usually a graphite tube
button flame ignition through activation of a small starter that is 1 to 3 cm in length and 3 to 8 mm in diameter.
flame. An optical sensor is often used to monitor the The furnace assembly is usually mounted on transla-
flame. If the flame extinguishes accidentally, the gas tional stages in place of the nebulizer-burner, and its
flows are automatically shut down. position is adjusted so that the radiation from the line
The burner is mounted on translational stages to source is directed through the tube. A hole in the top
allow the flame to be positioned so that the focused of the tube allows typically 5 to 50 pL of sample, blank,
line source radiation passes through the middle of the or standard solution to be injected manually with a
flame at the desired burner height to maximize the ab- syringe or with an automatic injector. Each end of the
sorbance. Usually, the burner can be rotated to reduce furnace tube is connected to a high-current, program-
the pathlength for cases in which the absorbance is too mable power supply through water-cooled contacts. Be-
large. cause the electrical resistance of the furnace tube is
small (i.e., milliohms), the power supply must be ca-
lectrothermal Atomizers pable of delivering several hundred amperes at 10 to
12 v.
With electrothermal atomizers, a discrete sample is de- Typically, the furnace is heated in three stages, as
posited, and the atomizer is electrically heated to pro- illustrated in Figure 10-2b, in which the temperature of
duce a transient cloud of atomic vapor. The absorption the furnace is progressively increased by passing larger
of this atomic vapor is probed to determine the amount currents through the atomizer tube. The first step is the
of analyte in the sample. In 1961, L'vov demonstrated drying or desolvation step, in which a sufficient current
that electrothermal atomizers in an A A spectrometer causes the furnace temperature to be increased and
could yield absolute detection limits in the picogram maintained at about 11O"C. During this stage, the sol-
range and concentration detection limits over 100 times vent is evaporated, leaving a solid residue in the fur-
better than achieved with flame AA. This potential for nace. The temperature is chosen to evaporate the sol-
extremely low detection limits provided the impetus to vent as rapidly as possible without spattering.
understand, develop, and refine electrothermal atom- The second step is the ash step, in which the power
izers for routine analytical use. supply current is increased so that the furnace temper-
The reason for the superior detectability provided ature is raised, typically to 350-1200•‹C. During this
by electrothermal atomizers is readily seen. For a typ- stage, organic matter in the sample is ashed or con-
ical flame atomizer with a chamber nebulization system, verted to H 2 0 and CO,, and volatile inorganic com-
the solution flow rate is about 5 mL minP1 and the ponents are vaporized. The temperature should ideally
nebulization efficiency (E,,) is 5%. Thus the actual rate be set high enough to remove all volatile components
of sample delivery to the flame is about 4 pL sP1. For without loss of the analyte.
a typical flame with a 10-L min-I total gas flow rate The final step is the atomization step. The current
and a gas expansion factor of 10, the analyte atomic through the atomizer tube is increased so that the tube
vapor produced from the sample is diluted in 100 L temperature is 2000 to 3000•‹C. During this period the
min-l or 1.6 L sP1 of hot flame gases. Thus every sample is vaporized and atomized to produce atomic
4 FL of sample is diluted in 1.6 L of flame gases. vapor that is probed by the source. The atomic vapor
By contrast, the bulk of a 5-FL sample is atomized is produced rapidly and diffuses out of the observation
in about 1 s into a furnace volume of about 2 mL in an zone to produce a transient, peak-shaped response.
electrothermal atomizer. Thus the dilution factor is about The current (related to the tube temperature) and
1000 times less or the atomic number density of the the duration of each step are adjustable so that con-
analyte produced from a given solution is about 1000 ditions can be optimized for a given element, type of
times greater in a furnace atomizer compared to a flame. sample, and sample size. Typically, the whole sequence
This increased number density comes with a pen- takes 45 to 90 s. The dry and ash steps are typically 20
alty; the concentration of other gaseous components to 45 s in duration, while the atomize step is 3 to 10 s
from the sample matrix is correspondingly larger. Thus in length. Sometimes a fourth "clean" step involving a
276 Chap. 10 1 Atomic Absorption Spectrophotometry
From To
HCL monochromator
Programmable
power supply
higher current and tube temperature is employed after ployed as construction materials. Many other shapes
the atomize step to remove any remaining sample res- and configurations for electrothermal atomizers have
idue in the tube. Alternatively, the atomize step can be been used. These include carbon cups, carbon filaments
repeated before the next sample is run. and braids, tantalum strips, and tungsten loops. Today,
During operation, the furnace tube is continually carbon furnace tubes are coated with pyrolytic carbon,
bathed in an inert gas such as argon or nitrogen. The which prevents the sample from soaking into the graph-
inert gas protects the atomizer surface from oxidation. ite and reduces sample loss due to diffusion through
The gas also transports the analyte atoms from the sur- porous graphite walls. A small amount of methane can
face to the center of the atomizer and removes all gas- be added to the sheath gas to reduce the rate of dete-
eous substances produced during the operational se- rioration of the coating.
quence. The vapor atmosphere inside the furnace tube The heating characteristics of the furnace are crit-
can be more inert than in flames due to the absence of ical for obtaining reliable analytical results. Faster heat-
flame gases and combustion products; oxide formation ing rates during the atomization step cause the atomic
can be reduced and atomization efficienciescan be higher. population to build up rapidly before the atomic vapor
As indicated above, the furnace is usually made diffuses out of the observation zone. This creates a
of some form of carbon in a tube shape. The original higher instantaneous atomic concentration and larger
L'vov design and some commercial designs are shown absorbances. In current commercial furnaces, the heat-
in Figure 10-3. Metals with a high melting point, such ing rate is in excess of 1000•‹Cs-'.
as tantalum, tungsten, or platinum, have also been em- The control of the temperature during the various
See. 10-7 1 Atomizers
Insulator
Electrical Insulator
connector
Water out
@)
rtz window
rele
con
I
constant load
spring
I
cycle steps is critical for reproducibility. In early furnace views the outside of the tube. This allows the user to
designs, constant power was provided in each portion choose the atomization temperature. With feedback
of the cycle. Constant power does not ensure a constant control, the preselected temperature is rapidly achieved
temperature. Also, for low final atomization temper- and held for the duration of the atomization cycle.
atures, less power is applied with a resulting slower, Although the power to the furnace is stepped es-
nonlinear rise to the final temperature. Many modern sentially instantaneously to its selected value during the
furnace designs use an optical temperature sensor that atomization step, it takes a finite time for the furnace
278 Chap. 10 1 Atomic Absorption Spectrophotometry
Loop
is pipetted into a sampling device (Figure 10-5c). The Nydride Techniques. The flame A A detection
sampling device can be a platinum wire loop, a tantalum limit for some elements, such as As and Se, is only on
boat or cup, or a stainless steel rod with a cavity. Usu- the order of 1 pg m L p l and not adequate for deter-
ally, the sample is dried at about 100•‹Cto drive off the mining environmental levels of these species. Elements
solvent, which leaves a solid residue in the sampling such as As, Se, Sb, , and Pb form hydrides
device. A mechanical positioning apparatus then places in acidic solutions For example, the re-
the sampling device into the flame, where the sample action of borohydride with As(IP1) to form arsine is
is atomized. The source radiation beam probes the tran- described by
sient cloud of atomic vapor that is produced above the
sampling device. Because the sampling device is at a
lower temperature than the flame, this technique is used
primarily for volatile elements such as Pb, Cd, As, Bi,
and Zn. Note that this technique is similar to electro-
thermal atomization except that the energy to heat the
sample is provided by a flame instead of by electrical The hydrides formed can be swept out of solution into
energy. The use of the sampling cup technique has de- an atomizer as shown in Figure 10-6; a peak-shaped
creased since the advent and acceptance of electrother- response is obtained. When basic borohydride is added
mal atomizers. to an acidic solution, excess hydrogen is also produced
Samples can also be introduced into the flame in according to
the gaseous state. Of course, the element must be con-
verted to a chemical form that is volatile at or near
room temperature. Elements such as As and Se are
easily converted in solution to hydrides that can then
be swept out of solution into the atomizer. Reaction of There are two primary advantages to the hydride
some metal ions with appropriate complexing agents generation technique. First, the analyte is removed from
produces volatile metal complexes. For example, Cu, the sample matrix, which reduces the potential for in-
Fe, Be, and Cr form volatile trifluroacetylacetonates. terferences. Second, detection limits are improved to
The volatile species can be introduced directly into the the ngImL level or below because all the analyte in a 1
flame through the nebulizer tube or a port into the spray to 50-mL sample is introduced into the atomizer in a
chamber. Alternatively, the vapor phase sample can be few seconds (i.e., the atomic number density
directed to a quartz tube heated by a flame or even into peak is much greater than with nebulization tech-
the furnace tube of an electrothermal atomizer. Some niques).
researchers have used a G C to separate and volatilize Various atomizers have been used with hydride
metal containing species with a flame A A apparatus as generation. These include cool combustion flames (e.g.,
a metal specific detector. &air or Ar-H,-entrained air) and heated quartz tubes.
280 Chap. 10 I Atomic Absorption Spectrophotometry
TABLE 70- 7
Signal and readout equations for flame AAS
0% T signal"
Total reference signalh
Total sample samplec
Measured transmittance
Line:
@,, = WHCLT,,B
Continuum:
a,, = WHCLT,,,sB,
Reference signal"
Line:
E, = mGR(X)WHiZT,,BT,
Continuum:
E, = mGR(X)WHRT,,,sB,T,
Sample signalf General:
E = rnG 1; B,Y(i)T,R(h) dh
Line:
E, = rnGR(A)WHflT,,,BT,
Continuum:
E, = rnGR(h)WHRT,,sB,T,
Sample transmission factor
Spectral absorptance
-
- - -
aMeasured with source radiation blocked and flame on; background fluorescence assumed negligible herc and for
total sample and reference measurements; with source modulation and synchronous detection the mean valuc of E,,,
is 0.
bMeasured with the source on and blank aspirating into the flame; with source modulation and synchronous detection
the mean value of E,, is E,.
CMeasuredwith the source on and an analyte solution aspirating into the flame; analyte fluoresccnce assumcd negligible;
with source modulation and synchronous detcction, the mean value of E,, is E,.
dY(X) and To, account for the optical efficiency of components in thc monochromator and of the external optical
components between the source and monochromator. In equation 10-6 and subsequent equations for a linc source,
To, is assumed to be constant over the source line width. In cquation 10-7 and subsequent equations for a continuum
source, To, and B, are assumed constant ovcr the spectral bandpass of thc monochromator.
eIn equations 10-9 and 10-10. T, and R(X) are assunlcd to he constant over the source line width or spectral bandpass.
respectively.
'In equation 10-12, T, and R(X) are assumed to be constant over the line source width.
measured transmittance is the true transmittance or the T = 0 when the lamp radiation is blocked). All com-
ratio of the lamp signal passed by the sample to that mercial A A spectrometers provide direct absorbance
passed by the blank T' = T = E,IE,, and the true readout as one readout option and in some cases as the
Bbsorbance A is obtained from the measured transmit- only readout option. Here the sample and reference
tance (A = -log T). It is important to realize that E d , signals are converted to sample and reference readout
E,,, and EE can be measured before synchronous de- signals displayed in absorbance units (see equations
tection, and noise in these signals
- is still present at the 2-32 to 2-34). Thus the reference absorbance signal with
modulation frequency. the blank aspirating (A,) and the sample absorbance
With modern A A spectrometers, the 0% T signal with the sample aspirating (A,) are used to calculate
is not directly measured (i.e., the signal modifier elec- the analyte absorbance (A = A, - A,).
tronics are internally adjusted so that E,, = 0 V or The radiant power passed by the monochromator
282 Chap. 10 1 Atomic Absorption Spectrophotometry
exit slit (@,) and impingent on the PMT when the flame
is off (typically, 10-lo to 10-l2 W) is given by equation Equation 10-17 may be found in the literature in
10-5. It is a function of the lamp spectral radiance and various forms. With the appropriate conversion equa-
the collection and transmission characteristics of the tions (see Appendix F), A,, and B, can be substituted
monochromator and of the external optical compo- for f,.Note that (Ah,,,)' is just the maximum value
nents. Equations 10-6 and 10-7 are the specific expres- of the absorption line profile (S,) or S,,. From equation
sions for line and continuum sources. 7-29, S,, = (S,,)J(n, 0), where (S,,), is the maxi-
Part of the incident radiant power is absorbed mum vaiue of a purely Doppler broadened profile
when blank or analyte solutions are aspirated into the (equation 7-26) and S(a, 0) is the value of the Voigt
flame. The general equations and specific equations integral at the wavelength of maximum absorption.
that apply with line and continuum sources for the ref- For most A A determinations, the transition from
erence and sample signals are given by equations 10-8 the ground state to the first excited state is employed
to 10-13. (i.e., i = 0, j = 1). Under these conditions, n, = no
The attenuation of the lamp signal with sample or and equation 7-9 can be used for no in equation 10-17,
blank aspirating is a function of the sample and refer- with the result that
ence transmission factors (T, and T,). The reference
transmission factor is less than 1 due to absorption by
flame species or blank species (primarily molecular spe-
cies), to scattering by particles, and to reflection losses
caused by refractive index differences between air and where c, is the molar concentration of the analyte and
the flame. The sample transmission factor is controlled it is assumed that Z(T) = go. If we substitute for fun-
by the same factors plus the analyte absorptance (a) as damental constants, the result is
shown in equation 10-14.
With a proper blank, T,. is the same for the sample
and blank measurements so that T = EJE, = TJT,. =
Sec. 10-2 1 Signal and Noise Expressions 2
where A,, and AX,,, are expressed in cm. Equation and rz, and hence analyte concentration is nonlinear
10-19 indicates how the measured absorbance depends because a,. K n;l2 K cZ2. AS the spectral bandpass is
on nebulizer-flame parameters (F, E,, I , Q, ef), the decreased, the linearity of the calibration curve im-
analyte (A,,,, f,,), and a combination of flame and an- proves.
alyte parameters (AX,,). It predicts a linear relationship The readout expressions presented can be adapted
between the analyte concentration and measured ab- for other atomizers and sample introduction techniques.
sorbance. As discussed in more detail in Section 10-4, With electrothermal atomizers, the absorption signal is
calibration curves become nonlinear at high absorb- time dependent because n,, and thus k(A), varies with
ances partially because k(A) does vary slightly over the time as the finite amount of sample is atomized.
finite source profile.
With a continuum source, the absorption coeffi-
Signal-to-Noise Expressions
cient and hence analyte spectral absorptance varies over
the wavelength range passed by the monochromator
The expressions for the relative standard deviation (RSD)
(A, + s). If the width of the absorption line profile is
in absorbance (uA/A) developed in Section 5-5 apply
less than this wavelength range, the absorbance with a
continuum source (A,) is given by to AAS. General and specific equations for AAS are
summarized in Table 10-2. Equations 10-21 to 10-23
relate the standard deviation (SD) in absorbance (a,)
A, = -log T = -log ( I - a,) (10-20) and @,/A to the SD in the total sample signal (us,).
Equation 10-24 shows that crx, is due to shot and
flicker noise in the transmitted source signal and 0% T
noise as discussed previously (equation 5-46), and in
where a, is the absorptance with a continuum source addition, to analyte emission noise (a,). The lamp sig-
given by equation 7-51 and A, is the integral absorption. nal shot and signal flicker noise are calculated from
Because A, involves an integral of 1 eck("" over the
- equations 10-25 and 10-26, respectively. The noise with
absorption profile with k(X) dependent on A, a simple the source turned off and the blank aspirating, the
expression cannot be derived. In the limit of small A , a, 0% T noise (u,,), is seen from equation 10-27 to be due
is proportional to n, (see Section 7-5) and equations 10- to background emission noise (cr,,), dark current noise
17 to 10-19 apply if s is substituted for AX,,. For small (a,), and amplifier-readout noise (u,,) . (See equation
absorbances, A, is less than A, by the factor Ah,,/s. 5-55 for an expanded form of this equation.)
For higher absorbances, the relationship between A, The RSD in E, due to signal flicker noise is spec-
TABLE 70-2
Signal-to-noise expressions for AASa
"Es, Samplc flicker factor (dimensionless); t,, source flicker factor (dimensionlcss); 5:. atomizer transmission
factor (dimensionless); E,, analyte absorption flicker factor (dimensionless); u,, analyte emission noise (V).
284 Chap. 70 I Atomic Absorption Spectrophotometry
ified by the sample flicker factor 6,. For AAS, the signal Equations 10-30 and 10-31 are the final expres-
flicker noise arises from three sources, as specified by sions for uA/A that account for all noise sources. Al-
three flicker factors (equation 10-28). Here 5, is the though these equations are written in terms of the sig-
source flicker factor and 5, is the flame or atomizer nals observed with transmittance readout, they apply
transmission flicker factor. The latter flicker factor ac- equally to direct absorbance readout since readout res-
counts for the fluctuating transmission properties of the olution and noise added by logarithmic conversion are
flame or other atomizer. The third flicker factor (5, or usually negligible.
5): is unique to AAS and is denoted the analyte ab- Figure 10-8 shows plots of uA/A vs. A for five
sorption flicker factor. It can be written in two forms limiting cases in which one of the five noise terms in
that are related according to equation 10-29. Analyte equation 10-30 is dominant. Curves a to c show how
absorption flicker noise is due to fluctuations in the precision varies with A for the three standard limiting
measured analyte neutral ground state number density. cases discussed in Section 5-5 in which 0% T noise,
It is believed to be due to fluctuations in sample intro- signal shot noise, or signal flicker noise (source or at-
duction or in atomization characteristics that cause no omizer transmission noise) are limiting. Analyte emis-
to vary. Thus cA is the RSD in no for a given time sion noise can be dominant at higher analyte concen-
constant or integration time. If A 3~ no 3~ C ,[A is expected trations or absorbances when the analyte emission signal
to be independent of A while a, due to analyte ab- is significant. The relative contribution of analyte emis-
sorption flicker noise is proportional to A . Note that sion noise (a,) to u,/A increases faster than 0% T noise
5; specifies the RSD in E, due to analyte absorption (curve d) because a, increases with analyte concentra-
flicker noise. tion, whereas a,, is independent of A . Curve e shows
10.0
U% I noise
Analyte emission noise
that analyte absorption flicker noise is usually dominant ally limiting and (, = 2 - 3 x lop3. The absolute rms
at moderate absorbances and the precision is often con- noise due to signal shot or flicker noise decreases with
stant. At higher absorbances, the contribution of ana- increasing A since these noises are proportional to TIi2
lyte absorption flicker noise to the RSD decreases be- and T, respectively.
cause of negative deviation in the calibration curve, as The absolute noise in E , due to analyte absorption
illustrated in Figure 10-9. Although a,lA decreases, flicker noise is proportional to A and often becomes
the RSD for determining the analyte concentration does dominant at A = 0.1 to 0.2 and remains so up to A =
not improve. 1.0 to 1.5. Typically, $A = 0.5 to 1.0% for a 1-s inte-
Although the limiting noise sources in a given gration time. Over this region, a,lA is relatively con-
analysis depend on the element, source, type of flame, stant if the calibration curve is relatively linear or con-
optical throughput, and other conditions, some general stantly decreases if the calibration curve bends off. The
statements can be made. For most elements, u,lA is analyte absorption flicker factor at higher absorbances
determined by different types of noise in different ab- can be estimated from
sorbance regions. From the detection limit to about
A = 0.1 to 0.2, the precision constantly improves with
increasing A and signal shot noise, source flicker noise,
or flame transmission noise are limiting. Often the mag-
nitudes of these three types of noise are within a factor
el
of 2 to 3 of each other and typically and 5, equal 2 where m is slope of calibration curve at A and $A is the
to 10 x However, for elements with resonance value of [, at A = 0.2. A t an absorbance of 0.2, 5,
lines at wavelengths shorter than 230 nm (e.g., As, Zn, can be easily evaluated because it is often the limiting
Cd, Pb, Se, Te, Sb, Sn, Bi, and Ir), where flame ab- noise source and calibration curves are usually linear.
sorption is significant, flame transmission noise is usu- Often for absorbances greater than 1 to 1.5,
0% T noise and analyte emission noise become domi-
nant and precision decreases. This region is not very
useful for analyses because nonlinearity in the calibra-
tion curve is often severe at absorbances greater than
1.0. The relative 0% T noise (a,,lE,) is typically 2 to
200 x With modern AA spectrophotometers dark
current noise and amplifier-readout noise are usually
negligible compared to background emission noise.
Digital readout with a resolution of 0.0001 A.U. pre-
vents readout resolution from being the factor limiting
precision. Thus 0% T noise, if significant, is usually
limited by background emission noise, which is larger
and more important at lower absorbances if the back-
ground emission signal is large at the analysis wave-
length (e.g., near the O H radical bands or in the visible
region) or where the lamp signal (i,) is small. Analyte
emission noise is most likely to be the limiting noise
source for elements with X > 350 nm such as alkali
metals, rare earths, some alkaline earths, and group 3B
FIGURE 10-9 Effect of calibration curve non- metals because the analyte emission signal is larger.
linearity on analyte absorption flicker noise. At Both background and analyte emission noise can be due
an absorbance of 0.2, a 10% relative fluctuation to shot or flicker noise. Because the analyte and back-
in the neutral ground-state number density (no) ground emission signals are not carried by the modu-
causes a 10% RSD in A and 5, = 0.1. When the lation waveform (see Section 5-61, the contribution from
analyte concentration is doubled, A increases only llf flicker noise in these signals over the measurement
to 0.32 because of nonlinearity. The same RSD
bandpass centered at the modulation frequency is re-
in no causes a smaller RSD in A because the cal-
ibration slope is one-half of the initial value. Thus
duced.
= 0.062 at this absorbance. [From N. W. Bower
Equation 10-31 indicates that the source radiance
and J. D. Ingle, Jr., Anal. Chem., 49,575 (1977), [e.g., lamp current or type of lamp (EDL vs. HCL)]
copyright 1977, with permission of the American and spectral bandpass should be increased wch that i,
Chemical Society .] is large enough that signal shot noise, a,, and a, are
286 Chap. 10 I Atomic Absorption Spectrophotometry
negligible at the highest absorbances to be measured. not due to the flame but to blackbody emission from
Note that i, is proportional to @, as described by equa- the furnace walls and usually important only in the vis-
tions 10-5 to 10-7. In addition, K (i.e., Af) should be ible region. The analyte absorption flicker factor may
reduced so that signal shot noise is negligible. The source be higher since it accounts for reproducibility in sample
flicker factor may depend on lamp current. Generally, introduction and atomization conditions. The time con-
a,,, a,, and 5, are less for cooler flames and leaner stant with electrothermal atomization must be smaller
flames. Typical precision plots are shown in Figure (e.g., 0.1 s) than with flame atomization to prevent
10-10. They show the limiting regions and the domi- distortion of the transient absorption signals. This in-
nance of analyte absorption flicker noise at moderate creases the absolute rms noise due to most noise sources
absorbances. compared to flame measurements. The smaller size of
The a,/A expressions can easily be modified for the furnace or extra apertures used to restrict blackbody
other atomizers or sample introduction techniques. With emission can reduce @, and i, and increase the relative
electrothermal atomizers, background emission noise is signal shot noise.
Copper Molybdenum
1 .o 2.0
Absorbance Absorbance
FIGURE 10-10 Typical precision curves for two elements in AAS. Curve
a in both figures represents the experimental values of u,/A measured
with analyte solutions aspirating. In the curves labeled b and c in both
figures, the absorbance is varied by placing filters in the light beam. In
curve b the blank is aspirating while in curve c. the flame is off. Clearly,
analyte absorption noise is dominant at moderate absorbances because
the RSD is worse at the same value of i, (i.e., A) when analyte is present
and responsible for the absorption. Comparison of curves b and c illus-
trates that flame background emission noise is greater than dark current
and amplifier-readout noise. The rapid decrease in precision for Cu at
high concentrations is due to analyte emission noise. (From N. W. Bower.
Ph.D. thesis, Oregon State University. 1978. with permission of author.)
Sec. 10-3 1 Instrumentation
of the experimental conditions, absorbance data, and With a double-beam system, the reference beam
statistical values. Video terminals or monitors provide does not pass through a "reference atomizer" to correct
faster display of this information as well as of calibration for blank absorption. Also, there is no compensation
curves. Video displays are especially useful with elec- for drift and noise in the flame transmission character-
trothermal atomization since the time profile of the istics. The added optical components reduce the meas-
absorbance peak, and in some cases the temperature ured photon signal levels (reduction in i,), which re-
profile, can be viewed. Changes in peak shape or po- duces precision if signal shot noise or 0% T noise is
sition can be indicative of interference problems. dominant. Finally, a DB system does not compensate
The use of flow injection techniques (see Sections for changes in the profile of the lamp emission line,
6-6 and 10-1) with flame A A detection is expected to which can influence the measured absorbance.
increase as a means of fast automated sampling. Flow
injection techniques can be used for on-line sample di- Background Correction
lution, for standard additions, or for introduction of
releasing agents and/or ionization buffers. In addition to the analyte, both the flame itself and
concomitant species introduced into the flame (or an
Double-Beam Systems electrothermal atomizer) can absorb the source radia-
tion. Flame absorption below 220 nm is serious, as shown
Most manufacturers of A A spectrophotometers provide in Figure 10-12. However, a water blank can compen-
double-beam (DB) models based on a design similar to sate for this flame absorption.
that shown in Figure 10-11. The primary advantage of Although absorption of the HCL analyte line by
a DB system is that it continually compensates for source other atomized elements is rare in AAS, scattering and
drift and flicker noise (lowers 5,). With a single-beam background absorption by molecular species originating
(SB) system, source drift between the sample and blank from the sample can occur, particularly with electro-
measurements can cause error. With a double-beam thermal atomizers. This can cause measured absorb-
system, the warm-up time for source lamps is also re- ances, and hence predicted analyte concentrations, to
duced since measurements can be made before the lamp be too high unless a proper blank can be prepared [i.e.,
stabilizes. However, many SB spectrophotometers pro- the reference transmission factor (T,) is different for
vide two to six separate HCL sockets and power sup- the samples and standards]. Absorption by molecular
plies, at added expense, so that additional HCLs are species such as NaCl occurs over a broad wavelength
already warmed-up when they are needed. range, as shown in Figure 10-13. Several molecular ab-
sorption background correction schemes have been de-
Rotating
veloped and incorporated into A A spectrophotometers.
mirror Most of these schemes involve measurement of the total
absorbance (A,), which accounts for analyte and back-
ground absorption, and the background correction ab-
Monochromator
7 sorbance (A,,), which ideally accounts for background
absorption at the analyte wavelength. The corrected
analyte absorbance (A,) is then calculated from
100
90
80
70
8 60
5
.*
*
4d
.
$ 50
c
E 40
30
FIGURE 10-12 Absorption by sev-
eral flames in the short-wavelength 20
region. The analysis wavelengths for
several elements in this wavelength 10
region are indicated. (Adapted from
J. Ramirez-Munoz, Atomic-Absorp- 0
180 190 200 210 220 230 240
tion Spectroscopy and Analysis by
Wavelength, nm
Atomic-Absorption Flame Photome-
try, Elsevier, Amsterdam, 1968, with
permission of publisher.)
400
is totally compensated by the background correction
WAVELENGTH h.m) absorbance measurement. The background absorbance
correction is in error if this condition is not true. The
-13 Molecular absorption spectra of
contribution of analyte absorption to A,, is some frac-
NaCl, NaF, and NaI in an electrothermal atom-
izer [5 pL of 0.1% (wiv) solutions]. (From B. R.
tion of A, or A: = fA,. Ideally, f = A:, = 0, so that
Culver and T. Surles, Anal. Chem., 47,920 (1975), A, = A, if A, = Ah and the corrected absorbance
copyright 1975, with permission of the American calibration curve is identical to the normal calibration
Chemical Society .) curve in the absence of background absorption. I f f is
significantly different from zero, but independent of
analyte concentration, the calibration curve for cor-
fect the accuracy of the correction and the shape and rected absorbance has the same shape as that of the
slope of the corrected absorbance calibration curve. normal calibration curve but is lowered by a fixed frac-
The total measured absorbance is the sum of the true tion (1 - f ) , as illustrated in Figure 10-14a. The lower
analyte absorbance (A,) and t e background absorb- calibration slope degrades the detection limit. I f f in-
ance (A,), or creases at higher analyte concentrations, the shape of
the calibration curve is changed and roll-over can occur,
as illustrated in Figure 10-14b. Thus two very different
analyte concentrations can yield the same absorbance
The measurement of the background correction ab- (i.e., the analytical curve is double valued).
sorbance yields an absorbance with contributions from e will now consider the background correction
290 Chap. 10 1 Atomic Absorption Spectrophotometry
the background absorbance is constant over the spectral rection can be erroneous since the background absorp-
bandpass and thus equal to the background absorbance tion probed by each source is different.
at the HCL wavelength (A, = Ah). It is also assumed The D, background correction method can work
that analyte absorption of the continuum radiation is with background absorbances greater than 1.0 and has
insignificant (Ah = 0) because the analyte absorption little effect on the calibration curve. Detection limits
profile is very narrow compared to the spectral band- are degraded by at least a factor of 2; the degradation
pass over which continuum source radiation is passed depends on the wavelength and amount of background
(typically, 0.1 to 1 nm). absorbance. With a single-beam instrument, the con-
Despite the success of the D, lamp correction sys- tinuum source correction can be implemented manually
tem, there are some limitations. First, the correction by sequentially measuring the absorbances with the HCL
can degrade detection limits because the radiance of D, lamp and D, lamp in position.
or H, lamps is weak at longer wavelengths (see Figure
4-5). The signal processing electronics usually require Zeeman Background Correction. Zeeman
that the reference signals for both sources be the same. atomic absorption (ZAA) correction is available with
Because Q,, for the D, lamp is often less than for the some commercial instruments. Here a magnetic field
analyte HCL, the lamp current for the analyte HCL is splits normally degenerate spectral lines into compo-
often reduced below its normal value. This degrades nents with different polarization characteristics. Ele-
precision if signal shot noise or 0% T noise becomes mental and background absorption are distinguished
significant. A tungsten lamp can be used in place of the based on the magnetic and polarization characteristics
D, lamp at higher wavelengths to reduce the problem. of the lines.
We could envisage a system in which the HCL and There are several ways to implement Z A A cor-
continuum lamp signals were separately processed in rection. Either the source or the atomizer can be placed
way that did not require reduction of the HCL intensity. in a strong magnetic field. The magnetic field can be
The precision of the background correction measure- parallel or perpendicular to the light path and generated
ment could still be poorer. with a permanent magnetic or an electromagnet (ac or
The correction can be in error if the background dc) .
absorption over the spectral bandpass is nonuniform or To illustrate the principle of the Z A A technique,
structured. For example, it is possible for atomic ab- we will first consider analyte-shifted Z A A correction,
sorption by lines from other elements within the spectral where the atomizer is placed in a transverse dc magnetic
bandpass to increase the background absorbance meas- field as shown in Figure 10-16. We will assume that the
ured even though this type of absorption does not affect normal Zeeman splitting applies (see Figure 7-12b).
the analyte absorbance measured with the HCL. Thus The analyte absorption lines are split into n and a com-
the method can overcorrect or undercorrect for back- ponents that are polarized parallel and perpendicular
ground absorption. to the magnetic field, respectively. The wavelength of
The alignment of two sources so that their beams the n component remains at the zero field wavelength
pass exactly through the same volume element of at- while the u components are shifted away from this cen-
omizer is tedious. If the lamps are misaligned, the cor- tral wavelength, as shown in Figure 10-17a. With one
Permanent
magnet
PMT, signal
Source Rotating
polarizer
Atomizer
Monochromator
orientation of the rotating polarizer, the lamp emission omizers require more space and thus a larger and more
n component is transmitted by the polarizer and is ab- powerful magnet to obtain the same field strength across
sorbed by the unshifted T component of the analyte. the atomization region. Longitudinal fields can be used
Thus A, or both analyte and background absorption at with electrothermal atomizers. In this case the absorp-
the normal analyte wavelength are measured. In the tion IT component is polarized along the optical axis and
second cycle, the source o component is transmitted does not absorb the n component of the source radia-
and A,, is measured. The lamp u component is not tion which is in another plane. Hence a linear polarizer
absorbed by the unshifted analyte n component because is not needed to isolate the source o component.
its polarization is different. The shifted analyte absorp- With source-shifted Z A A correction, the source is
tion a components (a- and u - ) are at different wave- placed in a dc magnetic field and a polarization mod-
lengths and ideally absorb no source radiation. as il- ulator (an electro-optic- or magneto-optic-based vari-
lustrated in Figure 10-17a. Thus under ideal circumstances able-phase retardation plate, see Section 3-3) is used to
only background absorption at the analyte wavelength rotate the plane of polarization of the two components
is measured by a source radiation (A, = At. and of the source beam back and forth by 90". The beam is
A: = 0 ) . Subtraction of the two absorbances measured then passed through a static linear polarizer. This method
in each cycle yields the corrected absorbance. of alternating the polarization component transmitted
If the atomizer is placed in an ac transverse mag- is often superior to the use of a rotating polarizer (e.g.,
netic field, a static linear polarizer is oriented between two linear polarizers mounted on a rotating wheel with
the atomizer and monochromator to transmit only the their polarization axes oriented 90" with respect to each
source u components for all measurements. As shown other). With a rotating polarizer, it can be difficult to
in Figure 10-17b, A, is measured when the field is off align both polarization components along the same op-
because the o absorption components are unshifted. tical axis. Often, additional optical components are
When the field is on, A,, is measured with ideally no required with a rotating polarizer to compensate for
analyte absorption by the shifted absorption u com- differences in the intensity of the two polarization
ponents. components.
With either ac or dc techniques, it is less conve- As illustrated in Figure 10-17c for source-shifted
nient to place a flame between the magnetic poles than Z A A , A, is measured when the T component has the
it is to place an electrothermal atomizer. Flame at- proper plane of rotation to be transmitted by the static
See. 10-3 l Instrumentation
linear polarizer. The unshifted source T component is noise source because the source intensity is reduced due
absorbed by the T component of the analyte and by to transmission losses in polarizer components and the
molecular species. When the plane of rotation is altered use of only a fraction of the field-free source intensity
90•‹, the wavelength-shifted source u components are for each measurement.
transmitted and ideally absorbed only by the back- The calibration slope for the corrected absorbance
ground species. may be reduced a factor of 2 to 3 over uncorrected
Source Zeeman correction can be used with any measurements (i.e., worse detection limits) and cali-
kind of atomizer. However, special magnetically sta- bration curves can exhibit greater nonlinearity and even
bilized lamps are required. Conventional HCLs cannot roll-over. With typical field strengths of about 1 tesla
be used. Often, EDLs or glow discharge lamps are em- (kilogauss), the shifts in polarization components are 1
ployed. to 10 m ~ The . wings of the a component profiles are
Many elements produce the anomalous Zeeman not totally shifted away from the T-component profiles,
effect (see Figure 7-12c), and this complicates Z A A and some analyte absorption can be seen in the meas-
correction because there are several u and IT compo- urement of A,, (i.e., A: f 0). This effect is worse with
nents. Both polarization components are split from the anomalous splitting, significant hyperfine structure, and
central wavelength with the u components undergoing isotope effects. Absorption in the line wings causes A,
wider splitting. With hyperfine structure and isotope to be less than A, and roll-over at absorbances of 0.5
effects, the u and profiles can be broadened consid- to 1.0 for most elements. The calibration sensitivity is
erably. The shift of the IT component away from the also reduced for cases where the IT component is shifted
field free central wavelength reduces the analyte ab- significantly from the zero-field wavelength. Detection
sorption coefficient (the calibration sensitivity) and is limits can be better or worse than those obtained with-
dependent on the field strength. This shift is not a prob- out background correction, depending on the element,
lem with analyte-shifted Z A A correction with an ac the limiting noise sources, and the configuration for
field since A, is measured when the field is zero. correction used.
There are several advantages of Zeeman tech-
niques over the continuum-source-based correction Pulsed Hollow Cathode Lamp Background
method. With a single source, normal source intensities Correction. The newest commercially available back-
can be used at all wavelengths without the alignment ground correction system is the Smith-Hieftje (S-H)
or differential drift problems experienced with two system. As shown in Figure 10-18a, a HCL is pulsed at
sources. In general, background correction is more ac- a normal current (6 to 20 mA) for 9.7 ms and a high
curate (A, = Ah) if the background absorption is struc- current (100 to 500 mA) for 0.3 ms during each period
tured over the spectral bandpass because the back- of the modulation cycle. During the low-current part
ground absorbance is measured exactly at or very near of the waveform, A, due to analyte and background
the wavelength that the analyte absorbance is meas- absorption is measured. When the short-duration high-
ured. current pulse is passed through the HCL, A,, is sam-
The Zeeman technique involves effectively a dou- pled. The WCL must be capable of stable operation
ble-beam in-time measurement. The difference in two when pulsed to high currents.
absorption measurements is continually calculated at The emission line from the HCL is considerably
the frequency that the polarization components are broadened and somewhat self-reversed at the higher
modulated. Thus the contributions from lif source flicker current, as shown in Figure 10-1%. Because the average
noise and atomizer transmission flicker noise are re- absorption coefficient over the line profile is much less,
duced, which can improve detection limits if these noise the measured analyte absorbance (A:) for a given an-
sources are limiting. alyte concentration is much smaller, as seen in Figure
There are some disadvantages of Z A A correction 10-18c. If the background absorbance is constant over
systems compared to conventional A A measurements the broadened emission profile, A, = Ah. The instru-
without correction. First, the implementation is rather ment automatically calculates the absorbance difference
complex and expensive. With analyte-shifted Zeeman (A, = Ar - Abc).
systems, the background absorption bands can also ex- As with Z A A correction systems, one HCL is
perience Zeeman shifts and reduce the accuracy of the employed so that the technique works at all wave-
correction (A; f A,). For optimized correction, the lengths, alignment is simple, and there is compensation
magnetic field strength should be adjusted for each ele- for source flicker noise and atomizer transmission flicker
ment, and this is not possible with some commercial noise. Additional optical components, such as beam
systems. The SIN and detection limit can be worse if splitter or polarizers, which reduce the measured source
signal shot noise or 0% T noise become the limiting intensity and can degrade the SIN are not required.
294 Chap. 10 I Atomic Absorption Spectrophotometry
+ t l -
+I
111~
2
Aa
h A,-A;
!
High current
+E
S: A',
P
Low current d
I I
Time
Am Concentration
(a 1 (b 1 (c1
FIGURE 10-18 Principles of the S-H background correction system. In
(a) the current waveform used to drive the HCL is shown. The time
between low-current and high-current segments ( t ) is only 4.5 ms, indi-
cating the temporal proximity of background correction. In (b) the ideal-
ized emission profiles of the HCL at low and high lamp current are shown.
In (c) the analyte absorbance calibration curves are shown for the low
and high current measurements. [Adapted from S. B. Smith, J r . , and
G . M. Hieftje. Appl. Spectrosc.. 37, 419 (1983). with permission of the
Society for Applied Spectroscopy.]
Overall the technique is relatively simple, inexpensive, bine the beams from several HCLs (i.e., wavelengths)
and can be used with any atomizer. into one beam.
The background correction accuracy is usually very A second problem is the limited linear dynamic
good (A; = A,), even for background absorbances range of AAS (approximately two to three orders of
greater than 2, unless the background absorbance over magnitude for a given analytical line). With one sample
the broadened emission profile (about 0.01 A) is struc- dilution, it may be difficult to adjust the concentration
tured. The primary disadvantage is a reduced calibra- of all elements to be determined to be both above the
tion sensitivity (10 to 8096, depending on the element) detection limit and in the linear range.
due to analyte absorption during the high-current pe- The optimum conditions (e.g., type of flame, fuel1
riod (A: 4 0). Roll-over in calibration curves at very oxidant ratio, burner position) for single-element de-
high absorbances can occur but is less severe than with terminations do vary. Thus a third problem is the need
Z A A systems. Detection limits are typically worse by to choose compromise conditions in which the detection
a factor of 2, and in some cases, five to six times worse limit and calibration curve linearity for some elements
than those obtained without correction. is degraded. This may be more critical with electro-
thermal atomizers, where optimum ashing and atomi-
Multielement Spectrophotometers zation temperatures vary considerably.
Of the three basic atomic spectrometric techniques, A A Line Source Multielement AA. Several designs
is the least well suited for simultaneous multielement for simultaneous multielement A A spectrometers with
measurements. With multiple single-element line sources, line sources for 2 to 10 elements have been proposed
all beams must be combined into one beam to pass which differ in the way the spectral signals at different
through the atomizer along the same optical axis. This wavelengths are resolved and detected. In one general
can be accomplished with beam splitters or with fiber approach, a direct-reader arrangement with a separate
optics. However, with very many line sources the align- slit and PMT for each element is employed. Alterna-
ment is tedious and the reduced intensity (50% loss per tively, the radiation transmitted through several exit
beam splitter) can ultimately reduce precision (increase slits in the focal plane can be directed to one PMT. The
the relative importance of signal shot noise and 0% T absorption signals from each element are distinguished
noise). Also, a separate power supply is needed for each with a time multiplex approach. Here the HCL for each
single-element HCL. These problems can be somewhat element is pulsed on at a separate time or the slits are
alleviated by the use of multielement HCLs or by the sequentially blocked and unblocked with a rotating wheel
use of a continuum source as discussed later in this with apertures. The data acquisition electronics are syn-
section. A grating can also be used in reverse to com- chronized to the lamp pulsing or slit openings.
Sec. 10-3 l Instrumentation 2
Some researchers have used a multichannel de- diode arrays, there is a trade-off between wavelength
tector (e.g., diode array) in the focal plane. There is coverage and resolution because of the size of the active
some compromise between wavelength coverage and detector element or array.
resolution due to the limited size of these detectors.
The limited dynamic range of these detectors also causes
problems. Ideally, the reference signal for each detector Continuum Source Multielernent AA. The most
element corresponding to a specific analyte line should successful design of a simultaneous multielement A A
be adjusted to be near the saturation value of the array spectrophotometer is based on a continuum source with
elements for the integration time used. In reality this an echelle polychromator. A high-intensity Xe arc lamp
is difficult because the source radiance of different HCLs is used as the single source, which alleviates the ex-
varies greatly. For weaker analysis lines, the relative pense, complexity, and source alignment problem ex-
amplifier-readout noise (a,,/E,) is greater because E, perienced with using multiple line sources. An echelle
is less and precision degrades. Different integration times monochromator or polychromator is required to obtain
can be used to alleviate this problem and different anal- a spectral bandpass on the order of the atomic absorp-
ysis lines can be used to extend the dynamic range. tion line width. For simultaneous analysis, a bank of
The only commercial simultaneous multielement PMTs are positioned behind appropriate exit slits in the
instruments available are dual-channel spectrophotom- two-dimensional focal plane of an echelle polychro-
eters. The radiation from two HCLs is passed through mator to intercept the transmitted source radiation cor-
the flame and then directed to a beam splitter. The responding to the analysis wavelength for each element.
transmitted and reflected components are directed to Sequential systems are based on a scanning echelle
two different monochromators. The radiant power iso- monochromator.
lated by each monochromator is detected and processed There are some disadvantages of continuum source
separately. Such instruments increase sample through- A A compared to conventional line source AAS. Be-
out by a factor of 2 and allow internal standard methods cause the spectral bandpass is still greater than HCL
to be used. Dual-line or continuum source background line profile widths (s > AX,,, see Section 10-2), the
absorption correction schemes can be implemented. In initial calibration slopes are less, and nonlinearity at
addition, two analysis lines can be used for one element higher absorbances is more pronounced. Characteristic
to yield an overall greater dynamic range. concentrations (i.e., the concentration yielding an ab-
Computerized commercial A A spectrophotome- sorbance of 0.0044) are typically two to four times poorer.
ters are available that allow automated sequential mul- The measured lamp radiant power (a,) can be
tielement analysis. The operator initially chooses new less because of a smaller lamp radiance over the ab-
or previously stored values for experimental conditions sorption line width compared to a line source or because
such as wavelength, slit width, lamp position, lamp cur- of lower optical throughput. The latter reason is be-
rent, integration time, and gas flow rates for each ele- lieved to be dominant. Typically, smaller slit widths and
ment. Once initiated, the instrument readies itself for particularly slit heights (100 pm) are used with echelle
determination of the first element and performs the monochromators and polychromators. Also, the source
requested absorption measurements. This sequence of radiation for a given wavelength is spread over many
setting up and measurement is automatically reported orders and only one order is intercepted by the exit slit.
in a sequential manner for each element (up to six ele- The lower measured radiant power and hence lower
ments) without operator assistance. reference signal (i,) increase the blank noise as signal
Although these instruments do not provide the shot noise or 0% T noise become more significant. The
high throughput and reduced sample consumption of a combined effect of reduced calibration sensitivity and
true simultaneous multielement instrument, they speed increased blank noise degrades detectability. Above 250
up the analysis and allow optimization of conditions for nm, detection limits are 2 to 5 times poorer and can be
each element. Normally, the burner position or type of 10 to 20 times poorer for analysis wavelengths shorter
burner head cannot be optimized automatically in com- than 250 nm.
mercial spectrophotometers. Most continuum source echelle A A spectropho-
Several researchers have developed sequential tometers are modified for wavelength modulation by
multielement A A spectrometers based on rapid-scan or placing a refractor plate behind the entrance slit (see
slew-scan monochromators or on an image dissector Figure 4-34). This provides background absorption cor-
tube (IDT) (see Section 4-4). In the latter case, the IDT rection and discrimination against lif source and at-
is placed in the focal plane of a spectrograph to select omizer transmission noise. The wavelength modulation
different wavelengths sequentially. As with the use of depth is typically about 0.1 A,and background correc-
296 Chap. 10 I Atomic Absorption Spectrophotometry
Emission
tion is accurate if the background absorption is constant
and unstructured over this wavelength interval.
With continuum source A A , it is also possible to
make measurements in the wings of the absorption pro-
file, where the absorption coefficient is less. This can
extend the dynamic range of the calibration curve to
higher concentrations.
Linearity
0 10 20 30 40 50 60 70 80 90 100
Cd conc., pg m ~ - '
FIGURE 10-20 Calibration curves for cadmium
for different HCL currents. The increase in non-
linearity with increasing HCL current is more se-
vere than for most elements. [Adapted with per-
mission from B. J. Russell, J. P. Shelton, and A.
Walsh, Spectrochim. Acta, 8 , 317 (1957), Per-
gamon Press, Ltd., Oxford.]
crease the initial calibration sensitivity, and reduce non- FIGURE 10-21 Effect of slit width on absorb-
linearity. Of course, unabsorbed lines at the analyte ance in AAS. In (a) the spectrum of a HCL lamp
wavelength or closer than the resolution of the mon- in the region near the intense analyte resonance
ochromator (0.2 to 0.005 nm) cannot be eliminated. line is shown. Several other less intense nonab-
sorbed lines are nearby. In (b) the absorbance
Also, for many HCLs, the line emission is superimposed
for a given analyte solution is seen to decrease if
on top of a weak continuum whose relative contribution the spectral bandpass is larger than 0.5 nm be-
(typically, 0.1 to 2%) can be minimized, but not elim- cause the nonabsorbed lines are passed by the
inated, by reducing the spectral bandpass. The effect monochromator. With elements such as Ce, Co,
of nonabsorbed lines and thc slit width on the amount Cd, Ir, La, Nb, Os, Pd, Sb, Tb, Ni, U, W, and
of stray radiation can depend on lamp current, which Rb, the proximity of nonabsorbed lines to the
can alter the relative intensities of the source analyte HCL resonance line makes adjustment of the
and unabsorbed lines. spectral bandpass more critical.
2 Chap. 90 / Atomic Absorption Spectrophofometry
1%. Equation 10-37 is often used as the fitting equation rates. Atomic absorption spectroplioton~etryis less sus-
for implementing curve correction. Alternatively. curve ccptible to tluctuations in excitation conditions (i.e.,
fitting based on a polynomial equation may be more the Roltzmann factor is lcss critical). This fact is often
accurate. overemphasized since temperature Iluctuations also af-
One assumption used in the derivation of Beer's fect the atomization efficiency (see Section 8-5).
law is that the concentration of the absorbers is uniform.
Thus a third major cause of nonlinearity in A A cali-
bration curves is variation of thc analytc atom density
and even the absorption profile both laterally and ver- 'The evCtluatlonol the accurxy of AA me,i\urcmcnt\ I \
tically over the cross section of the sourcc beam viewed \1rn11~1r
in m m y recpcch to t h t di\cu\vxl for A E (Scc-
by the monochromator. In the extreme, some of the tion 8-6) Under proper condition\, accuracy can be
sampled radiation could bypass the atomization zone hmted by rdnctom error dnd n o w to the range 0 5 to
(c.g., the flame) entirely. Also, the pathlengths of dif- 5 % Hlghe\t accuracy 15 o b t , ~ ~ n eby
d brxketrng s'lrnple
ferent rays on and off the optical axis can vary. Both mcd\urcn~ent\wlth mea~urenientsot \tdndards of ,lp-
effects cause negative deviations and arc minimized by proxiniately the \ame analyte concentration
using higher Fin optics and focusing the beam to a small
spot in the center of the Slame or electrothermal at-
omizer. It would appear that collimation of the incident
beam would be advantageous. This is uncommon be-
cause of the increased optical complexity.
As with other atomic spectrometric techniques (see
Section 7-l), nonlinearity occurs if the sample Slow ratc
(F)or the overall atomization efficiency (F,,) varies with
analytc concentration. With proper prccalihration, in-
strumental nonlinearity is usually negligible. With mod-
ulation, analyte cmission does not directly cause non-
linearity. However, high analyte cmission or hlackhody
radiation from clectrothcrnial atomizers can saturate
the PMT or electronics and yield erroncous rcsults.
Analyte tluoresccnce could cause nonlinearity at high
analyte concentrations, but the cffcct is usually negii-
gible because of quenching in the Slames used for AA
and the relatively high Fin collection optics.
recision
Systematic errors can arise due to analyte and solutions of high salt content, and are more likely with
spectral interferences, as discussed in Section 7-2. An- electrothermal atomization. Background absorption
alyte interferences, except for excitation interferences, correction is necessary in these cases.
affect flame AA and flame AE equally. When flame
AA is compared to AE in hotter, more inert, and elec- Characteristic Concentrations and Detection
tron-rich plasmas, AA is subject to worse volatilization, Limits
dissociation, and ionization interferences. Lateral dif-
fusion interferences are unique to flame AA with N,O- Typical characteristic concentrations and detection lim-
C2H2flames and are illustrated in Figure 10-22. its (DL) for flame and electrothermal AAS are listed
Generally, analyte interference effects are worse in Table 10-3. Clearly, most metals can be determined
and less well characterized with electrothermal atom- at concentrations lower than 1 kg mL-l. Electrother-
izers than with flames. The chemical form of the analyte mal atomizers are not hot enough to determine a few
after solvent evaporation (e.g., PbO or PbSO,) as well high-melting-point metals, such as W, Ta, and Nb.
as the salts in the matrix can greatly influence the atom- The initial calibration slope (m) is related to the
ization efficiency. The profile and the time of appear- characteristic concentration (m,) by m = 0.0044/mA
ance of the analyte absorption peak are also affected, (see Section 6-3). Thus mA can be used to compare the
as illustrated in Figure 10-23. Vaporization interfer- calibration slopes of different elements or of two atom-
ences can be worse than with flames because the salt ization techniques for the same element. It can be seen
particles formed are much larger than those formed in Table 10-3 that characteristic concentrations are usu-
from a nebulized droplet. Dissociation interferences can ally a factor of 10 to 500 better with electrothermal
be worse because of the higher number density of all atomization except for a few refractory elements (Al,
species. However, sample transport and ionization in- Mo) .
terferences are less important with furnaces. If the DL is defined as the concentration yielding
In AE, interference due to lines of other elements an absorbance equal to twice the blank standard de-
near or at the analyte emission wavelength are the ma- viation (sbk)in absorbance units (A.U.), DL = 2sbklm.
jor cause of spectral interference. By contrast, for AAS In terms of m,, DL = sb,mAl(2.2 x lop3). Thus
atomic spectral interferences due to overlap of atomic DL = m, if s,, = 2.2 x l o p 3 A.U. and DL =
absorption profiles is possible (e.g., between the 403.307- 0.1 m, if sbk = 2.2 x l o p 4 A.U. For flame AA, most
nm Mn line and the 403.298-nm Ga line), but rare. DLs reported in Table 10-3 are about h of m,, implying
However, spectral interference due to scattering and that a typical blank noise is about 2 x l o p 4 A.U. For
molecular absorption can be serious. Such interferences elements with resonance lines at wavelengths shorter
are expected for analytes with lines in the UV, with than 250 nm, such as Zn and Cd, the blank noise is
1 I 1 I , I I 1 I I I/ I 1 I I I
0 5 10" 0 5 10 15 0 5 10 15 20
TIME I N SECONDS
Typical detection limits and characteristic concentrations for flame and electrothermal AAS
Flame l~lcc~rothcrir~~~l"
generally greater due to higher flame transmission noise. Reported flame DLs by different manufacturers
Blank noise is also greater for elements with weak HCL are quite similar, probably becaus; of comparable neb-
emission lines, such as As, Se, and Te, because the ulizer-burner systems and sources. Flame A A DLs are
relative signal shot noise is greater. For those dements, less affected by the sample matrix than are flame or
EDLs are usually used in place of HCLs to overcome plasma AE D I s . In flame AA, the blank noise is limited
this effect. EDLs have 10 to 50 timcs the radiance of by sourcc and flamc transmission flicker noise or lamp
HCLs for some analysis lines. shot noise, which arc affected only slightly by the sam-
The flame DLs reported are obtained after ex- ple matrix. Also, depression of the calibration slope by
tensive optimization of instrumental parameters and with matrix components is rarely greater than a factor of 2
pure sample solutions, new lamps, and 3 to 10-s inte- for flame AAS.
gration times. With older lamps or smaller integration Detection limits achieved with electrothermal at-
times, the blank noise and hence DLs can be two or omizers are more variable between manufacturers be-
three times worse. cause they depend on the atomizer design and atomi-
Sec. 10-5 1 Methodology and Applications 301
zation conditions. Often, absolute or mass DLs and m, introduction or atomization technique may be required.
values are reported (e.g., in pg). Thus m, and the D L Separation procedures are sometimes necessary if in-
depend on the sample volume used and do not neces- terferences cause serious systematic errors that cannot
sarily improve linearly with sample volume. For most be eliminated by simpler means.
elements, the DLs with electrothermal atomization are
a factor of 10 to 500 better than achieved with flames, Flame Atomization
due to the larger calibration curve slopes (higher no).
The blank noise is usually slightly larger with electro- With flame A A most analyses are first attempted with
thermal atomization because smaller time constants are external standards. Selection of optimum experimental
employed to avoid distortion of the peak-shaped sig- conditions for flame A A is usually straightforward. All
nals. Detection limits in complex samples can be de- manufacturers provide a "cookbook" with suggested
graded significantly, due to reduced atomization effi- experimental conditions for most elements. The type
ciency (lower m) or higher atomizer transmission noise of flame, the type of source, the fuel-to-oxidant ratio,
in the case of significant background absorption. the analysis wavelength, the lamp current, the spectral
In general, electrothermal A A DLs are best for bandpass, and the observation height are specified. Fine
more volatile elements and often below 1 pg. For less adjustments may be necessary. Usually, the burner po-
volatile elements such as Al, Ba, alkaline earth metals, sition and the fuel-to-oxidant ratio are optimized by
and rare earth metals, p, is lower in a furnace and DLs maximizing the absorbance while aspirating a solution
are in the range 1 to 100 pg. Characteristic concentra- yielding an absorbance of about 0.5. This normally op-
tions and DLs are also higher for elements that form timizes precision at moderate analyte concentrations.
nonvolatile carbides (Ba, B, Ca, Mo, W, V, and Zr) For measurements near or at the DL, the ratio of the
with the carbon of the atomizer or carbon in the or- calibration sensitivity to the blank noise should be op-
ganics in the sample matrix. The furnace tube can be timized. Generally, the smallest HCL current and slit
coated with one of these carbide-forming metals to re- width that ensure that measurements are not signal shot
duce the loss in calibration sensitivity for other carbide- noise or background emission noise limited are best
forming elements. (i.e., a, is independent of Q,). Higher lamp currents
Some workers have demonstrated that furnaces or slit widths can cause lower calibration sensitivity and
are hot enough to excite emission from some of the nonlinearity for some elements, as discussed in Section
more easily excited elements (Al, Cr, Cu, Mn) with 10-4.
DLs near 1 pg, as with AAS. However, DLs for other Flame A A cookbooks generally list a number of
elements are significantly worse than with electro- possible lines that can be used for analysis. An example
thermal AA. is shown in Table 10-4 for Cu. Normally, the absorption
In general, flame A A DLs (Table 10-3) are better line corresponding to a transition from the ground state
than flame A E DLs (Table 8-6) except for easy-to- to the first excited state is used because it has the largest
excite elements (alkali metals). The Boltzmann factor transition probability (oscillator strength) and hence the
in flames is very small for elements with only UV res- smallest characteristic concentration or the largest cal-
onance lines. For many elements, flame A A DLs (Table ibration curve slope. Also, the corresponding transition
10-3) and ICP emission DLs (Table 8-8) are within an line from the source usually has the highest radiance.
order of magnitude but significantly worse than elec- Other less sensitive lines may yield better linearity at
trothermal A A DLs. The ICP does provide the best high concentrations, eliminate the need for excessive
DLs for many refractory elements and the multielement
capabilities and dynamic range of plasma emission tech-
niques must be considered. For As, Bi, Se, and Te, TABLE 70-4
hydride techniques provide the best DLs, while the cold Analysis lines for copper
vapor method yields the best D L for Hg.
dilution, increase the SIN due to less hackground cmis- Electrotlicrmal A A is useful when low DLs or
sion noise or flame transn~issionnoise, or rcducc sus- special sample considerations arc critical. Particularly
ceptibility to background ahsorption intcrfcrcnccs. for unalyte concentrations ncar or below 10 kg I , - ~ ' ,
Potential interferences for flamc AA arc identi- the precision o f flame AA is poor or the analyte con-
fied in A A cookbooks. With flamc AA, spcctr,'1 I ~nter-
' centration is below the DL. With elcclrothermal A A
fcrc~lccsare usually minimal. Background absorption these low analyte concentrations can often he measured
correction is sometimes required for elements with rcs- directly without time-consuming preconcentration steps.
onance lines below 300 nm or for solutions of high salt The ability to analyze small sample sizcs down to
content. I PI, is usef~ilfor sample-limited situations, such as in
Several options :Ire available i f analytc interfer- some clinical assays. Although special techniques are
ences that depress or enliancc the analytc absorbance available for introduction of submilliliter samples into
arc observed. First, tlic type of flame and flame con- flames (see Section 10-2), the flame AA DL in terms
ditions can be adjustcd to minimize analyte intcrfer- of concentration is generally worse than obtained with
cnces. For some clcnlents ionization buffers and re- continuous aspiration of 1 inL or more of sample.
leasing agents arc required; these can also improve Time-consuming sample preparation steps can be
linearity of calibration curves. Test solutions can be bypassed by placing viscous solutions (oil, blood) or
made to contain 1 mg inL ' of Cs or another casy to solid samples (plastics, paper) directly into the furnace
ionize element to prevent ionization interferences for atomizer. However, solid samples do present special
other elements, such as alkali metals, alkaline earth problems. It must be possible for the atomizer to melt
nletals, rare earth metals, Al, Au, aild Si. Lanthanum the sample to prevent blockage of the light beam and
at the 1-mg m L ' level is effective for minimizing solute to vaporize all the sample to prevent memory effccts.
vaporization intcrfcrcnces for sonic elcments, such as For the highest accuracy, solid stanclards similar to the
alkaline earth mctals, and is also a good ionization buffer.. sample in composition should be used. The atomization
Usually, the foregoing steps are adequate with characteristics for a givcn mass of analyte can be quite
flames. For special cases, standard addition procedures different if it is cieposited in solution form rather than
can be employed. They should be used with care be- in a solid matrix. Although it is casy to weigh small
cause many A A calibration curvcs exhibit subtle non- solid samples accurately, it is more difficult to transfer
linearity at moderate absorbanccs. Alternatively, ma- them faithfully to the atomizer. Also, sample liomo-
trix-matched standards or separation of interferences geneity is of concern for small sample sizcs.
may he required. For some elements the flame atom- For elcments such as Hg, Se, Sb, IJi, and Sn,
ization efficiency varies slightly when samplc solutions electrothermal atomizers provide hettcr AA DLs than
contain more than 1 % acid. Therefore, st;nidards should do flames. However, the cold vapor method is best for
be prepared in the same concentration of acid. Internal low Hg concentrations because the DL is lower and
standard methods arc rarely used and quitc inconve- losses of volatile mercury in the ashing step are avoided.
nient except with dual-channel instl-umcnts. They can Similarly, hydridc techniques arc usually preferred for
improve precision slightly if slow variations in atomi- low concentrations of As, Sc. Sb, IUi, and Sn because
zation conditions are limiting and can compensate for of better detection limits. These alternative AA tcch-
analyte interferences if an internal standard can be found niques do require sample volumes of 1 mL or more.
that suffers the same interference effects as the analyte. As for flame AA, cookhook settings are available
as an initial guide for setting the time interval and tem-
perature for the drying, ashing, and atomization steps.
Selection and optimization of the analysis wavelength,
Elcctrotlierm,~lA A cornplcmcntc 1l;lrne AA , ~ n dI \ m a t type of lamp, lamp current, and spectral handpass are
su~tahlewhen llarne AA DLs are ~n,rdequ,lteor \miplc very similar to flame AA. Fine tuning of thc drying,
volume\ are llmted Elcctrotherm,rl ,1tom17cr,'ire more ashing, and utomization conditions is quitc critical to
cxpeinivc than nebulltcr-bu~ner\,,~lthoughopcratmg achieve maximum atomization efficiency and minimal
cmt\ due to fl,nne gnw, 'ire le\s C o m p m A to fl,nne interference effects because the optimum settings are
AA mc,i\i~rements,clcctrothermdl A A ha\ wor\e p e - very matrix dependent.
cl\ion lor nie,isurcmcnt\ well '~bovcthe detection I~mit. Many potential interferences arc identified in
\utter\ from greater mterfercnce elfect\, require\ more electrothermal AA cookbooks. The major spectral in-
opcrnto~\k~ll,and h,r3 lower cample tliroughput Ad- terference is absorption by molecular species, which arc
dit~onaltime-con\uniiiig d~lutlon\tep\ r n q c~ctudllybe covolatilized with the analyte. This type of interference
~cquiredfoi come \drnplcc bec,lu\e t9t the cxccllcnt de- can be detected by nuking measurements with and
tcctdt7lllty without background correction. Altho~ighbackground
Sec. 70-5I Methodology and Applications 303
by acid digestion. Abundant eleanents ( N a , K , C a , Mg, b e aspirated into a flame. Usually, organomctallic
E;e) and many trace transition metals can usually be standards soluble in the solvent a r e used because t h e
determined directly with flame AA. However, to de- atomization efficiency, and hence the calibration slopc,
termine many metals which a r c present at conccntra- arc clifferenl between aqueous solutions of inorganic
tions lower than 10 p g E ', particularly in seawater, mctal starrdards and organic solutions of organic metal
electrother~nalA A andlor preconccntration steps (ion ackground absorption correction is often
exchange, solvent extraction) are necessary. nccdctl. These considerations makc the standard ad-
T h e use of A A S in the metallurgical and mining dition technique il useful tool and also apply to situa-
industries is common for analysis of metals, alloys, geo- tions whcrc metals a r c extracted from aqueous solutions
chemical samples, and electroplating solutions. Flame into a n organic solvent before aspiration into a flame.
AAS is adequate for determination of most clcrncnts For biological and clinical samples, it is critical to
at concentrations of 0.001%,(w/w) o r above in solid sam- remove t h e organic matrix hy digestion o r some other
ples after dissolution by acid digestion o r fusion. For mcthod before analysis. For example, serum protein
lower concentrations, clcctrothermal A A S may be rc- clogs burners and causes high background absorbance
quired. and residues with electrothermal atomizers. Deprotcin-
Analysis of petroleum products presents special ization with trichloroacctic acid is co
sample preparation difficulties. Oils and gasoliaic are simpler if detectability is adequate.
often diluted by a factor of 2 t o 10 with suitable solvcnts ical samples, a flame ionization buffer is nccdcd because
(heptane, xylcnc, methyl isobutyl kctone) so they can oi thc high content of Na and K.
Q
10-1. Multielement hollow cathode lamps are sometinics chromator factors, transducer factors, ;rmpli-
used in atomic absorption (AA) spcctropholo~lletry. ficr-readout factors) come into play and render
For example, a brass hollow cathode produces lines ;~bsolutemeasurcnicnts almost impossible.
of Zn and C h in addition to filler gas lines. Zinc is
often determined in AA using its 213.9-nm resonance The determinatiou of Pb ill a brass sample was carried
linc. In a I~rasshollow cathode. the Cu lines at 216.5 out with AA spcctrophotomctry and the mcthod of
and 217.9 nrn arc fairly near the Zn line. standard additions. 'H'llc original sample was diluted
(a) If the only rnonochrornator availahlc has a spec- to 50 mL after dissolution. This solution was intro-
tral bandpass of 5 nm, what effect will the Cu duced into the AA spectrometer and a transmittance
lines from the hollow cathode have o n AA cali- of 0.380 was obtained. To the original solution 20 yL
bration curves if (i) there is no Cu in the stantl- of a 10.0 kg mL-I 1'13 standard was then added. The
ards, and (ii) the standards contain a constant transmitt:rncc of this solution was 0.263. Find the
amount of Cu? concentration of Ph in the original sample. What as-
(b) If another nionochromator is available with a re- sumption has heen made in order to use n single
ciprocal linear dispersion of 2 nm 111m ' and slits standard addition'? Is there any built-in check on this
v;~riablcfrom 50 yrn to 2 mm, what slit width assumption in the procedure outlined above'?
should be used to avoid interference lrorn the CLI Compare and contrast the continuum sourcc, Zcc-
lines? man, and Smith-klicftjc methods of backgl-o11ndcor-
10-2. Is the following statcn~cnttroc orland false? (;riticiz~ rection in AA spcctrophotornetry. Consider in your
or dqfi.izcl the statement, hut begin with a definite djscussion thc range of applicability (i.e., number of
opinion. Use equations, diagrams, words 01-any other elements), the accuracy of the cor-rection, the case
information in the answc~-. of use, thc cupcnsc, the possibility of errors hcing
Flame atomic ahsorption (AA) spcctr-ometry introduced, and any other factors you dcenl impor-
would be a much better absolute technique than tant.
flame emission spectrometry. That is, you can Discuss the general applicability of AA spcctropho-
carry out determinations without standards. The tometry Cor multielcmcnt determinations. C'o~nparc
fraction absol-bed with a linc sowce a , is a sim- A A to TCP emission spcctrornct~-yfor the simulta-
ple function of concentration in AA since at low neous determination of multiple elements.
-
concentrations ( t x , k,,,l = Kn,, = K ' c ) , where
k,,, is the atomic absorption coefficient, 1 is the
In the c d y &rys ol AA 5pect1ophotomet1y v,uiou5
clmm a4 to ~nstrun~entA errors m d interterences were
pathlength, K and K' are constants, tz,, is the m d c It was often stated, f o ~example, thnt AAS I \
ground-state number density. and c is the so- le\\ su\ceptrhle to change5 in tlanc tcrnpcr'iturc thm
lution concentration. 'The constants K and M' A b S h e c , ~ ~ t the
w Ialgc ground-\t,~tcp o p u l , ~ t ~ o n
are known from theory. In flame emission, me,lwretl I n AAS chnngcs only sl~ghtlyw~thtcni-
however, all the instrumental factors (mono- per'rture, whle the cxc~tcd-shtcpopur,ktlon ch,mgc\
Chap. 10 1 References 305
drastically with temperature. For example, consider Why do hydride techniques for As and Se and the
1000 atoms in the ground state and 1 atom in the cold vapor technique for Hg yield better A A detec-
excited state. If the temperature changes such that 2 tion limits than conventional flame AAS with solu-
atoms are now in the excited state and 999 in the tion nebulization?
ground state, the AES signal should change by loo%, Why do different flames yield different A A calibra-
while the AAS signal will change by only 0.1%. Dis- tion slopes for a given element?
cuss why this argument is oversimplified and what Discuss the limiting noise sources in flame A A at
temperature-dependent factors it neglects. In actual low, moderate, and high absorbances. How is your
practice, are AAS and AES much different in their answer affected by the selection of the slit width and
behavior with temperature? HCL current in a given determination?
-7. One reason that combination flame emission/atomic Why is background absorption correction more im-
absorption instruments have not been great successes portant with electrothermal atomization than with
is that the monochromator requirements in the two flame atomization?
techniques differ substantially.
Why can the monochromator slit width affect the
(a) Discuss the function of the monochromator in the linearity of A A calibration curves? Consider both line
two techniques. What role does the monochro- and continuum sources in your answer.
mator play in reducing interferences in the two Consider the flame A A determination of Cu at 324.7
methods? nm. The solution flow rate is 6.5 mL min-I. e, =
(b) Your laboratory is considering the purchase of a 0.02. ef = 12, Q = 9.3 L min-', Z ( T ) = go = 2,
monochromator. Under consideration are an ex- g, = 4, Ah,, = 4 x 10-"m, 1 = 10 cm, and
pensive, high-resolution, variable-slit-width sys- A,, = 0.95 x lo8 s s l . The test solution is a 5.0-kg
tem (smallest spectral bandpass = 0.5 A) and a mL-I aqueous Cu solution. The flame temperature
less expensive, moderate-resolution, fixed-slit- is 2800 K. Find the absorption factor with a line source,
width system (spectral bandpass = 10 A). Which a,, and the absorbance A,.
system would you choose if your laboratory was Why is the ash step very critical with electrothermal
only going to do atomic absorption measure- atomizers?
ments? Why?
Contrast analyte-shifted and source-shifted Zeeman
10-8. What are the advantages and disadvantages of dou- A A correction techniques.
ble-beam Systems relative to single-beam Systems in 10-18. The flame A A detection limit for Cu is found to be
AAS? independent of the spectral bandpass over the band-
10-9. Discuss the major factors involved in the decision to pass range 0.2 to 1.0 nm. What noise sources can be
use flame or electrothermal atomization in AAS. limiting precision?
REFERENCES
The following books, chapters, articles, and reviews are gen- plication to Chemical Analysis, 2nd ed., Masson, Paris,
era1 references devoted entirely or substantially to atomic 1980.
absorption spectrophotometry. 8. C . Th. J. Alkemade and R. Hermann, Fundamentals of
W. J. Price, Spectrochemical Analysis by Atomic Ab- Analytical Flame Spectroscopy, Wiley, New York, 1979.
sorption, Heyden, London, 1979. 9. B. V. L'vov, Atomic Absorption Spectrochemical Anal-
L. Ebdon, A n Introduction to Atomic Absorption Spec- ysis, American Elsevier, New York, 1970.
troscopy: A Self-Teaching Approach, Heyden. London, 10. J. W. Robinson, Atomic Absorption Spectroscopy, 2nd
1982. ed., Marcel Dekker, New York, 1975.
"Atomic Absorption Spectroscopy, Past. Present. and 11. B. Welz, Atomic Absorption Spectroscopy, Verlag
Future," Spectrochim. Acta, 36B, (5 and 7 ) (1981). Two Chemie, Deerfield Beach, Fla., 1976.
issues devoted to AA. 12. A. Asha, CRC Handbook of Atomic Absorption Anal-
J. W. Robinson, "Atomic Absorption Spectroscopy," ysis, CRC Press, Boca Raton, Fla., 1984.
in Treatise on Analytical Chemistry, 2nd ed., vol. 1, pt. 13. A. Walsh, "Atomic Absorption Spectroscopy-Stag-
I, chap. 8, P. J. Elving and I. M. Kolthoff. eds., Wiley- nant or Pregnant," Anal. Chem., 49, 1269A (1977).
Interscience, New York, 1981. 14. J. E . Cantle, Atomic Absorption Spectrometry, Elsevier,
J. C. Van Loon, Analytical Atomic Absorption Spec- Amsterdam, 1982.
troscopy: Selected Methods, Academic Press, New York, 15. M. Salvin, Atomic Absorption Spectroscopy, 2nd ed.,
1980. Wiley, New York, 1979.
W. Slavin, "Atomic Absorption Spectroscopy-The 16. K. C. Thompson and R. J. Reynolds, Atomic Absorp-
Present, Past and Future," Anal. Chem., 54,685A (1982). tion, Fluorescence, and Flame Emission Spectroscopy:
M. Pinta, Atomic Absorption Spectrometry, Vol. 2. A p - A Practical Approach, 2nd ed., Wiley, New York, 1978.
Chap. 70 I Atomic Absorpt~onSpectrophotometry
17. G. F. Kirkbright and M. Sargenf, Atomic Absorpfion 24. J. J. Cotera and H. L. Kahn, "Background Chrrection
und Fluorc,.scrncc, Spc~c~tro.sc~op~~,
Academic Prcss, Lon- in AAS," Am. Lub., 193, Novcmbcr (1982).
don. 1974. 25. S. D. Brown, "Zeeman Effect-Based I3ackground Cor-
18. T. .I. Vickers. "Atomic Fluorescence and Atornic Ab- rection in Atomic Absorption Spectron~etry,"A n d .
sorption Spectroscopy," in Physical Mrt1locl.r in Modern Chem., 49, 1269A (1977).
C'l7crnicul Andysis, 'T'. Kuwana (ed.), vol. I , Academic 26. R. Stephens, "Zceman Modulated Atomic Absorption
Press, New York, 1978, p. 192. Spectroscopy," CRC C'rit. Rev. A n d . Chem., 9 , 167
( I 98O).
Thc following references are concerned \pec~l~cally
w ~ t helec-
trothelma1 AA. Articles on more specific topics in AA are given below.
19. S. R. Koirtyohann and M. L. Kaiser, "Furnace Atomic M. L. Parsons and P. M. McElfresh, "Comparison of
Absorption-A Method Approaching Maturity," Anal. Theoretical and Experimental Limits of Detection in
Clwn., 5 4 , 1115A (1982). Atomic Absorption Spectrometry Using Air-Acetylene
20. W. Slavin, C'ruphilc fitrnacc~ Source Hook, I'crkin- and Nitrous Oxide-Acetylene Flan~es," Aypl. Spcc-
Elmer Corp., Ridgcfield, Conn., 1984. tro.rc., 26, 972 (1 972).
21. R. E . Sturgeon, "Factors Affecting Atomization and M. L. Parsons, B. W. Smith, and P. M. McElfresh, "On
Measul-ement in Graphite Furnace Atomic Absorption the Selection of Analysis Lines in Atomic Absorption
Spectrometry ," Anal. Chem., 49, 1225A ( 1977). Spectromctry," Appl. Sprctrosc., 27, 971 (1973).
22. C. W. Fuller, Electrothermal Atomizution ,for Atomic N. W. Bower and J. D. Ingle, Jr., "Precision of Flame
Ahsorp~ionSpcc~lrometry,Royal Society of Chemistry, Atornic Absorption Measurements of Arsenic, Cad-
London. 1 977. miurn, Calcium, Cobalt, Iron, Magnesium, Molybde-
num, Nickel, and Zinc," Anal. Chrm., 49, 574 (1977).
Background absorption correction is discussed in the follow- T. C. O'Havcr, "Continuurn-Source Atomic-Absorp-
ing articles. tion Spectromctry: Past, Present, and Future Pros-
23. L. DeGalan and M. T. C. de 1,oos-Vollebregt, "Roll- pects," Analysl, 109, 21 1 (1984).
Over of Analytical Cilrvcc in Atomic A bsorption Spec- T. Nakahara, "Applications of Hydride Generation
trometry Arising from Background Correction with Pulsed Techniques in Atomic Absorption, Atomic Fluores-
Hollow-Cathode I,amps," Spcctrochiun. Actu., 34B, 1011 ccnce, and Plasma Atomic Emission Spectroscopy," Prog.
( 1984). Anal. At. Spcctrosc., 6, 163 (1983).
ic Fluorescence
Spectrometry
Atomic fluorescence (AF) involves the emission of pho- fraction of the excitation radiant power. This small an-
tons from an atomic vapor that has been excited by alyte fluorescence signal must be distinguished from
photon absorption. The phenomenon of atomic fluo- various background signals and noise.
rescence was studied in the late nineteenth and early In Section 11-1 we consider the types of fluores-
twentieth centuries by physicists who observed fluores- cence that can be observed. Next, instrumental design
cence from several elements (e.g., Na, Hg, Cd, T1) in and various instrumental configurations and compo-
heated cells and flames. Starting in 1956. Alkemade nents are discussed. The ideal characteristics of instru-
used A F to study physical and chemical processes in mental components are compared to the characteristics
flames and in 1962 suggested the use of A F in chemical of components actually used. This is followed by a re-
analysis. In 1964, Winefordner and Vickers proposed view of signal and noise expressions that provides in-
and demonstrated A F flame spectrometry as a new an- sight into how instrumental variables and atomic prop-
alytical method. After 1964, atomic fluorescence spec- erties affect the signals observed and the linearity and
trometry (AFS) was extensively studied and refined, precision of AFS. The phenomenon of saturated fluo-
particularly by Winefordner's group in the United States rescence is also presented. The chapter concludes by
and West's group in England. summarizing performance characteristics and applica-
Much of the research on AFS as an analytical tions of A F measurements.
technique has centered around the development of suit-
able intense sources. We have already seen (Section 7-5)
that at low analyte concentrations, a very small fraction 11-1 TYPES OF FLUORESCENCE
of the excitation source radiant power is absorbed. A
fraction of this absorbed radiant power is converted to There are five basic types of fluorescence: resonance
fluorescence as determined by the fluorescence power fluorescence, direct-line fluorescence, stepwise-line flu-
yield (Y). Finally. a small fraction of the fluorescence orescence, sensitized fluorescence, and multiphoton
emitted over 4 7 sr is actually collected and detected. fluorescence. These are summarized in Figure 11-1. Some
Thus, in many experiments. the fluorescence radiant specific examples are given in Table 11-1.
power detected may be to l o 9 or an even smaller In resonance fluorescence (Figure 11-la and b),
Chap, I I I Atomic Fluorescence Spectrometry
TABLE 7 7- 7 the same upper and lower levels arc involved in the
Examples of fluorescence types excitation-deexcitation process so that the absorption
and emission wavelengths are the same. Direct-line flu-
orescence (Figure 11-lc to f) results when the same
upper level is involved in excitation or emission, whereas
Kcwnancc uorescence (Figure 11-lg to l), differ-
Exciteti-state rcwnancc ent upper levels are involved in the excitation-deacti-
Stokes dircct-linc vation process. Both direct-line and stepwise-line flu-
Excitccl-\talc Stokes orescence are grouped in the category of nonresonance
dirccl-line
Anti-Stokcs dil-ccl-line
uorescence. Sensitized fluorescence (Figure 1 1-lm) oc-
Stohcs stepwise-linc curs when a donor species excited by photon absorption
Excitccl-state Stoke\ transfers energy to an acceptor atom which radiatively
stepwise-linc deactivates. Finally, multiphoton fluorescence (Figure
T l ~ e r ~ n a l la\sistetl
y anti- 11-111)results whcn two o r more photons promote an
Stokes stepwisc-line
atom to an excited state which then emits a photon.
Sec. 11-2 / Instrumentation 309
The intermediate levels may be virtual or real, and the with a mechanical chopper provides modulated exci-
excitation photon energies may be the same or differ- tation radiation.
ent. The selection rules for multiphoton excitation are Excitation optics focus the source radiation onto
different than for single-photon absorption (see Section the desired volume element in the atomizer. With con-
7-3). ventional sources, the collection efficiency of the ex-
Different types of resonance, direct-line, and step- citation optics is critical: the solid angle of collection
wise-line fluorescence can be distinguished. If the upper should be large. This consideration does not apply to
and lower states involved are both excited states, the lasers because of the directionality of the light beam.
fluorescence process is said to be excited-state fluores- A mirror can be placed on the opposite side of the
cence (Figure 11-lb, d , f, h, j , and 1). The lower state atomizer from the source to reflect transmitted exci-
is usually a low-lying metastable level that is thermally tation radiation back onto the atomizer.
populated. The adjectives Stokes and anti-Stokes de- The emission collection-detection system may be
note that the excitation wavelength is less than or greater dispersive (monochromator-based) or nondispersive
than the emission wavelength, respectively. If the ex- (filters or no filter). In either case, wavelength selection
citation process involves radiative excitation followed is usually required to isolate the analyte fluorescence
by further thermal excitation, the process is thermally line from background signals (emission and fluores-
assisted (Figure 11-lk and 1). cence from the atomizer and concomitants). For non-
Most analytical work has involved normal reso- resonance fluorescence, the wavelength selector allows
nance fluorescence because the transition probabilities the analyte fluorescence signal to be distinguished from
and the source radiances with conventional line sources the source scattering signal; for resonance fluorescence,
are often the greatest. Sensitized and multiphoton flu- such a distinction based on wavelength is not possible.
orescence have found little analytical use because of the A portion of the total fluorescence excited within the
low fluorescence radiances produced. With nonreso- atomizer is collected and imaged on the monochro-
nance fluorescence, wavelength selection can be used mator entrance slit or on an aperture preceding the
to resolve scattering from fluorescence because the ex- detector in nondispersive systems. The goal is to max-
citation and emission wavelengths are different. imize the solid angle of collection and the volume ele-
ment viewed. With dispersive systems, the Fin of the
collection optics should be matched to the Fin of the
11-2 INSTRUMENTATION monochromator. To minimize the collection of scat-
tered source radiation, the fluorescence is usually viewed
Figure 11-2 shows a block diagram of a typical single- at an angle of 90" with respect to the excitation axis.
element A F spectrometer or photometer. The atomizer The angle between the excitation and emission axes can
can be a flame, plasma, electrothermal device, or a be any angle except 180•‹,as this angle would result in
special-purpose atomizer (e.g., heated quartz cell). The direct viewing of the excitation source radiation. Ad-
excitation source can be a conventional line or contin- ditional baffles restrict the collection and detection of
uum source, a laser, or an ICP. Normally. a pulsed source radiation that is reflected or scattered from ex-
source (e.g., pulsed HCL or laser) or a continuous source ternal components.
Becauw ol the low hght levels to be detected, '1 Iluo~e\ccnce\1gn'11, can '~ctu;lllydccrc'l\e I he C d, LII,
I'MT 14 norn~,~lly the detector nlthough multichannel Hg, m d TI d~\ch,~rge l,nnp\ C I I the ~ mo\t u x l u l , m d
\oI~d-stdcdetector\ ' I I I ~ iin~tgcdl\\cctor tube\ (ID'I \) tl'mc AE dctect~o.lI~mrt\(DL,\) below I ng mL. ' h'ivc
hclvc been u\ed wlth cpcct1ogr~1ph5In some cclse\ ,I heen o b t , ~ ~ n e d
colur hhnd P M r (re\pon\e trom 180 to 320 nm) h,i\ Microw,~vc-poweredbDL5 h m e been the mo\t
been u\ed w~thout,I f~ltcr popular AT exc~t,itlon t o u ~ c c shcc,lu\e thcy povrde
The \rgn,ll proce\\or typlc,llly ~ncludc\e~thcr ,I ~el,rt~vcly clcm \pcctr,i nnd h~gherr;ldi,uice\ tli'ln most
syn~hronous, ~ cn m p l ~ f ~ e(lock-~n)
r ~f the soulce duty other conventiond \ouicc\ I h c ~ ru5c tor A F h,1\ dc-
cycle is SO%, or 1' gated ~ntcgr'rtol (boxcar ~ntcgr~itot) cre<r\ed in the lr~\tICW Y C ~ T \ Although bDL\ c m bc
lor pul\ed, low-duty cycle wulcec C'onvcnt~ondlr e d - prepxed to1 mo\t clement\, they ,ire ,~v,nLiblchorn
out dev1ce4 include drg~t,tlmeters, .ln,llog recortiers, coinrnelcr'~lsourue\ for only :I few elements T h c ~ prep- r
and pr~ntcr\. 'iratlon m d o p t l r n ~ ~ ~ i t 15
l o \till
n tedious m d d~tficult
7 he rel&lon\lnp between the ~ u c ~ i w r efluore\-
d Hollow c,lthode l ~ ~ n i phelve c long hccn u\ed m d
cencc 41gnal m d the ~n\trument,~l m d c~n;llytep a r m - \tdl u\cd '15 cxcitatton soi1ree5 even though the11
e t e n 1s d~\cussed111 d e t d In the next \ectlon along w ~ t h r,~d~ance\ ,ire u\u,illy lowc~t h m tlio\e prov~dcclhy other
SIN con\~deration\.To provde a Iramework to d~\cu<\ wurccc They <Ireavaldblc for most element\ 1or AbS,
the dlllcrent ~nstrumcnt~il coinponcnt\ dnd d o g m In IICL\ arc now u\ccl exclus~velyin 'I puked mode 111
this \ectlon, we fir41 note th,~t'it low m d y t e concen- wh~chthe p t k current I \ typ~c~illy 0 I to 1 A , and the
tr,ttion\ (see Scct~on7-5) the fluorc\cencc \~gn,llde- duty cycle I \ I to 10% Pul5cd opcr,ltlon mci e,l\cs the
tected 15 proport~on'll to the source r a d ~ m c c( ~ g n o r ~ n g peak r'idlance 10 to 100 times rel,itrvc to the \teady-
5aturdt1on eftect\ for now), the andytc number demity, \tntc r a d ~ , ~ n cwc~ t hdc current operatlon At high cur-
the tluorcscencc qudi~tumefflc~cncy,,lnd thc enll\\lon lent\, the lrnc prol~leI \ \rgn~f~c,lntly blo,~dencdso that
collect~oncffic~ency I'hus, t o maxlmuc the detected the I a h n t power h s o r b c d does not Increnw In d ~ ~ e c t
fluorc\cencc s~gnal,thew qu,~nt~tle\ 4hould be optln1- proportion to the ~ntcgr,ltedlme r a d ~ m c c Roo5tcd . out-
ized put ( h g h ~ n t e m ~ t yIIC'L\
) c m p~ovldeup to 10 time\
MC~x~mt7lng the tluorc\cencc \~gnaldoes not tell the r,ld~ance\ol norm'll \h~cldedIIC'L\
the whole \tory because detcct,~b~lity I\ ultlm~tely'I SIN A h ~ g hr,rd~:lnce contlnuum wurce would be idccll
p o b l c ~ ~The
i srnrlll fluore\ccnce \1gn,11 must he cl~\trn- tor AFS bcc,~u\eonly one wurce would be nccded for
gur\hcd t ~ o mb x k g ~ o u n d\ ~ g r ~ml \d n o w due to e m \ - ,111 clcnient\ tot smgle-clement 01 multlelerncnt mea5-
slon or fluorescence Iron1 other \peclcs or sc'ttterrng of urerncnt4 No prcwntly m d a b l e contlnuum wurce
the exctt,1tlon I cid~,tt~on pl ovldes \utf~clentlyh ~ g hr d r m c c over the e n t m w ~ l t e -
length rcglon needed tor AF Ihc mo5t w ~ t n h l ccon-
Excitation Sources trnuum cxc~tatlonwurce h,lr hecn 150 to 500-W Xe
high-prcssure arc I m ~ prun in ,I contrnuou5 mode Usu-
'The deal source for AFS would he stable dnd provdc ally. lamps w ~ t h.In ~ n t c g i dcll~pt~c,ll or par,~bol~c rc-
,In extremely hrgh rad~ance'lt the excrtat~onw,lvc- flcctor (e g , bIMAC, tcc Sect~on4-2) h'lve been em-
length5 lor the dements of ~ntcre\t.The mo\t populn~ ployed bec,lu\e thcy collect m d focu\ or coll~matemore
conventmnal h e \ource\ have been hollow c,~thodc than 2 n \r of the ~ ~ l d ~ ~ lemttcd t l o n from the lamp I he
lamps (HCL\), clectroclcle\\ d~\chargcI d m p @DL\), \pecttCil ~ x l i m c e1s lower t h m th,lt ol ,I c o n v c n t ~ o n ~ ~ l
and metal vapor d x h a r g c lamps No~m,rlly,,I \epdr,rte hne wurce. Ilowcvcr. the r'ld~ant powcr 1s cib\orbed
\ource 15 rcclu~redto^ each clement, dthough \omc mul- over the entlrc ,ib\orption prof~lcr 'ither t h m over lu\t
t~elcrnentlamp5 arc ,~va~lable. It the Iine \our ce p~ohlc the nnrrowel cnu\\lon polrle of the line source The
14 narrower t h m the ,innlyte ,ibsorpt~onprol~leIn the total ~ a c l t ~ ~ n~1Ixorbcd
cc l l y than th'it oh-
I \ g c n e ~ ~ ~le\\
atom17e1, ~t 1s des~lcdto m~~xiru17e the ~ n t e g r ~ ~ Iine
ted t'uned wrth m EDL There 15 a \ub\t,~nt~,ll dccrcc~\e111
i,td~ance. ~,ldi,~nce lor w;lvclength\ 41ortcr t h m 2 0 nm
Metal vapor lamp\ were used early in the dcvel- Lrt\cr\ excellent wurccs tor AFS In mnny re-
optncnt ot AFS, hut ale not L I much ~ tod,\y hec'iuse cpcch txxdu\e thcy c m provdc ~rr,rd~ancc\ many or-
they are c~va~lable tor only the no st v o h t ~ l eclement5 ders of ~mgnttudch~gherth'ln ,iv,i~l,~blc with convcn-
(Cd, Zn, Hg, G't, In, '11, Nck, and K) and they are t~onal\ourceb Both continuou\ wme ,ind puI\ed dye
d~ff~cult to pulse elcctron~cally When t h e l,lmp\ ,Ire l,i\ers have bccn u\ed to p~ovrdetunable rd1'1tlon over
operated at hrgh culrcnt4 to mcrease the t o t d Irne ra- the vis~blcI egmn Pulsed dye h w - 5 h v e found the mo\t
d ~ ~ t n cthe
e , \ource prolrle become\ \rgnit~cmtlybro,id- u\e becau\e they c,m hc tuned to wavelength5 a, low
cned and selt-rcverwd. The ,imo~lnt01 {ource r,rdr'~tion as -220 nm (N, h \ e r pump) or - 180 11m ( N d . VAG
cnntted anti abwrbed by the m,rlytc, ,rnd hence the I'tscr pump) wlth f~eclucncy doubhng The \pcctrd
Sec. 7 1-2 1 Instrumentation 31 1
bandwidth can be adjusted from 1 to less than 1 0 - h m atomizers by continuous sample introduction with
(much narrower than absorption line widths). and the pneumatic nebulizers (occasionally, ultrasonic nebuliz-
beam can be focused to a very small spot or expanded ers). Although early measurements were made with
to any cross section desired. There is a considerable total consumption burners, today only chamber-type
loss in source irradiance for small line widths. The very burners are employed because of more complete va-
high peak irradiances (lo4 to lo8 W cm-,) of pulsed porization and atomization of the smaller droplets in-
dye lasers can cause saturation, which provides added troduced. The burner head is usually of the Meker or
advantages, as discussed in Section 11-3. The high ir- capillary design (Section 8-2) with a circular or square
radiance allows the use of weaker nonresonance tran- cross section. Provision is often made for sheathing the
sitions, and thus wavelength discrimination against source flame with an inert gas such as Ar, which prevents
scattering. Unfortunately, the use of laser excitation in entrainment of the surrounding air. This provides a
routine applications is hindered by the cost and oper- separated flame in which the secondary reaction zone
ational complexity of pulsed, tunable lasers. is distinct from the interconal zone (see Section 8-1)
More recently an ICP has been used as a versatile and reduces background emission and quenching in some
excitation source in conjunction with another atomizer cases.
(e.g., flame or plasma). The excitation wavelength is Selection of the type of flame involves compro-
chosen and matched to the analyte by aspirating a high mises among atomization efficiency, background emis-
concentration of the analyte (e.g., 20 mg mL-l) into sion, and quenching characteristics. The general order
the source ICP. The ICP excitation source is charac- for quenching efficiency is A r < H, < H 2 0 < N2 <
terized by high radiance. long-term stability, narrow CO < 0, < CO,. In early work H2-based flames were
line widths, and freedom from self-reversal. As with used because of their reduced quenching efficiencies
lasers, the primary disadvantages are the cost and com- and low background emission. Detection limits can be
plexity of the source and its operation. improved by an order of magnitude by using a mixture
of 0, and Ar in place of air to reduce quenching from
N,. Unfortunately, these cooler flames provide inade-
Atomizers and Sample Introduction quate atomization efficiency for refractory elements,
and unvaporized solute particles can be a problem.
As for other atomic spectrometric techniques, the ideal The trend has been to go to air-C,H, and N,O-
atomizer would have a large and stable overall nebu- C,H, flames, as used in flame AAS. These increase the
lization-atomization efficiency (see equation 7-9) so that atomization efficiency for refractory elements and min-
no (or n, for excited-state fluorescence), and hence the imize analyte interferences and scattering. The loss in
fraction of absorbed radiation, is significant even for signal due to lower quantum efficiencies (i.e., quench-
small analyte concentrations. Thus minimal dilution of ing by CO, CO,) and the increase in background emis-
the atomic vapor in hot atomizer gases is desirable. The sion signals and noise compared to cooler H2 flames
atomization efficiency (E,) should ideally be unity and can be somewhat offset with higher radiance sources
independent of the sample matrix to prevent analyte such as lasers.
interferences. To reduce noise, the atomizer should cause More recently, the ICP has been used as an at-
minimal thermal excitation of emission from the analyte omizer for AFS. Plasmas generally provide better va-
and other species. porization and atomization efficiency and more free-
In particular for AF, the atomizer should provide dom from analyte interferences than flames. Quantum
an environment that maximizes the fluorescence power efficiencies are usually higher than in hydrocarbon flames
yield (minimal nonradiative deactivation of the excited because of the inert Ar atmosphere (lower quenching).
state). For excited-state or thermally assisted fluores- The background emission is highly dependent on the
cence, the atomizer should provide enough energy to viewing region, but generally is greater than in flames.
populate the appropriate state significantly. Measure- Usually, the viewing region is higher above the induc-
ments can be limited by noise in the signal due to scat- tion coils than used in AES to minimize background
tering of excitation radiation from unvaporized particles emission: excitation temperatures in the range 3000 to
in the atomizer. Thus the atomizer temperature and 3500 K exist in the region viewed. For some elements
residence times should be high enough to provide com- fluorescence from ions is observed. The higher tem-
plete vaporization. Rayleigh scattering from atoms or perature compared to flames enhances the usefulness
molecules cannot be eliminated and determines the fun- of excited-state and thermally assisted fluorescence.
damental background noise limit for resonance fluo- To date, electrothermal atomizers have been used
rescence measurements. sparingly in AFS. The advantages (e.g., small sample
Most A F measurements have been made with flame size, low absolute DLs) and disadvantages (e.g., sig-
Chap. I I 1 Atomic Fluorescence Spectrometry
BLE 77- F~gure1 1-5 In whrch thc lluor excnce 14 ~ollectcd,it 90"
Signal expressions for AFS"" to the cxut,il~on~1x14 I he volurnc elemcnt V In wh~ch
(I 1-1)
r1t0m4 ,Ire r,id~~it~on,illy exc~tcd14 clctcrmined by the
r c c t m g i h r cros5 teLtion ot '1 collnn'ltcd be,~rn( L x 11)
(1 1 2 )
and the p,ithlcngth of the atoinrzcr ( I ) No~mally,the
(11-7) emission o p t m restr~ctthe pathlength 'lncl the volume
( 1 I -I)
(11-5)
(I I 6 )
element ol c x e ~ t ~ ~ m
noted 1 ,lnd V , rcspcctlvely (1 .. 1, V -
t ~do no b s c ~ v ; i t ~ othew
the fraction of the emitted photons that are reabsorbed For a given element and fluorescence transition
before passing out of the viewed face of the atomizer. (specific excitation and emission wavelength), the flu-
It is understood that Y varies with the particular i and orescence signal varies with the instrumental configu-
j levels involved in the transition. ration. This is due primarily to the type of atomizer
The radiant power absorbed in V:' is given by (e.g., F, F,, Y), the type of source and excitation optics
equation 11-8. It is similar in form to equation 7-58 [e.g., E, or (E,),, h , L , and the solid angle of collection
except that the source intensity is expressed in terms of for conventional sources], and the type of emission col-
its spectral irradiance (E,),,, where hL(E,),, = (Q,),, lection system (e.g., I* and a ' ) . Both R ' and I* are
and the prefilter-effect factor. f,,, is added. The spectral considerably larger with nondispersive systems than with
absorptance in V" [a:'(X)] is given by equation 11-9. dispersive systems. For a given element, spectrometer,
The prefilter factor accounts for the fraction of the ex- and atomizer, the fluorescence signal varies with the
citation radiation that is absorbed before reaching the specific excitation and emission transitions utilized. From
viewed volume element and is calculated from equation equation 11-14, these differences are seen to be due
11-10. Thus the product of (E,), and A,,. is the source primarily to the magnitudes of E,,, n,, f,, and Y. With
spectral irradiance actually seen by atoms in V". We excited-state fluorescence, the number density of the
will assume that there is no absorption of the excitation lower excited state is usually lower than the ground-
or the fluorescence radiation by other species and that state number density unless this state is very close to
reflection or scattering losses at the atomizer-atmos- the ground state (e.g., Ga, T1, In, Pb. Sn). In the case
phere interface are negligible. Additional factors could of thermally assisted, stepwise fluorescence, only a frac-
be included to account for these effects or flame ab- tion of the number density of the upper state of exci-
sorption. which can be significant below 250 nm. tation is raised thermally to the state from which flu-
Equation 11-11is the final general form for the orescence occurs. When fluorescence occurs from higher
analyte fluorescence readout voltage obtained by com- excited states, there are numerous radiative and non-
bining equations 11-4 to 11-10. Equations 11-12 and radiative pathways for deexcitation. Thus fluorescence
11-13 are the limiting forms of equation 11-11 for low can occur at two or more wavelengths, and the fluo-
number densities with line and continuum source ex- rescence power yield for a particular transition may be
citation, respectively. Note that f,, = f, = 1 under these less than for resonance fluorescence from the first ex-
conditions. For line source excitation. the fraction of cited state. For elements with complex atomic spectra,
the source radiation absorbed is k,l" if it is assumed the fluorescence is distributed among wavelengths
that the absorption coefficient is constant over the source yielding low Y values for a particular transition. In some
profile and equal to the maximum value k,. Note that cases it is possible to monitor several emission wave-
E, is proportional to the volume element illuminated lengths simultaneously.
and viewed (V" = hL1"). For continuum source exci-
tation, the irradiance is assumed to be constant over
the absorption profile and equal to the value at the Linearity. In equation 11-11, three terms de-
wavelength of maximum absorption, (E,,),. In this case pend on the analyte number density or
the absorbed radiation in V" is equal to the product of
(E,,), and AT (the integral absorption in V*).
If we substitute equation 7-45 for k,, in equation
11-12, the result is equation 11-14, from which we see
that E,, is related to the number density of the state
from which excitation occurs (n,), the absorption oscil- Nonlinearity occurs when primary and secondary ab-
lator strength (fij), and the effective width of the ab- sorption effects become significant. Primary absorption
sorption profile (AX,,,). If we assume normal resonance effects are due to significant absorption of the excitation
fluorescence between the ground state and the first ex- radiation in the region before the viewed volume ele-
cited state (n, = no) and Z ( T ) = go, use equation 7-9 ment (i.e., the prefilter effect) and within the viewed
for n, in equation 11-14. and evaluate the constants, volume element V x .At high analyte concentrations, f,,
the result is equation 11-15. Expressions similar to equa- is less than unity. With continuum source excitation,
tions 11-14 and 11-15 can be written for continuum the integral of the spectral absorptance over the ab-
source excitation, if E,IAX,, is replaced by ( E,, ),,. Thus sorption profile is not proportional to no at high con-
at low analyte concentrations. the measured analyte centrations. Similarly, with line source excitation, the
fluorescence signal is proportional to the analyte con- absorptance over the source profile is no longer pro-
centration (c), instrumental factors portional to no at high analyte concentrations. TJse of
[mGR(X')R'T~,E,hLl'" , atomizer factors (F&,IQef), high lamp currents with HCLs or metal discharge lamps
and analyte spectroscopic factors (I:, fo,YIAX,,,). can broaden the source profile and enhance nonline-
Chap. 9 1 I Atomic Fluorescence Speci-rometry
,mty , % Y O ~ L ~ I I Y C(d15o/pt10/1 c / ~ cts( 'ire due t o sign~tic~rnt curve c,in he predicted only it ,111 gcometllc t x t o l \ '19
a b s o ~ptlon ot the fluorescence w1t111nV or 111the pmt- well '15 m'11ytc pcir,imetel\ (e g , the c~-p,ir,ni~eter) ,u-c
filter rcglon Rec,~useAE lines ,ire vely n'lrrow, scc- known. PIcllltcr cftects 'ire mlniniired by clecr c,~slng
ondary ,~bsotptlon I \ ,ilmost I ~ l ~ c iclue y \ to the ,ln,~lytc I,,,, by mclc~itlngthe Ir,~ctlon o f the cxc~tcdvolume
Ilcncc, ~t I \ cdllcd sclt-,lbw~ptlon Seconcl,iry abtor p- clclnent vlcwed (I + I ) , or by uvng I~ont-curlace11-
tlon cause\ f , to be less t h m unity ' ~ th ~ g h,in,rlytc n u n - l u m ~ n , ~ t ~(excltlng
on , ~ n dvlewlng Irom the same \ u r t x e
ber d e n s ~ t ~ c s of the dtomi7er) Self-,ih\olpt~oni\ negliglhle for non-
We now convdcr the ctlect ot e , ~ hof the lore- re\on,ince tluo~csccnce
gomg thrcc terms on I;, ,md on the c,llibratlon plot\
to1 re\onmcc fluorcsccncc FII\ t , Ignoring the ettect 01
/, and f,,, , we hnve ,ilrcndy noted In Section 7-5th,rt for
laige n,,, the fluore\cence \1gn,i1 cxponentldly ~ i p - Up to this point, we have assumed that the cxcit1' t'1011
plo,~che\ ,I Ilmltlng v,rlue with llnc tourcc c x ~ t ~ ~ t l o n sourcc docs not significantly alter the distribution o f
when '111 of the c w , ~ ~ l , ~r,idl,ition
ble 15 abtorbccl Wlth analyte states. With very high irradiances from focusccl
contin~~um wurce excltcltion, the ~ n t e g r dabsorpilon pulsed dye lascr beams, it is possihle to deplete signis-
and hence the fluorc\ccncc sign'il 'ire p ~ ~ p ~ r t i oto n ; ~ l icantly the population of the state from which excitation
n:,'l at high an,ilyte concentr,ltion\ hec,iu\e dbto~ptlon occurs.
stdl occur4 In 111c wlngs of the ibsorption piof~leeven To ~lnderstandthis effect, we will consitter the
though cltcctlvcly nll the sourcc ~ x l ~ ' l t r o,it n the mix- simple casc of resonance fluorescence between the ground
imurn oi lhc d x o r p t ~ o nprolilc has hecn L~bsorbedBn state of population n,, and first excited state of popu-
c ~ t h e rcase, ~t 1s ~ m p o r t m tto ~ c i n c n h cth'it~ E , 15 plo- lation 1 1 , , where the nunrber clensity of higher levcls
portlon,il to the mmunt oi ~ , i d ~ , ~ t,ib\orbecl ~on ((11 ,) (e.g., 11,) is not significant ( n , = n,, + n,). We shall
When n,, 15 I q e , cl,, ,ind hence 15, , I I n~o longer also assume the following: ( I ) quasi-continuum lascr
proportion,rl to / I , , excitation so that the irradiance is constant over the
A4 dl>() d ~ \ c ~ ~ \ 111\ e Section
d 7-5 (e~;u~ltron 7-65), analytc absorption profile; (2) the high source irradi-
1,for photon\ emltted ,inti ,tbsol bed w ~ t h mthe excrted ancc docs not affect the analyte absorption profile, the
m d v~ewettreglon 14 p r o p o r t ~ o n dt o n,,l tor large no temperature, or the velocity distribution in the atom-
T h ~ t coinlrlned
, with the clepenclence of the tr'ictlon ol izer; (3) the excitation bealn stimulates only absorption
'lbsorbed r c i d ~ , ~ t ~on o nn,,, cclu\e\ the c d ~ b r ~ i t ~curve
on (i.c., the lascr docs not enhance ionization); (4) the
t o 1011 over , ~ thigh n,, wrth l ~ n ewurce exclt,\tlon nnd beam intensity is uniform across its profile; and (5) the
t o r e x h a pldte,lu w ~ t hconmuurn sourcc excit,ltlon (\ee analyte number density is low enough that the self-
Flgure 7- 1 ti) Absorptron by ,Itoms In the nonlllumln-
dted postflltcr rcglon cause\ lurthcr attenuation of the
observed flirorcsccncc s ~ g n , m ~ ld allccts the magnltude
o f f , The rndnncr In w h ~ c hf, and the cal~br,ltioncurve
shdpe depend on the an;ilyte conccntr;ltlon 1 5 complex
and hlghly lntlucnccd by the geonletry ol ~llumm,~tlon
and vlewlng
Thc prctdtcr-cllcct i x t o r decrc,~sc\cxponcnt~dly
to relo as n,, 1s mcrcased with llnc tourcc excitation
[equation 11-10 w ~ t h/<(A) - k , , ] I hus 111 the Imlt ot
~ n h n ~ analytc
tc conccntrat~on,all the excitation rad~d-
tion I S absorbed betole renchlng the v~cwcdvolume
elerncnt Vv, and the ob\erved fluorescence tignal ,lp-
proachcs ~ c r oComhlned wlth the p~evlou\ two ciiccts, Concentration, p g / m L
the prcf~lrcl eitect cnu\e\ addlt~on,rl nonlinear~ty at
moderate andytc concentr,ition\ nnd ~ n c ~ e , ~ roll-over \ed FIGURE 11-6 Calibration plots for Cu with AA,
at h ~ g hconcentration\ Wlth continuum \ource cxcl- A E , and AF in an air-H, flame with HCL ex-
citation. The nonlinearity and rollover in the AF
tation, the plefilter effect c,iu\e\ \~tn~l,ir bel~avm-,al-
calibration plot arc duc to primary absorption
though the cllcct 15 Inore complex m d le\\ severe be- effects, including thc prcfiltcr effect, and second-
cause k(h) and f,, valy oven the , ~ b \ o r p t ~ oprof~lc n An ary absorption effects (self-absorption in this casc).
AF ccilibrctt~onplot 15 shown 111 Plgure 11-6. where ~t It [From N. W. Bower and J. D. Ingle, J r . , AppI.
compared to AA and A h c n l ~ b l ~ i t ~plot\ on Spectrosc., 35, 317 (1981), with permission of thc
In ,I r e d system, the e x x t \ h q e of the cnlibratmn Society for Applied Spectroscopy.]
Sec. 1 1-3 1 Signal and Noise Expressions 317
absorption effect, the prefilter effect, and any atten- on various constants, the total analyte number density,
uation of the source radiation over I' are negligible. and the source irradiance.
The equations that describe saturated fluores- For low source radiances (conventional sources).
cence for a two-level system are summarized in Table equation 11-22 simplifies to equation 11-23. which is
11-3. Equation 11-17 shows that the total fluorescence similar in form to the equations developed in Table 11-
radiant power emitted from V* (@,) is determined by 2. It predicts that @, is proportional to the source ra-
the number of atoms in the excited state (n,Vx), the diance, the fluorescence power yield, and the absorp-
Einstein coefficient for spontaneous emission (A,,,), and tion coefficient. As the source irradiance is increased,
the energy per photon (Izv). Under steady-state con- @, is no longer proportional to the irradiance and, in
ditions the rate of absorption and the rate of deacti- the limit (equation 11-24), 0, is independent of Y , (E,),,
vation by collisions, spontaneous emission, and stim- and B,,.
ulated emission are balanced, as shown in equation Experimentally, saturated A F has been demon-
11-18. Here we assume that thermal excitation is neg- strated with pulsed dye lasers, which provide irradi-
ligible and note that (E,),/c is the spectral energy den- ances from lo4 to 10' W cmP2 nm-l. An example for
sity (U,),. Equation 11-18 can be rearranged to solve thallium is shown in Figure 11-7. Measurements under
for the ratio of the number densities of the two states saturation can provide several advantages. The analyte
(equation 11-19). For high source irradiances, the sec- fluorescence signal is independent of source fluctuations
ond term on the right side of equation 11-19 vanishes, and quenching species. At higher number densities, lin-
which indicates that the maximum value of (g,n,/g,n,) earity is improved because primary absorption effects
is unity, the rates of stimulated absorption and stimu- and self-absorption are negligible. Self-absorption still
lated emission are balanced, and there is no net ab- occurs over the nonilluminated distance between the
sorption. We can define the saturation spectral irradi- viewed volume element and the edge of the atomizer.
ance, (E',),, to be the value of (E,), for which n, (or The exact dependence of @,on (E,), can be more
the measured fluorescence signal) is 50% of its maxi- complicated than presented here, particularly for small
mum value, as shown in equation 11-21. For go = gl, laser bandwidths, because some of the assumptions made
this saturation spectral irradiance corresponds to one- are not always valid (e.g., saturation only occurs in the
fourth of the total atoms in the excited state. For com-
plete saturation, one-half of the atoms are in the excited
state. Combining equations 11-17 and 11-19 to 11-21
yields equation 11-22, which predicts how 0, depends
TABLE 7 1-3
Equations for saturated fluorescence
@, = hvA,,n,V*
Bo1(E,iono
C
= [k,, + A,,, + q,,
Spectral irradiance, W ~ r n HZ-'
- ~ X 10'
TABLE 7 7-4
Signal-to-noise expressions for AFSa
ground emission noise because the average background 6-5) can be used to compensate for source intensity
emission signal relative to i, is reduced (see Section 5- variations as long as there is no saturation.
6). Source, wavelength, or sample modulation discrim- In the region of substantial calibration plot non-
inate against llf background emission flicker noise. linearity at high analyte concentrations. the SIN often
With high lamp radiances and large values of I* decreases, as shown in Figure 11-8. This is due to in-
and R',the excitation-source-dependent background creased analyte fluorescence flicker noise (increase in
signals become limiting and background fluorescence ),6 because of fluctuations in pre- and postfilter ab-
or scattering noise becomes dominant. Once the point sorption or decreased i, in the roll-over region with line
is reached that flicker noise in these signals is limiting, source excitation. In the visible region, analyte emission
further increases in the source radiance or emission noise can also become significant at high concentra-
collection efficiency provide no improvement or even tions. Its effect is reduced by increasing the source ra-
decrease the SIN. Background fluorescence can be due diance or by using low-duty-cycle, pulsed sources with
to molecular species (e.g., CH, O H , PH,, CaOH) or synchronous, gated detection. Source modulation pro-
atoms, particularly with continuum source excitation. vides discrimination against llf analyte emission flicker
Scattering of excitation radiation by small particles is noise.
proportional to and thus more likely in the U V Under conditions near or in saturation, scattering
region. With continuum source excitation, the scatter- or background fluorescence noise is usually limiting at
ing signal is proportional to W and llf noise in the low concentrations. At higher concentrations, source
background fluorescence and scattering signals can be flicker noise in the analyte fluorescence signal is re-
discriminated against with sample or wavelength mod- duced, (, due to fluctuations in the pre- or postfilter
ulation. absorption is reduced, and analyte emission is negligi-
Although resonance fluorescence is most com- ble. Very high source radiance can worsen the SIN be-
monly used, some of the other fluorescence types can- cause the scattering signal and noise increase in pro-
provide SIN advantages. In particular, scattering and portion to (E,),, whereas ,i becomes independent of
noise due to scatter can be eliminated if nonresonance (EdO.
fluorescence is used. Here it is possible to pick an emis-
sion wavelength where background emission noise is
low. In many cases, the SIN and the DL of excited-
state resonance fluorescence or various types of non-
resonance fluorescence can be poorer than that for nor-
mal resonance fluorescence if the signal decreases more
than the noise. The reasons for lower signals in these
cases were discussed earlier in this section.
At higher analyte concentrations, blank noise is
negligible and the noise carried by analyte signals is
dominant. Analyte fluorescence shot noise can be lim-
iting at moderate concentrations in which case i, should
be increased [larger hL(E,),, R', or I " ) or the electronic
bandwidth (K) should be reduced to improve precision.
At higher analyte concentrations, flicker noise in the
analyte fluorescence signal is usually limiting. With high
blank noise, the signal-shot-noise-limiting region is not
often observed (i.e., when the analytical signal-carried
noise becomes greater than the blank noise, signal shot
noise is negligible relative to signal flicker noise).
The total flicker noise in the analyte fluorescence FIGURE 11-8 Precision curve for AF. At low
signal is due to source flicker noise and analyte fluo- concentrations the SIN is limited by blank noise.
rescence flicker noise. With conventional sources, this As the analyte concentration and AF signal in-
crease, analyte fluorescence shot noise or flicker
latter noise source is usually dominant, and the preci-
noise become dominant. At very high analyte
sion is atomizer limited. Typically, 5, is 0.5 to 1% with concentrations, precision decreases due to ana-
a 1- to 10-s time constant or integration time, whereas lyte emission noise or noise due to fluctuations
6, is about lop3. With pulsed lasers, the pulse-to-pulse in pre- and postfilter absorption. (Adapted from
intensity variations (often 10%) significantly increase S. Ghaffari, Ph.D. thesis, Oregon State Univer-
tl.In this case, a double-beam system (see Section sity, 1984, with permission of author.)
Chap. I I I Atomic Fluorescence Spectrometry
TABLE 7 7-5
Detection limits for AFS (ng m L - ' )
Line Continuum
Element source.' source ' laser^' ICP" HCL
Conventional line excitation source. continuum excitation source. or laser excitation source \\ith tlamc atom-
ization: DLs in brackets with hydride o r cold vapor generation.
bICP as excitation source with flame atomization.
'Pulsed HCL excitation source with ICP atomization (Baird system).
has b e e n hampered by t h e lack of support by instrument atomizers a r e often much higher t h a n t h e best reported
companies until 1982. in cooler flames.
T h e most often cited advantages of A F S include I n terms of dynamic range a n d multielement ca-
low D L s , large linear dynamic ranges, multielement pability, atomic fluorescence spectrometry is superior
capabilities, simplicity, a n d freedom from spectral in- t o A A S . but comparable t o plasma A E S . Nondispersive
terferences. Overall D L s a r e not generally better a n d AF systems based o n conventional excitation sources
often worse t h a n those reported for flame AA, elec- can be relatively simple a n d inexpensive. However, AF
trothermal A A , o r plasma AE except for a few ele- systems based o n monochromators o r laser excitation
ments. I t is recognized in terms of minimizing analyte can b e a t least as expensive a n d complex as many A A S
a n d scattering interferences, C,H, flames o r plasmas o r A E S systems. Spectral interference problems a r e
a r e generally t h e best atomization sources for AFS as generally less severe for A F S t h a n for A E S , but com-
they a r e in A A S and A E S . Detection limits with these parable t o those experienced with flame A A S .
PROBLEMS
11-1. The following components are available for flame Compare the multielement capabilities of flame AFS
atomic spectroscopy experiments. with that of AAS and AES. If it is desired to construct
a single spectrometer capable of simultaneous or near-
Flame (nebulizerlburner system), F simultaneous determinations of most of the elements
Lenses (various focal lengths), L using two of the three flame techniques, which two
would you choose? Explain and justify. If you were
Hollow cathode lamp (high intensity). HCL
not limited to flames but could also pick plasma at-
Chopper (I-kHz mechanical chopper). C omizers. which two techniques would you choose,
Monochromator with photomultiplier tube at- and why?
tachment, MIPMT Discuss the potential interferences in AFS that are
Signal processing (ac) and readout system, SP not present in AAS and AES. Why is AFS considered
more susceptible to environmental effects (matrix and
Draw block diagrams of the setups you would put flame conditions) than the other techniques?
together for (a) flame atomic emission, (b) flame atomic Indicate the differences between the following types
absorption, and ( c ) flame atomic fluorescence. In- of atomic fluorescence: direct-line and stepwise-line,
dicate carefully in your diagrams the positions of the Stokes and anti-Stokes. normal and excited-state.
various components and the geometry of the source1 For a particular atom. only two levels need to be
flame1monochromator arrangement. considered for the resonance transition. The Ein-
11-2. Critically compare flame atomic absorption. atomic stein coefficient (A,,) for that atom is known to be
fluorescence. and atomic emission as to detection lim- 1.3 x lo8 s-', and the quantum efficiency in a certain
its for elements of various excitation energies. range flame is 0.09. What is the observed lifetime of the
of linearity of calibration curves, susceptibility to upper level?
spectral, chemical, and physical interferences, and Why is an excitation monochromator not normally
any other features. employed in A F measurements?
11-3. Consider the flame spectrometric techniques of atomic Filters are often used as wavelength selectors for A F
emission (AE). atomic absorption (AA). and atomic measurements. However, monochromators are al-
fluorescence (AF). The possible sources of noise in ways used for A A measurements and usually used
these techniques are flicker noise in the radiation for AE measurements. Why?
source, flame transmission flicker noise, flicker noise
Contrast the reasons for nonlinearity of calibration
in the flame background emission, analyte flicker noise,
curves at high analyte concentrations for A A , A E ,
shot noise in the PMT photoanodic current, Johnson
and AF.
noise in the load (feedback) resistor, amplifier noise,
dark current noise, and readout resolution uncer- Discuss which of the three flame spectrometric tech-
tainty. For each of the three techniques. one source niques (AE, AA. or AF) would be most suitable for
of noise tends to predominate near the limit of de- determination absolute concentrations in flames. Give
tection (SIN = 2). Give the most likely predominant your reasoning.
noise source(s) for A E , for A A , and for A F near the Discuss the advantages and disadvantages of satu-
detection limit (note that these are different for the rated A F measurements.
three techniques) and justify why the other sources For the Na ,P,,, -+ 2S,,2transition at 589.6 nm in a
can be neglected. H,-0, flame diluted with Ar, the fluorescence power
Chap. 7 7 I A t o m i c Fluorescence Spectrometry
J . D. WineSordncr, S. G . Schulman, and 7'. ('. O'Haver, 9. T. S. West. "Atomic-Fluorcscencc and Atomic-Absorp-
Lurnincsccwcr .Spcctrornctq in Anrtlytical (Ihc~rrzistry, tiou Spectrometry for ('hemica1 Analysis." Analyst, 99,
Wiley-lnterscicnce. New Y o r k , 1972. 880 (1974).
( i . IT. Kirkbright and M. Surgcnt, Atomic Ahsorption 10. K. F. Browner, "Atomic-Fltio~-esce~icc Spectrometry as
u n d Fluorr.sc.rncc~Spci,~ro.sc~)py,Academic Prccs, Lon- an Analytical 'I'cchniquc," Anuly,st, 99, 617 (1973).
don. 1974. 11. C'. '1'11. J . Alkcmade, T j . Hollander, W. Sncllernan, and
V. Sychra, V. Svoboda, and I . Rubeska, Atornic Fluo- P. J. T h . Zeegcrs, M r t d Vupor:s in Flarnc:~,Perganion
resccncc, Speclroscopy, Van Nostrand licinhold, Wok- Press, Oxford, 1982.
inghani. Bet-kshire, England. 1975. 12. T. J. Vickers, "Atomic Fluorescence and Atomic Ab-
J. D. Wincforclner, "Atomic Fluoresccncc Spectrome- sorption Spectroscopy," in I'hy.sicd Metllodr in Modrrrl
try: Past, Present, and Future," in Krccnt Advutzccs in C'hrtnicnl Analy.sis, 'I'. Kuwana, ed., vol. 1 , Academic
Annlyticul Spectroscopy, Pel-gamon Press, Oxford, 1982, Press, New York, 1978, p. 192.
PP. 151-164. 13. C'. T h . J. Alketnade and ti. Herrmann. fi~ndumcntrds
A . 11. IJlhnan, "Multielemcnt Atomic Fluoresccnce of Analyticnl Flunzc~Spec~tro.scopy,Wiley, New York,
Spectrometry ," Pwg. A m ! . A/. Spc,ctro.sc., 3, 187 (I 980). 1970.
N. Omenctto and J . D. Winei'ordncr, "Atomic Fluo- 14. R. Smith, "Flame Fluoresccnce Spectrometry," chap. 4
rescence Spcctromctry: Basic Principles anti Applica- in Spc~trochr~rzicalMc~hodsof Anczlysis, J . D . Winc-
tions," Prog. Arzal. At. ,S/~i~ctrmc., 2, 1 (1070). i'ordner, ed., Wiley-lnterscicnce, New York, 1971.
J . C . Van Loon, "Atomic Fluorescence Spectroscopy-- 15. N . W. Bower and J . D. Ingle, .IT., "Experimental and
Present Status and Futurc Prospects," h i d . Chcm., 53, 'Theoretical Comparison of the Precision o f Atomic Ab-
333A (1981). sorption, T~luoresccnce,and Emission Mcasurcments,"
J. D. Wincfortincr, "Principles, Methodologies, and , 317 ( 1981).
Appl. S p e c t r o , ~ .-1.5,
Applications of Atomic Fluorescencc Spectroscopy," .I.
Chrln. Educ.. , 55, 72-78 ( 1 W 8 ) .
Molecular spectroscopy differs from atomic spec- ever, this information is generally useful only for de-
troscopy in many aspects. With molecules, the bonding scribing line widths in pure rotational spectra in the gas
of atoms gives rise to rotational and vibrational energy phase; collisional broadening is usually dominant. Most
levels. Thus rotational and vibrational transitions occur molecular vibrational and electronic spectroscopy in an-
in addition to electronic transitions. These added tran- alytical chemistry is applied to solutions where molec-
sitions expand the wavelength range of interest from ular interactions cause rotational detail to be lost. The
the UV through the microwave region. The greater width of vibrational bands is determined by the large
number of excited states increases the complexity of range of rotational transitions possible between two vi-
spectra. Mixtures of molecules are more difficult to brational states. Similarly, the breadth of electronic ab-
determine by electronic spectral characteristics because sorption bands is determined by the multitude of vi-
of the breadth of molecular electronic bands compared brational transitions possible between two electronic
to atomic lines. The large number of bands or lines in states. In solution the vibrational detail is usually either
vibrational or rotational spectra also make determina- considerably blurred or not even observed.
tion of species in mixtures more difficult. In this chapter the origins of rotational and vi-
In many respects the variety possible in molecular brational spectra are first reviewed in terms of the en-
spectroscopy is greater because of the numerous dif- ergy levels and transitions possible. Next, factors that
ferent kinds of molecules compared to the approxi- determine the characteristics of electronic absorption
mately 100 different types of atoms. Also. molecular spectra of diatomic molecules are presented. This in-
spectroscopy can be conducted in the gas, liquid, or cludes a discussion of molecular orbital theory, molec-
solid phase, with spectral characteristics changing sig- ular quantum numbers, and term symbols, selection
nificantly with phase. The utilization of chemical re- rules, and band shapes. This discussion of electronic
actions to enhance selectivity plays a much larger role absorption spectra is then extended to polyatomic mol-
in molecular spectroscopy than in atomic spectroscopy. ecules with an emphasis on the symbolism used to de-
We can calculate natural. collisional. and Doppler note electronic states and on the types of spectra ob-
line widths for molecules in the gas phase as described served for different classes of molecules. Section 12-6
in Section 7-3 for atomic electronic transitions. How- reviews the origins and characteristics of luminescence
Chap. 12 1 Introduction to Molecular Spectroscopy
ecules, this nonrigid rotor model results in the subtrac- to more frequent collisions. In condensed phases, ro-
tion of a small term as shown in equation 12-3. Here tational structure is not observed. In liquids, rotational
the spacing between successive lines is seen to decrease energies become effectively nonquantized due to mo-
with increasing J. lecular collisions more frequent (1012 to 1013 s-') than
Application of an electric field splits and shifts the the period of rotation (10-lo s). In solids rotations are
degenerate rotational levels (the Stark effect). In this totally restricted. Hence microwave rotational spec-
case the angular momentum quantized along the direc- troscopy is little used for analytical purposes.
tion of the applied field must be considered and addi-
tional selection rules are applicable.
Hyperfine structure in rotational levels is caused 12-3 VIBRATIONAL SPECTRA
by the nuclear spin interacting with the angular mo-
mentum. Here the selection rules must take into ac- Pure Vibrational Transitions
count both J and nuclear spin quantum numbers.
For tetratomic molecules that are symmetric tops, Molecular vibrations of diatomic molecules may be de-
exact rotational energy levels can be calculated. En- scribed, as a first approximation, by the harmonic os-
ergy-level expressions are more complex and must con- cillator model. A simple harmonic oscillator is a me-
sider the angular momentum contributions along dif- chanical system consisting of a point mass m connected
ferent axes. Selection rules must be broadened to include to a massless spring. The mass is under action of a
hJ = 0 in addition to hJ = ? 1. Also, additional se- restoring force proportional to the displacement of the
lection rules exist for the angular momentum along a particle from its equilibrium position and the force con-
unique axis. Molecules of analytical interest are usually stant k of the spring. The potential energy V is pro-
more complex. However, the equations derived for sim- portional to the square of the displacement. The fre-
ple molecules provide a basis to describe the rotational quency of vibration v is given by v = (k/m)1'2/(2.rr).
transitions observed. Likewise for a diatomic molecule, V is a function
In the gas phase at pressures above 1 torr, colli- of the displacement q of the internuclear distance r of
sional broadening determines the line shape and half- the atoms from their equilibrium position re (or of a
width of rotational lines. The line shape is Lorentzian particle of reduced mass y from the center of gravity).
with typical half-widths of and 10-j cm-I This potential energy function is parabolic in shape as
at 0.1, 0.01, and 0.001 atm, respectively. For gases at shown in equation 12-4. (Refer to Table 12-2 for all
higher pressures. the rotational energies are blurred due equations in this discussion.) Use of this potential en-
TABLE 72-2
Equations for vibrational spectroscopya
Reduced mass:
Rotational-vibration transitions
E,,=C(v+$) +B,J(J+ 1) f o r v = 0 , 1 , 2 . . . . a n d J = 0 . 1 , 2 , . . . (12-10)
Force constant (K m-')(typically, 200 to 2000): q = r - r , , change In internuclear distance from
its equilibrium value (cm): r. internuclear distance (cm); re. equilibrium internuclear distance (cm);
v, vibrational quantum number, 0. 1. 2. . . . ; 6. molecular constant (cm-I): X.
anharmonicity constant
(dimensionless): typicall). 0.002 to 0.02; Be,dissociation energy (cm-I): Bt. rotational constant de-
pendent on r (cm-I).
Chap. 92 I Introduction to Molecular Spectroscopy
ative io Lhc xcro-point vihrational cnergy. polyatomics havc 3iV - 0 normal moclcs. Each lnocle
Sec. 12-4 / Electronic Absorption Spectra of Diatomic Molecules 329
lectrsnic States
spect to a reflection in the plane of symmetry axis or A, S, and 0 are the same, but the overall state is an
antisymmetric (2- state) with respect to this reflection. odd since an even 1s state and odd 2p state are com-
(u orbitals give only + states, while .rr and 6 orbitals bined. Thus the term symbols are and 3C,+.If the
can lead to either + or - states.) excited electron is promoted to the 2p.rr orbital, the
The ground-state configuration of homonuclear ~ . A = 1,
electronic configuration is ( I s ~ , ) ~ ( 2 p n , )Now
diatomic molecules can be found in a manner similar and S = 0 or 1. The possible term symbols are In[, and
to that for the configurations of many-electron atoms. 3n,,.
-
With atoms, electrons are put into hydrogen-like AOs, The electronic states for heteronuclear diatomics
paying attention to the Pauli principle and Hund's rules. are obtained in a similar fashion, although the sym-
For homonuclear diatomic molecules, the electrons are bolism and energy ordering of the MOs may be differ-
put into I-&+ -like MO's formed hypothetically from AOs. ent. For diatomics with similar nuclei, the Mulliken
Again the Pauli principle must be followed, and Hund's symbolism is often used. For example, CO (14 elec-
rules tell us the lowest-energy state is that of the highest trons) has a ground-state electronic configuration of
multiplicity. KK(~u)~((yu)~(w.rr)~(x.rr)~ with a lC + ground electronic
For H,, for example, both electrons go into the state. The m, xu, and w.rr MOs are bonding MOs and
lsu, orbital with opposite spins. The electron config- the yu MO is a nonbonding MO yielding a bond order
uration for the ground state of H, is thus ( 1 s ~ ~Since )~. of 3 (a triple bond).
A = 0 and S = 0, the ground-state term symbol is If the atomic numbers of the atoms differ by more
]Xi. The bond order is 1. For Li, (6 electrons), the than two units, the electronic energy is determined largely
electron configuration is ( l s ~ , ) ~ ( l s o ~ ) ~ ( 2 s uAgain
,)~. by the heavier atom. The orbitals formed from inner-
A = 0, S = 0, and the ground state is 'C,C and the shell electrons are primarily atomic in character. Thus
bond order is 1. For B, (10 electrons) the ground-state not all valence-shell electrons contribute significantly
configuration is (lsu,)2(lsu,*)2(2su,)2(2su~)2(2p~u)2or to the MOs. Usually, the numbered MO symbolism is
K K L L (2p.rrJ2. In the latter case, K K and LL indicate employed.
filled lus and 2us bonding and antibonding MOs. For For HF (10 electrons) the ground-state electron
+
this configuration A = 1 1 = 2 or A = 1 - 1 = 0. configuration is ( 1 ~ ) ~ ( 2 ~ ) ~ ( 3 u ) ~ (and
l . r rthe
) ~ , term
For the C states (A = O), the electrons can be opposite symbol is l X +. Here the 1u orbital contains atomic in-
spins or the same spins, giving rise to the two states ner-shell electrons, and the 20 and l.rr orbitals are non-
IC,+ and 3Z;. The A state (A = 2) is forced to be a bonding AOs. Only the 3u orbital is an effective MO
singlet state by the Pauli principle. It is, in fact, a 'A, resulting in the HF single bond.
state. The ground state, 3C;, is the state of maximum The term symbols resulting from different com-
multiplicity. binations of equivalent electrons (i.e., same n and I
Electronic term symbols for excited-state electron quantum numbers) and nonequivalent electrons are
configurations of homonuclear diatomics are deter- summarized in Table 12-3.
mined in a similar manner. For example, consider
excited H, in which one electron has been raised Electronic Transitions
from the ground state to a higher bonding MO. For the
(ISU,)~(~SU,)~ electronic configuration, A = 0 and S = In electronic absorption transitions, electrons are pro-
0 or 1, which yields term symbols '2; and 3C,+for the moted to empty or partially filled MOs. These transi-
singlet and triplet states, respectively. For the (lsu,)l tions between ground and excited electronic states are
(2pu,)l electronic configuration, the possible values of symbolized by specifying the term symbols of the elec-
TABLE 12-3
Electronic term symbols resulting from equivalent and nonequivalent
electrons
Equivalent Electronic Nonequivalent
electrons terms electrons Electronic terms
332 Chap. 12 I Introduction to Molecular Spectroscopy
Homonuclear diatomics
(Laportc rule)
Intensity +
Because one atom can be bonded to two or more Transitions from bonding or nonbonding orbitals to very
atoms in polyatomics, MOs are often formed from com- high energy states near the ionization limit are called
binations of three or more AOs. The familiar concept Rydberg transitions (denoted A'-+ R). These transitions
of hybridization (e.g., sp, sp2. sp3) is often employed. are observed in the vacuum UV region with conven-
Although not considered in our previous discussion, it tional single-photon spectroscopy.
is often necessary even for some diatomics to consider Other symbols are used to represent electronic
mixing of AOs before formation of MOs. states in polyatomics. These include Platt symbols, for
which A denotes the ground state, B excited states in-
Electronic States and Transitions volved in highly allowed transitions, and L and C ex-
cited states involved in partially forbidden transitions.
For many polyatomic molecules, it is still possible to Particularly for organic molecules, it is also common to
consider that transitions involve a and .rr bonding and represent electronic states as singlet states (So, S,,
antibonding MOs and nonbonding (n) MOs as shown S,, . . .) and triplet states ( T I , T,. . . .), where the
in Figure 12-6. Typical characteristics and Mulliken's subscript indicates the energy ordering. Usually, the
symbolism for these transitions are given in Table ground electronic state is the lowest-lying singlet state
12-5. In Mulliken's symbolism, N denotes the ground (So) because electrons are all paired in the ground-state
state and V. Q , and R denote excited states. As shown electronic configuration.
in Table 12-5, N -+ V transitions occur from bonding Finally, electronic states can be specified with group
to antibonding orbitals and are strongly allowed. The theory notation. Here it is necessary to identify the
N -+ Q transitions involve promotion of an electron in point group to which the molecule belongs. The symbols
a nonbonding orbital to an antibonding orbital and are for the symmetry species or irreducible representations
usually weaker than N -+ V transitions. Of the N -+ Q of the point group are then used to denote the electronic
transitions, the n -+ a x transitions are generally more states. For example, water belongs to the C,, point
intense than n -+ .rrA transitions. Both N -+ Q and group and the electronic states are denoted A,, A,. B,,
N -+ V transitions are called sub-Rydberg transitions. and B,. The MOs are specified with the corresponding
lowercase symbols. Thus the electron configuration for
H 2 0 is (1a,)2(2a,)2(1b,)2(3a,)2(lbl)2.
The group theory notation is even used for dia-
t o m i c ~which belong to the C,, point group. Here the
1
t t
u* (antibonding) 2 + ,C-, II,A , and @ term symbols normally used cor-
j respond directly to the A,. A,. El. E,. and E, symmetry
n* (antibonding)
t t species, respectively.
A further complication arises in polyatomics be-
cause the ground-state and excited-state molecular con-
n (nonbonding) figurations may be different and described by different
s (bonding) point groups. For example, CS, is linear (D,,) in the
ground electronic state but is bent (C,,) in the first two
1 1 o (bonding)
excited states.
Once the electronic states are identified. the al-
FIGURE 12-6 Electronic transitions between u. lowed transitions can be determined by group theory
T . and n orbitals. techniques. Because A is no longer a good quantum
TABLE 72-5
Characteristics of electronic transitions between a, n, and T orbitals
hlultiken's
Transit~on designation A (nm) E (L mol-I cm ' ) Examples
u N- V <200 Saturated
-
--t J,<: -
hydrocarbons
~i --t T' A' V 200-500 = 10-' Alkenes. alkynes.
aromatics
-
n --t u"' ,Y--, Q 160-260 10:- 101 H 2 0 . CH,OH.
CH3C1. CH,NH,
17 --t 7:. .V Q 250- 600 10'- 10' Carbon>ls. nitro.
nitrate. carhoxyl
3 Chap. 72 I Introduction to Molecular Spectroscopy
numher, selection rules bascd on 12 arc n o longer valid. Group\ that '\re re\pons~blelor absorptwn arc cdlcd
The selection rule bascd on spin (AS = 0) is still good ores and ,ire ottcn ~solatcddouble boj~d\The
as a first approximation. Many violations to this rule absorpt~onchar,icter~\t~c\ of common chromophorc\ arc
occur because of appreciable spin-orbit coupling. In l~stcdIn Table 12-0.
general, thcrc must bc a change in symmetry for the 'Ihc M O approxh I \ \ t ~ l lw e d III \ome cases but
two clcctronic states involved in a transition. it 15 u\ually locallzcd to part~cularbond\ In the mole-
cule Por example, for H,CO the ground-\tate electron
configuration can bc wrlttcn as
u A I
Group Exm~plc (10' cm ') (nm) ( L inol ' cm ')
II
e s CH,-CCH, 22
N O , CHI-NO, 36
47.5
28.8
'-38.5
(i.
39
55.5
58
49
38.8
4'). 7
55
07
43
46.5
52.5
44
46.5
40.3
48
Sec. 12-5 I Electronic Absorption Spectra of Polyatomic Molecules 337
nonbonding orbitals of 0 . The three o bonds to the with crystal field theory or ligand field theory. Gen-
carbon atom are formed from sp2 trigonal hybrid or- erally, transition metal complex absorption transitions
bitals and the .rr bond is formed from the remaining p are in the visible region of the spectrum. An example
orbital of C and an 0 p orbital. Only the valence elec- is the blue Cu(NH,)?+ complex.
trons in the rC0and no orbitals are involved in tran- Lanthanide and actinide ions have narrow ab-
sitions to empty .rr& and a$, antibonding orbitals. sorption bands due to transitions to empty f orbitals.
The absorption characteristics of a molecule can The spectra are little affected by the solvent or anions
often be predicted from the chromophores present in present because the f orbitals are well shielded.
a molecule. Similarly, the absorption spectrum of an Charge-transfer absorption occurs with many in-
unknown can be used to identify the presence of certain organic ions where the components of the complex have
functional groups in a molecule. Chromophores sepa- donor-acceptor properties. These. transitions have very
rated by a -CH2- behave almost independently. If high molar absorptivities in the visible region and occur
there are two chromophores in a given molecule which when an electron in an orbital associated with a donor
absorb at the same wavelength, the total molar ab- which has low electron affinity is transferred to an or-
sorptivity is approximately the sum of the individual bital associated with an acceptor of high electron affin-
molar absorptivities. The absence of absorption bands ity. An example is the Fe(II1) thiocyanate complex where,
in the region 200 to 800 nm is a good indication of the in the excited state, Fe(I1) and SCN are the predom-
absence of chromophores and thus a saturated organic inant species. Common anions which undergo charge-
molecule or an inorganic molecule with no double bonds. transfer transitions are SO:-, NO;, I,, CrOi- . and
Shifts in the wavelength of absorption of chro- MnO;. For the latter two anions, the absorption tran-
mophores to longer or shorter wavelengths are denoted sition involves a transfer of an 0 electron to an empty
bathochromic and hypsochromic shifts, respectively. An metal orbital.
increase or decrease in molar absorptivity is denoted a
hyperchromic or hypochromic effect, respectively. Con-
jugation e C==C-C=C, C=C-C==O,
Ph-C=C) usually causes a hyperchromic effect and a Electronic Band Shapes and Intensities
bathochromic shift.
Auxophores are groups which do not absorb them- Molecular absorption electronic spectra can be com-
selves, but when conjugated to chromophores cause a posed of one band or several bands, depending on the
bathochromic shift and a hyperchromic effect by in- types and number of chromophores in a molecule. Sep-
ductive or resonance effects. Examples are -OH, -Br.
and -NH2. Usually. they have at least one pair of n
electrons which interact to lower the energy of the n:'
orbital associated with the chromophore.
In solution, solvent-solute interactions often af-
- -
arate absorption bands can often be observed due to
transitions from the ground state to different electronic
excited states (e.g., So S,, So 3,). As for diatomic
molecules (Section 12-4), the shapes and intensities of
bands are determined by the transition moment and the
fect the absorption wavelengths of chromophores. Peaks Franck-Condon factors.
from n -+ 7 " transitions often suffer a hypsochromic Experimentally. the absorption transition strength
shift as the polarity of the solvent increases. This be- at a particular wavelength is evaluated from the molar
havior is attributed to the increased solvation of the absorptivity (e). The molar absorptivity or the inte-
nonbonding pair, which lowers the energy of the n or- grated molar absorptivity over the whole band (E) can
bital. In contrast, a bathochromic shift with increased be used to calculate oscillator strengths, transition prob-
solvent polarity is often observed for n -+ .rriT:" transi- abilities, and Einstein coefficients as detailed in Ap-
tions. pendix F. An oscillator strength (f) value of 1 corre-
Absorption by inorganic molecules is often due to sponds t o a very strong absorption with a molar
transitions involving unfilled d and f orbitals on the absorptivity at the wavelength of maximum absorption
metals or to charge-transfer transitions. In transition (E,,,)of about 10' L mol-l cmA1.Typical values off are
metal complexes, the bonding between d orbitals of 1 0 - q o l o p 6 for the rare earth ions. 10-"or aquo
the metal and orbitals from the ligand breaks the de- complexes of transition metal ions. 0.03 for MnO, , and
generacy of the five d orbitals. Thus transitions are nearly unity for organic dyestuffs.
between lower-energy filled d orbitals and higher-en- The particle in a box or free electron model has
ergy unfilled d orbitals. The type of splitting of d orbitals also been applied to electronic transitions. Consider the
depends on the molecular geometry (e.g., octahedral, electron. which undergoes a transition to be a particle
tetrahedral) and the magnitude of the splitting depends in a box in which the positive charge is symmetrically
on the particular ligand and is quantitatively treated distributed about the midpoint. It can be shown that
Chap. 72 1 Introduction to Molecular Spectroscopy
the transition probability (K') in .l cm' is given by converted to translational, rotational, or vibrational en-
ergy with no emission of radiation. In contrast, the
radiative dissipation process involves emission of a pho-
ton. Most analytical luminescence determinations are
carried out in solutions or frozen solids, and the Sol-
where e 1 i the charge ot an electron (1.6 x 1 V ' " C) lowing discussion is directed to these applic-at'~ o n s .
and u equali the width of box or w e oi molecule In
meters. Substitut~onof equation 12-15 Into equatlon
F-39 y ~ e l d j
Part of the energy-level diagram for a hypothetical ar-
omatic molecule is shown in a Jablonski diagram in
Figure 12-7. The various activation and deactivation
processes are indicated. 'The absorption or excitation
where N is Avogadro's number. This equation indicates process (a) is very rapid and on the order of the period
that E is related to the size of the molecule or more of oscillation of the electric field of a visible photon
correctly to the part of the molecule over which the (10 l 5 s). Absorption transitions from the ground vi-
electron involved in the transition resides. This explains brational level of the ground singlet electronic state ( S o )
the increased molar absorptivity in conjugated systems. to different vibrational levels in the first and second
'The model also predicts the hathochromic shift oh- excited electronic states (S, and S,) arc shown.
served with conjugation. Molecules in excited vibrational states rapidly dis-
sipate their excess vibrational energy and relax to the
ground vibrational level in a given electronic state. The
INE A energy goes into thermal or vibrational motion of
the solvent molecules in condensed phases. This non-
Up to this point we have been concerned about how radiational process is dcnoted vibrational relaxation.
~noleculesare promoted to excited electronic states by Normally, vibrational relaxation proceeds in a stepwise
absorption o f electromagnetic radiation. Now we will fashion (A'u = 1) in which one vibrational quantum is
investigate the ways in which an excited molecule can lost per collision. This typically takes l W J 1t o 10-"'s.
dissipate its excess energy and return to the ground Since a typical vibrational period is 1 P " s, many vi-
electronic state. The two general modes of deactivation brations occur before the exccss vibrational energy is
involve nonradiative and radiative processes. In non- lost.
radiative clcactivation, the excess electronic energy is The crossover between two states of the same mul-
tiplicity is a nonradiative electron state transition called After intersystem crossing. a molecule in the T,
internal conversion. This is likely to occur when the state deactivates by vibrational relaxation to the ground
potential energy curves for two electronic states cross vibrational level of T,. Normally, the triplet state deac-
such that the lower vibrational levels of the higher elec- tivates by external conversion or intersystem crossing
tronic state are approximately the same energy as higher to the ground state ( T , + S,). The triplet state can also
vibrational levels of the lower electronic singlet state. deactivate by emission of a photon. This radiational
Internal conversion ultimately results in the conversion deactivation process between electronic states of dif-
of excess electronic energy to excess vibrational energy. ferent multiplicity is called phosphorescence (typically,
Internal conversion can occur between excited states T, -+ So + hv). Usually, phosphorescence takes
(e.g., S, -+ S,) or between the first excited electronic to lo4 s to occur because the process is spin forbidden.
state and the ground electronic state (S, + 3,). Gen- Thus phosphorescence is usually observed only if ex-
erally, internal conversion between excited electronic ternal conversion is reduced by cooling or other tech-
states is rapid (10-I, s). Internal conversion from the niques. The wavelengths of phosphorescence for a given
S, to the So state depends on the molecule but is often molecule are generally longer than those for fluores-
less efficient if there is a wide energy separation be- cence because the energy of T, is less than S,, due to
tween S, and So. so that there is no overlap of the the electrons being unpaired and in different molecular
potential energy wells. After internal conversion, the orbitals.
excess vibrational energy is rapidly dissipated through Fluorescence caused by direct excitation to the S,
vibrational relaxation to the ground vibrational level of state or internal conversion to the S , state is called more
the lower electronic state. precisely prompt fluorescence. Delayed fluorescence has
Fluorescence is a radiational transition between a longer lifetime than prompt fluorescence because S,
electronic states of the same multiplicity. For most mol- is populated by indirect mechanisms. For example, in
ecules, the electrons are paired in the ground state so E-type delayed fluorescence, S, is populated by ther-
that fluorescence involves a singlet-singlet transition. mally assisted intersystem crossing (TI -+ S,) back from
Because internal conversion to S, and vibrational re- a triplet state originally derived from an S , state. P-type
laxation are more rapid processes than fluorescence, delayed fluorescence involves a bimolecular reaction be-
fluorescence usually occurs from the ground vibrational tween triplet states followed by triplet-triplet annihi-
state of S, to various vibrational levels in So (S, + +
lation (T, + T I + So S,). The term photolumines-
+
So hv). For this reason, only one fluorescence band cence is used to describe any emission of photons after
is normally observed even if absorption to different photon excitation. For molecules, photoluminescence
excited singlet states occurs. Typically. fluorescence re- includes prompt and delayed fluorescence as well as
quires 10-lo to s to occur. Fluorescence usually phosphorescence.
appears at longer wavelengths than absorption because If the energy of the excitation photon is greater
absorption transitions are to higher excited electronic than the convergence limit of the excited electronic state,
states or to higher vibrational levels in the S, manifold. a bond is ruptured after absorption. This process is
This is illustrated in Figure 12-7. Fluorescence can occur called dissociation. If a bond is ruptured after internal
from higher electronic states in rare instances. Azulene conversion the process is called predissociation. Pre-
and its derivatives exhibit S, -+ So fluorescence. dissociation or dissociation is more likely in molecules
The term external conversion refers to nonradia- that absorb at wavelengths shorter than 200 nm (200
tive processes in which excited states transfer their ex- nm corresponds to 140 kcal mol-l).
cess energy to other species, such as solvent or solute Excited singlet or triplet states can also be de-
molecules. One primary mechanism of external con- populated by photochemical reactions. This includes
version is dynamic quenching. It involves the nonra- reactions of excited states with solvent or solute mol-
diational transfer of energy from excited species to other ecules. Excited states are distinctly different from the
molecules during collisions. Therefore, the rate of dy- ground state in polarity and reactivity.
namic quenching is reduced by cooling the sample.
Absorption transitions to triplet states are forbid- Quantum Efficiencies and Power Yields
den by symmetry (AS = O), although weak absorption
is possible in some molecules. The triplet state can also The efficiencies and hence intensities of fluorescence
be populated from excited singlet states by a process and phosphorescence depend on the relative competi-
denoted intersystem crossing, which is a crossover be- tion between radiative and nonradiative routes of deac-
tween electronic states similar to internal conversion tivation. Different ways of expressing luminescence ef-
except that the states have different multiplicities (usu- ficiencies are summarized in Table 12-7. From equation
ally S, -+ T,). 12-17, the luminescence quantum efficiency or lumi-
Chap. 12 I Introduction to Molecular Spectroscopy
BLE 72-7
Expressions for luminescence efficienciesa
'I~,.,,,
Phosphorevxncc q u ; m t u m elliciency: dl,, = ---
'I) 4 .I,
nescence quantum (+,) is the ratio of the lurni- The corresponding luminescence
ncsccnce radiant p to the absorbed radiant power (Y,,,) is the ratio of radiant power
whcrc the radiant powers arc expressed in photons per over frequency interval v to v + dv to a),, as shown
second. Thus +, indicates the fraction of the absorbed in equation 12-22. If excitation occurs over a small fre-
photons which are converted to luminescence photons quency interval centcred at v and emission is observed
0 5 1 , 5 I). Eql tions 12-18 and 12-19 are specific at v'.
expressions for the antum efficiency and
the phosphorcscen ciency, respectively.
Here and elsewhere we will use the subscripts b' and P
to distinguish fluorescence from phosphorescence. The
luminescence power yield (equation 12-20) is defined in With this relationship and equation 12-21, we see that
an analogous fashion to the lumincscence quantum ef-
ficiency (equation 12-17) except that the radiant power
of absorbed and emitted radiation is expressed in terms
of photons per secon
1 lurni~~escenceyuantu
The total luminescence power yicld for a given exci-
is the fraction of the absorbed photons that results in
tation frequency is the integral of equation 12-24 over
luminescence over a frequency interval v to v + dv, as
the emission band. or
defined in equation 12-21. This equation also shows that
the spectral lumincscence quantum efficiency can be
expressed in terms of the luminescence quantum effi-
ciency and the emission hand profile (SI)which is nor-
malized so that J S:, dv' = I . In this discussion a prime
is used to distinguish an emission frequency or function As a rough approximation,
fronl excitation quantities. Note that
Note that YL < 6 , and that Y, always depends is small, and detection of fluorescence is difficult. The
on the excitation frequency (see equation 12-25). Even quantum efficiency of a molecule is appreciable only if
if +, = 1, some of the excess electronic energy is con- k, is comparable to or greater than k,,, since this in-
verted to heat before luminescence occurs by internal creases the probability of emission before nonradiative
conversion and vibrational relaxation. As the excitation deactivation.
frequency is increased, Y, decreases because a greater Typically. k , is lo6 to lo9 s-l. To understand the
fraction of the absorbed radiant power is converted to factors that affect the magnitude of kt,,, we can break
heat. it up into its components:
ulatcct by intersystem crossing and the rate of deacti- TI -t S,,trnn\rtlon is spm forbrdcten. Thus temperature
vation 7', . Note that (<\I,,+ (11,) y2 I . 15 uwally 'I more c~rt~calv'u~ablc for phmphore\cencc
k,',, = h:, +A,',, (12-38) Usually, I<,..is 105 to 10% I , so that (T,.), is 10 ' to
10 5 .Because lz,, < 10' s , ( T ) , L O ' s. For organic
whcrc k,', and I<,',, 'ile thc f~r\t-orderr,ite con\t,lnt\ foi systcms k,,'s are rarely larger than 10' s I . whereas for
these procc\\cs 7 yp~cdly,k,', /> k,',,, so thdt pho\- inorganic complexes, k,, is most commonly 104 to
phore\cencc I\ u\u,~llyobserved only ~f I<:, 14 \ u b \ t ~ u ~ - 1U2 s p ', but can be as low as 1 (Ip' s I .
t~allyreduced by cooling the \cnnple to ,I r~gidgl,ls\ in 'The equations above indicate that nonradiative
Iquid N, Sub\tant~,dreductwn of h i , 15 rcqu~rcclbe- dccay clecr-eases the steady-state l'luorescencc and phos-
c ~ s typrcally
c h,, < 10' \ ', due to the lact that the phorescence intensities and lifetimes hy the same fac-
Sec. 12-6 1 Luminescence Spectra 343
K, [Ql
Quenching and Excited-State Reactions + ~ =
l (+!)-I + - (12-48)
4:
The luminescence signals observed can be reduced by
the presence of concomitants through several mecha- Since the observed fluorescence signal is proportional
nisms. Reabsorption of emitted radiation by other spe- to the fluorescence quantum efficiency, a plot of the
cies (or even other analyte molecules) is sometimes reciprocal of the observed fluorescence signal vs. the
called trivial quenching. We will refer to this process quencher concentration should yield a straight line. From
as secondary absorption to distinguish it from mecha- the intercept and slope. the Stern-Volmer constant can
nisms that cause nonradiative deactivation before pho- be determined. Deviations from the ideal behavior are
ton emission as discussed below. sometimes observed when the extent of quenching is
large.
Note that K, is equal to the quencher concentra-
Dynamic Quenching. Quenching normally re- tion where the fluorescence quantum efficiency or flu-
fers to nonradiative energy transfer from excited species orescence intensity is reduced by a factor of 2 relative
to other molecules. Dynamic quenching or collisional to the value with a quencher concentration of zero.
quenching requires contact between the excited lumo- Also, the fluorescence quantum efficiency and lifetime
phore and the quenching species, the quencher (Q). are reduced the same amount by dynamic quenching.
The rate of quenching is diffusion controlled and de- In fact, K, is given by
pends on the temperature and viscosity of the solution.
The quencher concentration must be high enough that
the probability of collision between the analyte and
quencher is significant during the lifetime of the excited
species. where 7: is the fluorescence lifetime in the absence of
If external conversion is controlled by collisions the quencher. Similar Stern-Volmer expressions apply
with a single quencher. a second-order rate process. the to phosphorescence measurements where +,replaces
rate constant for external conversion. is given by 4,.
In aqueous solutions at room temperature, the
bimolecular collision rate is about 101•‹L mol-' s c l .
Hence, if all collisional encounters result in quenching,
where k, is the second-order rate constant in L m o l l we can estimate that the maximum value of k, is about
s-' for quenching and self quenching is assumed neg- 101•‹L mol-' s-'. For a fluorescence lifetime of 1 ns,
ligible. From equations 12-31 and 12-32, the fluores- equation 12-49 shows that the Stern-Volmer constant
cence quantum efficiency in the absence of quenching is approximately 10 L m o l ' . Substituting this result in
(6:) is given by equation 12-47 shows that dynamic quenching of fluo-
rescence is usually negligible when the quencher con-
centration is below 1 mM. With a lifetime of 1 ms. such
6: kF
as is typical for phosphorescence, K, can be as high as
=
k, + k,,. + k,, lo7 L mol-'. Clearly. phosphorescence is totally
quenched by an efficient quencher at 1 mM concentra-
whereas with quenching
tions.
Dissolved oxygen is a very efficient quencher for
k~ (12-46) long-lived triplet states, and its equilibrium concentra-
= kF + k,, + k,,, + k,no tion is typically about 1 mM in many solvents. For this
reason it is usually necessary to deoxygenate solutions
By ratioing equations 12-45 and 12-46 we obtain the solidify the sample to eliminate diffusion for sue-
Stern-Volmer equation cessful phosphorescence measurements. Oxygen is also
a good quencher for some fluorophores. such as aro-
6; matic hydrocarbons, particularly if the fluorescence life-
- = 1 + K,[Q] (12-47)
+F time is greater than 10 ns.
Chap. 12 I Introduction to Molecular Spectroscopy
--
50 A and can be as I'rrge ,I\ 100 A btt~cientlong-lmgc
cause the energy difference between 0 and or' =
energy tlmster 14 f,worecl in \ituCrt~on\ where the e m s -
slon \pectriun of the donor ,ind the ,tb\orpt~onspectrum 2 is less than between 71" -0 and
11'
11'
Tlicre are many exceptions to the mirror image
2.
of the 'receptor ove11,tp ,rncl the m o l x ,ib\o~pt~vlty 01
the donoi I \ rel,~t~vely h ~ g hIn tlie over1,rp I eglon Ihc rule such as when the molecular geometry of P;,, and S,
I ,rte o f energy t~,tn\ter incl c,i\cs w ~ t h.Icceptol concen- arc cliffcrcnt or if the fluorcscence is from an cxcirncr.
tl,rtion ,t\ the .Iver,rge d~,t,lncc between molecules de- Also, more than one fluorcsccncc band is observed in
cfe,t\e\ 7 11e dependcncc o i the degrcc of q~~enclnng nnolcculcs such as biphenyl, wl~crcfluorcsccncc origi-
o n the qucnche~conccntr ation I, complex m d doe\ not nates from ctilfcrcnl parts of the rnolccule.
tollow the Stern-Volrncr model Usually, one phosphoresccncc band is observed
I n both dyn,rmrc and long-~angcqucnch~ng,the and the breadth is determined by the range of prohable
qircncher I \ promoted to an exc~tcd\ l < ~ t cOltcn, the transitions from the ground vibrational level of T , to
qucnchcr ~ l c ~ l c t ~ v aby t c snonr,id~nt~onaI procosc\ different vihrationnl levels in S,,. 'l'lie mirror image rule
I lowcvcr, In some c , w \ , the clucnchcr c,in i ~ u n ~ n c \ - does not apply hecause of the very different nature of
ccncc Thi\ lun~~ne\cence 15 telmeti sendtize singlet and triplet states.
cence The overall qu,lntum eff~c~ency ot w r n s ~ t ~ ~lu-
ed
mlncsccncc c,in be g~e'rter th,in the lum~nesccncc
~ L I ~ ~ cll~c~ency
L I ~ I ot tlie ongin,rl lumopliore ~t the cl-
11c1cncy01 energy trmsfel 1s h ~ g hm c l the l~unlne\cence 'I'he lum~ne\cenceetf~c~ency ot a molecule depend4 on
cll~c~cncy ol the quenclier 14 h~ghcith'rn t l ~ r tot the 1t5 5tructure m d o n the env~ronmcnt111 wlnch the lu-
\pec~c\or~grn,rllycxc~teclby plioton ,ibwrption rumcscence 14 mensu~etl Although it I \ ditt~cultto p e -
Sec. 12-6 I Luminescence Spectra 345
sitions.)
Luminescence is rarely observed from nona-
romatic hydrocarbons that have some double
bonds (T electrons). Weak fluorescence in the
UV is observed for some aliphatic carbonyl
compounds involving n-T" transitions. Some
highly conjugated, but nonaromatic hydro-
carbons (e.g., p-carotene and vitamin A) do
exhibit substantial fluorescence due to n - n A
transitions. Diacetyl exhibits significant phos-
FIGURE 12-8 Relationship between absorption phorescence even at room temperature.
and fluorescence. The absorption and fluores- 3. Many aromatic hydrocarbons are intensely
cence emission spectra are represented by the fluorescent because they possess a low-lying
solid and dashed lines, respectively. The fluores-
cence transitions occur generally at longer wave- n - n X singlet state. The energy required for
lengths because the energy differences are less. excitation is often low enough to prevent bond
In solution the vibronic detail is often not present disruption. Phosphorescence is less likely
and only a broad band is observed. without atoms providing n electrons or sub-
stitutent groups that promote intersystem
crossing.
dict if a molecule exhibits luminescence, some general 4. Phosphorescence is often favorable in aro-
rules can be stated. It should be noted that there are matic molecules containing carbonyl groups
always exceptions to these rules. (e.g., benzophenones) or heteroatoms such as
The nature of the lowest-lying excited singlet (S,) nitrogen (e.g., pyrimidine, pyrazine). These
is critical in determining the luminescence behavior of groups introduce nonbonding electrons and
a molecule because fluorescence and intersystem cross- the possibility of n-IT' transitions. The re-
ing usually occur from this state. In organic molecules, sulting increased rate of intersystem crossing
the transitions between S, and S , can involve n-n'" generally reduces fluorescence intensities.
transitions IT,^:' excited states) or n-T* transitions Strong fluorescence is observed for some het-
(n,n'%xcited states). Similarly, the triplet state (T,) erocyclic molecules (tryptophan and others that
can be a T,T" or an n . ~ *excited state. include the indole ring moiety in which nitro-
The most efficient fluorescence usually involves gen is not part of the aromatic ring system)
T-IT" transitions because the transition probability is because the T,T' state is lower in energy than
high (i.e., large e , k,, and k, and small 7,). For n,n" the n . n X state.
states, the fluorescence transition probability is lower 5 . Substituents attached to aromatic rings can
because of the low degree of overlap between n and n'" dramatically influence quantum efficiencies
orbitals. This makes fluorescence less favorable. How- and emission wavelengths as summarized in
ever, the rate of intersystem crossing is usually en- Table 12-8. The groups often influence the
hanced because the energy difference between the sin- nature of the lowest-lying excited state (n,nA
glet and triplet is smaller and the degree of spin-orbit or T,T-). In general, all groups except those
coupling is greater. This can result in high phospho- having little effect on +, cause a red (bath-
rescence yields. The rate of intersystem crossing is gen- ochromic) shift of the fluorescence emission.
erally 1000 times faster between states of different elec- The table shows that electron-donating groups
tronic origin (S,(n,T") + T 1 ( n , ~ * )or S 1 ( ~ , n g+
) (ortho-para directing groups) such as -OH
T,(n,.rr")). Phosphorescence from an n, T" triplet tends in general increase +, relative to the parent
to be short lived and more efficient than transitions from compound. By contrast, electron-withdraw-
a T,IT* triplet. ing groups (metadirecting groups) such as
Chap. 12 I Introduction to Molecular Spectroscopy
TABLE 12-10
Fluorescence ofdinear aromatics in EPA at 77Ka
Naphthalene
Anthracene
m 0.46 365 400 <0.01 0.04
10. Fluorescence from metals usually occurs only acteristics are critical because they affect the nature of
in organometallic complexes, particularly rigid the excited state. There is often a rapid (10-l1 to 10-l2 s)
metal chelates. Often, the ligand is nonflu- reorientation of solvent molecules around the excited
rescent. The complex exhibits fluorescence species that occurs before photon emission. Thus the
if the lowest-lying singlet state of the ligand energies of the excited state during emission and the
is changed from an n , + t o a T,T"state. ground state immediately following emission can be dif-
Sometimes fluorescence involves charge- ferent than they were at the time of absorption. For
transfer transitions with metal d electrons and IT-.rr4 transitions, the excited state is often more polar
ligand orbitals. In some rare earths, lumi- and basic than the ground state. Increasing the solvent
nescence is due to f-f transitions of metal polarity or protic nature decreases the energy of the
electrons. excited state more so than that of the ground state. with
a resultant red shift in the wavelengths of luminescence.
Environmental Effects By contrast, the excited state is less polar than the ground
state for n-n* transitions and a blue shift occurs with
Environmental factors such a temperature, solvent, pH, increasing solvent polarity or hydrogen-bond-forming
and the presence of other species can have a profound capability. In polar or protic solvents, the lowest-lying
affect on the luminescence characteristics of a given singlet state can switch from being n , ~ to ' r,7iX if these
molecule. These factors can affect the rate constants of states are relatively close in energy. This explains why
luminescence and nonradiational deactivation or the some heterocyclic or carbonyl-containing compounds
nature of the lowest-lying excited state. fluoresce weakly or not at all in nonpolar, aprotic sol-
vents, but appreciably in polar. protic solvents. The
Temperature. In general, increasing the tem- overall luminescence quantum efficiency is often less in
perature decreases luminescence efficiencies because hydrogen-bonding solvents, due to an increased rate of
the rate of dynamic quenching is increased. The effect S, + So internal conversion.
is much more dramatic for phosphorescence measure-
ments. Delayed fluorescence (E-type) intensities can p H Effects. The pH of solutions in protic sol-
actually increase at higher temperatures because the vents can be critical for aromatic molecules with acidic
rate of thermally assisted T, + S, intersystem crossing or basic functional groups (e.g.. phenols, amines). In
is enhanced. some cases, only the protonated or unprotonated form
of the acid or base may be fluorescent. For example,
Solvent. The viscosity. polarity, and hydrogen- many phenols are fluorescent only in the nonionized
bonding characteristics of the solvent can significantly form. The fluorescence of amine containing compounds
affect luminescence characteristics. In some cases, lu- can decrease in acidic solution as -NH,' forms and
minescence efficiencies increase with solvent viscosity withdraws electrons from the ring system.
due to the reduced rate of bimolecular collisions and The pK, of the excited state can be a factor of 4
the rate of dynamic quenching. to 9 lower than that of the ground state. For example,
The solvent polarity and hydrogen-bonding char- the pK, of the ground state of 2-naphthol is 9.5, while
Chap. 72 I Introduction to Molecular Spectroscopy
1
- =
I + -
-
T,KT
--
r r,, qVro
weight of the molecule (g mol-I). Thus the @, component increases relative to the @ com-
ponent. Both @, and @, decrease with time also with
the same exponential form as equation 12-39, whereas
the difference D(t) = @ ( t ) - @,(t) decreases as
PROBLEMS
For a molecule the rate constants for fluorescence data: h,,, = 209 nm. E,,, = 15 L mol-I cm-', and
and nonradiative decay are 1.00 x 10"nd 9 x 10' AT = 600 cm- ' (see Appendix F).
s-I. respectively. Calculate the fluorescence quantum 12-5. Determine the electronic configuration and bond or-
efficiency and lifetime. Also calculate the radiative der for the ground state of N2.What are the possible
lifetime. term symbols?
From the results of problem 12-1, calculate the flu- 12-6. Why do the half-widths of electronic absorption bands
orescence power yield if the wavelengths of maximum differ?
exatation and emission are 300 and 400 nm, respec- 12-7. The fluorescence lifetime without quenching is 6.0 ns
tively. while with a quencher concentration of 1.00 x lo-'
Calculate the frequencies of the two lowest rotational M, the fluorescence lifetme is 2.0 ns. Assume the
transitions of IC1 in MHz and cm-'. Assume that the rate constant for intersystem crossing is 1.00 X 10"
equilibrium internuclear distance is 2.32 A and that s-I and that internal conversion from S, to So is neg-
centrifugal distortion is negligible. ligible. Calculate the Stern-Volmer quenching con-
Calculate the oscillator strength, the integrated molar stant ( K , ) ,the rate constant for fluorescence, and the
absorptivity. and the transition probability for the quenching rate constant (k,).
n -+ 7iXtransition for acetaldehyde from the following 12-8. For a molecule, k , = 1.0 x 1 0 9 - I , k,,, =
Chap. 12 / introduction to Molecular Spectroscopy
und Phosphorescence, Wiley-Interscience. New York, 17. C. A . Parker. Plzotolunzinescence of Solutions, Elsevier.
1969. Amsterdam, 1968.
16. W. R. Seitz, '.Luminescence Spectrometry (Fluorimetry 18. S. Udenfriend, Fluorescence Assay of Biology and Med-
and Phosphorimetry)," in Treatise on Analyticul Chem- icine, vol. 11, chap. 1. Academic Press. New York. 1969.
istry, 2nd ed., 1. M. Kolthoff and P. J . Elying, eds., Pt. 19. J. R. Lakowicz. Principles of Fluorescence Spectroscopy,
I, vol. 7. Wiley-Interscience. New York. 1981. pp. 161- Plenum Press. New York. 1983.
204.
Liltraviolet-visible molecular absorption spectropho- used instcad of spcctrophotometric. The introduction
tometry (olten called light absorption spcctrophotom- of thc Beckman DU spectropliotometer with U V ca-
ctry or just UV-visible spectrophotonietry) has a long pability and double-beam recording spectrophotomc-
a d continuing history of use in analytical chemistry. ters by Gary in the 1940s stimulated the dcvelopnient
This technique is based on measuring the absorption of of new spectrophohxnetric mcthods.
near-UV or visible radiation by molccules. Radiation In this chapter we first consider the gcneral design
in this wavelength region causes electronic transitions characteristics of BJV-visible spectrophotorneter-s in-
at wavelengths characteristic of the molecular structure cluding sonlc commercial examples. Next signal and
of the molecule, as detailed in Chapter 12. SIN expressiolls are introduced to provide an under-
The term spectrophotometry, rather than sprc- standing of the instrumental and chemical factors that
tromptry, is used for absorption measurements because affect the measurement accuracy and precision. This is
they involve measuring, either simultaneously or se- Sollowecl by a discussion o l the causes of nonlinear cnl-
quentially, the ratio o f the radiant power transmitted ibration curves and the methodology and performance
through a sample solution to that transmittccl by a blank char;lcteristics. 'I'he chapter concludes with a survey of
solution. This ratio. the transmittance (T), is used to the l9road range of applications in which UV-visiblc
calculate the absorhancc (A = l o g a'). Under suitable spectrophotometry is used.
conditions, the absorbance is directly proportional to
the analyte concentration through Beer's law (A -
Quantitative molecular absorption measurements
ah).
Signal processor
source and readout
Shutter Sample Detector
cell
(a)
Detector
Rotating
sectored mirror ~
I
and readout
Reference cell
---=-ISample cell
Shutter
Spectrograph
U
-'H-.-.I
Multichannel Signal processor
REFERENCE
Tungsten
lamp - - -
of the sample by radiation at wavelengths not moni- 13-2 employ interference filters for wavelength selec-
tored. If these effects are not significant, the wavelength tion. Provision is made for easy changing of the filters.
selector can be placed after the sample cell. With mul- Inexpensive photometers are useful for routine analysis
tichannel spectrophotometers, the multichannel detec- in which a large spectral range can be tolerated. Often
tor must be located at the focal plane of the spectro- a set of filters with half-widths of 10 to 20 nm is provided
graph; the sample cell is placed before the wavelength with central wavelengths corresponding to the analysis
selector and illuminated with white radiation. wavelengths of species commonly determined.
Simple photometers such as that shown in Figure In single-detector spectrophotometers. an image of
Reference L,~hr
,p\ Field lens
phototube sate
/ ' Entrance silt Objective lens
HIGH EFFICIENCY
COLLIMATOR MIRROR ENTRANCE SLIT
EXIT MIRROR
FRONT OF
INSTRUMENT
UV-VIS-NIR
(b)
FIGURE 13-3 Two examples of simple single-beam spectrophotometers.
(a) The Spectronic 20. A vee-shaped slit (light control) is used to adjust
the radiant power incident on the sample cell. The occluder blocks the
light beam when the sample cell is removed. (b) The Turner 320. The
monochromator is based on the Ebert design, and the lamp current is
varied to adjust the radiant power incident on the sample cell. [(a) Cour-
tesy of Milton Roy Co., Analytical Division. Rochester, NY. (b) courtesy
of Sequoia-Turner Corp., Mt. View, Calif .]
Visible source - - ..
- -----
tungsten-halogen lamp 1
e Reference
Electronics
Deuterium
lamu
Reference
Lead
\f?u
Beam
splitter
sulfide
detector
Q
the source is focused on the entrance slit of the mono- wavelength setting (see Figure 13-4a). Sometimes a
chromator with an Fln that matches that of the disper- double monochromator (Figure 13-4b) or a double-pass
sive device. A low-dispersion monochromator with a monochromator (Figure 1 3 4 ) is used to reduce stray
fixed slit width (typically, a 2- to 20-nm spectral band- radiation.
pass) and manual wavelength adjustment is used for With multichannel spectrophotometers (Figure
such simple single-beam spectrophotorneters as those 13-5). the wavelength resolution is determined by the
shown in Figure 13-3. Higher-quality SB and DB spec- size of the pixels in the diode array. Therefore, the
trophotometers are based on medium-dispersion mono- entrance slit of the spectrograph is fixed at a slit width
chromators. Depending on the instrument, the spectral approximately equal to the width of an individual diode
bandpass can be varied from 10 nm down to as low as element; resolution of the order of 1 to 2 nm is common.
0.05 nm. For instruments intended for only fixed-wave-
length measurements, slew scanning is often provided Sample Compartment and Sample Cells. The
to allow rapid adjustment of the analysis wavelength. sample compartment is a light-tight compartment with
Scanning spectrophotometers usually provide a wide a lid that has provision for securely holding one or more
range of scanning rates (from 1 up to as large as 2000 sample cells. The radiant power transmitted by the
nm minP1). Some typical commercial double-beam wavelength selector (or from the source in multichannel
scanning spectrophotometers are shown in Figure spectrometers) is directed to the sample cell(s) in the
13-4. Rapid-scan spectrophotometers based on an os- sample compartment. In some spectrophotometers, ad-
cillating optical component in the monochromator (see ditional optics are used to collimate the light beam, to
Section 4-6) are available to acquire spectra in 1 s or focus the light in the center of the sample cell, or in the
less. case of double-beam instruments, to split and direct the
Many fixed-wavelength and scanning spectropho- light beam to the sample and reference cells. The change
tometers allow manual or automatic placement of filters in the width of the beam across the sample cell should
in the optical train to reduce stray radiation at wave- be minimal, so that all rays travel approximately the
lengths significantly different from the monochromator same pathlength. Often, additional lenses or mirrors
Chap. 13 1 Ultraviolet and Visible Molecular Absorption Spectrophotometry
holder. Alternatively, thermoelectric devices are used lengths up to 10 cm, often of cylindrical shape (Figure
for temperature control. A magnetic stirrer below the 13-6c), are provided for absorption measurements of
sample cell allows mixing of solutions added to the sam- weakly absorbing solutions or gases. In simple spec-
ple cell. trometers or photometers, an inexpensive test tube-
A wide variety of sample cell types (i.e.. construc- shaped sample cell (Figure 13-6d) is often used.
tion materials, pathlengths, and shapes) are available. With standard sample cells, test solutions are added
Common construction materials include normal glass, to the cell manually, or in some cases, automatically
quartz, fused silica, or various plastics. (See Appendix with electrically controlled dispensers. Different sample
B for transmission characteristics.) The more expensive cells can be used for different solutions. The sample
quartz or fused silica sample cells are required for meas- cell must be emptied or evacuated and then rinsed be-
urements at wavelengths shorter than 300 nm, where fore the next sample is added.
normal glass and most plastics absorb significantly. Sample throughput can be increased by using var-
Some common cell shapes and designs are shown ious types of flow cells (Figure 13-6e) in which the filling
in Figure 13-6. The standard spectrophotometer cell and cleaning operations are easily automated. Test and
shown in Figure 13-6a has a 1-cm internal pathlength rinse solutions are pumped to the cell or drawn through
and a 4.5-mL internal volume. Semimicro and micro the cell and then stopped for the absorption measure-
cells (Figure 13-6b) with a 1-cm pathlength but a smaller ment. The pumping time is adjusted to ensure that the
internal volume are available for sample-limited situ- previous solution has been completely removed and
ations. Sample cells can be purchased with pathlengths replaced by the next sample solution. This approach is
from 1 mm to 5 cm. Short pathlengths are useful for used with automatic samplers to provide a highly au-
highly absorbing solutions. Sample cells with path- tomated sequential batch analyzer (see Section 6-6).
From
column
/, Quartz
,,,fwindows
UV source Detector
+
I
TO waste
(f
Signal processing with noncomputerized double- where k' is the conversion factor in volts per absorbance
beam spectrophotorneters is more complex. The sample unit (A.U., a dimensionless quantity). Here the dark
and reference signal amplitudes are first extracted from signal must be negligible, subtracted, or otherwise com-
the modulated signal waveform. Then the ratio or log- pensated before logarithmic conversion.
arithm of the ratio of the sample and reference signals In all spectrophotometers and photometers. there
is computed. For scanning instruments, the spectral data is provision for compensating for the dark signal and.
(A or T vs. X) are plotted in real time on an analog in most, for adjusting the magnitude of the reference
recorder. signal. The reference signal is wavelength dependent
With microcomputer-based spectrophotometers, because the lamp radiance, wavelength selector trans-
most of the computation steps discussed above (cal- mission efficiency, and detector responsivity vary
culation of T and A and extraction of amplitude infor- with wavelength. Accuracy can be limited by resolution
mation from modulated waveforms) are performed on if the reference signal is significantly less than the full-
digitized and stored reference and sample signal data scale capability of the readout device in the transmit-
by software. The readout device can be a digital meter. tance mode or of the ADC in computerized instru-
plotter, a printer, a video monitor. or a combination of ments. Also. the accuracy of analog computational
these devices. circuits (ratio, logarithmic) is often best over a certain
In all spectrophotorneters, the noise equivalent range of input voltages. Therefore, the reference signal
bandpass ( A f ) is controlled by some limiting time con- at each wavelength is usually manually or automatically
stant or integration time which is set to about 1 s on adjusted to a value that uses the full resolution capa-
simple instruments or is user controllable over the range bilities of the readout device and provides good com-
0.01 to 10 s with more sophisticated instruments. With putational accuracy. The adjustment of the reference
multichannel spectrophotometers, Af is usually deter- signal and the compensation for the dark signal are
mined by the integration time set by the manufacturer, implemented by different means in different instru-
which ensures near saturation of the diodes experienc- ments. as we shall see.
ing the highest intensity.
Fixed- Wavelength Spectrophotometers. With
Readout Considerations simple spectrophotometers or photometers of single-
beam design, the readout scale is calibrated in trans-
The reference signal (El) and sample signal (E,) must mittance or absorbance units. Thus the readout signals
be extracted from the readout signals to calculate the are manually adjusted to define and utilize these scales
transmittance or absorbance. The transmittance can be once the wavelength is selected.
calculated from the three readout signals defined in For transmittance readout. the signals E,, and E,,
Table 13-1 and equation 2-27, which is repeated here: are adjusted to correspond to the full-scale reading (Ef3)
and zero reading of the readout d e ~ i c erespectively,
. as
shown in Figure 13-7. A 0% T control knob is first used
to adjust the readout signal to zero on the scale when
Absorbance Readout Transmittance
At room temperature blackbody emission from readout signals readout
the sample cell or solutions in the UV-visible region is
negligible and analyte and background luminescence
signals are usually insignificant compared to the lamp
signals (E, and E,). Thus subtraction of E,,, or Ed in
this case, from the total reference and sample signals
yields E l and E,, respectively. and allows calculation of
the true transmittance (T' = E,IE, = T) and absorb-
ance (A = - log T).
For instruments that use analog logarithmic cir-
cuitry to provide direct absorbance readout, the ab- FIGURE 13-7 Adjustment of the readout scales.
sorbance is calculated from the sample and reference With transmittance readout, the total reference
signal is adjusted to the full-scale setting of the
logarithmic readout signals defined in Table 13-1 and
readout device ( E f i )and the 0% T signal is ad-
the equation justed to the zero setting of the readout device.
With direct absorbance readout, the logarithm of
the reference signal is adjusted to the zero setting
of the readout device.
2 Chap. 13 I Ultra violet and Visible Molecular Absorption Spectrophotornetry
A is not directly calculated and E, varies during scan- the optical path for the reference and sample beams is
ning. To generate a point-by-point spectrum. it would identical except for the differences in transn~issionchar-
be necessary to adjust the readout to 100% T o r 0 A . U . acteristics of the cells.
with blank solution, replace the blank solution with Computerization of scanning spectrophotometers
sample solution, read T or A , change the wavelength. enhances their performance and capabilities. The es-
and repeat the reference adjustment and sample read- sential feature is that the reference intensity spectrum
ing procedure for each wavelength. Alternatively, a (E, vs. A) and the sample intensity spectrum ( E , vs, A)
separate scan of E,. vs. A with the blank solution and of are digitized and stored. This allows spectra to be ob-
E, vs. A with the analyte solution could be obtained and tained conveniently with single-beam instruments of
followed by a point-by-point calculation of T o r A from simpler optical design. Because the reference and sam-
the values of E, and E, at each wavelength. Here the ple intensity spectra are obtained at separate times, it
accuracy can be limited by readout resolution at wave- is critical that the light-source radiance remain constant.
lengths where E, is small due to a lower lamp radiance Computer calculation of T or A essentially eliminates
or detector responsivity. computational error and nonlinearity. Very high scan
Traditionally. absorption spectra have been ob- rates (e.g., 1000 nmlmin) can be used with such spec-
tained with scanning double-beam in-time spectrome- trophotometers because the data can be digitized rap-
ters (often called ratio-recording spectrophotometers). idly and displayed later. With conventional scanning
During scanning, T or A is continually displayed, and instruments, the response time of the computational
the reference voltage is maintained at a constant value circuitry or the analog recorder limits the maximum
for best readout resolution and computational accuracy. scan rate. For baseline compensation a spectrum of
In the auto slit mode. the slit width is adjusted during blank versus blank in double-beam instruments can be
the scan to maintain the incident radiant power at a stored and later subtracted from the sample absorption
constant value by feedback circuitry or a slit program. spectrum to account for differences in the wavelength
This mode of operation results in a constant dependence of the optical efficiencies of the optical
SIN at all wavelengths but a variable wavelength res- components in the sample and reference paths.
olution. In the auto gain mode. the PMT gain is con- As with noncomputerized scanning instruments,
tinually decreased or increased as the wavelength is there is usually some provision for adjusting E, to a
changed to maintain a constant photoanodic current. relatively constant value during the scan. Either auto
Although the resolution remains constant throughout slit or auto gain modes can be used. In one commercial
the spectrum. the SIN is degraded where the lamp ra- single-beam scanning instrument, filters with different
diance or detector responsivity is lower. transmission characteristics are automatically posi-
Ideally, a spectrum with blank solution in both tioned into the optical path during the scan to maintain
the sample and reference cells would yield zero ab- the incident radiant power at a relatively constant value.
sorbance at all wavelengths once the absorbance is set Both SB and D B multichannel spectrophotome-
to zero at one wavelength. In reality, there is some ters provide the information and calculation options of
variation in the baseline absorbance due to differences scanning computerized single-detector spectropho-
in the wavelength dependence of the reflectance and tometers. However. the spectral data can be obtained
transmission characteristics of the different optical com- much more rapidly because all wavelengths are meas-
ponents and the sample cells in the sample and refer- ured simultaneously. Mechanical simplicity and wave-
ence beams. Thus some type of baseline compensation length accuracy are added advantages. particularly in
scheme is often provided. In older instruments. a mul- SB designs, where an automatic shutter is the only mov-
tipot system was used to adjust the baseline to zero in ing part.
a number of separate wavelength regions.
Although not common, some manufacturers have Other Features
used a sample modulation approach for double-beam
measurements (see Section 4-6). Here the sample and Display Options. With both fixed-wavelength
reference cells are secured in an oscillating carriage and scanning instruments, provision is often made for
which alternately places the sample and reference cells displaying a portion of the transmittance or absorbance
into the optical path. The detector output is similar to range on the readout device (e.g., 100 to 90% T or 0.4
that of double-beam in-time spectrophotometers (see to 0.5 A.U.) to enhance the readout resolution. With
Figure 13-8) although the modulation frequency is typ- noncomputerized instruments. the display range must
ically 2 to 4 Hz. The sample modulation design provfdes be selected before the scan is made. In contrast, com-
double-beam information without having to split the puterized instruments allow the user to display at leisure
original optical beam into two separate beams. Thus the acquired and stored data in different formats. Thus
Chap. 13 I Ultraviolet and Visible Molecular Absorption Spectrophotometry
,my p.~rticul,u wavelength reglon of the totd qxxtrum Derivative Spectra. A I'irst or higher derivativc
,~ccluiredc m be dlspl,~yedwith m y dcwed readout \c,lle of the absorbance spectrum (c.g., dA/rlX, d2A/riX') is
Scde e x p m \ ~ o n15 u\eful up to the polnt t11,tt the dr\pl,iy often useful for accentuating spectral details such as
15 Irm~tedby w,ivclcngth ~ e w l u t ~ o the n , rewlut~onof absorption band maxima and shoulders on bands, as
the ADC, or the nolw in the d ~ g ~ t l t c\ign,11\
d C'o111- discussed in Section 13-5. With noncomputcrizcd in-
putcr-b,~\edin\trumcnt\ d \ o dlow succe\uve \pcctr,1 struments, the first derivative is ohtailled with clec-
to he stored m d l,~tel\upcnrnpo\ed lor conip,irl\on 01 tronic tlerivativc circuits which output a response pro-
,idded, ruhtr,~cted. m d norm,~lizedbclore di\pl,iy A portional to dA/dt during a scan. The time derivative
plot oi log A v\ A c m also be generatcd c,~silyw ~ t h is proportional to dA1rfA through the dependence of A
dppropnalc \oltw,~re 11115 type of d,~t,lprcscntntlon o n the time t as dctcrrnincd by the wavelength scan
weight4 the rcl,it~vcchmge in ,lbwrbmce w ~ t hwme- ratc. A derivative spectrum can also be obtained with
length 'it low and h ~ g h,~b\orb,incc\equally lil Beer'\ a D W spcctrophotoineter by scanning both wavelengths
I'IW 15followed, log A spcctr't d~ilcrcntconcentr'ltion\
drc p'1ral1el and \h~ltcdfrom e,lch other by log tx
A concmfratroii rcudout mode I\ av,ul,~ble wlth
-
si~nultancouslywith a constant wavclcngth dilfcrcncc
[AAIAh tlAItih if Ah is small] or by wavclcngth mod-
ulation during a scan (see Figure 4-344. Finally, com-
many In\trument\ as a convenience te'tturc Th1\ I\ rccllly puterized instrumcnts allow derivative spectra to be cal-
,i d ~ l e c absorbmce
t readout mode in which the voltngc culated from the stored absorption spectrum with
or number coiie\pond~ngto the cib\orbn~lccI \ multi- appropriate software.
plied by ;1 sultdblc t x t o r before di\pl,iy. A standard i \ Taking the derivativc also accentuates the noise
u\ed to adlu\t the r e d o u t to corre\pond to the m,rlyte as dAldX due to noise is oftcn greater than that due to
concentldl~on,d t c r which the andyte conccntr,~t~on ot the change in absorptivity with wavclcngth. Thcrcforc.
\'imples 14 dmctly reported. I h e mult~plic,~tlon txtor smoothing routines are often built into computcrizcd
i \ d l u \ t e d w ~ t h'1 potentiometer In noncomputcr17ed instrumcnts.
in\trunIents or softw'lre-\elected In coi11putcr17cd In-
struments
Probe-Type Photometers. Normally, the sam-
ple is brought to the sarnplc cell in the instrument. Fiber
Dual- Wa velength Spectrophotometers. The
optics arc making possibl~a new generation of instru-
d~fiercnccIn the ,~bsorbance\mensurcd 'lt two wave-
ments for in situ measurements. Fiber optics can be
lengths [ A A = A ( h , ) - A(X,)j i \ 'i u\eful qu,ml~tyIn
used t o direct light between rhc sample and the instru-
\(>me appl~cdtion\, d~scu\\cd111 SectJon 11-5. Spc-
ment. Figure 13-9 shows an example of a fiber optic
a A ~ e dud-wclvelength
d (DW) spectrophotomete~sale
pro!~ephotornctcr that allows absorption measurements
a v d ~ l ~ ~ to m d d1sp1'1y A A They ~ l r econ-
b l ccdlc~il~ite
directly in sample containers.
f~guredto pass ihc radl,~tionat the two wnvclength\
,iltern,itely through the \ample cell (1 e , double-c11,in-
nel 111-tme) The s~gnal\,ire processed much hkc a dou- M~crocomputerEnhancements. We h,wc '11-
ble-beam ~n-timespcctrophoton~ctcr,except that AA ready noted t l ~ mlcrocomputcr-b:1scc1
t instrument\ li,~ve
1s c'~lcul,~tedWavelength \elect~onIS based on a con- profoundly cnhmccd the powei and c a x 01 uw ol spec-
vcnt~on,ilmonochrom,~to~ wlth 'I v~hrnt~ng retrxtor plate trophotornete~\~n term\ of cf&i , I ~ ~ L I I \ I ~\to~,igc,IOII,
located ncar the exit \11t (Figure 4-14) 01 ,I d i ~ ~ ~ l - g r d l n g 'ind dl\pl,iy In d d d ~ t l o ~contr01
l, m d c,~lcul,itiono p t ~ o m
monochromator A d i d - g r a t ~ n grnonot hrourzator con- ,Ire gre'itly ~ncre a x d Oltcn, 1' k e y h ~ r ~JIow\ ~ d u\er
t,iin\ two s t x k c d gi'iting\ th,tt arc ~ndependcntlycon sclcctlon of w,~veleirgth,tcallnrng ym-meter\ (e g , \cm
trolled 'I he r,~di~itron selected by c x h g r , ~ t ~ npnsxs
g ratc and r'lngc), 411t width, type 01 lanip, tlmc con\tmt
th~ough,I dlttcrcnt portion ot the u m ~ cexit \ht or c x ~ t s or ~ntegrat~on tlme, 01 number 01 rcpctrt~vcrncawrc-
at 'I sl~ghtlyditterent angle A mechm~calchopper 1s merit\ 01 scan\. P r o g ~ m ~ r n i n(lnd
g \tol,rgc of lepctltrvc
conf~guredto ~nterceptthe b c m ~ \uch \ th,lt ' ~ tI' glvell or db401bd~lccv,llue\ dt one 01 more w,iveIength\
t m c , one beam 14 blocked w h k the other I\ d~rected plovlde\ SIN enhmccmcnt m d ,~llow\m u m . \t,rnd,ird
to the sample cell T o generate ,I DW \pectrum, elthe1 dcv~'~tions, and other \tC~tl\trcnl tlat,l to he ccllculated
one wavclcngth 1s sc'lnned with the o t l w w,ivclcngtli It 1s oftcn po\\ible to rnc~isurcm d \tore the ab-
f ~ x e dor both w~~vclcngth\ 'ire \canned \imultaneously \orbmcc d,lta 11oni ,I \erIe\ of \t,lndard\ along w ~ t hthe
w ~ t h'I con\tant wdv~lcngthditterencc W ~ t hmicrocorn- \t,~ndurtlconccntr<ltlons The c , l l ~ b ~ ~ l d,~t,i
t ~ o n,ire then
puter-ba\ed rca~~~l~ng"~n\trumetit\ or multrchdnnel \pee- fltted to gener'ltc n cAbr'~tion fuiiction wh~chI \ \ub-
trophotomete~\,A A or '1 DW \pectluin c,ln be c,rlcu- 4eyuently used to calculate ,~ndreport ,m,llyte concen-
L~tcclf ~ o mthe \tored \pectrd c~b\orh,inccdat<i tr,lt~on\from the \tored \,irnplc cibsorhmce\ The ab-
Sec. 13-2 1 Signal and Noise Expressions 365
Photodiode
Readout Expressions
amplifier1
Important signal expressions for molecular absorption
measurements are summarized in Table 13-2. Although
most expressions are written for conventional spectro-
photometers with PMT detection, they can easily be
adapted to other situations.
The source spectral radiant power [(@,),I and to-
tal radiant power (@,) incident on the sample cell(s)
depend on the source radiance (B,) and the collection
and transmission characteristics of the wavelength se-
lector and other optical components [Y(A)], as shown
in equations 13-8 and 13-9. Equations 13-10 and 13-11
apply when a monochromator is the wavelength selec-
tor; in the latter equation, B, is assumed to be constant
over A, I s.
Absorption, reflection, and scattering by the sam-
ple cell and its contents attenuate the incident radiation,
as illustrated in Figure 13-10. The attenuation by a sam-
ple cell filled with blank solution at a given wavelength
Probe is characterized by the reference transmission factor
tip
(T,). From equation 13-12, Tr is seen to be due to the
reflectance losses at the two cell walls [(I - pJ2], ab-
sorption and scattering within the cell walls (1 - a,),,
and absorption by the solvent and nonanalyte species
(1 - 4 .
The reflection loss at one cell wall is determined
by the reflectance at the air-cell wall (p,) and the cell
FIGURE 13-9 Probe-type photometer. Light
wall-solution interfaces (p,) (equation 13-13). For a
from a tungsten lamp is directed into the sample
solution through a fiber optic. The light passes typical glass or silica (quartz) cell filled with an aqueous
through the solution and is reflected by a mirror solution, the Fresnel equation (equation 3-8) indicates
back to a second fiber optic. This returning light that p, and p, are typically 4 and 0.370, respectively.
passes through an interference filter to a detector Thus Tr, considering only reflection losses, is about 91.5%
where the signal is processed as in a conventional (we will ignore multiple reflections for the time being).
photometer. (Courtesy of Brinkmann Instru- The exact value depends on the wavelength-dependent
ments, Inc., Westbury, N.Y.) refractive indices of the cell wall material and solution.
The absorption and scattering by the cell walls
sorbance data at a number of preselected wavelengths depend on the type, thickness, and quality (e.g., im-
may be printed or used in fitting routines for multicom- perfections) of the cell wall material (see Appendix B).
ponent analysis. Normally, a, is insignificant in the visible region and
Many software packages are also available for spe- becomes significant in the UV or NIR region at a wave-
cialized applications such as kinetic studies or chro- length dependent on the cell material.
matographic detection. More detail is presented in Sec- Absorption and scattering by the blank solution
tion 13-5. are wavelength dependent and determined by the cell
pathlength, the solvent absorptivity, the concentration
and absorptivity of all nonanalyte species, and the size
distribution and number of scatters. For wavelengths
13-2 SIGNAL AND NOISE EXPRESSIONS shorter than 190 nm. a factor accounting for absorption
by 0, should be added to the equations. Purging the
In this section we use and adapt equations developed entire optical path with N, can eliminate this absorption
in Chapters 2 to 5 to express quantitatively the rela- effect.
tionship of absorption readout signals and SIN to in- Equation 13-14 shows that the transmission factor
strumental and analyte parameters. with analyte solution in the sample cell (T,) is deter-
TABLE 13-2
Signal expressions for molecular absorption spectrophotometry"~"
I n c i d e n t u;ttli;ult p o w e r
S a m p l e transmission Cactor
K c f c r c n c c r e a d o u t signal
S a m p l e r e a d o u t signal
With m o n o c h r o m a t o ~
"Y(X) and T;,,,accoont for- the optical cllicicncy ot :ill component\ wed 101 wavclc~igthselection and ol the extcrn;il optical component\ bctwccn the source
:lnd the dctccto~-.Thcrclmc. the incident ( y x c t i a l ) r;rdi;int powcr 111 thew equ;ition\ actually repr-cscnt\ the iadiant power tnc~dcnton the detector when
tlrc soiirple cell i s removed. When the wavelength sclcctor 15 hefurc Ihc sample cell. the r;rdiant powcr incldent on the sanlple cell can he greater bccauw
ol lo\scs due to optical cornpr~nentshctwcci~the \;rinplc ccll and tletecto. I n ccluatioils 13-11. 13-1s. ;rnd 13-21, ;ill w;1vclcngtl~-tlcpc1ic1ei1tquanlitic\ aic
as.;umed to be con\tant tlvcr A,, i s .
Sec. 13-2 I Signal and Noise Expressions 367
Abs:rption and scattering by cell wall (e.g., W) and gains ( m G ) ,E, varies typically about two
orders of magnitude over the range 200 to 800 nm be-
cause of the dependence of To, and T,. and especially
B, and R(X). on wavelength.
Equation 13-22 defines the measured transmit-
tance as the ratio of equation 3-16 to equation 3-19,
where m and G are assumed to remain constant for the
sample and reference measurements. For a monochro-
FIGURE 13-10 Radiation losses in a sample cell. mator-based instrument. equation 13-23 applies where
The incident radiation (@,),, is attenuated at both W, H, and are assumed to remain constant for both
cell walls by reflection at the air-wall and wall- measurements. Finally, in equation 13-24, all wave-
solution interfaces and by scattering and absorp- length-dependent quantities [B,, Top, R(A). T,] except
tion within the cell wall. Inside the cell walls, €(A) and t(A) are assumed to be constant over A, i s
losses occur due to scattering and absorption by and identical for the sample and reference measure-
the solvent and blank species, and for standard ments.
or sample solutions, also because of absorption If €(A) is constant over the wavelength range passed
by the analyte. The radiant power transmitted by by the wavelength selector (e.g., A, t s for a mono-
the reference solution (a,), and the sample so-
lution (@,), is determined from equations 13-12 chromator-based system), equations 13-22 to 13-24 sim-
and 13-14, respectively. The incident ray is not plify to equation 13-25 and A is proportional to c. Strictly,
drawn perpendicular to the cell wall. as it nor- this is valid only if the incident radiation is monochro-
mally is, to illustrate the reflections that occur. matic (i.e.. infinitesimally small spectral bandpass). Be-
The effect of the multiple reflected rays (a) is cause the bandpass is finite in real instruments, the
discussed in Section 13-3. measured T o r A only approximates the spectral trans-
mittance or absorbance at the wavelength setting (A,)
mined by the same factors that affect T, plus naturally of the monchromator. For further discussions we will
the analyte spectral absorptance [a(A)]. The analyte drop the spectral designation (A) and write Beer's law,
spectral absorptance at a given wavelength is related to in the normal form (A = -log T = ~ b c ) In . doing so
the analyte molar concentration and molar absorptivity we realize that A, T, and E are effective values deter-
[€(A)] at that wavelength with equation 13-15. Here mined by a weighted average of the corresponding spec-
T(A) and A(h) are the spectral transmittance and ab- tral quantities [T(A), A(A), and E (A)] over A, t s. More-
sorbance due solely to the analyte (T, = I), respec- over. the proportionality between A and c is only an
tively. Often in the literature. the transmittance of the approximation, as we discuss in more detail in Section
solution, ignoring cell wall effects, is called the internal 13-3.
transmittance and is equal to T(A)(l - a,). The equations in Table 13-2 apply to multichannel
The reference readout signal (E,) is determined spectrophotometers with only minor modifications. For
by the spectral radiant power passed by the reference a diode array instrument, the sample and reference sig-
solution [(@,),I, the PMT responsivity, the PMT gain, nal expressions are applicable to a given detector ele-
and the signal processor gain, as shown in equation 13- ment in the array, where the wavelength range observed
16. Equations 13-17 and 13-18 are the more specific is determined by the width of a diode-array pixel and
forms that apply when a monochromator is used. In the dispersion of the spectrograph. As with vacuum
equation 13-18, all wavelength-dependent quantities are phototubes and photodiode detectors, the detector gain
assumed to be constant over A, +
s. Equations 13-19 (m) is unity. Because wavelength selection occurs after
to 13-21 are the corresponding equations that relate the the continuum radiation passes through the sample cell,
sample readout signal to the radiant power passed by (Oh), represents the effective incident spectral radiant
an analyte solution [(@,),I. In these equations, T, is power (i.e., the spectral radiant power incident on a
replaced by T,. detector element with the sample cell removed).
Equations 13-16 to 13-18 make it clear why the
reference signal is highly instrument and wavelength Signal-to-Noise Expressions
dependent. For a given analysis wavelength, the ref-
erence signal is instrument dependent because of dif- In this section we discuss the factors that affect the
ferences in the lamp radiance, the optical efficiency of precision with which the absorbance of a given solution
the wavelength selector (WHRTops), the detector re- can be measured. In other words, if we make repetitive
sponsivity [R(A)], and the gains (mG) among instru- absorption measurements on the same solution, what
ments. For a given instrument with a constant bandpass factors affect the measured standard deviation? We will
3 Chap. 13 1 Ultraviolet and Visible Molecular Absorption Spectrophotometry
see that noisc, readout resolut~on,a ~ l dcell positioning The RSD in E,, due to signal flicker noisc is spec-
reproducibility must all be con\idered. ified by the samplc flicker factor E,. Signal flicker noisc
arises from two sources as specified by two flicker fac-
Theory. The expressions for the relative stand- tors (equation 13-33). IIcre t, is thc source flicker fuctor
ard deviation (RSD) in absorbance (tr,/A) developed and <,
is the cell trunsrnission flicker factor.
in Section 5-5 apply to molecular absorption spectro- Cell transmission noisc is not noise in the normal
photometry. General and specific cquations are surn- sense, as it is not observed as Sluctuations during the
marized in Table 13-3. Equations 13-26 to 13-28 relate mcasurcmcnt oS a given test solution in a conventional
the standard deviation (SD) in absorbance (u,,) and samplc cell. The transmission flicker factor is a measure
tr,/A to the SD in the total sample signal (a,,). In this of random variation in the reference signal due to the
discussion, we will ncglcct the small contribution o f the sample cell being removed and replaced in thc optical
precision of the blank or 0% T measurements to the path between measurements. Thus it accounts for var-
magnitude of cr,/A. iations in the reference transmission factor (7;.) due t o
Equation 13-29 shows that u,, is due to shot and samplc cell placement. A random variation is observed
Slicker noise in the transmitted source sig~laland 0% T because the incident light beam is imaged onto a slightly
noise, as discussed previously (equation 5-46). The lamp diHere11t portion of the cell walls every time the cell is
signal shot noise and signal tlicker noise are calculated moved; the reflection, transmission, and scattering
from equations 13-30 and 13-31, respectively. The noise characteristics of the sample ccll vary spatially. The
with the source radiation blocked or 0%) T noise (a,,,) position of thc samplc ccll can also affect the portion
is seen from equation 13-32 to be due to dark current of the active surface of the detector that is illuminated
noise (u,,) and an amplifier-readout noisc (tr,,). and hence the effective detector responsivity for each
TABLE 73-3
Signal-to-noise expressions for molecular absorption spectrophotometrya
measurement. With flow cells, transmission flicker noise and E, are constantly ratioed. Formally, source flicker
can be caused by pump pulsations, bubbles. tempera- noise is due to nonfundamental fluctuations in the source
ture-dependent refractive index variations. or incom- radiance. The effective source flicker noise can be greater,
plete mixing of the components of the mobile phase. due to mechanical vibrations or varying magnetic fields
Equations 13-34 to 13-36 are the final expressions due to choppers and motors such as those used in dou-
for oAiA that account for all noise sources. In all these ble-beam instruments. Vibrations or nonreproducible
equations, the three terms in the brackets represent the imaging of the sample or reference beams onto the
contributions from signal shot noise, signal flicker noise, detector active surface results in additional noise if the
and 0% T noise. respectively. Equation 13-36 takes into detector responsivity in spatially dependent.
account that the gains (mG) are usually adjusted so that The cell transmission flicker factor varies from
E, equals the full-scale setting (E,) of the readout de- to It depends on the quality of the sample
vice or of the ADC in computerized instruments. cell, the condition of the sample cell (e.g., scratches,
If direct absorbance readout is provided by analog cleanliness of the surfaces), and the ability to position
logarithmic circuitry. the same equations apply in most the sample cell reproducibly. Generally, t2is larger with
cases. Here Efi represents the input voltage to the com- test tube-type cells, where the rotation of the cell is not
putational circuitry. which corresponds to 0 A . U . If easily controlled. Dual-wavelength measurement tech-
measurements are limited by readout resolution. a term niques compensate for both cell transmission and source
accounting for the uncertainty in absorbance units due flicker noise.
to readout resolution is added (see equations 5-59 and With low-resolution readout devices, 0% T noise
5-60), is often limited by readout resolution (u,, = u,). The
The blank noise (a,,)in absorbance units can be relative readout resolution is dependent on the char-
estimated with equation 13-37 (repeat of equation 6- acteristics of the readout device or A D C and can vary
13). The detection limit (DL) can be estimated from from 0.2% T o r 0.002 A.U. with analog readout (meters
this value and D L = ko,,/m, where lz is the confidence or recorders) to 0.001% T or 10-j A . U . with high-
factor and m is the calibration slope ( ~ b ) . resolution digital readout or ADCs. In modern spec-
trophotometers with sufficient readout resolution, the
Effect of Variables. The limiting noise sources limiting 0% T noise is usually dark current noise (a, >
in a given absorption measurement depend on the in- o,). Dark current noise is more likely to be the limiting
strument, the instrumental parameters, the sample cell, noise source under conditions where i, is smaller.
and the blank and analyte absorbances. Equation 13- Source modulation allows discrimination against lif dark
36 makes it clear that the measurement precision is current noise or amplifier-readout noise.
critically dependent on the reference photocathodic The shapes of the plots of uAiAvs. A for the three
current (i,), the source and cell transmission flicker fac- limiting cases (one type of noise term dominant) were
tors ( t l and t2), the relative 0% T noise (a,,/Ef,), and discussed previously (Figure 5-7). In all cases, u,/A
the transmittance. decreases with increasing A up to an absorbance of
The reference photocathodic current can vary typ- about 0.43, after which a,lA continually increases if
ically from 10-l2 to A . depending on the instru- 0% T noise is dominant. If signal shot noise is limiting,
ment and the instrumental conditions (A,, W ) chosen the increase in u,/A is delayed until the absorbance is
for a given analysis. For a given instrument, i, can easily about 0.87 and is more gradual. Precision continually
vary two to four orders of magnitude with changes in improves with increasing A if signal flicker noise is dom-
the slit width or analysis wavelength. Among instru- inant. The latter behavior is also observed if readout
ments set to a given analysis wavelength. i, varies greatly resolution is limiting in instruments that provide direct
due to differences in the incident radiant power as de- absorbance readout with analog logarithmic circuitry.
termined by the specific characteristics of the light source For real instruments, one or more noise sources
(e.g., B,), the wavelength selector and imaging optics may be dominant in different absorbance regions and
(e.g., WHCkT,,s), and the detector [e.g., R(A)].The the optimum absorbance with respect to precision usu-
transmission characteristics of the specific sample cell ally lies between about 0.43 and 1to 3. The contribution
and blank solution also influence the magnitude of the of signal shot noise and 0% T noise to @,/A vary as
reference signal. T - I * and T - l , respectively. Therefore, if signal flicker
The source flicker factor is typically lo-' to 10-5 noise is dominant at low absorbances, one or both of
and dependent on the type and age of the lamp and the these two types of noise will limit the precision at higher
quality of the lamp power supply. It is often nearer the absorbances and ultimately cause the precision to de-
lower limit for single-beam instruments using optical grade. Noise independent of the lamp signal (0% T
feedback or for double-beam instruments because E, noise) always eventually becomes limiting at some higher
Chap. 13 I Ultraviolet and Visible Molecular Absorption Spectrophotometry
absorbance where t h c lamp signal and hence noisc sources nificimt ( c . g . , signal flickcr a n d s h o t noise o r signal s h o t
related t o t h e lamp signal b e c o m e small. T h e s e limiting noise a n d 0%) T noise).
situations m a y n o t b e o b s e r v e d because t h e m a x i m u m Simple single-beam spectrophotornctcrs a n d p h o -
a b s o r b a n c e o l ~ t a i n a b l cis limitcd b y t h c display r a n g e t o m e t e r s often h a v e a large spectral bandpass, a n d t h u s
of t h e r e a d o u t device or b y excessive nonlinearity d u e provide a large light t h r o u g h p u t (large a,, a n d i,); they
t o stray radiation o r o t h e r factors (see Section 13-3). often u s e lower-resolution r e a d o u t devices (0.1 (% T o r
N o t e also t h a l is always a transition region between t h e 0.001 A.U. or worse). F o r these cases, noise m a y not
limiting regions o v e r which t w o types of noise a r e sig- b e obvious such that nleasurements a r c r e a d o u t rcso-
~ ~
(:I) (tl)
FIGURE 13-11 Precision plots for two spcctrophotometcrs. In (a) the
dependence of the precision (ts,lA) on ;~bsorbanccfor a simple spectro-
photometer (Turner 330) is shown. In curve 1, the precision is limited by
the readout resolution o f the spectroplioton~etcr'sanalog metcl- (0.2%
T). To obtain curves 2 and 3, a digital meter with a resolution of 0.001 %,
T w a s employed so that readout resolution is not limiting. In curve 2, the
sample ccll was moved between mcasurelncnts and precision is totally
limited by cell trmsmission flicker noise. For curve 3, the ccll was kept
stationary. I n this case source flickcr noise limits precision up to about A
= 1 after which 0% T becomes dominant. Signal shot noise is insignificant
a t a11 absorbanccs. Thc precision plot for a high quality double hcam
spectrophotomcter (Cary 118C) is shown in ( h ) . Hcrc sonrcc flickcr rmise
is limiting at low absorba~ices.As the absorbance iiicreases, source flicker
noisc and signal shot noise hccome comparable in ~n;lgnitudearid cven-
tually signal shot noisc heco~ncsdominant at rnodcrate a1xo1-banccs. With
further increases in absorbance, the signal shot noise decreases further in
magnitude and becomes comparahlc anti eventually insignificant com-
pared to the dark current noise at high ahsorbances. ((a) Adaptcd f'rorn
J . D. Inglc, Jr., A d . ('11inz. Acta, 88. 131 (1977), with pe~missiono f
Elsevlcr Science I'ublisbcrs.]
Sec. 13-2 / Signal and Noise Expressions 371
lution limited (a 0% T noise) at all absorbances. The should be improved until noise is observed. This can
precision characteristics of a simple spectrophotometer be accomplished by substituting a high-resolution read-
are illustrated in Figure 13-lla. out device (smaller a,) or by using scale expansion tech-
With higher-quality instruments. the light through- niques.
put is often less (i.e., smaller it). and the readout reso- Before the advent of high-resolution digital read-
lution is T o r A.U. or better. With such spectro- out devices and ADCs, scale expansion was a popular
photometers. noise fluctuations or cell positioning technique for increasing precision. It is still useful for
imprecision can normally be observed. Often source or some fixed-wavelength instruments and for displaying
transmission flicker noise is limiting at the detection spectra. Scale expansion allows a portion of the trans-
limit and lower absorbances, while signal shot noise or mittance scale to be displayed on a readout device. This
0 % T noise become dominant at moderate or high ab- is often accomplished by calibrated multiplication and
sorbance~.The absorbance at which the precision starts suppression of the signal processor output signal before
to decrease is largely a function of the reference pho- it is sent to the readout device. For example, consider
tocathodic current. At wavelengths where the source the case where a 1-V full-scale meter with a resolution
radiance and detector responsivity are high (e.g., 400 of 1 mV is used to display the range 0 to 100% T . The
to 600 nm), i, is large and signal flicker noise dominates relative readout resolution is 10-3/1 = or
up to high absorbances. The reference photocathodic 0.1% T . If the readout signal is amplified by a factor
current becomes small at short wavelengths near 200 of exactly 10 and then 9 V is subtracted before display,
nm because the source radiance is low and often at long the readout scale now represents 100 to 90% T, E is
wavelengths (greater than 650 nm) because the detector
fs
effectively increased to 10 V , and the readout resolution
responsivity of many PMTs is low. Thus, for these is 0.01% T. If 8 V is subtracted, the readout scale rep-
wavelength regions, signal shot noise or dark current resents 90 to 80% T. Note that o,/A is proportional to
noise is more likely to dominate, particularly at high u,lEf,only if readout resolution is limiting. In the ex-
absorbances. This behavior also occurs when i, is small ample above, the precision at a given absorbance is
due to narrow slit widths or a large blank absorbance improved by a factor of 10 only if readout resolution is
( T , << 1). The precision characteristics of a double- limiting before and after expansion.
beam spectrophotometer are illustrated in Figure For instruments that use logarithmic circuitry to
13-llb. provide direct absorbance readout, similar electronic
For NIR measurements with PbS detectors, the manipulation can be used to improve readout resolution
detector noise is considerably higher than for PMTs, by displaying a smaller portion of the absorbance scale
PTs, or PDs. such that 0% T noise limits the precision (e.g., 1.0 to 1.5 A . U . instead of 0.0 to 3.0 A.U.). In
at all absorbances.With multichannel detectors 0% T some instruments, the signal processor gain is auto-
noise is often limiting at all absorbances and is due to matically increased by a factor of 10 when the trans-
electronic readout noise. The relative 0% T noise mittance falls below 10% T to enhance the readout
(oo,lEfi) is typically 0.01 to 0.05% for diode signals near resolution for small sample signals. With computer-based
saturation. or multichannel spectrophotometers. one can choose to
For instruments in which the slit width can be display a portion of the transmittance or absorbance
controlled. precision is improved by increasing the slit scale to accentuate detail; however, the uncertainty due
width to increase it until measurements become limited to readout is actually limited by the readout resolution
by signal flicker noise or readout resolution or the wave- of the ADC.
length resolution is insufficient. Source flicker noise, In the past it was common to use analyte solutions
signal shot noise, and dark current noise can be de- of known concentration and hence transmittance to set
creased by increasing the time constant or integration one or both ends of a transmittance readout scale. For
time (i.e., reducing K or Af). The l/f nature of source example, with a standard solution of 90% T i n the sam-
flicker noise precludes reducing its magnitude signifi- ple cell, the 100% T control can be used to adjust the
cantly for noise bandwidths smaller than 0.1 Hz. readout to 100% T. Next. the 0% T control can be used
to adjust the readout to 0% T with a standard solution
normally yielding 80% '0. Now sample solutions with
Scale Expansion. With low-resolution readout transmittances in the range 90 to 80% T can be meas-
devices. the readout resolution often limits the precision ured with 10 times the readout resolution under normal
[o,lA = (a, /Ef,)/(- T ln T)]. which causes a large un- conditions. Thus this chemical scale expansion tech-
certainty in the measured absorbance for low absorb- nique (often called differential or high-precision spec-
ances and also for high absorbances with transmittance trophotometry) can also provide the same advantages
readout. To utilize the inherent capabilities of an in- as electronic scale expansion. However. this tedious
strument fully, the relative readout resolution (cr,/Ei,) technique is rarely used in modern instruments with
Chap. 13 I Ultraviolet and Visible Mo/ecular Absorption Spectrophotometry
high-rewlution d~gitalrcadout, clrct~onicscale expan- decreasing in a single-beam instrument and the blank
slon, andlor chrect abwrbance readout. mcasurerncnt is always made before the standard meas-
urement, the tr;insmittancc calculated will always be
too low; this yields a positive intercept corresponding
to the absorbance change due t o drift between the blank
and sarnpie measurement.
Random error can also result in a nonzero inter-
From Bccr's law, a calibration p!ot of A vs. c from cept. Whcn calibration data arc fit by least-squares
~ncasuremcntson a series of standards should be linear methods, the intercept is normally not zero. Errors in
with an intercept of zero. Jllowevcr, calibration curves preparation of standards can cause similar effects. Sta-
are sonietirncs found to be nonlinear or have a nonzcro tistical mcthods can be used to evaluate the uncertainty
intercept. These effects are rarely due to Beer's law in the intercept and determine if it differs significantly
being invalid, but rather are a consequence of meas- from zero.
uremcnt conditions in which the assumptions used to
derive Bccr's law (see Tahle 3-1) are not valid. The
eviatiola and negative eviation from lin-
earity arc ~rsedto clcscribc nonlinear calibration curves If several species absorb at a given wavelength, the
that bend toward or away from the concentration axis, add~tivcform ol Beer'\ law can be u\ed:
respectively. fience in the case of a negative deviation,
the calibration slope decreases with increasing analyte
concentration. We now discuss the specific instrumental
and chemical effects that cause such apparent deviations
from Beer's law.
where the subscript i identifies that quantity for the it11
absorbing species. For the moment we will assume that
the blank compensates for the absorbance due to con-
A nonzero intercept is usually due to a n improper blank comitants. but consider that the analyte can exist in
rneasurcment or adjustment or to nonequivalent meas- several chemical forms in solution, which may be in
urement conditions for the blank and standard solu- ey uilibriurn.
tions. The blank mcasurerncnt should account for all If only one of the chemical forms absorbs at the
the attenuation of the incident bearn by the sample cell monitored wavelength, we can rewrite Beer's law as
filled with standard solution except for that due to the
analyte. This implies that the reference transmission
factor T,(equation 13-12) should he identical for the
standard and blank measurements. Differences can oc- where c, is the total or analytical concentration (the
cur because of a11 improper blank [i.c.. the composition total concentration of analyte placed into solution) and
of thc blank solution is difi'crent from the matrix of the f is the fraction of c,, that exists in the absorbing form.
standard solutions such that the absorption by nonan- Thus we see that the "c" in Beer's law is not necessarily
alyte species ( a , ) or the refractive index which affects the analyte concentration but more precisely the con-
the reflection Ioss (p,) is different]. centration of the absorbing form of the analyte. Clearly,
If different and nonequivalent sample cells are a linear calibration curvc is observed only i f f is inde-
used for blank and standard measurements, differences pendent of the analytical concentration (c,,). A negative
in cell wall absorption and scattering (a,.) or reflection or positive deviation in the calibration curvc is observed
properties ((I,.) can cause a nonzero interccpt. With dou- if J' decreases or increases with increasing c,, respcc-
ble-beam instruments, differences in cell properties are tively.
adjusted by zeroing the absorbance with blank in both The situation becomes more complex if two or
cclls. In single-beam instruments, either one sample cell more chemical forms of the analytc absorb at the anal-
should be used for all solutions or n measurement should ysis wavelength. Consider an equilibrium between a
be made with the blank solution in each cell. Correction monomer (M) and a dimer (D) in solution (2M + D)
factors can be developed to account for sample cclls with an equilibrium constant K ( K = c , l c ~ ) . The total
with different transmission charactcristicx. Drifts in the absorbance due to both species from equation 13-38 is
lamp radiance, PM'I' or signal processor gain, or dc- given hy
teclor responsivity can also cause apparent nonzero i n -
tercepts. For example, if the lamp radiance is constandy
Sec. 13-3 I Apparent Deviations from Beer's Law 373
The analytical concentration (c,) is found from libria. the pH can be adjusted or buffered to force the
equilibrium predominantly in one direction. For ex-
ample, at high pH (> 12). Cr(VI) exists primarily as
yellow chromate, and a linear calibration curve is ob-
If equation 13-41 is solved for c, and c, and the results served at an analysis wavelength of 370 nm. At very
are substituted into equation 13-40, we obtain low pHs, nonlinearity is observed at most wavelengths
because of equilibria between several absorbing species
(e.g., HCr20;, H,CrO,, chromium sulfate com-
plexes).
Equilibria involving metal complexes or the acid
and base forms of a species are also pH dependent. The
latter case is illustrated in Figure 13-12. Absorption
If E D = EM, the second term vanishes and a linear spectra of the indicator phenol red are shown as a func-
calibration curve is obtained. In other cases, the con- tion of pH. The plot of absorbance vs. total indicator
tribution of the second term in equation 13-42 to the concentration can be made linear by adjusting the p H
total absorbance decreases with increasing analytical of all solutions to a value that results in the indicator
concentration and causes a positive or negative devia- being totally unprotonated or protonated or that main-
tion depending on the sign of the quantity ( 2 ~ , - c,). tains a constant ratio for the concentration of the two
The fraction of the total analytical concentration in the forms for any total indicator concentration.
dimer form (c,/ca) increases with increasing c,. Thus if Note in Figure 13-12 that the absorbance is in-
e, > kM, a positive deviation is observed because the dependent of pH at about 495 nm. This wavelength is
decrease in the absorbance due to the loss of two mon- called the isobestic or isoabsorptive point. An isobestic
omer molecules is more than compensated by the gain point is observed when two absorbing species in equi-
in absorbance due to the formation of one dimer mol- librium have the same molar absorptivity at one wave-
ecule. By contrast, a negative deviation occurs if length. In fact, the observation of an isobestic point is
ED < 2~~ because the increase in dimer absorbance a criterion to prove the existence of two interconvertible
does not compensate for the loss in monomer absorb- absorbing forms of a species with overlapping spectra.
ance. Negative and positive deviations due to dimer The isobestic point is an analytically useful wavelength
formation are observed with methylene blue when the because a linear calibration curve is obtained at this
absorbance is monitored at 664 and 600 nm. respec- wavelength without controlling such solution conditions
tively. as pH.
Many types of equilibria involving the analyte are
possible, and the shape of the calibration plot depends Nonlinearity due to Other Chemical Effects
on the particular case. For example, if a metal is de-
termined by the absorbance of a metal complex, the Nonlinearity can also occur if the analyte molar ab-
total absorbance can be due to a number of complexes sorptivity is dependent on the analyte concentration.
involving different numbers of ligands associated with Such effects are usually minor and occur at relatively
the metal or polynuclear complexes. Each complex can high concentrations (> M). Differences in solute-
have a different spectrum and thus different molar ab- solvent interactions, solute-solute interactions, or hy-
sorptivities at a given wavelength. The relative fraction drogen bonding at high concentrations can change the
of each metal complex can vary with the total metal chemical or electrostatic environment and hence the
concentration. Sometimes it is possible to choose the absorptivity of the analyte.
initial ligand concentration so that only one metal com- Changes in the solution refractive index (T) with
plex forms in a significant amount. Alternatively, with analyte concentration can also cause nonlinearity by
a sufficient excess of ligand, the fraction of each com- altering the position. size, or solid angle of the image
plex formed can be independent of the initial metal transmitted to the detector. the reflectance loss at the
concentration. cell wall-solution interface (p,), or the analyte molar
Many equilibria involving analyte species are pH absorptivity. In the last case. c in Beer's law is replaced
dependent. A classic example is Cr(V1) in solution by cql(q2 t 21,.
(Cr20,2- + H20 2Hf + 2Cr0,2-). The relative
fraction of yellow chromate to orange dichromate varies
with the Cr(V1) concentration and the pH; positive or
negative deviations can be observed depending on the We have repeatedly emphasized that Beer's law is valid
monitored wavelength. For such pH-dependent equi- only for monochromatic radiation. To illustrate the ef-
74 Chap. 73 / Ultraviolet and Visible Molecular Absorption Spectrophotometry
lcct ot nonnionochromntic, Iieterochromnt~c,or poly- eracc tran\mlssion lac to^ , or detector re\ponslvity at thc
chrom&c ~'id~ktion rn r e d mc'i\urerncnt\, wc f m t con- two wavelength\. From equation 13-44, the abwrhnncc
s d c r 'I \ ~ n ~ p lexample
e in which the \~rrnplc cell I \ 15 glvctl by
~lli~rnin,~ted w ~ t htwo riionoch~onint~cline\ of wavc-
length\ iland A, In thls c'r\e, the ~ntegrdl\In ccpltlon
13-22 reduce to \umm'itlon\ 01 the c l u m t i t ~ o'it two
wavclength~ I1 we ,i\\umc th,it Beer'\ law I \ followed
tor each wavclcngth (eqi~,ition13-25), c q i ~ ~ i t ~13-22 on
\lmpl~f~e\ to If th14 cqu,rt~on14 drtfe~entl'itedwlth re\pcct t o h c , we
find that the cffectlvc cih\orpt~v~ty( t ) 15 glvcn by
1' =
1()
----
"I"
x [I + r x 10'" ")"']
(13-44) F =
t , + rt,
1 + r 1 t r
-
Note tli,lt r 14 the w e ~ g l l t ~t a~c~t og ~for the i~ntle\ired and linearity is obscrved with an cl'fcctive molar ah-
cont~lbutionand account\ to1 d~ttercncc\In the \ o u ~ c c sorptivity that is a weighted average of a , and s,. For
r'id~ancc,wavelength \elector optical citiciency. I efer- large absorbance ( h c 3c-) with F , >> F,, we find that
Sec. 13-3 I Apparent Deviations from Beer's Law 375
E = e2. Hence the least absorbed wavelength controls the slit function is modeled as a triangular function
the calibration slope because effectively all the radiation (equation 3-60), and the absorption band shape [de-
at X, is absorbed, and further increases can only atten- pendence of &(A)on A] is described as a Gaussian func-
uate the radiation at h,. tion (equation A-3). The results of such studies are now
These results pinpoint the two major effects of summarized.
polychromatic radiation. First, the calibration slope at First consider the case where the monochromator
low analyte concentrations is lower than it would be at wavelength (A,) is set to the wavelength of maximum
the wavelength of highest molar absorptivity (i.e., el is absorption (A,) of an absorption band of true half-width
greater than e given by equation 13-47). Second, neg- AX. With an infinitesimally small spectral bandpass or
ative deviations occur at larger absorbances (i.e., e de- monochromatic radiation. we can measure the absorb-
creases with increasing c from the value given by equa- ance at the maximum (A,,), calculate the molar ab-
tion 13-47 to e,). The magnitude of these two effects sorptivity at the maximum (E,) determine the true half-
depends on the difference between e, and E, and the width. and obtain a linear calibration curve. We are
value of r as illustrated in Figure 13-13. concerned with how a finite spectral bandpass (s) affects
The same type of treatment can be extended to the observed absorbance (A), the calculated effective
an infinite number of wavelengths as would be the case molar absorptivity (E),and the linearity. It is convenient
with most instruments that use a continuum source. to express the results in terms of how AIA, or ele,
Here the wavelength range incident on the sample is vary with the bandwidth ratio (siAh). As seen in Figure
determined by the transmission characteristics of the 13-14a, the observed absorbance with a finite bandpass
wavelength selector. To evaluate the integrals in equa- is less than A,. However, the calibration curve is rea-
tion 13-22, it is necessary to describe functionally the sonably linear up to 1.5 A . U . if s1AX a 0.43. With
wavelength dependence of all quantities in the integrals sIhX = 0.43, the effective molar absorptivity is about
(e.g., lamp radiance. molar absorptivity). Even if this 92% of em.For a spectral bandpass such that s1AX < 0.1,
difficult task is accomplished, it is not usually possible the observed absorbance is within 0.4% of A, as shown
to obtain an analytical function for the dependence of in Figure 13-14b. Figure 13-14 also illustrates that the
A on c or other variables. However, qualitatively, the observed absorbance is much less than A, and serious
effects of polychromatic radiation on the shape of the negative deviations occur when the spectral bandpass
calibration curve are the same as described for the sim- approaches the true half-width or is greater.
ple two-wavelength example. From these results we can develop some practical
Numerical integrations have been carried out with rules of thumb. The monochromator spectral bandpass
equations similar to equation 13-24 where only the or the filter half-width should be less than 1110 of the
wavelength dependencies of the molar absorptivity and true half-width of the absorption band when the wave-
the monochromator slit function are considered. Often. length is set to the peak maximum. This ensures good
No stray ladiatmn
present
within the bandpass. In this case. at higher absorbances traverse the same thickness of the absorbing medium.
the absorbance measured can be too high and a positive This assumption is false if the cell walls are not parallel
deviation occurs. or flat or if the incident beam is not collimated. To
Standard procedures are available to evaluate the illustrate this effect. consider the simple case where half
stray radiation readout fraction. Often they are based of the rays in the incident beam experience pathlength
on measuring the absorbance of a concentrated solution b, and the other half experience b,. The measured trans-
or a cutoff filter at a wavelength where the true ab- mittance is given by
sorbance is large enough that T << r. The observed
absorbance is used to calculate r (A' = -log r). For
example, NaNO, (50 g L-l). NaI (10 g LP1), or NaBr
(10 g L-') are used to evaluate the stray radiation at
340, 220. and 200 nm, respectively. Such procedures Equation 13-54 is very similar in form to equation 13-
assume that stray radiation is the only cause of nonlin- 43, which describes the case of two wavelengths being
earity and usually underestimate the effective stray ra- absorbed with different molar absorptivities. Thus the
diation in a particular analysis because the test solution measured T or A values are between those that would
or filter absorbs more of the stray radiation than the be measured separately with each pathlength. If the
analyte of interest. difference in the pathlengths is great, a negative devia-
Note that r evaluated with the procedures above tion is observed at higher absorbances where the rays
is different than the stray radiation fraction f because traveling the shorter pathlength dominate the trans-
r depends on the differences in the reference transmis- mitted radiant power.
sion factor and the detector responsivity for radiation Pathlength effects are usually most important for
within the bandpass [(a,),,]and outside the bandpass test tube-shaped cells, as illustrated in Figure 13-17a.
[(O,),,]. Consider a simple case where the source emits Even if the width of the incident beam is one-half the
at only two wavelengths (A, and A,) with equal radiance. cell diameter. the effective pathlength and measured
If A, is selected for analysis and f = 1'36, then 1% of absorbances are only about 2.5% low relative to that
the radiation at A, is incident on the sample cell as the expected from the sample cell diameter and nonline-
stray radiation component. If the detector responsivity arity is insignificant even up to absorbances near 2. With
is 10 times greater at A,, r = 0.1. rectangular and cylindrical (flat window) sample cells,
Stray radiation is more significant at analysis the pathlength for different rays varies primarily be-
wavelengths where the product of the source radiance cause the incident beam is never perfectly collimated,
and detector responsivity [B,R(A)] is low compared to
the average for the rest of the wavelength range. Nor-
mally, r is largest from 200 to 220 nm with a D, lamp
and 340 to 400 nm with a tungsten lamp because the
stray radiation arises from wavelength regions where
the lamp radiance is greater. The stray radiation read-
out fraction is often greater in the NIR region than the
visible region because R(A) is smaller for longer wave-
lengths. Stray radiation can be reduced by using ab-
sorption band filters in conjunction with the mono- FlGURE 13-17 Examples of variability in path-
chromator, double monochromators, or holographic length. Top views are shown of the incident ra-
gratings. The first two options cannot be used with diode diation entering and traversing a test tube-shaped
array-based spectrophotometers. for which stray ra- sample cell in (a) and a standard spectrophotom-
diation effects can be particularly troublesome because eter cell in (b). For the test tube-shaped sample
of the large aperture in the focal plane. cell, the pathlength varies across the beam cross
section even if the incident beam is collimated.
Other Instrumental Causes of Nonlinearity In addition, rays impingent on different portions
of the sample cell are refracted differently and
experience different pathlengths. If the rays in
There are numerous minor causes of nonlinearity. Mosl the incident beam are not parallel as shown in
are significant only for applications requiring very high (b), the effective pathlength is greater than b and
accuracy. varies with the angle of the incident ray. The
maximum angle of divergence (8) of an incident
Variability in Pathlength. Beer's law is based ray with respect to the normal to the cell wall is
on the assumntion that all ravs in the incident beam shown.
Chap. 73 I Ultraviolet and Visible Molecular Absorption Spectrophotometry
as chown in E ~ g i i ~13-1717
e However, even with :I mocl-
eratc d~vergc~lcc mgle 01 13 5" lor the ~nclclentbc'rm,
the cl tcct~veptthlength and nieasurccl ,rhsotlm~cc,Ire
only O 5% too h g h '~ndnonl~ne,rnty14 ~ n \ ~ g n ~ t i c a n t
Miscellaneous Causes. Nonideal performance version of the analyte to its absorbing form as discussed
of the detector (e.g., hysteresis), the signal processor, in Section 13-5. Often separation steps (e.g., filtering,
or the readout device can affect linearity. Thus the char- solvent extraction) are necessary to eliminate interfer-
acteristics of these components must be such that the ents, and in some applications. spectrophotometry is
transmittance readout signals are directly proportional used as the detection technique for chromatography
to the radiant power incident on the detector. With (see Section 13-5).
direct absorbance readout, the accuracy and linearity
of logarithmic conversion can be limiting. Incorrect set- Choice of Solution Conditions. In analytical
ting of 0% T on the readout scale or improper sub- procedures, one must first determine the absorbing form
traction of the dark current signal from the total readout of the analyte to be used. The nature of the absorbing
signals can also cause nonlinearity. species greatly influences the range of concentrations
to be measured, the calibration sensitivity, the detection
limit, the precision, the accuracy, and the linearity. For
13-4 METHODOLOGY AND PERFORMANCE trace analysis, the absorptivity of the absorbing species
CHARACTERISTICS must be large enough to yield a good detectability and
precision. The chemical nature of the absorbing species
In this section we explore the factors that influence the affects how it interacts with potential interferents or
use and performance characteristics of quantitative mo- takes part in equilibria that can cause nonlinearity. Sim-
lecular absorption spectrophotometry. Usually the an- ilarly, the spectral characteristics of the absorbing spe-
alyte concentration in the analytical sample is evaluated cies determine the potential for spectral overlap with
from the sample absorbance and a calibration curve. other species.
The calibration curve is most often generated from ab- The solvent and such solution conditions as pH,
sorption measurements on external standards. ionic strength, and temperature should be optimized
and controlled to minimize interferences and nonline-
General Considerations in Quantitative arity due to equilibria and to maximize the time stability
Analysis of the absorbing form (i.e., to prevent decomposition
due to oxidation. photodecomposition, or other mech-
Sample Treatment, Sample treatment varies anisms). When an analyticai reaction is used to form
considerably with the application. Because most deter- an absorbing derivative, the reagent concentration(s)
minations are carried out on solution samples. solids must also be optimized to achieve the goals noted above.
must be dissolved. Sometimes the analyte can be de- Additionally, the time and temperature required to form
termined directly from the absorbance of its original the absorbing product, the order of addition of re-
chemical form. More commonly, derivatization is nec- agents, and the reaction of concomitants with the re-
essary because the absorption bands of the analyte in agent must be considered. Masking reagents are some-
its original chemical form possess inadequate absorp- times added to block the reaction of concomitants with
tivity or are in an unsuitable wavelength range. De- the analytical reagent. Ideally, the conversion of the
rivatization involves converting the analyte to a strongly analyte to the measured absorbing form should be quan-
absorbing form by a selective analytical reaction with titative. Otherwise, the efficiency of conversion should
appropriate reagents. The absorbance of a reaction be constant irrespective of the concentration of the an-
product is related to the original analyte concentration alyte or the concomitants. When solvent extraction is
by external calibration. Usually, an excess of the re- employed, the effect of the type of solvent on the ex-
agent(~)is added, and the analytical reaction is allowed traction efficiency for the analyte and concomitants, the
to proceed to equilibrium before measurements are made. molar absorptivity, and the spectral positions of the
Alternatively, the rate of the chemical reaction can be absorption bands should be evaluated.
related to the analyte concentration, or the analyte can
be titrated with a standard reagent that reacts with it Choice of Wavelength and Spectral Bandpass.
with known stoichiometry. The reaction between the For standard quantitative procedures, the absorption
analytical sample or standards and reagents can be car- spectrum of the absorbing form of the analyte is avail-
ried out directly in the sample cell or in another suitable able in the literature, and the wavelength (A,), a suitable
container. In the latter case, an aliquot of the reaction spectral bandpass. and solution conditions (e.g., re-
mixture is transferred to the sample cell after the re- agent concentrations) are specified. In developing a
action is complete. In such specialized applications as quantitative method for determining the analyte con-
titrations and kinetics studies and determinations, ab- centration, the choice of the absorbing species and ul-
sorption measurements are made prior to complete con- timately the particular absorption band and wavelength
382 Chap. 13 I Ultraviolet and Visible Molecular Absorption Spectrophotometry
TABLE 73-4
Spectrophotometer specifications
Specification Value"
A.U. This precision is maintained up to 1 to 3 A.kJ., realistic estimate of the practical UI, I~ecilusethe ma--
clepcnding on the limiting noise sources. b'or a given Pyte is ideally isolated from other absorbing species be-
analysis wavelength, the slit width is the primary vari- fore delcction.
able that the user can vary to change the shape of rhc
precision curve
bxccpt tor mccnuremcnts ,it low ,ibsorb,rnccs n e x
the detectlor1 hinlt, the ovc~,dlprccis~onol .I spect~o-
photometric deterrn~n~ition is often Iimltcd by the re-
produc~hilityof the sample xqulsitlon. sample t r c d -
ment, s m ~ p l cplcicenlcnl, 01 c,ihbration steps The ove~all
KSD can range from O 3 t o 596,depending on the \lt-
udtlon
analytical sample, but absent from the standards, that cause systematic errors include calibration errors (e.g.,
absorb or scatter at the analysis wavelength (i.e.. that improper standards), sampling errors, contamination
affect a,), that fluoresce significantly. that change the and loss, decomposition, and different measurement
reflection loss as the cell wall-solution interface (i.e., conditions (e.g., pH, temperature, sample cells with
that affect p,), or that alter the focusing of the beam different pathlengths or transmission characteristics) for
image on the detector. When an analytical reaction is the samples and standards.
used to convert the analyte to a strongly absorbing form,
blank spectral interference also arises from species in Absolute Accuracy, U p to this point, we have
the sample, which do not necessarily absorb themselves, been concerned primarily with the accuracy of deter-
but which react with the analytical reagent(s) to form minations with external calibration. A determination
a species absorbing at the analysis wavelength. can be accurate even if the measured absorbance is not
Most analyte interferences are nonspectral in na- the true absorbance as long as the calibration accounts
ture and caused by species in the sample, but not the for nonideal behavior. In several applications it is nec-
standards. which affect the concentration of the ab- essary to know the true absorbance. The instrumental
sorbing form of the analyte or the molar absorptivity specification of photometric accuracy is a measure of
of the absorbing form. Thus nonspectral interference the ability of the instrument to measure absorbance
occurs when the sample matrix causes any of the factors values accurately.
which affect the linearity or slope of the calibration From Beer's law (A = ~ b c )either
, F, b, or c can
curve (see Section 13-3) to be different between the be determined if the absolute absorbance is measured
standards and the sample. Concomitants can affect accurately and two of the other three quantities are
the equilibria between different forms of the analyte, known. For example, the molar absorptivity can be
the fraction of the analyte in the absorbing form, the determined from the absorbance if the pathlength and
electrostatic environment of the analyte, and thus the analyte concentration are accurately known. An accu-
molar absorptivity, and the relative contribution of po- rate value of the molar absorptivity is needed for cal-
lychromatic or stray radiation to the measurement. Also, ibration of b or c or for fundamental studies relating
the conversion efficiency of the analytical reaction can measured absorption parameters to fundamental quan-
be altered by species that interact with the analyte and tities (e.g., transition probability) or molecular struc-
prevent its reaction with the reagents or that consume ture. Similarly, the pathlength of a sample cell can be
a significant fraction of the reagents. determined from the absorbance if a solution with a
Interference effects are reduced by a careful choice known analyte concentration and molar absorptivity can
of the analysis wavelength, spectral bandpass, andlor be prepared. Sample cells with pathlengths accurate to
analytical reaction to achieve selectiv~tyfor the analyte. cm are available from the NBS. To determine b
Careful control and choice of such solution conditions or E accurately, extreme care must be exercised in pre-
as reagent concentrations, the type of solvent. pH, and paring solutions of known analyte concentration.
ionic strength can enhance selectivity and prevent shifts In analytical applications, the sample analyte con-
in equilibria due to species only in the sample. Sepa- centration can be directly calculated from the measured
ration of the analyte from some interferents by solvent sample absorbance and known values of h and F without
extraction or another technique is sometimes essential. resorting to external standardization. This is common
Filtering is often recommended to reduce potential scat- where the purity of compounds must be assayed or with
tering interferences. An internal blank is occasionally clinical enzymatic assays for which pure standards for
useful when analytical reactions are employed. Here enzymes or proteins are not readily available.
the absorption due to species in the original sample When absolute measurements are made, experi-
which do not react with the analytical reagent(s) can mental conditions must be adjusted to ensure linearity.
be compensated with a blank that is the sample reaction For very high accuracy work, the more subtle causes of
mixture minus a critical reagent needed for the analyt- nonlinearity and systematic errors such as multiple re-
ical reaction to proceed. In some cases, an internal flections and variable pathlengths must be considered.
blank can be made by adding another reagent which An angle of incidence 3% from the normal causes a
blocks the formation of the absorbing product. Stand- 0.1% positive error. The preparation of the blank so-
ard addition techniques can be used when analyte in- lution and measurement of the blank must be conducted
terferences affect the slope of the calibration plot. Ad- in a way that .the reference transmission factor (T,) is
ditional selectivity can be gained by using titration, the same for the sample and blank solutions. Thus a , ,
reaction-rate, derivative, o r dual-wavelength tech- p,, and a, in equation 13-12 must be the same for the
niques, as discussed in Section 13-5. reference and sample measurements. Additionally, the
Factors other than interference effects that can wavelength setting of the monochromator must be ac-
Chap. 13 1 Ultraviolet and Visible Molecular Absorption Spectrophotometry
the maximum absorbance Xi,, is related to the stoichi- eters have become powerful tools for kinetics studies
ometric ratio in the complex by llm = X,,/(l - X,,,). because complete absorption spectra can be recorded
In both the molar ratio and continuous variation meth- throughout the course of the reaction.
ods, negligible dissociation of the complex is often as- Some other fundamental applications include the
sumed; extrapolation techniques can be used if the dis- determination of molecular parameters and the molec-
sociation is not excessive as shown in the figure. ular weight. The shape. width. and wavelength of the
The Mollard method involves the measurement of absorption bands of a pure compound can be used to
the absorbance of only two solutions. One measure- elucidate the energy-level structure of the molecule, as
ment is made with metal ion analytical concentration discussed in Section 12-4. The integrated absorbance
c, and a large excess of ligand to yield absorbance A,. can be used to calculate fundamental quantities such as
The second absorption measurement is made on a so- the transition probability. as reviewed in Appendix F.
lution with an excess of metal ion compared to the If a compound is derivatized with a reagent possessing
ligand analytical concentration c, to yield absorbance a chromophore of known molar absorptivity that is not
A,. The amount of excess reactant in both cases is affected by the coupling, the molecular weight (MW)
adjusted so that dissociation of the complex is negligible can be calculated from the measured absorbance and
and the concentration of the metal complex is deter- the formula
mined by the concentration of limiting species. Under
these conditions. llm = (c,A,)I(c,A,). ecb
A variation of the Mollard method is the slope MW = -
A
ratio method, in which a series of solutions with excess
L and different c, and with excess M and different c, where c is concentration of the species in g L - '
are prepared. The ratio of the slopes of plots of A, vs.
c, and AL vs. cL is equal to llm. Curvature of the plots Conventional Quantitative Determinations
indicates that dissociation of the complex is significant.
Numerous schemes have been proposed to deter- As discussed in Section 13-4. the analyte is usually de-
mine equilibrium constants from the data acquired with termined by measuring the absorbance of an absorbing
the foregoing methods. For example, consider the con- product after equilibrium has been reached for the re-
tinuous variations method (Figure 13-18b) applied to a action of the analyte with selective reagents. Many an-
simple 1 : l metal complex (I = m = 1). If A, and A, alytical reactions have been proposed for a host of spe-
are the extrapolated and measured absorbances at the cies, and often numerous derivatization reactions are
maximum, respectively. the equilibrium concentrations available for a given analyte. Only a small percentage
(denoted by brackets) are given by of all developed procedures are accepted and used rou-
tinely.
A, In certain applications, the absorbance of the an-
[ML] = C, -
A1 alyte is measured directly provided that it is the pre-
TABLE 73-5
Spectrophotometric determination of inorganic anions
Spcc~cs Re'lgenrs
Determination of Organic Species. The ana- hydrazine to trap the pyruvate formed through hydra-
lytical reactions used for the determination of many zone formation. Although the reaction is quite selec-
organic species can be organized around the specific tive. such acids as glycolic and L-lactic acid can also be
functional group in the molecule that reacts with the oxidized and cause interference at high concentrations.
reagent(s). Table 13-6 lists some common reagents used Some clinical assays are based on the enzyme-
for different functional groups and typical molar ab- catalyzed reduction of the substrate with IVADH, in
sorptivities for the absorbing products that are meas- which case the decrease in absorbance due to the oxi-
ured. Many types of reactions are used including sub- dation of NADH to N A D + is monitored. It is also
stitution. addition, elimination. and rearrangement possible to couple the NADH formed in one reaction
reactions. The specificity and efficiency of the analytical to another reaction to form a product which has a higher
reaction and the molar absorptivity depend on the type molar absorptivity than NADH or to monitor other
and placement of other functional groups in the mol- reaction products. If the NAD'INADH couple is not
ecule. Because many of these reactions are selective for directly involved in the substrate reaction, one or more
only a type of functional group, they are used when the enzymatic reactions can be coupled to the original sub-
analyte is the primary species in the sample with the strate reaction such that NADH is eventually produced.
reactive moiety. For example, the determination of glucose is often based
In the clinical area, organics are often determined on the following reaction sequence:
as the substrate in very selective enzymatic reactions.
Many assays involve the oxidation of the substrate by glucose + ATP =
HK
glucose 6-phosphate + A D P
the coenzyme nicotinamide adenine dinucleotide
(13-60)
( N A D + ) . The concentration of the substrate is deter-
mined from the absorbance measured at 340 nm due to G-6PDH
TABLE 73-6
Spectrophotometric determination of organic species based on the type of function
group
Functional Molar absorptivit) Absorption
group Reagent (L mol- ' cm- ') maxima (nm)
Acid Pinacyanol
Carbonyl Dinitrophenylhydrazine
Chromotropic acid
3-Methyl-2-benzothiazolone
hydrazone
Ester Hydroxamic acid
Olefin Phenyl azide
(cyclic)
Peroxide Ferrous thiocyanate
Phenol 4-Aminoantipyrine
Pyrrole Dimethylamino-
benzaldehyde
Sulfonate Methylene blue
390 Chap. 73 i Ultraviolet and Visible Molecular Absorption Spectrophotometry
Optical a p c r t u r z
Wavelength
..-, d~al
Light shield
Fisher photometric
accessory Sot- automatic photometric
titrations. Notc that fiber optics arc
uscd lo direct thc light beam through
t h e titration vcsszl. (Courtesy of
Fisher Scientific Go.)
ance, or a related quantity, vcrsus the analyte concen- initial inlcrfering reaction, the analytical rate measure-
tration of standarcis. ment cam be made after the interfering reaction has
Reaction-rate methods offer several unique ad- reached equilibrium. Because of this greater specificity,
vantages compared to conventional equilibrium-based kinetics measurements can he faster than equilibrium
measurements in which the analytical reaction is 211- measurements i f tim--consumingseparations steps can
lowed to proceed to equilibrium before measurements be avoided. If the reaction is quite specific and there
are made. 'I'hcrc can be a considerable time savings for is no rate when the analyte is absent, further time sav-
reactions that require several minutes or more to reach ings can Ijc realized by omitting frequent blank meas-
equilibrium. Kinetics methods can hc morc selective urements. Nonstoichiometry, unstable reaction prod-
(greater freedom from intcrferenccs) for two primary ucts or reagents, or side reactions make some analytical
reasons. First, a kinetics measi~rcmcntis a relative reactions unsuitable or difficult to use for equilibrium-
measurement a n d only changes in the reaction monitor based determinations. Hlowevcr, the initial ratcs of such
signal are measured. Thus nonreacting spccies or in- reactions can often be related to the analyte conccn-
strurncntal factors that contribute to the absolute mag- tration. Finally, catalysts can be ~miquclydctcrmincd
nitude of the reaction monitor signal (e.g., the absolute since they affect the rate b ~ l tnot the cyuilibrium con-
absorbance), and that may vary from sample to sample, centrations.
but not during the reaction, do not interfere as t h ~ y The limitations of kinetics-hased methods should
would in an equilibrium-based method. Second, con-- also he untlerstood. The reaction rate must be in a
ditions can often be arranged so that thc kinetic con- suitable range and a half-life of a Scw milliseconds to
tribution of only one specics is significant over thc meas- tens of minutes is typicai. The lower limit is determined
urement period. For example, in the caw of a very fast Iry the rime necessary to mix the reagents, whercas the
See. 13-5 i Applications 393
upper limit is restricted by slow drifts in the instrumental software, the region over which the rate is constant
monitoring system and the need to have a reasonable (pseudo-zero order) can be deterrniried and the initial
analysis time. Reaction conditions such as temperature, rate calculated and reported. Several other rate com-
pH, and reagent concentrations must be carefully con- putational approaches are utilized and can be imple-
trolled as they often have a greater effect on the rate mented through software or analog or digital circuitry.
than on the final equilibrium concentrations. Detection In the variable-time method. the amount of time re-
limits and the SIN can be lower compared to equilib- quired for the reaction-monitor signal to change by a
rium-based methods because only a portion of the re- predetermined amount (AS) is measured. The change
action is used in the measurement (i.e., the change in in the reaction monitor signal in a predetermined time
absorbance measured is much less than the final ab- interval (At) is measured in the fixed-time method. In
sorbance at equilibrium). Also, the noise equivalent the derivative method, the instantaneous slope of the
bandpass must be large enough to prevent distortion of reaction monitor signal is measured. The fixed-time and
the changing reaction monitor signal. However, the in- variable-time methods are integral methods in which
creased selectivity against variable and uncompensated the measured quantity (ASlAt) approximates the true
absorbance by other species can result in practical de- instantaneous rate (dSIdt). If At is sufficiently small and
tection limits comparable to or even better than those the measurement is made very early in the reaction
achieved with equilibrium-based methods. Concomi- (pseudo-zero order conditions), the initial rate is meas-
tants which alter the rate of reaction cause analyte in- ured. Even if there is significant curvature in the re-
terference. Species which react with the reagents at action monitor signal over the measurement time At,
rates comparable to the analyte to form products that ASIAt is still linearly related to the analyte concentration
absorb at the monitored wavelength result in a blank under suitable conditions (e.g., first order for the an-
interference unless a suitable blank rate can be deter- alyte for fixed-time methods).
mined. Several techniques have been developed to com-
Reaction and measurement conditions are often pensate for imprecision in rate measurements caused
arranged so that the rate law is simple and the measured by variations in the rate constant due to run-to-run
rate, or another appropriate quantity, is directly pro- changes in the magnitude of variables that affect the
portional to the analyte concentration. Most commonly, rate constant. For first-order reactions, precision can
the reagent concentrations are in a sufficient excess that be improved by making rate measurements near the
the reaction is pseudo-first order with respect to the half-life, by computing the ratio of the rate measured
analyte in the initial stages of the reaction. at two different times during the reaction, or by esti-
To make rate measurements reproducible and mating the equilibrium position from reaction monitor
convenient, a spectrophotometer should have provision data taken over several half-lifes. It is also possible to
for mixing the sample and reagents in the sample cell, correct for changes in variable values by measuring the
temperature control. and suitable hardware andlor soft- magnitude of the variable for a given run and applying
ware for timing the acquisition of absorption data and a suitable correction factor.
computing the rate or a related quantity. Mixing in the Kinetics-based measurements with absorption
sample cell with magnetic stirring bars is suitable for monitoring are usually the method of choice for assay-
measuring rates of reactions with half-lifes of a few ing enzymes in clinical samples. For example, the en-
minutes or longer. Specialized accessories or instru- zymes L-lactate dehydrogenase and hexokinase are de-
ments based on stopped-flow mixing extend the range termined by measuring the rate of increase in absorbance
down to half-lifes on the order of milliseconds. Pres- due to the formation of NADH or NADPH in reactions
sure-jump, temperature-jump, or flash photolysis tech- described by equation 13-59 and equations 13-60 and
niques can be used for even faster reactions. Some in- 13-61, respectively. In this case, the substrate concen-
struments allow the reactions in several sample cells to trations are adjusted to be large enough that pseudo-
be monitored simultaneously. The sample cells are se- zero-order kinetics prevail. These same reactions are
quentially positioned in the light beam in a repetitive the basis for the determination of the substrates L-lac-
manner at a sufficient rate to allow absorbance vs. time tate and glucose. For substrate determinations, condi-
data to be acquired in a faithful manner for all cells. tions are arranged such that the substrate concentration
Highly automated instruments are available for high- is low enough that the reaction is first order in the
sample-throughput routine measurements. substrate.
Rate information is extracted from the time-vary- Inorganic and organic species are determined by
ing reaction monitor signal in several ways. With mod- measuring the rate of formation of absorbing products
ern computer-based instruments, curve-fitting tech- in noncatalytic reactions which are often used for equi-
niques are applied to the stored data. With suitable librium-based measurements. For example, phosphate
Chap. 13 / Ultraviolet and Visible Molecular Absorption Spectrophotornetry
can be determined by mc;lwring the 1nlt1,11,ib\orh:~ncc urcmcnt\ mu\t often be irnde 111 cloudy or turbid \us-
change due to the hrmttlon ot ~nolybtlenuniblue (the pen\lon\ I t tl~tt~cult to o l x ~ v ea \mall mdlyte &-
reduct~onof 12-MI'A formed In re'ict~on d o c r ~ b c dIn \orption band In ,i convcntwndl ah\orption spcct~urn
equ'itlon 13-58) I he ienction I \ peudo-11rst order In when ~t15 \upcr~mpo\edon top of LI Idlge ,lnd ~clntrvcly
pho\ph,~tc11 the reagent concentr,ltlon\ ,Ire suil~c~ently constant bxkgrounct h \ o r b m c c ba\eline In kinetlcs
Lrrge lnorganlc c,~tdly\t\m,ly bc detcrmlned w ~ t h\lm- s t u d m or niea\urcment\, the reference wnvelength e m
ildr procedure\ be \ct to 'I v,due lor which the absorhmce doe\ not
change duling the course of the r e x t m n Dual-wdve-
ther Quantitative length mcasur elnents c m even Lornpensatc lor conconi-
~ t m abwrptron
t when the m d y t e and concomlrnnl bands
Derivative Techniques. Some spectropho- are ot 'ibout the u m e brc'ldth and \eriou\ly overlap,
tometers allow the first derivative or higher-order dc- 1' s {liown In P~gurc13-22
rivatives of the absorption spectrum to be calculated In A i ~ t ~ oton di\cr~nmatlon'lg,tin\t jpcctral In-
and displayed (e.g., dAIriX vs. A) by the techniques tertcrence4, D W tcchn~quec,~lJord' ~ d d ~ t ~ oadvan-nd
discussect in Section 13-1. As shown in Figure 13-21, tages There 1 4 mnlc compen\,\tmn ior s'lrnple cell cr-
derivative spectra can he used to emphasize and distin- rors The light benms for both wavelength\ tollow
guish overlapping bands, to locate the maximum of a ldent~calhght path\ through one sample cell m d strike
broad hand, or to extract quantitative information when the \,me portlon of the photocathode of thc PMT Thu\
the analyte absorption band is overlapped by a con- sy\tem,rtic elrors due to d~tfercncesin the tran\mlsslon
comitant absorption band that is not compensated with characten\tlc\ 01 d ~ f l e ~ e cells,
nt the positioning ot the
the blank measurement. cell, or the refritct~vc~ n d e xof dillcrcnt s'mple\ are
Derivative spectrophotornetry is particularly ad- compcns,lted to the cxtcnt t h d they arc ~ndependent
vantageous when a narrow analyte absorption band is of wavelength Random error due to \ample cell p l x e -
superimposed on top of a relatively flat or gradually merit c,rn dl40 be reduced
changing absorbance due to turbidity or overlap with Dependmg on the \ituation, the precl\ion of D W
the side of a broad interferent absorption band. The me;isurenient\ can be poorer t h m convent~on,tl4111glc-
derivative spectrum emphasizes the contribution from wavelength measurement\ A l w , strdy r a d ~ , ~ t ~ and oii
the analyte for which the change in absorbance with polychron~~tt~c rad~atmnc m c m \ e more scr~ousnonlin-
wavelength is much greater than that for the hack- earity and even roll-over In the cahbrat~oncurve
ground (i.e., the contribution from the background is
:I tlat baseline). The difference hetween the niaximuin
and minimum values of the derivative is proportional Difference Spectra Techn~ques. In d ~ f t e ~ e n c c
to the analyte concentration cvcn in the presence o f an spcctroscopy. the d~iferencebetwccn spcctr'i 14 dl\-
uncompensated and variable background absorbance. played or the c~h\orb,incc vdluet 'it a spcc~flcwc~velength
are cubtrdcted for two sample\ of \lightly diftcrent com-
Dual-Wavelength Techntques. Some spcctro- pos~tlonor phy\lcal statc Recau\e common fcc~turerIn
photometers c m be configured to output the diiicrcnce the two \pcctrd cmcel, the cl~tference\pectrum x c c n -
In '~bsorbdnce(AA) between two w~weleiigth\,nt dls- h a t e \ the subtle d~tfelenccsIn ,ih\orpt~ondue to var-
cussed in Sect~on13-1 Thi\ du;il-w,~vclength(DW) op- idtion\ III solvent, p H , tempcrdture, or c~n,ilyteor rc-
tlon 1s part~culdrlyu\eful when it 15 drlhcult to prepare dgent concentration For cxdmple, the d ~ f i e r c n c c
a blank that compen\dte\ for cth\orpt~onby other \pe- \pectrum oi some bdrl~tur~ltes In (1 45
cles or scattcrmg The he,m\ corrc\pondlng to A , and pH 10 butfel over the rmge 230 to 280 nm is highly
A, are normally \clcctcd to be the \<inlplcmcl icference c h d ~ a c t e r ~ \ t When
~c only one component is prewnt,
be,\m\, re\pect~vcly Oltcn, X, i \ d l u \ t e d to a wavc- the d~ffelencein absorb,tnce at 260 nm I \ useful for
length corre\pondlng to annlytc dhsorption whde A, 1s quant~t'ltlvedetermm'itions
\et to ,I wavelength where the m d y t c doe\ n o t h s o r b D~llercncespectra c m be obtnmed by placrng the
(e g , the b,~\eot the an,tlyte absorption bnnd) 01 ,In two test wlution\ 111 the rctcrence m d \,imple cell\ ot
isobe\tlc point A \pectrum 14 obta~rlcdby keep~ngA , double-beam ~pectrophotometeror hy tnk~ngthe drl-
constdnt m d scmnlng only A, over the analytc d b \ o r p ference betwccn two \pectra tli'1t 'tre \uccewvelq \tored
tron hand. Thus the \drnple I \ u\ed ,i\ n blank m d AA Bec,lu\e m a l l a l m r h n c e dlftcrcnces ,ire otten nieii\-
1s used ds the dndlyt~cdsrgn'rl proport~on,llto the ,In- used, the \pectrophotonieter lnust be c'1phle ot hlgh
dytc conccntrdtwn photometr~cp r e c ~ o nm d ,~ccu~;icy, s,lmplc cell\ mu\t
Dud-wnvclcngth nlcns~~rcnient\ tind w~de\prc,d be closely inntched, m d \ample ccll plxcmcnt must be
use ~nb~ochem~cdl applrcat~onswhere db\orptlon mens- rcpr oduc~ble
First
spectrum
3i0 io
Wavelength, n m Wavelength, n m
(a 0)
Normal
S~ectra
First
Derivatives
to polychromatic radiation. The radiance provided by tometers because the lower throughput would reduce
other mercury lines, especially phosphors. is lower than the radiant power to a level that would increase the
that at 254 nm; the baseline noise is often greater. Other baseline noise. The wavelength versatility provides sev-
line sources such as Cd lamps (229 and 326 nm) and eral advantages. The wavelength of optimum response
Zn lamps (208 and 214 nm) are used in particular where for the compound(s) can be found and then used in
the compounds of interest only absorb at wavelengths determinations. The wavelength is chosen both to max-
shorter than 254 nm. imize the absorbance and to minimize the response from
In dual-channel absorption detectors, radiation other species whose chromatographic peaks overlap the
from two wavelengths emitted by a line source passes analyte peak. Many compounds only absorb strongly
through the flow cell and is detected simultaneously by near 200 nm. On the other hand, spectral interference
separate filter-detector modules. Monitoring two wave- is often less at longer wavelengths. If chromatographic
lengths simultaneously (a common wavelength pair is peaks are sufficiently separated in time, it is possible
254 and 280 nm) is useful for cases in which monitoring to program the wavelength to the value that is optimal
at only one wavelength does not provide adequate ab- for each peak during the chromatographic run. In some
sorption for all the compounds of interest. The ratio of cases, the flow can be stopped and an absorption spec-
the absorbance at the two wavelengths can be calculated trum obtained.
and displayed as the chromatographic signal. This re- Many commercial diode array absorption detec-
sponse ratio is useful for confirming peak identity be- tors are now available. Spectra can be obtained in as
cause it is independent of the analyte concentration. as little time as 0.01 s during the chromatographic run.
shown in Figure 13-24. The obvious advantage of such detectors is that ab-
More expensive and complex absorption detectors sorption data at all wavelengths are taken and stored.
are based on monochromators and conventional con- The optimum wavelength(s) for plotting can be eval-
tinuum sources (e.g., tungsten and D, lamps). The spec- uated after the chromatographic run. With appropriate
tral bandpass is typically in the range 2 to 10 nm: it software, the user can display multidimensional ab-
cannot be as low as for some conventional spectropho- sorption data, as shown in Figure 13-25. A more con-
ventional chromatogram is obtained by plotting the ab-
sorbance at one wavelength vs. time. Alternatively, the
absorbance plotted can be for different wavelengths in
different portions of the chromatogram to optimize the
Time -
FIGURE 13-24 The response ratio technique.
calibration slope and selectivity for each compound.
If the compounds of interest do not absorb suf-
ficiently, a post-column reaction system may be needed.
Chromatograms are shown as a function of the The eluted compounds are mixed with a reagent that
absorbance at 254 and 280 nm and as the ratio of converts the species to a strongly absorbing product
absorbances at these two wavelengths. A square before the eluant is sent to the detector. Ninhydrin is
peak indicates a constant ratio. Lack of a square a common post-column reagent for amino acids. and
peak indicates a response from two or more com- the absorbance is monitored at 440 and/or 570 nm. Metal
pounds.
Corticosterone
Dexamethasone h
FIGURE 13-25 Absorption spectra
taken with a diode array absorption
detector during a chromatographic
run of three steroids. The spectra are
taken 5 s apart. (Courtesy of Hew-
lett-Packard Company.) Wavelength, nm
3 Chap. 13 I Ultraviolet and Vlslble Molecular Absorption Spectrophotometry
@ 3 @- ,
I
1 Sample tube
I
I
Feedback
Signal processor
Source - Monochromator + \
-- PMT - and
readout device
Polarizer Analyzer
radiation can also be achieved with an electro-optic In single-beam in-time instruments, the axis of the
polarizer such as a Pockels cell (see Figure 3-15) placed linear polarizer or the retarder is mechanically rotated
between the polarizer and analyzer and driven with an between two orientations that are 90" different to il-
ac voltage. In some spectropolarimeters, another elec- luminate the sample cell in an alternate fashion with d
tro-optic polarizer is employed to adjust the plane of and 1 circularly polarized radiation. The instrument au-
polarization for the null signal. The dc voltage applied tomatically calculates the difference in absorbance be-
to the device is related to the optical rotation of the tween the two components. It is often necessary to com-
sample. pensate for the difference in the incident intensities of
Measurement of the circular dichroism (CD) is the two polarization components. The rotation of the
quite similar to conventional absorption measurements. plane of the linear polarized beam can also be electri-
The primary difference is that the radiation incident on cally modulated with an electro-optic device such as a
the sample cell must be circularly polarized: a circular Pockels cell or a magneto-optic device such as a Faraday
polarizer is placed between the monochromator and the cell. Double-beam in-space designs are often based on
sample cell and is sometimes available as an attachment splitting the incident beam into both polarization com-
for spectrophotometers. A circular polarizer is con- ponents, which are each separately directed through
structed from the combination of a linear polarizer and two equivalent sample cells.
a 90" retarder such as quarter-wave plate or a Fresnel Both C D and O R D measurements are employed
rhomb (see Figure 3-13). It is necessary to measure the primary in fundamental studies to provide structural
absorbance with both d- and I-circularly polarized ra- information about optically active compounds. Most
diation and calculate the difference. This difference is applications are in the biochemical area (e.g., amino
in turn used to calculate the molar ellipicity (see equa- acids, proteins).
tion 3-28), which is the normal measure of CD.
PROBLEMS
13-1. Describe the major differences between a nonline- (a) The nonlinearity due to polychromatic radiation
arity due to stray radiation and that due to poly- is usually negligible if the spectral bandpass s is
chromatic radiation. What are the major causes of much smaller than the full width at half maximum
stray radiation in a monochromator? What steps are (FWHM) of an absorption band. As a rule of
taken in many monochromators to reduce the stray thumb s < 0.1 FWHM for negligible deviations.
radiation figure? The absorption band of Pr3- at 482 nm has a
13-2. The slit width of the monochromator in a UV-visible FWHM of 3.0 nm. What slit width should be used
spectrophotometer is extremely important since it de- to avoid a nonlinearity with a monochromator
termines throughput, and thus signal-to-noise ratio that has a R, value of 4.0 nm mm-I?
(SIN), as well as spectral bandpass. and thus reso- (b) A different absorption band of Pr7- has a FWHM
lution. value of 13.0 nm. For the same monochromator
Chap. 13 I Uitravioief and Visible Molecular Absorption Spectrophotornetry
changes the reaction mechanism. It is noted that calculation that K = 4.0 x 10-'% and mG = 1.0 x
absorbance is nearly independent of concentra- 1012V A - ' .
tion at these absorbances. 13-16. A solution contains 35.3 k g mL- Cr. The Cr is pres-
13-9. Give the major reasons that the following methods ent only as dichromate (Cr,0,2-) in 1.0 M H2S0,.
or techniques are becoming increasingly useful and The transmittance of this solution was 0.762 at 440
popular in instrumental analysis. Give the major ad- nm in a 1-cm-pathlength cell. What is the molar ab-
vantages over alternatives. sorptivity of dichromate at 440 nm?
(a) Enzyme-catalyzed reactions to determine sub- 13-17. If the relative standard deviation in concentration is
strates. given by
(b) UV-visible spectrophotometers with diode array
detectors. g c -
- - UT
(c) Double-beam spectrophotometers with an option c TlnT
for slit programming or photomultiplier voltage
programming. where a, is the standard deviation in transmittance,
(d) Double monochromators for UV-visible spectro- show that the optimum precision occurs at 36.8% T.
photometers. Assume that a, is constant and independent of con-
( e ) Flow injection analysis with colorimetric detec- centration.
tion. 13-18. If absorption measurements are made under signal-
13-10. The simultaneous determination of titanium and va- shot-noise-limited conditions, the relative standard
nadium, each as their peroxide complex, can be done deviation in concentration is given by
in steel. Preliminary measurements indicated that 1.00
mg of Ti in 50 mL with H 2 0 , gave an absorbance of
0.269 at 400 nm and 0.134 at 460 nm in a 1.00-cm
cell. Under similar conditions, 1.00 mg of V in 50 mL
gave an absorbance of 0.057 at 400 nm and 0.091 at where k is a constant for a given reference current
460 nm. For a 1.000-g steel sample treated with H,O, and noise equivalent bandpass. Show that the opti-
and ultimately diluted to 50 mL, the absorbance at mum precision occurs at 13.5% T in this case.
400 nm was 0.172 and that at 460 nm was 0.116 in 13-19. In a flow injection analysis experiment, a sample of
the same 1.00-cm cell. Calculate the percent titanium 1 . 5 x 1 0 - W Mu2+ is injected into a flowing stream
and vanadium from these absorbance values. containing a buffered reagent. As the sample is car-
13-11. Calculate the measured absorbance for 2.00 x lo-' ried downstream, it mixes and reacts with the reagent
M solution of an analyte molecule with a molar ab- in a reaction coil. The product is then detected spec-
sorptivity of 1.OO x lo5 L m o l cm- in a 1.00-cm- trophotometrically. Identify as many variables of the
pathlength cell. The stray radiation readout fraction system as you can. Indicate those which it would be
(r) is 2 % . difficult to vary experimentally and any you would
13-12. Consider a solution of a weak acid H A which has a not wish to vary.
dissociation constant K,. The undissociated acid H A 13-20. Several replicate experiments are carried out using
has a molar absorptivity of E,, and the anion A - has the FIA system described in problem 13-19. The av-
a molar absorptivity of E , - . The analytical concen- erage height of the resultant peaks is h and the stand-
tration of H A is designed as c,. Derlve an expression ard deviation is a .
for the absorbance of a solution of this acid in terms (a) What would be the effect of optimizing the re-
of c,, K,, [H'], E ! ! , , and .,8 . Indicate how the Beer's sponse function R = hla?
law plot depends on the relative magnitude of E,, (b) Would it be better to optimize R = h / ( u + I)?
compared to E , _ . the p H , and c,. Assume that the 13-21. From the following data, indicate whether the non-
hydrogen ion concentration is independently ad- linearity in the calibration curve is due to instrumen-
justed by another buffer. tal causes or chemical equilibria involving the ana-
13-13. For a given analyte solution, indicate what a plot of lyte.
a,lA versus the slit width W of the monochromator (a) The absorbance at which nonlinearity occurs
would look like. Justify the shape of the curve. As- changes from 0.5 to 1.0 when the cell pathlength
sume that the slit width is varied from 0 to some large is doubled.
value. (b) The absorbance at which nonlinearity occurs is
13-14. Calculate the effective pathlength of a cell of width independent of cell pathlength.
b in which the angle of incidence is l o from the nor- 13-22. Radiation at two wavelengths is incident on a sample.
mal. The reference signal for both wavelengths is identical,
13-15. For a molecular absorption spectrophotometer. I , = but the molar absorptivities are 1.0 x lo4 and
1.00 x ~ O - ~ ) A , = X ,1.00 x 1 0 - ~ , g , ,=, (a,),, = 1.0 x 10j L mol-I cm-I. Calculate the absorbance
1 x 10 l 4 A . Calculate the detection h i t for species at analyte concentrations of 1, 10; 20, and 30 kM in
X where ~b = 1.00 x 10% mol-I. Assume in your a 1-cm-pathlength cell.
lnlrared (1R) \pcctrometry deals w ~ t hthe lnteractlon can be quite con~plexbecause of the many possible
of mtn,rred r d i a t ~ o nw ~ t hm ~ t t e rThe IW spectrum of vibrational transitions and the existence of overtones
a co~npounctc m provide inp port ant informatmi about and sum and difference bands. However, IR absorption
tt\ chemical nature and molecular structure Most com- bands are often quite sharp and characteristic of certain
monly, the Ypcctrum I \ obta~nedby mca\unng the ab- groups within the molecule. The IR spectrum for a given
sorption ot IR ratl~ation,,11though 1nt1arcd emismn and inolcculc is also unique to that moleculc and thus highly
reflection ,\re alw used. Infrnred \pearornetry hnds ~ t \ useful in compound identification. Detailed analysis of
wdest cippllcabllity In the m,~lysisot organrc materids, I R absorption bands can be of great assistance in de-
hut ~t 1s J s o useful for polyatom~cmorganlc moleculc~ termining the inolecular structure of a compound, par-
.ind lor organometalhc con~pounds. ticularly when used in conjunction with another struc-
The mfrared regon ot the electrorn,ignetlc spec- ture-sensitive technique, such as nuclear magnetic
t ~ u r n1s generally comldeled t o lie In the wclvclength resonance spectrometry or mass spectrometry.
r'lnge lrom 770 nm to 10OO pm, the corresponding w'ive Infrared spectrometry is also useful for quantita-
number rmge 1s from 12,900 to 10 cm ' (see Tdble tive analysis. although it is considerably more difficult
1-1) R,idimt energies in the IR I egion ,,re on the order to achieve accurate and precise results with IR spec-
ot the cnelgles of vibrationd tr,tn\lt~on\ Hence IR trometry than with UV-visible methods. Beer's law pro-
qxctroccopy 15 one branch of v ~ bdtiond l spcctro\copy vides the basis for quantitative IR methods as it docs
The 1R Icglon 1s otten further subdrvlcted Into three in UV-visible spcctrophotomctry. However, nonlinear-
sub^ egions I he near-infrare regmn (nearest to the ities and the presence of overlapping absorption bands
visible) extends 11om 770 nm 2 5 pm (12,900 to 4000 can Icad to severe pmblems in quantitative applications
cm I ) , the mid-infrare rcglon from 2 5 to 50 pm (4000
- of IR absorption.
t o 200 cmP1), and the far-inli-ared reglon from 50 t o This chapter is concerned primarily with infrared
1000 Frn (200 to 10 cm ') absorption spectrometry, because of its widespread util-
Ne,i~ly all inolcculec ,~hsorb~iifrnrcdI adi,rtion, the ity compared t o emission and reflection spectrometry.
cxccpt~on\benig homonuckdi d ~ ~ i t o r n ~such c s as O , , We begin our study by considering the basis of IR ab-
N,,m d H2 The IK spect~umot a poly,ltomic rniolccul,ir sorption, the selection rules and the types of transitions
Sec. 14-7 1 Basis of Infrared Absorption 05
ism that is uccd to label tbc vibrational energy Syninietr~chtrctching Asy mmctric stretching
levels.
shows three f~~nttnmcntal absorption bands in the IR
region.
Carbon dioxide, on the other hand, is a linear
~noleculeand thus has four fundamental vibrational
modes, as illustrated in Figtire 14-lh. The symmetric
strctch in C 0 2 does not give rise to :I dipole moment
change and thus is infrared-inactive. 'I'hc other three
niodcs are infrared-active, but the two hcnding modes
are degenerate and absorb at the same frequency. Thus
C 0 2 shows two fundamental absorption bands in the
IR region, one due to the asymmetric strctch and the
other to the degenerate bending niodcs.
Also introdi~cedin Figure 14- 1 is the v,, symbolism
that is used to label the frequencies of the various fun-
damental vibrations. By convention, the highest fre-
quency totally symmctric vihration is labeled v , , the
next-highest frequency symmctric vihration v,, and so
on. After assigning a11 the symmctric vibrations, the
highest-frequency assymmetric vihration is counted next. FIGURE 14-2 V~brnt~onnl motle~for a meth-
Thcn the rest of the asymmetric vibrations are num- ylene group (a) and breatlimg vrlx'it~onfor ,irmg
bcrcd in thc order of decreasing frequency. The bending cornpoi~rid(b)
Sec. 14-1 I Basis of Infrared Absorption 407
I --
--
0 H and N
-
H stretch
-
C = 0 stretch C - 0 stretch
I C - H stre1 '
- - C = N stretch C - K stretch
I i = C and C =
N stretch C = C stretch
-
-
C - C stretch
K - H bend
N
-
-
H bend
C H bend C - H bend
0 - H bend
Wavelength. pm
group absorption is only slightly affected by the c o n - mcchan~cdterms the re\onmcc mter,iction mlxcs the
position of the rest of the molecule in thc group fre- cncrgles of the two i n ~ t ~ d\tcitcs
l The \ep<il,ltion and
quency rcgion.
-
In the region from 1300 to 400 cm ', vibration;~l
frcqucncics are affected by the entire molecule, as the
ielatlvc Inten\~tlesof the h m d \ clcpcnd o n ne,rrne\\ ol
the two unperturbed lrcq~iencie\.The inter,ictlon of the
( H \trctch In Jdehydes at dbout 2800 c m l w ~ t hthe
broader ranges for group absorptions in Figure 14-3 t m t overtonc of the 1400 cm ' ~n-plancC'H bending
illustrate. Since absorption in this fin vibr tion on 15 mother exmqAc ot bernr~resonance Th14
characteristic of the molecule as a whole, this region n to 2900
interdctlon givcs a doublet In the ~ e g ~ o2700
finds widespread use for identification purposes by com- C111
parison with library spectra, as discussed in Section Fe1n11re\onmce I \ not alw,~ysed\y to detect It
14-4. \hould be su\pected wheneve1 ,I normdly single hand
dppear\ '1s ,i cloublct In complex molecule\, ~t cnn
\ornetme\ be detected by dcutercltlon or by u\mg vdr-
IOU\ wlvcnts In compounds ~ o i i t ~ r ~ methyl
n ~ n g group,
The energy of a particular vibrational mode, and thus l~rt1.11cleuterat~onher\ been uscd t o c i cumvent
~ Fcr mi
its frcqucncy, may be influenced by the presence of rcson'tnce between the C'H dclorrndtion overtone '1nd
other vibrations in the molecule through vibrational the CIf st~etchmgv ~ b r ~ i t ~ 1ohe
n ilcqucncles ale \hifted
coupling. Strong coupling can occur between two enough by the isotope effect that Fcrru~resonmce no
stretching vibrations, for example, if the two vibrations longer occurs.
have an atom in common. Interaction also occurs bc-
tween two bending vibrations with a common bond bc-
tween the groups. Coupling is strongest when the cncr-
gies of the isolated vibrations arc approximately equal.
If the groups are separated by two or more bonds, little Three distinct types of instruments employed for IR
or no coupling occurs. absorption spectrometry are considered in this section.
As a result of vibrational coupling, the position Dispersive instrumcnts with a monochromator are used
of an absorption band cannot be specil'iccl exactly. For in the mid-1K region for spectral scanning and quan-
cxanrplc, the coupling of the C - 0 stretching vibration titative analysis. Fourier transform IR systems are widely
with nearby C-C and C-kl vibrations leads to a dif- applied in the far-IR region and becoming quite popular
ference of 20 cm ' in the C-O stretch in methanol for mid-IR spectrometry. Nondispersive instrurncnts that
and ethanol. Although coupling leads to uncertainty in use filters for wavelength selection or an infrarcd-ab-
group vibrational frequencies, it also makes the infrared sorbing gas in the detection system are often uscd for
spectrum unique for a given molcculc and is one of the gas analysis at specific wavelengths.
rna,jor reasons that IK spectrometry is so valuable in
qualitative analysis.
In order for vibrational coupling to occur several
conditions must he f~ilfilled.First, both vibrations must Modern dispersive infrared spectrophotomctcrs are in-
have the same symmetry and approximately the same variably double-beam instruments, but many allow sin-
frequency. Second, there must he an apprcciablc in- gle-beam operation via a front-panel switch. Single-bcam
teraction between the groups responsible. This in turn instruments are not very practical in the 1K region bc-
implies that the two groups are near to each other in cause of the absorption of IR radiation by atmospheric
the molecule and that a vibrational energy transfer H,O and CO,, as shown in Figure 14-4. Double-beam
tncchanism exists. operation compensates for atn~osphericabsorption, for
When an overtone or combination frequency in- the wavelength dependence of the source spectral ra-
teracts with a fundamental vibration, the interaction is diance, the optical efficiency of the mirrors and grating,
called Fermi resonance. Fcrmi first proposed such a and the detector rcsponsivity, and for source and de-
mechanism to explain the splitting of the 1340-cmp' tector instability, which can he serious in the TR region.
fundamental of CO,. In this case Ferrni resonance givcs Also, all solvents absorb in the infrared rcgion, which
rise to two bands in the Raman spectrum where only makes single-beam operation somewhat impractical even
one is predicted. For COL, an overtone of the degcn- with storage of the reference spectrum.
erate bcncling vibrations (2v2) is at 2 x 066 crn ' =
1332 c m '. This frequency is very close to that of Ihc Dispers~veSpectrophotometer Des~gns. A
symmetric stretching vibration (v,) at 1340 cm ' . The malcpr~tyof the d ~ \ p e r \ ~ vIR
c spectrophororneter\ ac of
resonance interaction raises the frequency of one of the t h ~ \wrrtlng u\e the optrc,~lnull p ~ i n c ~ p illu\tratcd
le In
vibrations and lowers that of the oil~er.Isr quanlunin F~gurei4-5. R,mo rccordrng \y\tcnnrs, sum~l,~ito tPme
Sec. 14-2 l instrumentation 409
Double-beam
Wavenumber. cni-' mode
4000 2500
50
100
90
2 3 4 5 6 I 8 9 10 11 12 13 14 15 16
Wavelength, p m
used in the UV and visible regions, are used with IR fiers and their reduction in price, these systems have
spectrophotorneters, but until recently were found only gained in popularity in recent years. Optical null sys-
in the more expensive research instruments. Several tems read out directly in transmittance. Ratio recording
differences can be noted when the schematic of Figure spectrometers can read out in transmittance or absorb-
14-5 is compared to the corresponding diagram for a ance. Because of the present popularity of FTIR sys-
UV-visible spectrophotometer (see Figure 13-4). First, tems, not many dispersive spectrometers have been in-
with infrared spectrophotorneters, the sample cell is troduced recently.
usually placed in front of the monochromator to min- The radiant power emitted by infrared sources
imize the effects of infrared emission and stray radiation varies dramatically over the wavelength region covered
from the cell compartment. In the UV-visible region, by most spectrophotorneters. Slit programming is used
the sample cell is often placed after the monochromator in modern IR instruments to compensate for these var-
to minimize photodecomposition of the sample. The iations and thus maintain a nearly constant radiant power
sample and reference beams are usually chopped at level at the detector. Either a cam is used to vary the
relatively low frequencies (5 to 30 Hz) with IR spec- slit width as a function of wavelength in a predetermined
trophotometers because of the slow response of most manner or the reference beam detector signal is mon-
IR detectors. Modulation also allows discrimination itored and compared to a preset desired level. In the
against IR emission by materials near the detector and latter case, any difference is amplified and used to drive
detector noise with a llf nature. a slit control motor, which opens or closes the slits to
In inexpensive infrared spectrophotometers, the minimize the difference signal. The cam method is in-
recorder pen and the attenuator are mechanically cou- expensive, but not very flexible since the program can-
pled as shown in Figure 14-5a. In more versatile sys- not be varied as components age or a new source is
tems, the pen is driven by a separate servomechanism. installed. The slit servo method allows the instrument
A potentiometer with a fixed voltage across it is coupled to adapt to changing conditions.
to the attenuator so that the attenuator position controls Because of slit programming, the resolution of IR
the wiper of the potentiometer. The voltage on the spectrophotorneters degrades in wavelength regions
wiper is proportional to the transmittance. This voltage where the source intensity is low or where strong ab-
is the input to a normal servo recorder. By using several sorption occurs in the reference beam. Regions of high
different fixed voltages across the potentiometer. any solvent absorption. for example, are often accompanied
portion of the transmittance scale may be expanded to by wide slits and low resolution. Care must often be
full scale on the recorder. taken in interpreting IR spectra in such wavelength re-
Ratio recording systems are less frequently used gions.
in IR spectrophotorneters because of the slow response
times of the thermal detectors used and the high gains Components of Dispersive infrared Spectro-
required. With the improvements in electronic ampli- photometers. Sources and detectors for IR spec-
Chap. 94 I Infrared Spectrometry
Reference
coupling
18
Sample
8.
I -p*-Chopper
synchronously demodulated (b). The
sign of the dc output of the synchro-
nous demodulator (lock-in amplifier)
depends o n the phase of the ac dc-
ccll modulator- input, which in turn dc-
pends o n which beam is more in-
tense. The dc output can be furthcr
amplified t o drive a dc motor which
is connected to a beam attcnuator and
the pen on the recorder. Altcl-na-
tivcly, the dc signal can he rcmodu-
lated by an electrical chopper and ap-
plied to one winding of an ac motor.
The other winding is attached to the
same signal that dl-ives the electrical
chopper. In either case, if the rcf-
erencc beam intensity is higher than
the sample beam intensity, as in the
lower nc signal in (b), the motor turns
in the appropriate direction to move
the attenuator further into the ref-
erence beam and reduce its intensity.
b, < 4
Similarly, if the sample beam inten-
sity is higher, as in the upper ac signal
Demoduldtcd Filtcrccl Modulated in (b), the attenuntor is pulled back
so as to increase the reference bcam
intensity.
trometry werc dl\cussed in Section\ 4-2 and 4-4, rc- more cfficient. In the near-IK region, photon energies
\pcctively Ncrnst glowel\ and glob'm ( x c T,thle 4-2) are high cnough that photon detectors can be used.
are typc.illy u\cd as source\ for gene1(11-purpowmld- Photoconductive cells, such as CdS, PbS, and PbSe, are
IR ~nstruinent\and cdn d \ o be used 111 the 1'11-I R reglon often employed in this rcgion. A multichannel IR spec-
to wdvelength5 01 = 50 pm Some Inexpenwe mid-IR trophotomcter has been constructed that uses an 1K
rn\tr uinent\ u\e n ~ c onie
h or i h o d ~ ~wire
~ n i tource\ elcc- spectrograph and a linear pyroelectric dctcctor array.
trrcdlly hecited to = 1100 I< I hcsc ,u e less ~ n t e n wthan At this time such a systctn is still quite expensive.
the globdr or Ncrn\t glower, but they have longel hie- I'risms were colnrnonly used in older infrared in-
t l m o Tung\ten f~lamcntlamp5 can he uwd In the near- struments for wavelength isolation. Glass and quartz
IR region In thc very far - 1 K region. high-prcswre mer- cannot be used because of excessive absorption at wave-
cury arc Limp\ cao be u\ed to wavelengths of = 300 lengths longcr than 3 pm. I Jcnce prisms werc made o f
pm, cilthough they provde ielatrvcly low radimce ionic crystals such as NaC1, LiF or CaF2. Ilnfortunatcly,
'I hermocouple\ dre the mo\t widely u\ed detec- with NaCl great care has to be taken to protect the
tor\ In d ~ r p c r s ~ vm~d-lnfiarcd
c nntruments, '~lthough prism from moisture since it is highly hygroscopic. Mod-
bolometers and Gol,iy cell\ ale wrnetlmcs ~ l \ e d .For ern IR spectrophoton~eterscruploy reflection gratings
the iar-TR rcgion, the G o h y (pneum;it~c)detector 15 that are similar t o those used i n the UV-visible region,
Sec. 14-2 1 Instrumentation 41 1
although the groove spacing is greater (e.g., 120 grooves Crystalline alkali halides, such as NaC1, KBr, CsBr,
mm-l). Reciprocal linear dispersion values in the tens and CsI, are most widely employed as window materials
of nanometers per millimeter range are common. Rep- (see Appendix B) because of their high transmittances
lica gratings are now available made from various plas- in the IR region. The long-wavelength limits for many
tics. To.reduce the effect of overlapping orders and stray IR materials are listed in Table B-3. In the very far-IR
radiation, filters or a preceding prism are employed. region, quartz becomes transparent again at wave-
Because a blazed grating is only efficient over a limited lengths greater than 50 Fm. The Irtran series of window
wavelength range, two or more gratings are often used materials (see Appendix B) is prepared by sintering
with several filters to scan a wide region. various types of powders under high pressure and tem-
Lenses are rarely used in the IR region. Instead, perature. These have nearly identical optical properties
mirrors are used to focus and collimate the radiation. as single crystals of the same substance. Since water is
Mirrors are generally made from Pyrex or another ma- present in nearly every organic solvent and as a vapor
terial with a low coefficient of thermal expansion. Front in the laboratory environment, the solubility of the win-
surfaces are usually coated with a vacuum-deposited dow material in water is of particular interest. Water is
thin metal film of aluminum, silver, or gold. Aluminum also a desirable solvent for many IR studies. The water
is the most widely used material, although silver and solubilities of common window materials is also listed
gold have higher reflectivities. Aluminized mirrors re- in Table B-3.
tain their reflectivities for long periods of time, while
silver surfaces tend to tarnish when exposed to sub- Features of Dispersive Spectrophotometers.
stances normally found in laboratory atmospheres. Dispersive IR spectrophotometers are available with a
Window materials are a particular problem in I R wide range of performance features and prices. Inex-
spectrometry. Windows are used for sample cells and pensive instruments that cover the mid-IR region may
to permit various compartments of the spectropho- have resolution figures in the range 1 to 3 cm- l . More
tometer to be isolated from the environment. Window expensive systems often have options available for ex-
materials must be transparent to IR radiation over the tending the range to the far-infrared region and may
wavelength range of the instrument, and inert to the have resolution figures in the range 0.1 to 0.5 cm-l.
various chemicals they will contact. They must be ca- The optics of a commercially available, inexpensive in-
pable of being shaped, ground, and polished to the frared spectrophotometer are shown in Figure 14-6.
desired optical quality. Newer I R spectrophotometers contain microcom-
Thermocouple
detector
0
Nichrome
wire
source
L----_I
Sample compartment
%L
Collimating
mirror
FIGURE 14-6 Optical diagram of Beckman Acculab infrared spectro-
photometer. This system is of the optical null type. Radiation from the
nichrome wire source is split into two beams and sent through the sample
and the reference cells. The reference beam attenuator is controlled so
that the reference beam intensity matches that of the sample beam. The
two beams are recombined and alternately pass through the monochro-
mator to a thermocouple detector. The recorder pen is mechanically linked
to the reference beam attenuator. (Courtesy of Beckman Instruments,
Inc.)
Chap. 74 I Infrared Spectrometry
Mirror wtdl
ccnter hole
for laser heam
(a)
ferometer discussed in Section 3-6. The reader may wish gion is illustrated in Figure 14-8. The mirrors directing
to review this section before proceeding. We consider the interferometer beam through the sample and ref-
here some of the design and performance features of erence cells are oscillated rapidly compared to the
FTIR spectrometers. movement of the interferometer mirror so that sample
and reference information can be obtained at each mir-
Interferometer Systems. Instruments for far- ror position. The double-beam design thus provides
IR absorption spectrometry are most often single-beam compensation for source and detector drifts.
designs, while mid-IR spectrometers are most often Moving the interferometer mirror without any tilt
double-beam systems. Figure 14-7 shows a typical ar- is a particular problem in FTIR systems. In the far-IR
rangement for a single-beam FTIR system. Note that region, where wavelengths are long, mirror drive mech-
the sample is placed after the Michelson interferometer. anisms can be relatively crude. Often the mirror is po-
Because of the modulation characteristics of interfer- sitioned by means of a motor-driven piston which moves
ometry, infrared emission and stray radiation from the through a cylinder with normal grease as a lubricant.
sample compartment appear as a dc component of the For mid- and near-IR operation, however, a more pre-
interferogram and are inherently discriminated against. cise and sophisticated drive mechanism is required.
Since a computer system is a necessary part of the FTIR Usually, the mirror mount is floated on an air cushion
spectrometer, single-beam systems acquire. calculate, held within stainless steel sleeves and driven by a voice
and store the reference spectrum, usually versus air as coil similar to that in a loudspeaker. Very high reso-
a blank. Later the sample spectrum is obtained and true lution systems require tilt-compensation devices and some
sample absorbances are then calculated. even stop the mirror at each sampling point to adjust
A double-beam spectrometer for the mid-IR re- its planarity prior to measurement.
Interferometer
Fixed
Movable -
mirror
Sample
compartment
/
IR source
... h
detector /
Modern F T I R systems also pvovide for automatic roelectnc detector hci\ \utt~c~cnt
re\pome t m e for r'lprd-
location of the zero path difl'crcnce position and for \cannmg ,~ppllcat~on\
sampling of the intcrfcrogram at prcciscly spaced in- Pyroelectr~cdetector\ are ,tl\o w~delyuscd 111mrcl-
tervals. 'I'his is accomplished by the three-interfcr- rntrared ~n\trumcnt\Although thcrmocouplc\ h m e 1)
omctcr system shown in Figure 14-9. The triple intcr- v'~luc\ dn order ol magnrtude la-gel t h m pyroelectr~c
fcrometer design yields very high precision in determining detector\ (\cc Tnble 4-5) at low modul,ltion trequcn-
1R wavelengths. Data collection a t exactly the same e m , the TGS clctecto~h,~\better le,ponwrty at h ~ g h
mirror position on every scan allows coherent signal modul,~t~on irequenc~es(c g., I kFb) Mercury-doped
averaging for SIN improvcmcnt. germmrum bolomete15 and vlrcon bolometer\ have been
uwd 101 very hrgh 4cn\rt1vrty mrd-rnfrd~cd mea\iIre-
nienl4
Components of FTIR Spectrometers. Source4 In the far-mfrxed r eglon, b e m \pl~tter\arc mo5t
tor FTIR arc dentred to thosc used In clr\pe~\~ve 1K commonly m;rde trom thrn him\ ot Myla1 (polyethylene
\pectrophotometer, Thermal detector5 'ire, however, te~ephth~llate)Unfortunately, Mylar \trongly abwrbs
too slow 111 I e5ponsc f o 1 the 1110dul~1tronf~equenc~c, In the m d 1 K reglon and I \ not mechc~mcally4trong
normally oht'uncd. For lal-IK \pectlo\copy, the Gol'iy enough to m,ikc Into t1lm4 of the necessary th~ckncss
pncumat~cdetector and the tr~glycmewlt'~te (TGS) (.;I km) Thu\ n11d-rntrxcd bc'im \pl~tter\ 'Ire nor-
pyroelectr~cdctcclo~are mo4t common Only the py- rnally t h f~lrncof germanrum or \~llcondepo,itcd on
From 111
Linear drive motor sourcc
Sl
HeNe laser
Laser
and
detector
flat alkali halide substrates. A layer of Fe,O, on quartz (see the next section) and various reflectance attach-
or CaF, has been used in the near-IR region. ments (see Section 14-7). Some even have accessories
Computer system requirements for FTIR spec- for such new techniques as FTIR photoacoustic spec-
trometers vary dramatically with the wavelength cov- trometry (see Section 17-1).
erage and the resolution desired. Low-resolution (5 cm- ')
far-IR spectrometers normally acquire less than 1000 Nondispersive IR Instruments
points per interferogram, whereas moderate resolution
(0.5 cm-l) mid-IR systems may acquire as many as There are many routine applications of infrared spec-
32,000 points. The recent dramatic decrease in the price trometry where it is desired to obtain quantitative in-
of computer memory and disk storage has reduced the formation about one constituent or a few species in a
expense of the computer system for FTIR spectrome- mixture instead of obtaining a complete spectral scan.
ters. Such applications include monitoring a single constit-
uent in a process stream, and atmospheric gas analyses
Features of Commercial FTIR Systems. where a few important gases are routinely determined.
There has been a clear trend in recent years toward the For these types of determinations, the dispersive spec-
development of low-cost FTIR instrumentation. The trophotometers and FTIR systems just described are
least expensive FTIR systems are intended for use in overly complex, too expensive, and often not rugged
one spectral region. such as the mid-IR or far-IR. These enough. Single-beam, nondispersive spectrometers can
are often priced to compete with the lower-priced dis- be used in such applications for accurate quantitative
persive systems (e.g., $12,000 to 15,000). Typically, these determinations. Among the nondispersive spectrome-
benchtop systems provide resolution values of a few ters used for these purposes are filter photometers, fil-
wavenumbers. More versatile, and more expensive, in- ter-wedge spectrophotometers, and infrared gas ana-
struments are available to cover a wider wavelength lyzers. We consider here each of these nondispersive
range and provide higher resolution. One commercial spectrometer types.
instrument system, for example, can produce spectra
from the UV to the far-IR region with a few changes Filter Photometers. Several simple IR photom-
in sources, detectors, and beam splitters. The resolution eters are available that use filters for wavelength selec-
provided varies from 0.5 cm-I with moderate expense tion. Figure 14-10 shows an example of a commercial
systems to less than 0.1 cm-I with research-quality in- IR gas analyzer designed around a filter photometer.
struments.
In the far-IR region, Fourier transform spectrom-
eters are clearly superior to conventional. dispersive
ReferenceBoard
systems in terms of SIN, resolution, and scanning time.
In the mid-IR region, FTIR systems allow the spectrum
to be obtained with higher SIN for comparable analysis
time compared to dispersive systems. Conversely, the
spectrum can be obtained in a much shorter time with
nearly equivalent SIN. For example, a moderate reso-
lution (few wavenumbers) spectrum can be obtained
with comparable SIN to a conventional IR spectrum in
a few seconds. Fourier transform IR clearly shows su-
perior characteristics for applications where transient
Cell windows
species or time-varying samples are involved (kinetic
studies or GCIIR), for measurements where energy is Chopper
limited (far-IR region or highly absorbing samples), for
measurements at very high resolution, and for meas- FIGURE 14-10 Portable, infrared filter pho-
urements where excellent wavenumber reproducibility tometer for gas analysis. A nichrome wire source
is used with a pyroelectric detector. Interchange-
and accuracy are required. Wavenumber reproducibil- able interference filters are available for the de-
ity is particularly important for accurate background termination of specific gases. Gaseous samples
subtraction. are introduced into the cell by means of a battery-
In recent years, there has been an emphasis on powered pump. A folded-pathlength design al-
increasing the versatility of FTIR systems through pro- lows the cell pathlength to be set from 0.5 to 13.5
viding new accessories for spectrometers. Most FTIR m at the factory. (Courtesy of the Foxboro Com-
manufacturers now provide microsampling accessories pany.)
Chap. 74 I Infrared Spectrometry
Intcrterencc t~lter\ale civ,illd>le at w,welcngth\ t h i t Jnfmred Gas Analyzers. I he1 c are several pho-
corre\pond to trmsit~on\In ~ndmtr~'llly important g,r\cs tometer\ crv,~~lable t h d employ no w~lvelengthselection
Filter photometer\ w ~ t hlong-pathlength cell\, ,ilch devrcc ,it Al, ,15 ~llu-,tlated ~n 141gu1c 14- 12 'I hcsc non-
au the instrument \hewn In Figure 14-10, can detect '1 di\pcrslve ga\ a r r d y ~ e l \,Lrc vely s~inplc,tugged, and
Icw p d ~t\ pen rnillmn of the wlccted compound Arrlong ~nexpcn\~vc Selectwlty 14 ,tchlevcd by t ~ l l ~ nthe
g detcc-
the g d x \ thLlt can he detected with rc,rd~ly Llv,ul,hle tor compartment with thc ,m~lyec8'1s to that the de-
l~lter, 'ire phwgene, hyctrogcn cyan~dc,;ccrylonitnIe, tectol c ~ l ~ w r' bI \I ~le\pond\ only t o radl'ttlon ~n the
c n ~ b o nmonox~de,m d chlor~n~tted hydrocClrhon\ The w'lvelength r egron d b m bed by the malyte Sd\.
lnstrurnent 15 battery oper'itcd ,tnd wc~gh\less t h m 30 Anothet type of g,l\ ,~n,ily~er U X ~'I double-bem~
lb Smce the fdter, are e x p e n w e m d tomcwhat Incon- ~irrmgcment Rat-liat~onin the retcrence b e a n p,14\es
venient to chmgc, t h ~ sinttrument 15 best culled fol through both the \anple cell and ,i ~efercncecell f~lled
dcterminatmn of a unglc conmtucnt or for infreyucntly w ~ t hthe 'lr~alyte gas R,~dl;lt~on ru thc sample beam
mon~toring1' few constituent\ pa\scs through thc \ample cell only The two beam\
s t r ~ k bolometers
c that ~ i arms
c ol a Whcahtone brdge
'I'hc null balance point ol the bndge 14 rel,ltetl to the
Dielectric Filter Spectrometers. Circular wedge analyte concentratlon. Interfercnt\ can he troublesome
dielectric filters are available (see Appendix C) in which IJI the latter approach il they hnve ab\orptmu spectra
the transmitted frequency varies continuously and uni- that ovcrlnp that of the amlytc 111tcrlerence eftcctt can
formly on a disk-shaped substrate. These dielectric fil- be min1rn17ed by putting the intelle~irigsub\tance at
ters can be used like a monochromator to select wave- h ~ g hconcentratlon 111 a cell h i t precede\ the tnmplc
Icngths. The spectrum can he scanned by slowly rotating cell. Th14 rc111ove5 r;~dlrttlonat the wavelengths that the
the disk. A spectrometer based on such a filter is shown mterlerent can abwr b
in Figtire 14-11. With microprocessor control over
wavelength selection and data collection, the spectrom-
eter of Figure 14-11 can be programnicct to determine Special-Purpose Spectrometers. N o t surpris-
absorbances at multiple wavclengths. Calibration data ingly, tunal~leIK lasers have been used to makc special-
(ahsorptivitics, slopcs of working curves, etc.) can he purpose BR spectrometers. The lasers that have been
stored in computer memory and used to determine the used at present arc diode lasers and parametric oscil-
concentrations o f the various components automati- lators (see Section 4--3).Diode lasers have very narrow
cally. Note that the sample area allows solid, liquid, or bandwidths (e.g., 1 to 10 " cm ') and relatively high
gaseous samples to be used. Such spectrometers are irradianccs, but their tuning ranges are quite narrow
considerably more expensive than fixed-wavelength fil- (20 to SO e m p 1 ) .'I'hc parametric oscillator i s tunable
ter instruments. but much more versatile. over a wides range, but proctuces a broader spectral
FILTER WHEEL
DETECTOR I Stainless
Potentiometer--
(a)
Back
plate
14-13 Demountable l ~ q -
u ~ dcell tor mtl ared cpect~ometiy.
Teflon spaccrc are w e d to vary the
cell pathlength In the r m g e 0.015 to
1 mm.
pure liquid between two NaCl platcj whlch arc thcn materials. Organic solvcnts must also he dry if water-
clamped together In a den~ount~tblc cell wch as that soluble window materials arc employed. When it is nec-
shown In Flgurc 14-13, Spacer5 can be u\ed to Increae essary to use aqueous solutions, as it often is with in-
the pathlcngth over t l ~ lormed
t by capllary actlon organic and biological samples, thc insoluble window
Obvlou\ly, thc th~cknc\\cannot be very prccrecly con- materials listed in Table 63-3 ~ n ~ i 1x2
s t uscd. Fourier
trolled by this technque, and hence ~t n used only for transform IK techniques are often uscd with aqueous
qual~tativc\ t u d w . solutions, because cxcellent compensation for solvent
absorption can be obtained.
Solution Samples. The first problem when us- To obtain spectra for the qualitative analysis of
ing solution samples for JIK spectrometry is to find a nonvolatile solutions, a demountable sample cell, such
suitable solvent. As shown in Figure 14-14, all common as that shown in Figure 14-13, is often uscd. For volatile
solvents absorb in the IR rcgion. but there are wavc- samples or when the cell thickness rnust be controlled
length regions wherc each solvent is somewhat trans- as in quantitative methods, sealed sanlplc cclls are ncc-
parent. 01' course, solvcnt transparcncy is not the only essary. Sealed cclls arc either of fixed pathlcngth design
concern. 'The sample must also be soluble to the desired or variable in pathlength. Sealcd cells arc available with
Extent in the solvent and thc solvent must bc chcn~ically pathlengths of 0.1 to 1 mm. For quantitative analysis
lution concentrations -
incrt toward thc sample. For qualitative analyses so-
10%) are usually satisfactory.
The most commonly used solvents arc carbon tet-
this allows the determination of many species in the
concentration range 0.05 to 10%.
The thicknesses of narrow path YR cclls are often
rachloride, carbon disulfide, and chloroform. Water is determined by an intcrfercnce fringe nlethod. The
infrequently employed, not only because it is a strong transmittance spectrum of an empty cell is recorded
infrared ahsorher in certain regions (see Figure 14-4), against air in the reference beam. The reflected radia-
hut also becausc it dissolves the most cornmon window tion Srom the two walls of the ccll interferes with the
transmitted beam to produce an interference pattern
Wavenumber, cn-' similar to that shown in Figure 14-15; note that the ccll
5000 2500 1667 1250 1000 833 714 625 is cssentinlly a Fahry-Pcrot cavity. If n interference
Carbon disulfide maxima are ohscrved in a wavcnumber region AE, the
Carbon tetrachloride cell thickness h can bc calculated from
Tetrachloroethylene
Wavenumber, cm-' Satisfactory mulls are often obtained with a good deal
of practice.
4000cn-1 :
4000 crn 1 3
PR beam through a small apcrturc. Chmmcrcial bcam- through the microscope. Most infrared microscopes al-
condensers are available. The sinlplest arrangement uses low collection of data in either the transmittance mode
KBr or AgCl Icnscs mounted on a bed that can be or the reflectance mode (see Section 14-7). Spectra can
inserted into the sampling area of a normal spectrom- be obtained on samples, such as fibers, with diameters
eter. An off-axis elliptical mirror microscope is also as small as 10 bm.
available commercially. Mirror systems avoid the chro-
matic alxrrations that plague lens systems.
attachments, solid microsamples can bc hand
paring minute pressed disks. ellets as small as 0.5 lnln
in diameter have been uscd to obtain spectra. Nonvol-
atile liquids can be dissolved in a volatile solvent and The milor uses oi I R spectrometry dre for qualltat~ve
the solution ~ ~ s etod wet finely ground KBr powder-. identlf~c~~tlonof compounds m d tor the dctermmat~on
The volatile solvent is thcn removed and a microdisk ol molecu1,rr structune. Both of the5e applic,lt~onsrely
is pressed. Volatile liquids arc transferred into suitable on q u t e \im~larmethodology. In ctructure dcterm~na-
microcclls that fit in the beam-condensing system. tlons, ~t IS Important that the compound b e ~ n gstudled
is a technique that is u11- be ,I pure compound so th,lt the correct mterprct,rtion
r remaining stagnant for about of thc IR 4pcctrun~can be made Intolmat~onfrom IK
25 years. Ncarly every F I I R manufacturer makes a spectrometry 1% normally combincd w ~ t hthdt from sev-
microscope attachment for thcir spectrometer. In ad- cl'tl other techniques (e g., m,~\sspectrometry, NMK
dition, stand-alone systems arc available based on ctis- spectrometry) to deduce the structure Fol q ~ ~ l l ~ t a t ~ v e
persive spectrometers. ost of these allow visible light- identificat1on5, sample\ need not be pure compound\,
ing of the sample so that precise alig~mcntcan he done but c m be rcl,~t~velyclinple mixtures Inlor mnt~oncon-
Sec. 14-3 1 Sample Preparation Techniques 421
Ill
i I I I , I / I I 1 , I I I 1 I , , , / I I
2.50 1im 2 75 3.00 3 25 3 50 3.75 4.00 45 5.0 5 5 60 65 70 75 8.0 90 I0 11 12 1 3 14 IC 20 25
cerning functional groups in the unknown molecule can numbers or wavelengths of the absorption maxima are
be obtained from the IR spectrum and used in con- identified. A correlation chart or a similar aid is then
junction with spectra of known compounds to aid in used to attempt to identify the groups responsible for
identifying the unknown compound. Since the proce- the bands. After the strongly absorbing bands have been
dures used in identifying compounds and in obtaining assigned, identification of the moderate and weak ab-
structural information are essentially identical, both ap- sorption bands is then attempted. A complete assign-
plications are considered here. ment of all the bands in the spectrum, even for pure
compounds, is usually impossible because of the pres-
orrelation Charts ence of combination and overtone bands.
For qualitative analysis, final identification is most
Qualitative applications of IR spectrometry begin with often done by comparison of the spectrum obtained to
identifying the functional groups in molecules respon- reference spectra available in library collections (see
sible for infrared absorption. The group frequency re- below). Any additional information, such as boiling point,
gion (see Figure 14-3) is the most useful for this pur- melting point, and other spectroscopic data. is fre-
pose. Because of the widespread use of IR spectrometry quently of great assistance in an unambiguous identi-
for qualitative purposes, a good deal of empirical in- fication. For structural determinations, the IR spectrum
formation is available concerning group frequencies. is often only one piece of evidence among several. In-
The most comprehensive of these are correlation frared spectral evidence is frequently strengthened by
charts such as that shown in Figure 14-16. This type of calculations of the number of expected infrared active
chart not only contains all the group frequency infor- modes based on assumed structures. Such calculations
mation, but also is organized according to the type of use the symmetry of the molecule and the techniques
compound in which the group is found. of group theory or normal coordinate analysis. Any
Usually, after the spectrum is obtained, the ambiguities that arise in interpretations can often be
strongest absorption bands are selected, and the wave- resolved with the aid of mass spectrometry, NMR spec-
Chap. 14 / Infrared Spectrometry
The cell for most GC-IR experiments is a long (25 to nately, at higher pressures such lines are substantially
100 cm), narrow (few mm). gold-coated, flow-through broadened and can be made to merge into a broad
gas cell, known as a light pipe. The infrared beam envelope. With some quantitative IR techniques for
undergoes several reflections as it traverses the cell. The gases, higher pressures are intentionally used to broaden
cell volume must be small in order to prevent mixing the lines in order to obtain adherence to Beer's law.
of the separated components in the light pipe. The long In the liquid phase. absorption bands are seldom
pathlength is necessary to obtain adequate absorbances narrower than 3 or 4 cm-I and often wider than 10
for the low concentrations measured. Infrared methods cm-'. As a rule of thumb recall from Section 13-3 that
have also been used to monitor liquid chromatographic the spectrophotometer spectral bandpass s should be
effluents. less than one-tenth the FWHM of the absorption band
for 0.5% accuracy. With modern grating IR spectro-
photometers and many FTIR spectrometers, spectral
14-5 QUANTITATIVE INFRARED bandpasses are often small enough to measure absorb-
ABSORPTION SPECTROPHOTOMETRY ances accurately. However, older prism instruments often
possessed insufficient resolution and produced nonlin-
Although IR spectrophotometry is used most often for earities due to polychromatic radiation. Even with mod-
qualitative analysis and structural determinations, ern spectrophotometers, however, double-beam sys-
quantitative IR measurements are, nonetheless, quite tems with automatic slit servo mechanisms can give rise
important. We shall first consider the characteristics of to such deviations in spectral regions where the source
IR absorption and IR instrumentation that combine to intensity is low or the detector is unresponsive. Thus
make routine quantitative uses more difficult than rou- in quantitative infrared spectrometry, even with mod-
tine uses of UV-visible spectrophotometry. These in- ern instruments, molar absorptivity data obtained on
clude the need for frequent calibrations, nonlinearities, one instrument may not transfer accurately to another,
and the difficulty in measuring the analyte absorbance. and calibration curves made with external standards are
Next, the precision of IR absorption measurements is almost always necessary.
considered briefly. This section concludes with several Stray radiation, extraneous radiation, and scat-
examples of quantitative infrared absorption measure- tered radiation can also be serious problems in IR spec-
ments. trophotometry, particularly with dispersive spectrom-
Quantitative applications of IR spectrometry are eters. Stray radiation inside the monochromator poses
based on Beer's law. Because of the extremely thin cells an identical problem in IR spectrophotometers as with
used and their deterioration with age, frequent pathlength UV-visible instruments. However, because of atmos-
calibrations are necessary when using liquid cells for quan- pheric absorption in the IR region, the effects of stray
titative IR methods. Such calibrations are often done by radiation may appear even though the sample absorb-
measuring interference fringes (see Figure 14-15). To avoid ance itself is small. For this reason it is difficult to obtain
such pathlength calibrations. external standards can be exact cancellation for atmospheric absorption with some
used in the same cell used to measured analyte absorb- samples even though compensation is achieved without
ance; the pathlength is then taken into account in the the sample. Dispersive spectrometers usually use filter-
slope of the resulting calibration curve. Again, because grating arrangements to minimize stray radiation. With
liquid cells deteriorate and the pathlength varies with time, FTIR systems. the stray radiation arises from aliasing
more frequent calibrations must be made than with UV- which can be effectively avoided by electronic filtering.
visible spectrophotometry. Usually, gas cells are of suf- Extraneous radiation is also more often a problem
ficient pathlength that a direct pathlength measurement in the infrared region than in the UV-visible region,
is possible. especially with dispersive spectrometers. Here, any IR
radiation not emanating from the primary absorption
Nonlinearities source is considered extraneous radiation. Although the
modulation schemes of modern IR spectrophotometers
Nonlinearities arise from similar causes to those con- are designed to reduce the effects of IR emission and
sidered in Section 13-3 for UV-visible spectrophotome- other sources of extraneous radiation, total elimination
try. Nonlinearity due to polychromatic radiation is much of these effects is not possible. Hence measurement of
more common in IR spectrophotometry than in the UV- high absorbances (A > 2) is not recommended with
visible region. This is particularly true when the sample dispersive systems. With FTIR spectrometers, extra-
is in the gas phase at reduced pressure. and rotational neous radiation merely increases the dc level of the
fine structure is present. Such absorption lines can be interferogram and is effectively discriminated against
as narrow as a few tenths of a wavenumber. Fortu- in the data processing. With FTIR and solvent absorp-
Chap. 14 I Infrared Spectrometry
L
0.500
b m d . U\u,illy, the method ~lluttratcdin Flgur e 14-18 - 0.030
I \ uvxl to e5tnnnte the l ~ \ e l ~ n Iehe
. cho~ccof the po1nt5 0.530
of nnnnnuni ,tbtorption I \ often nnclcnr , p,~rt~cul,lrly 1 .o
w ~ t hoverliqqxng b m d \ The m,\lor ~ L I I C ~ Iprjnciple
II~
to try to ,Ipproximcite the b~i\eImeto he the ;~bsorhmce
I\ :::
1.5
"c (1))
at the m,tlytic,il wmelength if the m~ilytcband were 8 Baseline neth hod for measuring
dbsent ahsorhanccs in quantitative IR spcctro~netry.The
The batelme method ~l\\urne\th'tt the backg~ound 1005% 1 (a) or 0 A (b) reading is established by
vanes linearly with w,wclength In the leglon of the ab- drawing a straight linc belwccn points of mini-
wrpt10n band Tlns 'ltsumpt~onI \ mo\t l~kelyto hold mum absorption surr-ountlingthe al~sorptionband.
when there ax lcw ovcrlqplng bmcl\ p ~ e \ e n t ,m d the In some automatcci systems readings arc taken at
chosen baselme 1s nearly ho~l/ont'il If 'I h n d flom '1 scvcral wavelengths 011 either side of thc absorp-
concomitant d ~ ~ c c t loverlaps
y thc m'ilyte b m d , the tion hand, and a lineal. regression is uscd to obtain
the absorb;~ncco r transmittance at the analytical
b,~schncmcthod doe\ not work Whcn \uch sevele
wavelength.
background abtorption 14 prctcnt, ~t 1 4 often wi\e to
choose mother analyl~c,~l wnvclcngth whcic thi\ '14-
\urnptmn may be vdid S i r n ~ l d y ,~t 15 p ~ e f c r ~ ~ to
ble over okhcr neth hods employed in I R spectro~net~ y. First,
work w ~ t hlow absorbance b,i\clmcs t h m with those the baseline is established with the same cell and thc
drdwn at h ~ g h enb\or
~ bancc\ Oltcn. the dnJy\t mu\t same solution that is used for the analyte absorption.
resort to t~ral-md-errormethod\ 011 \ynthct~crnlxtu~e\ This compensates for- cell absorption arid cell reflection
in order to choose m nppropr1,itc I,,iscl~nc losses as well as background absorption hy the sample.
Thcrc arc scvcral ,iclvnntclgcso l the h,nehnc method Second, since measurements arc made o n a spectrum
Sec. 14-5 I Quantitative Infrared Absorption Spectrophotometry 425
obtained by scanning, there are no critical wavelength equal to the detector noise. This equation assumes that
settings or adjustments to be made. Finally, the method there is no uncertainty in measuring the 0 or 100% T
is quite rapid and relatively easy to automate with com- signals and a constant uncertainty in measuring the
puter-controlled spectrometers. transmittance, a, = a,,IE,, that is independent of the
transmittance. Under these conditions the optimum
Measurement of Integrated Absorbance. % T for minimizing the RSD in absorbance is 36.8
Quantitative applications of IR spectrometry are nor- (A = 0.434), and the RSD in absorbance follows curve
mally based on measurements made at the absorption a of Figure 5-7. Since the minimum RSD in absorbance
maximum as just described. However, there are appli- is a rather broad function of T, transmittances between
cations that use the integrated absorbance defined be- 20 and 60% provide nearly optimum results.
low: In actuality, because of background absorption
and overlapping bands there may be a considerable
uncertainty in the 100% T value used to calculate the
sample absorbance as shown in the baseline method
described previously. Thus in practice infrared meas-
The integrated absorbance and the integrated absorp- urements often show RSD values in the range 5 to l o % ,
tion coefficient are related to the total probability for particularly with complex sample matrices.
absorption as discussed in Appendix F. For example, Equation 14-8 also gives us some guidance in im-
consider the measurement of the total aliphatic C-H proving the SIN for a given IR absorption measurement.
content of a sample from the 2900-cm-I C-H band. Since a,, is usually constant for a given IR detector, the
The shape and position of this band vary from com- SIN in measuring a given T (AlcA) can be improved by
pound to compound, but the integrated absorbance is reducing the relative 0% T noise by increasing E,. This
proportional to the number of C-H groups in the mol- is normally accomplished by increasing the slit width.
ecule. The integrated absorbance is also much less sen- Equation 13-18 shows that E, is proportional to W'
sitive to changes in the monochromator spectral band- since s is proportional to W. iXormally. E, is set equal
pass than the peak absorbance. to the full-scale setting (100% T) of the instrument.
With earlier instruments it was necessary to carry When W is increased, the gain of the electronics G is
out the integration graphically or by planimetry. With decreased to keep E, constant. In any case the SIN
modern computer-controlled systems. the calculation of usually improves with W 2 with dispersive IR spectro-
integrated absorbance is often included in the software photometers. It should be noted that although the SIN
or firmware that accompanies the instrument. improves with increasing slit width, the resolution, of
course, degrades as the slits are opened. The through-
Precision of IR Absorption Measurements put can be much larger with filter-based, nondispersive
spectrometers, which allows lower values of electronic
The precision of IR absorption measurements can be gain to be used. Baseline noise levels as low as
treated in a similar manner to that of UV-visible ab- A . U . are possible.
sorption (see Sections 5-5 and 13-2) measurements. For Fourier transform IR spectrometers show the mul-
dispersive IR spectrometers, signal expressions are the tiplex advantage where the SIN of spectra taken with
same as those given in Table 13-2, while signal-to-noise the same resolution and same measurement time is
ratio expressions are identical to those in Table 13-3. n1 times greater than that obtained on a grating in-
Because of the lower radiance of IR sources and the strument, where n is the number of resolution elements
higher noise levels of IR thermal detectors compared (see Section 5-6). Michelson interferometers also show
to UV-visible sources and detectors, signal-carried shot increased throughput over grating spectrometers (Jac-
or flicker noise is not observed. Thus the limiting source quinot's advantage). Let us now briefly estimate the
of noise is usually detector noise (Johnson noise). which magnitude of the throughput advantage. The through-
is independent of the illumination level. Hence the RSD put factor at a particular uavelength Y (cm2 sr) is the
in absorbance, aAIA, often conforms to equation 5-53, product of the solid angle of the spectrometer (Q), the
which is repeated here: area of the source viewed (A,) and the transmission
factor (Top) of the optical system (see Section 3-6) as
UA -
- -
(T0r x - - 1 shown by
A E,. T ln T
throughput factor ol the ~ntertcrornctc~ V, 15 then mmat~onwere 795 crn (A due nlmtly to p-xylenc),
768 cm I (m-xylene), and 74 1 c m ' (o-xylene). Spcctrd
-
simultaneous equations result from the additivity of ab- 14-6 NEAR-INFRARED AND FAR-INFRARED
sorbance~: ABSORPTION
A,,, = 1 . 5 0 6 ~+~0.048c,,,
+ 0.000~~ The techniques and applications of near-infrared (NIR)
and far-infrared (FIR) spectrometry are quite different
A,,, = 0 . 0 2 5 ~+~ 1.440cn, + 0 . 0 0 0 ~ ~
from those discussed above for conventional. mid-IR
A,,, = 0.032c,, + 0 . 0 3 3 ~+~2 . 4 0 5 ~ ~ spectrometry. We briefly discuss here a few of the fea-
tures that make these spectral regions quite useful to
where the numbers are ah values in L g-' at each fre- the chemist.
quency and the c values are concentrations in g L - l .
As shown in Figure 14-19, the baseline method is Near-Infrared Spectrometry
used to obtain the absorbances at each frequency. With
these measured absorbances, the simultaneous equa- Near-infrared spectrometry shows some similarities to
tions can be solved for the three concentrations to yield UV-visible spectrophotometry and some to mid-IR
c, = 0.095 g LP1, c, = 0.118 g L P 1 , and c, = 0.180 spectrometry. Indeed, as was discussed in Chapter 13,
g L-'. The accuracy of the simultaneous determination the spectrophotometers used in this region (0.77 to 2.5
is better than 10%. km) are often combined UV-visible-NIR spectropho-
tometers. Although a few electronic transitions can oc-
cur in the wavelength range 0.77 to 1.0 pm, the majority
Reaction-Rate Studies. If a reacting molecule of the absorption bands observed are due to overtones
or product has an absorption band that is separated (or combinations) of fundamental bands that occur in
cleanly from other bands, IR absorption can be ex- the region 3 to 6 Fm, usually hydrogen-stretching vi-
tremely valuable in kinetics studies. Unfortunately, be- brations. Hence the region bears a strong relationship
cause of the special thin cells usually used in conven- spectrally to the mid-IR.
tional IR methods, reactions are not conveniently carried
out in the cell as they can be in UV-visible studies. Instrumentation and Techniques. The instru-
In one study. the reaction between phenyl iso- mentation used in the NIR region has been described
cyanate and various alcohols was investigated. The re- in Chapter 13. Tungsten lamps are used as sources, and
action was carried out in a vessel suspended in a con- the PbS photoconductive cell is the usual detector. Sev-
stant-temperature bath. At the desired times, a sample eral commercial instruments intended primarily for the
was removed from the mixture with a syringe and trans- UV-visible region allow near-infrared operation.
ferred to an IR cell of known thickness. The absorbance Sample preparation and handling techniques are
of the isocyanate band at 2260 to 2270 c m l was meas- not nearly as troublesome as those used in the mid-IR
ured by scanning a narrow region surrounding the band. region and are often identical to the techniques used in
Several such samples were withdrawn at various times the UV-visible region. Similarly, sample cells are usu-
during the 5- to 7-h period over which the reaction was ally the same quartz or fused silica cells used in the
studied. A calibration curve had previously been pre- region 200 to 770 nm. Cell pathlengths vary between
pared from solutions of known phenyl isocyanate con- 0.1 and 10 cm. Because there are few instrumental or
centration. The absorbancs values were then converted sample handling difficulties. NIR molar absorptivities
to concentration and used to test various rate laws; the are much more reliable than molar absorptivities ob-
reaction was found to be first order in phenyl isocyanate tained in the mid-IR region. Consequently, data ob-
and first order in the alcohol. tained in one laboratory can often be used reliably by
Obviously, the reactions that can be studied by another.
procedures similar to that described must be very slow Several solvents are suitable for use in the NIR
so that changes do not occur during the solution transfer region, although solvents containing 0 - H , N-H, and
and scanning step. Thus conventional IR spectrometry C-H groups are usually avoided. Figure 14-20 shows
is not very convenient for rapid reactions. Also. it is the regions that are useful for several common solvents.
difficult to thermostat the cell compartments in most Note that CCI, is transparent throughout the NIR re-
conventional IR instruments. Often, the geometry used gion, and CS, absorbs only weakly in a narrow region
in conventional IR systems (sample cell in front of the near 2.22 pm.
monochromator) leads to considerable heating of the
sample unless precautions are taken. Fourier transform Spectral Information. Because of the nature of
IR spectrometry has been used successfully to study the absorption bands observed in the NIR region, the
fairly rapid reactions, as discussed previously. spectral information obtained is not nearly as charac-
4 Chap. I4 / Infrared Spectrometry
Wavelength, pin
1.0 1.2 14 1.0 1.8 2.0 2.1-
('arhorr tetrachloi-idc
Carbon disulfitlc
C'hlorofornr
(ethanol stabilized)
Mcthylenc ~ h l o r ~ t l c
1)ioxanc
111( T I - l x ~ t yether
l) -20 Characteristics of
Yriethylenc gy col
scveral common solvcnts in thc near-
diinethyl ether infrared region. The solid lines in-
dicate useful regions of low solvcnt
Hcptani: absorption; the numbers iridicatc the
maximum dcsir;ihlepathlcngths in cm.
Uen~ene (Kcprinted with permission from I<.
Aczton~trilc
F. Goddu, "Near Infrared Spectro-
photometry," in Advuncr.~irz A m -
U~nletl~ylforinai~~i~le ly/icul C h w ~ i s ~ rund
y lm/run~en~~~-
tion, C . N . Reillcy, ed., vol. I , Wiley,
Dimethyl sulfoxide New York, 1960. Copyright 0 John
- Wiley (YL Sons, Inc., 1960.)
teristic of a particular n~oleculeas information obtained n near 2 0 kni. and 1' s~nglecib\orptio~l
b ~ n a t ~ oband band
in the fingerprint region of the mid-llx. Hence N I R n e x 1 0 pm Second,rry 'Lrnlnes have only vnglct hand\
spectrometry is not as generally useful for qualitative m d no bands near 2 0 pin Tcrl~ary,imines, 01 course,
analysis as is mid-IR spectrometry. The greatest utility h ~ none c ot the N-H band\ rnentloned above, dl-
of NIR spectra is in the detection and subsequent quan- though they \how NIR absorption clue to C-H 'ind
titative determination of functional groups that contain other hydrogen-containrng group\ Near-mtr,u ed spcc-
unique hydrogen atoms. trometry thus prov~dcsa very c l e a way to d14tlngulsh
The functional group correlations normally made the three ,lmine types
in the NlR region are summarized in Figure 14-21. Note Qu,mtlt,~t~vc applic,ttlon\ ot NIR yxxtromctry In-
that molar absorptivities arc also shown for many of clude the dcterm~mtionot alcohol\, phenols, hydro-
the functional groups. Because of the reliability of mo- perox~dcs,organlc m d \ , and water through measure-
lar absorptivity data in this region, it is often used, in ment ol the fundamentdl 0-11 ttretchlng v~brationIn
conjunction with band location, for qualitative identi- the rcglon 2 7 to 3.0 ~ I I Idnd the detelni~natronof e\ter\,
fication of the absorbing group as well as for quanti- ketones, ;lnd m d s through me,\wlenicnt of the tlrst
tative analysis. overtone of the carbonyl group nt 2 8 to 3 0 pm Dc-
tcctlon hmits to1 con~pounct\with "rcaton,rble" rnolal
nbsorptivit~es(c g , > 1 0 L niolk' em ') nre usually
Appiicatmns. In contrast to rn~d-IR spcctro- lev, than 0 1%
photometry, NIR spectrophotometry I\ most w~delyused
lor guantrtatzw orgdnlc lunc~tonnl-groupanalysit How-
ever, the NIR reglon has d t o been used for yunhtat~ve
ancllyses and \tudics oi hydrogen-bond~og solute-sol- The tar ~nfr'lred rcglon (50 to 1000 pm) h , ~ \long hecn
vent intcract~ons,organonietdlllc cornpou~ids.nntl in- cons~dered,I ~ ~ 1 1 ~ 1 ~a be gl el o nto1 obt,~inmgc h e m ~ c m-
~~l
orgmlc compounds tormation In the ~ ' 1 s thow ~ eve^, tal-JR \pectr omctry
As an example, cons~derthe qualit,rtive ~dentlfi- wat trought w ~ t hcxpcrinientd dltllcultre\ Thcte are
c,ttion of primary, secont-lary, m d tcrtl'lry milnes As very few sources that c,ln be used In t h ~ sregion, m d
c m be seen from F ~ g u r e14-21, thctc m m e \ h m e qurze thew ,uc notormu\ly weclk rn Intensity Gr'ltmg Imtru-
d~fterentNIR spcctr,~ P n m a ~ yamme\ h m c two stlong ments h,ivc ~ c l a t ~ v e low l y throughput in thc Idr-IK le-
d x o r p t ~ o nb'~ndt between 2 85 ,111d 1 05 p m , due to glon, p,u t ~ c u l a ~ wl y~ t hthe order-\olt~ngt~ltcrsth,rt d~e
fundamental N-M stretch~ngv ~ b r a t ~ o na\ ,corrcspond- necesw y to plevent n c ~ d ~ a t ~d~ftr,ictcd
on lrorn h~ghcr
Ing overtone doublet between 1 45 and 1 55 km. a com- gr'ltlrig o r d e ~ ts ~ o m~c,uAnrngthe detector Pt I \ not sur-
Sec. 14-7 1 lnfrared Reflection and Emission 42
prising then that FTIR spectrometry was first applied tering background is much less than seen in the mid-
in the far-IR region. The high throughput of interfer- IR region with mulls.
ometers and the rather low mechanical tolerances re- Far-infrared spectrometry has been widely used
quired in this region have made it relatively easy to in structural studies of organosulfur. organophospho-
obtain FTIR spectra with an adequate signal-to-noise rus, and organometallic compounds. In addition, the
ratio. far-IR region has been used in qualitative analysis as a
supplement to mid-IR studies. Many closely related
Instrumentation and Techniques. Almost all
compounds, such as isomers and polymers of different
FIR studies are now carried out with FTIR spectrom- chain lengths, have quite similar mid-IR spectra. but
eters (see Section 14-2). Sources in the far-IR region markedly different FIR spectra.
are typically mercury arc lamps, and detectors are usu- It is clear that far-infrared spectrometry can pro-
vide unique and valuable spectrochemical information.
ally the Golay pneumatic detector or the TGS pyro-
The widespread use of FTIR instruments is leading to
electric detector.
an explosion of interest in this spectral region and a
Solution sampling in the far-IR region is ham-
pered by the lack of suitable solvents. Since absorptiv- reevaluation of its relative importance.
ities are low in this region, it is desirable to use long-
pathlength cells, which only compounds the problem of
solvent transparency. Hydrocarbons, such as cyclo-
hexane and benzene, are among the most useful, while
CCl,, CS,, and CHC1, are also used. Sample cells are
often made with polyethylene windows, but quartz has Infrared reflection techniques, particularly the atten-
been used in the region 50 to 330 p.m. uated total reflectance (ATR) method, are widely ap-
Solid samples are generally preferred to solution plicable. The ATR method is frequently used to obtain
samples in the far-IR region. Scattering is not nearly as spectra of difficult samples. such as polymer films, rub-
serious in this region as it is in the mid-IR since the ber, food, and various resins. Although the ATR phe-
scattered radiation intensity depends inversely on the nomenon was first observed by Newton, it was not widely
fourth power of the wavelength. Spectra have been ob- applied to IR spectroscopy until the 1960s. Specular
tained on samples dispersed in mulls and in polyeth- and diffuse reflection methods are also used in the in-
ylene and wax matrices. Pressed polyethylene plates are frared region. Infrared emission spectrometry is not
commonly used with solids. used nearly as often or as routinely as IR absorption
spectrometry. However. IR emission can provide some
unique information and is quite useful for certain stud-
Spectral Information and Applications. The ies and samples. In this section we first consider IR
far-IR region can provide unique information. For ex- reflection methods and ATR in particular and then dis-
ample, the fundamental vibrations of many organo- cuss the principles of IR emission methods.
metallic and inorganic molecules fall in this region due
to the heavy atoms and weak bonds in these molecules. lnfrared Reflectance Methods
In addition, lattice vibrations of crystalline materials
occur in this region, and electron valencelconduction Most routine IR spectroscopy is carried out in the ab-
band transitions in semiconductors often correspond to sorption mode, on the beam transmitted through the
far-IR wavelengths. sample. However, information about the sample can
Far-infrared spectrometry has been applied to many also be obtained by studying the radiation reflected
different chemical problems. For example. low-fre- from it.
quency metal-ligand vibrational modes have been stud- We will consider here the three general types of
ied by FIR techniques. Because such low-frequency vi- reflectance phenomena that are used: specular, diffuse.
brations were difficult to study in the past, the spectra and internal reflection. This last method provides more
of many complexes were incorrectly assigned. Such chemical information, while specular and diffuse re-
studies have now become almost routine with FTIR flectance can provide optical information.
instruments and have allowed inorganic chemists to as-
sign such low-frequency modes correctly. As an illus- Specular a n d Diffuse Reflectance. As dis-
tration of the utility of FIR spectrometry in studying cussed in Section 3-1, specular reflectance occurs from
inorganic complexes, Figure 14-22 shows the FIR spec- smooth surfaces, while diffuse reflectance occurs from
trum of a copper complex obtained in a Nujol mull surfaces with irregularities. With many solid surfaces it
matrix. The absorption bands are sharp and the scat- is desirable to obtain reflectance information as a func-
Chap. 14 i Infrared Spectrometry
Wavenumber, cm-1
tlon ol w,ivclength, lncldent angle, artd inc~clcntbeam of incident angle can be made with an optical arrange-
polan~ntion. ment that rotates the source and source mirror relative
A \irnplc reflectance attachment of the type wall- to the sample axis. Insertion of polarizcrs into the beam
able commercmlly for ungle b e a n IR \pectrophc)tom- allows reflectance to be obtained as a function of the
cter\ 1s ~llustrntedIn bigure 14-23 The spectrum rs le- incident beam polarization.
corded w ~ t hthe \ample rn place and then w~eha reference Reflectance measurements are also ~ ~ s ctod de-
m l r o r suhtltuted lor the sample The ratio ot the two termine the thickness of thin films dcpositcd on solids
spectra I \ the letlectancc of the sample leldt~vcto thc or grown o n solid surfaces. Here rays reflcctcci directly
reference. Reflectmce me,jsurementr can J \ o be m d c from the surface interfere with those reflected from the
wlth double-beam rpectrophotometcrr by plncmg an interface between the film and the substrate. 'The num-
~ d e n t ~ c a lt t d u n e n t w ~ t hthe reference mlrror In the ber of intcrferei~cefringes in a certain wavenumber
retercnce be^^. This allev~atcsthe nced to d o pomt- interval is uscd to obtain the film thickness in a manner
by-pant ratlo calculatmn, to ohtam the reflectance similar to that uscd to obtain sample cell pathlengths
yxctrum. M~rcroreflectanccdttachment\ have dso been (see Figure 14- 15).
con\tluctcd with ~ u l t d d eoptics for condenung the beam Diffuse reflectance methods are used to study ir-
Spccular reflcctdncc me,r\urcments as ,I tunctlon regular surfaces. With such surfaces the reflected ra-
diance is nearly independent of the viewing angle. Such
measurements are often nlade with an integrating s
similar to that shown in Figure 14-24. Usually, with this
technique the radiant power reflected from the sample
is obtained as a function of the wavelength of the IR
beam. A reference reflector is then placed in the sphere
and its spcctrum obtained. The reflectance spectrum of
the sample relative to the reference is then calculated
MI R.1,
by ratioing. In the past few years, diffuse reflectance
ITTIR spectrometry has been shown to be quite useful
-23 Speculn~ reflectance m e ~ ~e- su~
for thc analysis of solids.
mentz w ~ t hn srngle-bc'm qxct~ophotometcr.
Mlrror M , deflect\ the beam to the reflectmg w ~ l -
ple S or to .I rcfelence n~~rrcsr. M , M ~ ~ r oM,
r Internal Reflection Methods. The phenome-
returns tlw bemm to i t s orlgin:d p t h nolr o"iotLiP m t e ~ n ~lcfleciion
d wa\ cQiscu\rcdill S e ~ t i o n
See. 14-7 1 Infrared Reflection and Emission 433
distorts the spectrum unless the sample thickness is less based to a large extent on BR emission measurements.
than the penetration depth of the evanescent wave. 'I'he Following this period, however, IR emission spectrom-
wavelength dependence of d,, has an effect on the spec- etry was nearly dormant until after World War 11, when
trum in that longer-wavelength absorption bands are scientists and engineers again became interested in ther-
more intense than in the corresponding absorption spec- mal radiation from rocket exhausts, flames, hot gases,
trum. and other sources. In recent years, the widespread avail-
Note that the penetration depth can he changed ability of FTIR instruments has led t o a resurgence of
by changing the crystal material, the angle of incidc~lce, interest in IR emission, and new applications are now
or both. I t is thus possible to obtain a depth profile of appearing regularly. Today I R emission is a useful tcch-
a surface using ATK spectroscopy. I n practice a mul- nique for gaseous samples, for some solids and for spec-
tireflection crystal with a 45" incidence angle can ac- era1 distribution studies of new materials and reference
commodate most routine samples. IK sources. Thc lumincsccnce of materials has also been
One of the maJor features of ATR spectroscopy measured in the IR region (IR fluorescence). However,
is that absorption spectra can he obtained on a wide lurnincsccncc measurements are quitc difficult because
variety of sample types with minimal preparation. I11 the long lifetimes enhance the probability of radiation-
addition to polished solid slabs, samples of fabrics, less decay processes.
threads, yard, or fibers can be pressed against the ATR
crystal. Powders, pastes, or suspensions can also be
Studies of lR Sources. Infrared emission spec-
accommodated. Viscous liquids can be spread o n the
trometry is still used in studying the emittance of 1R
ATR crystal, and cells arc available for obtaining ATR
radiation sources and the cmissivity of various mate-
spectra of less viscous liquids. There are even ATR flow
rials. In these studies radiation from the emitting sample
cells available. Water solutions can be used if the crystal
is collected and imaged on the entrance slit of an IR
is water insoluble.
spectrometer. Usually, the radiometric quantity desired
Because the attenuation in ATR takes place in a
is the spectral einittancc or spectral cmissivity. In order
very thin layer, surface layers and gases adsorbed on
to obtain the true cmittance spectrum, a calibrated spec-
surfaces can readily be studied. If the samplc thickness
trometer must bc used or the spectrum obtained must
exceeds the beam penetration depth, variations in thick-
be compared to that of a blackbody or calibrated source
ness do not affect the intensity of the bands obtained.
at the same temperature and under the same instru-
However, the ATK band intensity is proportional t o
mental conditions. Care must be taken to correct for
concentration in accordance with eer's law, so that
absorption by atmospheric H,O and CO,.
useful quantitative measurements can be made.
Double-beam methods have been used to measure
The spcctra obtained with ATK methods are quitc
spectral crnissivities of materials directly. In a true dou-
similar to IR absorption spectra except as noted above.
ble-beam system, the source under study is placed in
'I'hus standard I R spcctra and correlation charts can be
one beam of an optical null spectrometer and a rcfer-
used for qualitative analysis. I h e r e are. however, some
encc source or blackbody is placed in the other. The
significant differcnccs in the peak shapes and relative
two beams are initially balanced with the same source
intensities obtained in ATR and those obtained by IR
in both beams. In the semi-double-beam method, spec-
absorption methods. In ATR the peak shape is not only
tra of the reference and the sample are recorded in turn
a function of absorption coefficient, but also a function
relative to the usual source of an optical null spectrom-
of refractive index. This can introduce asyinnletries in
eter and then compared. In both cases computation is
the band profile. Also. the orientation of the samplc
simplified over single-beam nlethods because absorp-
on the ATR crystal can influence the band shapcs and
tion clue to atmospheric gases is canceled.
relative intensities.
The ATK method has been applied to many sanl-
ple types. It is convenient, and A'I'K attachments are Solid, Liquid, and Gaseous Samples. Upon
relatively inexpensive. They are available for nearly 31 thermal excitation, molecules that absorb in the IR re-
commercial IiK spectrophotometers. gion can also emit IR radiation. Because transitions can
be observed between excited vibrational levels, I K
rare emission spectrometry can provide information that is
different from that provided hy IR ahsorption spec-
I n l l ~ i ~ ecinl\\ion
d spcctlo\copy waj quite important 111 trometry. The major drawbacks to analytical 1K emis-
the e,rlly part of the twent~ethcentury In me,~\iircn~cnt\ sion spectrometry are the low radiances emitted by many
of the \pcctr al d ~ \ tI ibut~onol blxkbody I , ~ d i a t ~ oIn he
. samples at reasonable temperatures and the difficulty
cxpe~lmentalverit~cat~on 01 Plmck'\ 1,1w wcls, 111 tctit, in extracting the desired analyte emission from the
Chap. 14 / Problems 435
background IR radiation emitted by concomitants in hundred degrees are used to prevent extensive evapo-
the sample and the sample surroundings. ration. The liquid sample can be placed in a heated
The use of FTIR instruments is particularly at- sample cell of the type usually used for IR absorption.
tractive for the study of weak IR emitters or of samples Because of low radiances, the FTIR approach is usually
at low temperatures because of the throughput and mul- used for IR emission studies of liquids. Fourier trans-
tiplex advantages. In fact, FTIR systems have proven form IR emission spectrometry has also been applied
very useful in IR emission studies of planetary and stel- as a detector for constituents separated by liquid chro-
lar radiation sources, which are very weak emitters. matography.
With such sources, multichannel averaging techniques With gaseous samples the IR emission spectra can
are usually employed for further SIN improvement. With sometimes be obtained without heating, particularly with
modulation techniques FTIR systems can compensate FTIR methods. Ammonia gas, for example, has been
for IR emission by the sample surroundings (cells, hold- detected in clouds. Similarly, air pollutants have been
ers, etc.) With computers it is relatively straightforward detected remotely in plumes from industrial plants. In
to correct emission spectra for atmospheric absorption one case an interferometer was mounted on a reflecting
or emission from concomitants. telescope, and CO, and SO, were detected several
Conducting solid samples can be heated for IR hundred feet from an industrial smokestack.
emission studies by an electrical current. An inert at- Rapid scanning FTIR emission measurements have
mosphere is used to prevent oxidation. Alternatively, also been used to study transient species in shock tubes,
solids can be dissolved in suitable solvents which are electrical discharges, rocket exhausts and flames. GC
evaporated onto a salt plate. The plate is then heated effluents have also been detected. Although such spe-
electrically near the spectrometer entrance slit, and the cies can be studied by I R absorption spectrometry, IR
IR emission is monitored. In one interesting applica- emission is simpler because it requires no external IR
tion, pesticides such as malathion, DDT, and dieldrin radiation source, and difficult optical alignment is
were identified by IR emission on salt plates in quan- avoided. Because of its simplicity and its potential for
tities in the range 1 to 10 kg. obtaining valuable information, IR emission, particu-
The IR thermal emission spectra of liquids can larly with FTIR methods, is certain to be more widely
also be obtained. Usually, temperatures below a few used in the future.
PROBLEMS
14-1. Find the wavelengths (ym) and the wavenumbers lengths of the cells if the number of fringes between
(cm-I) for each of the following infrared frequencies: 1200 and 1000 cm-' was:
(a) 7.031 x loi3 Hz. (a) 13.
(b) 1.034 x 1014 Hz. (b) 6.
(c) 2.897 x 1013 Hz. (c) 22.
14-2. The mid-infrared region of the spectrum is usually 14-6. The following are typical reasons for instrumental
considered to be 2.5 to 50 y m . What frequency range errors in IR spectrophotometry. Some can be cor-
in Hz does this region encompass? rected by proper instrument adjustment. some are
14-3. What frequencies (Hz) are found in the far-infrared compensated in double-beam systems, and others can
region (50 to 1000 km)? be eliminated by appropriate calibrations. For each
potential source of error suggest first how it could be
14-4. Use the simple harmonic oscillator model for the vi-
detected and second how it could be corrected.
bration between two atoms and calculate the vibra-
(a) Amplifier gain is set too high.
tional frequencies (cm-') and wavelengths (ym) ex-
(b) Beam attenuator (optical null system) is nonlin-
pected for the following bonds.
ear.
(a) The C-C bond in ethane (k = 4.5 N cm-I). (c) Stray radiation levels are high in some wave-
(b) The C-C bond in benzene (k = 7.6 N cm-I). length regions.
(c) The C-N bond in CH,C?J (k = 17.5 N cm-I). (d) The solvent absorbs strongly in a certain region
(d) The C-H bond in ethane (k = 5.1 N cm-I). of the spectrum.
Compare the calculations with the group frequency (e) The spectrum is scanned too rapidly.
ranges given in Figure 14-3. 14-7. A n infrared spectrum was acquired for a high-boiling
14-5. Interference fringes were obtained for three empty liquid with an empirical formula of C,H,,O. Band
cells to determine their pathlengths. Find the path- intensities were described as weak (w), medium (m),
Chap. 74 I Infrared S p e c t r o m e t r y
,mi stlong (s) Mnlor h o 1 ption I,md\ were tound Contrast the use of niulls a n d salt pellets fol- the in-
, ~ tZOO0 (w), I690 (\), 1610 (w), 1575 (w). 1 Z6O (s), frared analysis of solids.
I ZOO (m), 1250 (\), 1077 (w). 104s (w), 980 (m), 700 'I'he mid-IR region is the most common region uscd
(s), m c l 725 (ni) un D r , ~ w1' s m m y ~ o n ~ l u \ ~ ,is
ons in I R spectrophoton~ctry.Discuss the unicpc infor-
l e , ~ \ ~ h l,I\e to the ~ d e n t ~ of
t y the co~npound mation provided and spccial instrumental rcquirc-
c s the rnlrrol hc tlrlvcn In an FTIR
What d ~ s t m ~must ments for measurements in the near-IR and far-IR
spectrometer In o r d c ~to ,lch~cvc0 05-, 0 5-. m d 4 0- regions.
cni ' resolut~on'j What is the purpose of as1 integrating sphere in diffuse
reflectance measurements?
Criticize or defend the following quotation. Use as
many specifics as you can to justify whct1ic1-or not Contrast conventional 1R absorption measurements
you agree with the statcinent. with attenuated total reflection measurements.
Over most o f the mid-infrared spectral range, Fourier In an A T R mcasurcmcnt, the sample has a refractive
transform instruments appear to have signal-to-noise index of 1.37 at 3.9 km. 'lhc A'I'Ii crystal was AgCI,
ratios that are better tlian those of ;I good-cjuality which has a rcfractivc index of 2.00 at this wavc-
dispersive instrument by niorc tlian an order of mag- length. If the anglc of incidence is 4S0,what is the
nitude. However, the Fourier transform method of- effective penetration depth of the cvancsccnt wavc?
fers little or no adv;mtagc (other than shortened anal- How would this change if tlie angle of incidence was
ysis time) over a good-quality grating spectro- 60•‹?
photomctcr for routine qualitative applications in the The X-H fundamental vibration in a n~olcculecon-
mid-infrared region. Furthern~ore,it suflers by com- taining the X-H functional group occurs at 3010
pxison in terms of high initial cost and suhst;intial cm I . If deuterium were substituted for hydrogen,
n-saintenance problems. The latter arisc h c c ; ~ ~ sthe
c at what wavcnumbc~-would you expect the X-D
quality of a Fourier transform spectrum dcgr;ldcs niucli vibration'?
more rapidly with instrument maladjustnicnt than docs Single-beam instruments are quite useful for UV-vis-
a spectrum produced by a grating instrument. ible absorption measurcmcnts, but are not often used
In scanning the infrared absorption spectrum of a in the I R region. Explain.
solution t c identify a ncw product in a reaction mix- 'I'he C=N stretching vibration in HCN occurs at 2006
ture, the slits open very wide in the spectral I-egion cm- '. What is the force constant of the C-N bond?
in which the product should :~bsorb.The source is In the mid-lIi region, the SIN of absorption mcas-
still quite intense in this region and the detector is uremcnts is rarely limited by signal shot n o w as it
still quite responsive. Givc possible reasons why this often is in the UV-visible region. Discuss the reasons
occurs and what errors it might cause. State how the for this.
problem might be corl-ectcd. In I R spectrophotomctcrs, thc source is usunlly niod-
Why are nondispcrsive I R instruments often used for ulnted nt 30 H7 or 15 137 DISCUS\ the ndvant'lges nnd
tlie determination of gases instead of dispersive 1K the disddvdntdgcs i>t usmg thcsc modulntion h e -
spectrophotomcters? quencies
The tollowing are gencrd references dc'ilmg with ~ n i r a l e d 1970 An cxcellcrt book on I R m\trumentat~on,~nclud-
spectroscopy Ing clcctronio nnd optlcs
I . W. J. Potts, C1~rnic.dInJrarcd Spectroxo~py, vol . 1 , Wilcy , 5. D. N. Kendall. Applied Injkrred Spcctroscopy, Rein-
New York, 1963. A classic reference filled with practical hold, L,ondon, 1966.
information. 6. G. M. Barrow, Introduc,ion to Molecr~lrcrSprctroscopy,
2. A . I,. Smith. "Infrared Spectroscopy," in Trcatisc o n McGraw-FTill, New York, 1902.
Atzalylical C'l~rmistry,2nd ed., pt. 1, vol. 7. chap. 5, 7 . N. B. Colthup, I,. H. Daly, and S. E. Wibcrlcy, Irztro-
1. M. Kolthoff and P. J. Elvinc. eds.. Wilev-lntcrscicncc. ducfion to InJrared and Ranzun Sprc~troscopy,2nd ed.,
New York, 1981 A f'r~rly recent treatment of I I i Acadenilc Press. New York, 1975.
methods. 8. K. Conlcy, Irqrarcld Spc~ctro.scol~y. 2nd cd., Allyn and
3. A . L. Smith, Applied Infrared Sprctroscop~~, Wiley, New Bacon, Boston, 19.72.
York, 1979. One of the newer books with comprehensive 9. G . 1{errberg, Molcculur Sl~ectraand Molec~~lar Structure
I R covel-agc. Il: Inficrred and Rurnurz Spectra o f Pol~~ufornic
Mol~~culcs,
4. J . E . Stewart, Irfrarrrl .";l,c~c~troscopy:Experinzenfal 'Van Nostrand, New York, 1945.
Methods und 7idmiyur.s, Marcel Dekker, New York, 10. A . I,. Smith, "Infrared Spectroscopy," in Handhook oJ
Chap. 74 / References 437
Spectroscopy, vol. 11: J . W. Robinson, ed. : CRC Press, Netherlands. 1980. This book presents the proceedings
Boca Raton, Fla., 1974. of a NATA study institute in 1979.
The following references deal specifically with Fourier trans- 18. G . A . Vanasse, ed., Spectrometric Techniques, vols. I-
form infrared methods. IV, Academic Press, New York. 197771985, There are
chapters in this set that deal with interferometers, FTIR,
P. R . Griffiths, ed.. Transform Techniques in Chemistry, diode lasers for IR, and many other topics of interest.
Plenum Press, New York, 1978. Covers FTIR as well as
FT mass spectrometry, FT NMR, and other transform
The following references deal with specific topics of concern
methods.
in this chapter.
P. R . Griffiths and J . A. deHaseth, Fourier Transform
Infrared Spectroscopy, Wiley. New York, 1986. A com- 19. J. F. Butler, K. W. Nill, A. W. Mantz, and R . S. Eng,
prehensive treatment of FTIR theory and practice. "Applications of Tunable-Diode Laser IR Spectroscopy
P. R. Griffiths, Chemical Infrared Fourier Transform to Chemical Analysis," in New Applications of Lasers
Spectroscopy, Wiley, New York, 1975. to Chemistry, G . M. Hieftje. ed., American Chemical
R . J. Bell, Introductory Fourier Transform Spectros- Society, Washington, D.C.. 1978.
copy. Academic Press, New York. 1972. 20. N. J. Harrick, Internal Reflection Spectroscopy, Wiley-
J. Ferraro and L. Basile, eds.. Four~erTransform In- Interscience, New York, 1967.
frared Spectroscopy, Academic Press, New York, 1979. 21. G. Kortum. Reflectance Spectroscopy, Springer, New
A. E . Martin, "Infrared Interferometric Spectrome- York, 1969.
ters," in Vibrational Spectra and Structure, vol. 8, J . R. 22. D . L. Wetzel, "Near-Infrared Reflectance Analysis,"
Durig, ed., Elsevier, Amsterdam, 1980. Anal. Chem., 55, 1165A (1983).
J. R. Durig, Analytical Applications of FT-IR to Molec- 23. B. R. Buchanan and D . E . Honigs, "Advances in Near-
ular and Biological Systems, D . Reidel, Dordrecht, The Infrared Spectroscopy," Spectroscopy, l(7) (1986).
Molecular luminescence spectronietl y 14 concerned with in I955 by the Americm Intlr~irncntConipmy Since
the me,l\uremcnt of photon cmJssion from molecule\ th'it tlme, inst~unientntronha\ been cont~nu~dly nn-
Andyt~calIum~nc\ccnce\pectrometry c m be b,~\ecton proved, and the number ot ,ippl~catronsh'ls grown enor-
p h o / o l u ~ n u ~ e ~ ~or
e n(chernrl~~m~nc~,(
c~ O Ke t.luor rsc-cnic mously 'I he exccllcnt detect1011Imrts, slmpl~city., ~ n d
m d yho,phore\cmtc spectrometry 'Ire type\ o f pho- \cIcct~v~ty of fluorc\ccnce h,r\ spurred the dcvclopinent
tolunime\cence \pectronietry In whrch cxcitdt~onoccur\ and adoption ot iluorometry ,i\ ,I nn~ilortechnrquc In
by ,~lxorpt~on of photons Fluo~e\ccnccinvolvo ern{\- chemistry, blochern~\try,biology and rned~cmc
sron between \tdte\ 01 ihc wnie mult~plicity.u\u,dly Phosphore\cencc W'I\ tint recognl~edIn 1568 by
slnglet t o \~nglet,ghosphorc\cence rnvolve\ i , ~ d ~ a t m n , ~ l Ccll~niThe concept thnt pho\pliore\ceiice u~volvcsab-
tr,~n\~tioii\ hetween \tatc\ of dlffeieni multipl~c~ty, u,u- wrptlon m d rernl\\lon ol hght W,I\ e\tcibl~\hedin the
ally trlplet to \inglet. In chcmilum~netccncc4pectrorn- ~ e v e ~ l t e c ncentury,
th about two ccnturie\ before the rc-
ctry, cmlssion occur\ from cxcitecl \t,~tc\produced by , ~ l l ~ , ~ t that
l o n ~tdpphed to tluorcxcnt ni'1te11~11\
'I\ well
,I chcmlccil I e x t i o n In the mnctccnth century, tdmond Rccqucrcl developed
I he phenomenon ot tluorc\cence ha\ bccn known the f ~ r \ pkosphoroscope,
t ,I niech,~n~c,ll
dcvlcc allownig
tor mmy c e n t u ~ ~ eIn\ 1852, Stoke\ introduced the term the \eyuent~alcxcrt'ltion m d vlewlng of phospliore\c-
"fluorc~ccncc" '~nd1,lter identllled wild 15 now known encc Thi\ cdlowccl the dur ,~tloii,ind exponent~,llclccay
a5 the Stoke\ sliitt, in which the cnirttcd ~adr'lt~on 15 ot of phosphore\cencc to bc o l ~ ~ r v eInd 1x94, .Tame\ Dc-
,I longer wavclcngth t11'1n t l i ~ ol t the emtlng r x l ~ , ~ t ~ o n war ob\cr vcd pho\phorcsccncc tr o m \elution\ cooled by
He even u\cd two colored tiltel\ to \elect thc cxclt,ition liqu~dM . L>cw~\m d K,)\ha In 1944 propo\ed th,it phw-
m d emr\\ron wdvclengtli\ In the IS(%)\,Stoke\ pro- phoresccncc could be used to rdcntity organlc c o n -
po\ed and dcmon\trdtett fluoromctric m,llysrs By the pounds. The f m t p'qxr on quantltatrvc pho\phollmetry
1930s, phototube\ and photomultiplier\ ~eplaccdpho- byK1er5, B r ~ t tm , d Wentworthd~dnot 'lppc'lr unt~l1957
togrqhlc phtcs as r ' l d ~ ~ ~ tcietcctol
~ o n s ,lid Jlowcd more I n i p o ~ t m tadvances In pho\phorctccncc lnstrurnentc1-
convenient and lower-hglit-level mc'nur cment\ The t m t tion were ~ e p o l t c dby Wlnefordner'\ group In the 1960s
cominercd spcct~olluoroii~eter, b,~\cclo n ,I dcslgn by More recently. thcrc h,l\ been con\~cler;lblcactlvrty rn
Bowman, C,lull~cld,mcl Ilclenfrrcnd, w,~\rntroduced the <tre,iot room-tcmpc~dture pho\pho~c\cencc.
See. 15-7 I Instrumentation 439
The use of chemiluminescence as an analytical ally, only a fraction of the wavelength range of emission
technique has emerged relatively recently compared to is monitored.
photoluminescence techniques. Chemiluminescence from For many of the reasons noted above. chemilu-
living organisms such as fireflies (bioluminescence) has minescence signals are also quite small. However. for
been observed since antiquity. Chemiluminescence due this type of luminescence, the efficiency of the chem-
to oxidation by oxygen or peroxides of phophorous, iluminescence reaction. rather than the source intensity,
halogens, and organic species was observed by many determines the number of excited molecules produced.
workers late in the eighteenth century and the nine- In this discussion of luminescence instrumenta-
teenth century. The first uses of chemiluminescence in tion, we emphasize molecular fluorescence spectrom-
analytical chemistry date back to the 1930s, in which etry, the most widely used luminescence technique.
chemiluminescent species were used as endpoint indi- However, many of the design principles apply equally
cators in titration reactions. Quantitative techniques to phosphorescence and chemiluminescence spectrom-
based on relating the chemiluminescence intensity to etry. The special instrumental requirements for the lat-
the analyte concentration have only appeared in the last ter techniques are discussed in Sections 15-4 and 15-5.
two decades. The basic component schematic for photolumi-
This chapter begins with a discussion of molecular nescence measurements is shown in Figure 15-1. The
luminescence instrumentation. Next signal and SIN source and excitation wavelength selector and the aux-
expressions are developed and used to discuss optimi- iliary optics define the characteristics of the excitation
zation of measurement conditions. The next three sec- or primary beam incident on the sample. The emission
tions separately address the special considerations, optics and wavelength selector determine the charac-
methodology, performance characteristics. and appli- teristics of the emission or secondary beam that is in-
cations of fluorescence, phosphorescence, and chem- cident on the detector. Note that the emission is nor-
iluminescence measurements. Section 15-6 deals with mally collected at 90" with respect to the excitation axis.
more specialized applications of luminescence meas- Other configurations are discussed later in this section.
urements, including lifetime and polarization measure- For chemiluminescence measurements, the excitation
ments. source and wavelength selector are not required.
15-1 INSTRUMENTATION
-.- .~ Light-ahsorbin@
surface
Fluorescence
Readout device
(a)
Excitation tnonochromator
stable, high radiance or irradiance at the excitation cence or scattering flicker noise is dominant at the de-
wavelengths of interest. For conventional sources, ap- tection limit. Compared to conventional sources, lasers
propriate lenses or mirrors are used to maximize the are more expensive, more complex, more difficult to
solid angle and the area of source radiation collected maintain (i.e., more downtime), and often less stable.
and to focus an image of the source onto an aperture Additionally, photodecomposition can be a problem
or into the sample cell. Elliptical reflectors provide the with high excitation irradiance. Pulsed lasers and Xe
most efficient radiation collection (over ~ ' T Tsr), although lamps required more complex and expensive signal
the lamp image magnification is large. The reflector and processors (e.g., boxcar integrators).
lamp can be separate units or one integral unit (see Laser-based spectrofluorometers have been con-
Figure 4-4e). structed, however, in many research laboratories. The
For simple fluorometers, a low-pressure mercury most popular lasers are the pulsed nitrogen laser (337.1-
arc lamp is the most common source. The intense lines nm line), the argon laser (488.8- and 514.6-nm lines),
at 254, 312, and 365 nm are suitable excitation wave- and tunable dye lasers. The wavelength provided by the
lengths for many molecules. These lamps can also be N, laser is suitable for excitation of many species. The
coated with phosphors to shift the wavelength to longer wavelengths provided by the Ar ion laser are somewhat
values (e.g., 254 to 280 nm is common) with a significant limiting, as the fraction of molecules that are excited
decrease in radiance. to fluorescence at these wavelengths is small. Dye lasers
In commercial spectrofluorometers, the most are the most versatile laser source because the excita-
common source is a 75- to 450-W high-pressure Xe arc tion wavelength can be optimized for the molecule of
lamp (see Figure 4-4). High-pressure Hg and Xe-Hg interest. Pulsed dye lasers with pulse widths less than
lamps are also employed. The radiance at the Hg emis- 1 ns are well suited for lifetime measurements.
sion wavelengths is much greater than that produced
by Xe at these wavelengths (see Figure 4-6). Arc lamps Wavelength-Selection Devices
require a large power supply capable of providing 15
to 30 V at 5 to 20 A. Often the supply is configured to The central wavelength and bandpass of the excitation
adjust the voltage across the arc to maintain a constant and emission wavelength selectors (e.g., filters or
arc current and a more stable radiance. In arc lamps, monochromators) are chosen to maximize the fluores-
there is a tendency, particularly during warm-up, for cence signal and to minimize the background signal. To
the arc to change position between the electrodes; this prevent the direct viewing of elastically scattered source
is denoted arc wander. Arc wandering can cause sudden radiation, the wavelength ranges passed by the exci-
variations in the observed luminescence signal, espe- tation and emission wavelength selectors should not
cially if the image of the arc is focused on a small slit overlap. Increasing the excitation bandpass with con-
aperture. Sometimes arc wander can be reduced by tinuum sources has the benefit of increasing the incident
placing the arc lamp in a varying magnetic field (e.g., radiant power. Often, the excitation bandpass is larger
from a magnetic stirrer). To prevent danger from toxic than one-tenth the half-width of the absorption band.
ozone generated by high-pressure lamps, the lamp At first thought, one might expect this condition to
housing should be sealed or have provision for de- cause nonlinearity due to polychromatic radiation as it
ozonating or venting the contents out of the laboratory. does in absorption spectrometry. However, as we will
Pulsed Xe arc lamps are gaining popularity as lumi- see in the next section, the amount of radiation power
nescence excitation sources. They provide a larger rel- absorbed by the analyte for sufficiently low analyte con-
ative radiance below 300 nm compared to arc lamps centrations is proportional to the analyte concentration
operated in the dc mode and can be used for time- even if the molar absorptivity varies over the excitation
resolved phosphorescence measurements. Deuterium bandpass. Small excitation bandpasses can provide more
lamps can be employed for excitation at wavelengths selectivity by restricting the excitation of fluorescence
lower than 300 nm, where the radiance of dc powered from concomitants.
Xe and Hg arc lamps falls off dramatically. Because the emission band is usually broad (20 to
It might be expected that the much greater irra- 100 nm), increasing the emission bandpass results in a
diance of lasers compared to conventional sources would larger fluorescence signal. This is tempered by the need
be ideal for molecular photoluminescence measure- to distinguish the analyte fluorescence from background
ments. Additionally, no excitation wavelength selector fluorescence and Raman scattering. At low analyte con-
is required. However, lasers are presently not used in centrations or in the absence of self-absorption, the
commercial systems except those designed for lifetime profile of a fluorescence band is independent of con-
measurements. Detection limits with laser excitation centration. Therefore, a large emission bandpass does
are often no better than those obtained with high-in- not cause nonlinearity even though the fluorescence
tensity conventional sources if background lumines- profile varies over the bandpass.
Chap. 75 1 Molecular Luminescence Spectrometry
Also in this case, cooling the I'MT to reduce the dark a5 1llu5tratecl rn F~gulc 15-4. bor chern~lun~~ne\cence,
current signal and noisc can furtlicr enhance the SIN. only the emlwon \pcctr,l ,ire relevant
I11 some spectrofluoro~~ieters,
thc cxcit,'I t'lon sourcc
radiation is modulated with a mechanical clioppcr and Total Luminescence Spectra. A more com-
pr~msscclwith synchrono~lsdctcciion electronics. This plctc picture of the spectral characteristics of a lumi-
processing discriminates against the dark signal and nescence molecule can be obtained by plotting a total
[(/'dark current and amplifier noisc. Other spccializetl ectrum, io shown in Figure 15-5. It is
signal processing tecliniqucs arc discussed in later sec- seen to be a three-dimcnsiond plot or a contour map
tions. of the luminescencc signal as a f ~ n c t i o nof both the
As for spectrophotorneters, incorporation of mi- excitation and emission wavelengths. The data acquired
crocomputers has greatly enhanced tlic capabilities and are often termed the excitation-emission matrix.
convenience of niolccular l~nnincsccncemeasurements. The acquisition and display of such a plot can be
Kcyhoard control of scanning parameters (scan rate and accomplished with a computerized single-channel spec-
wavelength range), slit widths, and time constants or trofluoromcter. This procedure requires acquiring and
integration times is often provided. Repetitive meas- storing a large number of emission scans with the ex-
urements or scans can bc programmed for SIN en- citation wavelength incrcnicnted to a new value for each
l~inccment.Common data manipulation and display scan. The data acquisition time can be reduced by using
options applied to the stored luminescence data include an cmission spectrograph and a multichannel detector
blank signal subtraction, calculation and display of dif- to obtain each emission spectrurn more rapidly. Tlic
ference spectra or derivaiivc spectra, fitting of calibra- video fhorometer design shown in Figure 15-6 provides
tion data and calculation of the analytc concentration a rapid and novcl means o f acquiring the data for total
in samples, calculation of statistical information, and luminescence spectra. Once the data for a thrce-di-
smoothing of spectra. Because the data arc stored, high mcnsional spcctrum are acquired, a conventional ex-
scan rates can be used without distortion by the rc- citation or emission spectrum can be plotted by sclccting
spun" time of analog recorders. Later any desired por- the portion of the data to display.
tion (wavelength interval) of the spectrum can he dis-
played with the desired scale expansion. Software Synchronous Spectra. Some spectrofluo-
packages arc available for specialized applications, in- rometers allow both the excitation and cmission mono-
cluding kinetic studies and measurcments, H P I X de-
tection, and analysis of mixtures.
I Ovalene
Scattered Anthracene
light I
Detector
surface
Emission wavelength. nm +
Source Intensity Compensation. The drift and to a sample cell filled with a standard solution of the
flicker noise of arc lamps can be significant. To correct analyte of interest. The resulting fluorescence is de-
for these nonfundamental temporal changes in lamp tected and used to generate a reference signal. A ref-
radiance, the single-beam instrumentation described erence signal can also be obtained by monitoring the
earlier in this section is upgraded to a double-beam transmitted radiant power. This technique is appropri-
design. Several possible arrangements are illustrated in ate if sample absorption is negligible.
Figure 15-9. They are all based on obtaining a reference Most commonly the sample fluorescence signal is
signal that tracks the source intensity. For the designs continually ratioed to the reference signal to compen-
shown in Figure 15-9a and b, the reference signal is sate for source fluctuations and drift. Analog ratio cir-
obtained by directly monitoring a portion of the exci- cuitry or division by a computer can be employed. Al-
tation radiation with a separate detector or the sample ternatively, the reference signal is compared to a preset
fluorescence detector. Another approach (Figure 15- voltage. The difference signal is used in a feedback
9c) involves diverting a portion of the excitation beam network to correct for source radiance variations. Either
\ '
Beam
splitter
Excitation
Source
A-selector - sample
Reference Emission
h-selector
Signal processor
FIGURE 15-9 Double-beam de- and readout
signs for source compensation or cor-
rection. In (a) a double-beam in-space
design is shown. A small portion of
the radiation passed by the excitation Excitation
monochromator is reflected by a beam h-selector
splitter to a reference detector (e.g.,
PMT, P D , or PT). In some instru- Source Sample
ments, the beam splitter is placed be- - I
fore the excitation wavelength selec- t
tor, or the source is directly viewed Attenuator Emission
h-selector
from a different angle by the refer-
ence detector. The latter approach r
cence spectral quantum efficiency over 610 to 620 nm optics and of the responsivity of the PMT. The shape
is constant to i 2 % for excitation wavelengths in the of a properly corrected emission spectrum reflects only
range 350 to 600 nm and to + 5% down to 250 nm. how the luminescence efficiency varies with the emis-
The difference between a corrected and uncor- sion wavelength, as shown in Figure 15-11. Correction
rected excitation spectrum is shown in Figure 15-11. A is more difficult than for excitation spectra because a
corrected excitation spectrum and the absorption spec- real-time correction signal cannot be obtained. Nor-
trum are often identical under proper conditions. Ide- mally, the transfer function of the emission mono-
ally, the corrected excitation spectrum reflects the chromator-PMT combination is determined with a cal-
wavelength dependence of excitation efficiency which ibrated light source of known spectral radiance (see
is proportional to the molar absorptivity. The excitation Section 4-1) or with a compound or series of compounds
and absorption spectra will differ if significant self-ab- with known corrected emission spectra. Alternatively,
sorption occurs or if the quantum efficiency is depend- the output spectral radiant power of the excitation source
ent. on the excitation wavelength. For the highest-ac- and monochromator system can be calibrated with a
curacy corrections, factors must be included to account quantum counter. After calibration, a reflector or scat-
for the wavelength dependence of the transmittance and terer, with a wavelength-independent reflectivity, is
reflectance of the beam splitter and the transmittance placed in the sample position. The excitation and emis-
of the cell walls of the quantum counter and the sample sion monochromator are set to the same wavelength
cell. and scanned simultaneously at the same rate to deter-
mine the calibration function. The uncorrected emission
Corrected Emission Spectra. Uncorrected spectral data for each resolution element are multiplied
emission spectra of a given compound differ among before display by the predetermined correction factor
instruments because of the wavelength dependence of for that spectral element. The correction is easily ac-
the efficiency of the emission wavelength selector and complished by software in computerized instruments
where the correction factors for each wavelength are
stored in memory. In older instruments, the correction
factors were programmed into a multitap potentiometer
ganged to the emission monochromator wavelength drive.
TABLE 75-1
Signal expressions for photoluminescence spectrometrya
"O,i)r,. Luminescence spectral radiant po\ier incident on the PLIT (N'): B,, . lumlncscencc spectral radiance obscr\cd
from V ' (W cm-' s r r l nm-I); l ( h ) and l ( h ) . excitation and emission throughput factor, rcspccti\el) (cm' 5s): 7;,(X)
and TL,(X'). excitation and emission transmission iactor. respecti\el) ( d m e n s ~ o n l e s s ) f: f , and f!'. solid angle of
collection of source excitation radiation and of luminescence from the sample cell. rcspccti\cl) (sr): T, and T:.
excitation and emission cell \%alltransmission. reapectivel) (dimensionlcss): a, and O,,,radiant po\\er and specttal
radiant power absorbed by the analyte in V ' . respccti\,cly (W, or W nm-I): Y,!. lum~nescenccspectral poucr h ~ c l d
( n m - ' ) : f,. secondary absorption iactor (dimensionless): p, and p:. reflectance of cell wall at excitation and em~ssion
i i a ~ e l e n g t h s respectively
, (dimensionless): a, and a;.absorptance of c d l wall at excitation and emiss~onua\clcn_eths
(dimensionless): A y ( h ) and A , @ ) . total solution absorbance at an excitation vmelength in the prefilter region and
In V ' . respectively (dimensionless): A ' ; ( h ) and Al;'(h'). total solution absorbance at an cmssion aa\elength in V
and in the postfilter region, respectively (dimensionless): ( a , ) , , . sourcc spectral radiant po\\cr mcidcnt on cell ndll
(\?I nm-I): f,,. prefilter factor (dimensionless): J , , fraction of excitation radiation absorbed b) the a n a l ~ t c(dimcn-
sionless); B,. source spectral radiance (Wcm-' s r l nm-I): A , . source area Liewed (cm'): R.,.solid ;inglc of collcct~on
of excitation monochromator (sr): i.V and U", slit a i d t h of excitat~onand emiss~onmonochromator. ropect~vsl! (em):
H and H', slit height of excitation and emission monochromator. rcipcctivel) ( c m ) : s and 5 ' . spectral bandpass of
excitation and emission monochromator. respectively, ( n m ) : A , ( X ) . analyte absorbance o \ c r b (dimensionless): T,,,
and T<&.transmission eificienc) of optical components In the excitation beam (betiiecn the source and the ccll) m d
the emission beam (between the cell and the detector). respectively (dimensionless): Ah', half-width of the em~ssion
band (nm): E . analyte molar absorpticitg (L mol ' c m - ): b,, and h,,.. prefilter and postfilter pathlcngth. respect~\cl)
(cm): b " . pathlength of illumlnat~onand observation (cm): 1 % . uidth of the excitation h e a n In the center of the ccll
(cm); V . , volume element of illumination and obserlation (cn~:).
fluorescence. With source modulation, the mean value spectral radiant powers. Here and elsewhere a prime is
of Ed is zero. Longer-lived phosphorescence signals are used to distinguish the emission wavelengths or quan-
distinguished from fluorescence and scattering signals tities related to the emission wavelength from excitation
by temporally separating the excitation and observation wavelengths or related quantities.
steps. Hence with time discrimination, E,, = 0,EL =
E p , and EbL = E,,, where the subscript P denotes Initial Assumptions. Now we derive equations
phosphorescence. In chemiluminescence (CL) meas- relating the luminescence radiant power seen by the
urements, there is no external excitation source, no detector [(@,,),I to instrumental and solution variables.
fluorescence, phosphorescence, or scattering; thus First, we calculate the radiant power absorbed by the
EL = EcL, and EbL = EbcL. analyte. Then we find the fraction of the absorbed ra-
Equation 15-4 shows that EL is determined by the diant power that is converted to luminescence and col-
PMT and signal modifier gains, the PMT responsivity, lected by the emission wavelength selector. We consider
and the analyte luminescence spectral radiant power the idealized model of Figure 15-12, in which the lu-
incident on the PMT [(B,,),]. Similar expressions apply minescence is collected at 90" with respect to the ex-
for the relationship between different components of citation axis. The volume element V in which molecules
the background readout signals and their respective are radiationally excited is determined by the rectan-
Chap. 15 / Molecular Luminescence Spectrometry
incident sourcc spectral radiant power [(a,),,], the ex- r'rnge ot wavelengths over whlch equatroii 15-13 IS In-
citation cell wall transmission factor (T,.), the prcl'ilter- tegrated IS determmed by the excitation spectral band-
efl'ect factor (A,,.), the total spectral absorptancc (aj::), pdss (lo i ,) The excitation throughput factor (equa-
and the fraction o f the total spectral radiant power ab- tlon 15-41 c m be cpecrf~callywr~tten (we equatmn
sorbed by the analyte (,f,,). 3-70) d\ Y(X) = WHl1,,T,,,t(h) We a\\umc that the
For a conventional source, the excitation spectral wurce entlancc o p t m 'Ire properly confrgurcd to match
radiant power is a function ( c q ~ ~ a t i o15-7)
n of the sourcc thc Eln (or whd angle a,,) of the excitation monoch-
spectral radiance (6,) and the throughput factor for thc lomitor (see Flgure 15-13) and t11,lr the image of the
excitation wavelength sclcctor and auxiliary optics [Y(hj]. source kocuwd on the cntrnncc 41t 1s I q c r than the
The excitation throughput l'actor is given by equation { l ~ dt ~ m c n s ~ o nIn
s that ca\e the p o d u c t A,L4 is equdl
15-8, where A , is the sourcc area viewed and imagcd to W1Il2,, It the sourcc lm~lglng14 1 1, A , = W H and
Sec. 75-2 1 Signal and Noise Expressions 453
I I
Source Excitation Sample
monochromator cell
FIGURE 15-13 Excitation considerations. A lens or mirror or combi-
nation of optical components collects the source radiation with solid angle
R, and focuses an image of the source onto the entrance slit of the ex-
citation monochromator. The imaging solid angle matches the solid angle
of collection of the monochromator (R,,). Overfilling the monochromator
would increase the stray radiation and decrease the optical throughput.
The two solid angles are related by 0 , = m2n,, where rn is the magni-
fication. The area of the source image focused at the entrance slit is m'
times the original source area. Often the dimensions of the source arc
image are larger than the entrance slit dimensions. and the source area
seen by the entrance slit (A,) is given by WHIm2.Thus for a given mon-
ochromator and entrance slit dimensions, 0 4 , = n,,WH. Hence col-
lecting the source radiation at a larger solid angle than the acceptance
solid angle of the monochromator generally provides no greater radiation
throughput because the area of the source viewed is reduced correspond-
ingly. If the source dimensions are smaller than the entrance slit dimen-
sions. the throughput can be increased by choosing R , > Cl,, up to the
point that the entrance slit is filled with the magnified source image. When
an elliptical reflector is employed to collect the source radiation (the source
at one focus and the entrance slit at the other focus), the collection solid
angle can be greater than 2 ~However. the image is highly magnified such
that a significant increase in throughput is not realized. Similar consid-
erations apply to imaging the radiation leaving the exit slit onto the cell.
As shown, the solid angle of incidence onto the cell (0,) is often made
less than Re, to reduce beam divergence and create a beam that is more
nearly collimated. In this case. the size of the image in the cell is magnified
and greater than WH.
R, = a,,. The dimensions of the source arc or its image for all imaging optics between the source and the sample
at the entrance slit or some other aperture (an aperture cell and Tf(h) is the spectral transmittance of the ex-
mask at the cell wall) may limit the maximum magnitude citation filter. Typically, a low-pressure Hg arc lamp is
of A,. the source. The combination of some limiting excitation
As is also shown in Figure 15-13, the cross-sec- aperture between the source and sample cell and the
tional area of the image in the viewed volume element type of imaging (i.e., magnification) determines the ac-
(wh) depends on the imaging between the excitation tual source area (A,) that is viewed and imaged into
monochromator exit slit and the sample cell. To achieve the sample cell and the cross-sectional area of the beam
a more collimated beam within the cell, the Fin of in- within the cell. The source radiation is usually collected
cidence is often increased, with the result that the image with fast Fll or F12 optics so that the solid angle of
size in the cell is greater than WH. collection (R,) is relatively large. Normally, the exci-
Equation 15-14 is a limiting form of equation tation filter isolates a single source line. In this case the
15-13 which applies when an excitation monochron~ator integration in equation 15-13 is not necessary, B, is
is employed and all wavelength-dependent quantities replaced by the line radiance B, and all other quantities
are constant over the excitation spectral bandpass. Be- are evaluated at the source line wavelength, as shown
cause the excitation bandpass s is proportional to W, in equation 15-15. If two or more source lines are trans-
the absorbed radiant power is proportional to W2. mitted, @; is determined by evaluating the right side
For a fluorometer, the source radiation is col- of equation 15-15 for each excitation wavelength and
lected and imaged into the sample cell with optical com- then summing the results. With a phosphor-coated lamp,
ponents. The excitation transmission factor is given by equation 15-13 applies, and the limits of integration are
T&) = T,Tf(X), where T, is the transmission factor determined by the combination of the wavelength range
4 Chap. 15 1 Molecular Luminescence Spectrometry
of the source cml\slon prot~le,rnd the \pcctr,rl trms- lum~ne\cenccrnclinnt power ~ n c ~ d c nont the detector
mrttmce 01 the cxc~tdt~on flltel When thr\ result is sub\t~tutedmto equatron 15-4, we
With lajcr exc~t'tt~on,'111 w,~vclcngth-clcpcnderlt oht'r~n cquat~on15-22 We will now use th14 ccluat~on
q i ~ ~ l n t ~ tcrm
c su\u,rlly be con\~dercdto hc constmt ovel to ~ I \ C L I S \emr\slon parmietcrs
the llnc w ~ d t h ,m d thc totd absorbed powcr 1s detel- The most crrt~cdlenuss~on~nstrun~ental p,tr~lmc-
mlncd by 'in equat~ons~mrlarto c ~ l u ~ ~ 15-15 t ~ o n In t h ~ s tcrr ale h , T,,,,(hl), , ~ n dti' I he pathlength h I \ c r ~ t ~ c a l
caw we repl,~ccI2,A,RI,I,(A) by (I),,, the totd rad~ant bccauw ~t allcctr the mcigmtudc ol @, or the s v c ot
powel medsurcd at the cell w'rll the volume clement from wh~chlum~ncscence15 col-
lected The latte~two tactors rnlluencc w l ~ t tractmnt ol
Emission Variables. We can consider the vol- the lumlnevxncc cmtted ovcr a h r o d wavelength rmge
ume clcment V:' as a source of luminescence charac- m d rn all drrect~on\1s ~rctu;iIIycollected r~ndob\ervcd
terized by analyte spectral radiance H,, . This spcctral Wlth an eml\\lon monochrom'rtor, the crnlsvon
radiance, referenced to just outside the vicwcd cell wall, trnnsrnls\lon t x t o r 111 eclucitlon 15-20 1s glven by
is calculated from equation 15-16. The product of /,,,,(hr) = l,,,,t(Xr) The eml\sron monochrormto~\pee-
and the luminescence spcctral powcr yield ( Y , , )equals tral bdndpls\ ( s t ) dctermmes the ~ ~ i v e l c n grmgc
t h ovel
thc total spectral lurnincsccncc radiant power emitted whlch t ( h l )is I m t e , equatwn 15-22 15 ~ntegr'ltcd,,rnd
in V:::. This radiant power is converted into a radiance cm~s\lon1s v~cwed As lor the excltat~onmonochro-
by dividing by 4nhh'Y The secondary absorption factor mator (see Flgu~e 15- 13). the Eln or ~ j h dmglc (f2, ,,,)
,f, accounts for the absorption of the photons between of the e m w o n monoch~om,rtorshould he matched For
their point of enlission in V" and the viewed cell face. ' g~vcnm,tgnrfrc'~tlon (rn') of the opt~csbetween the
1
Finally, T: is the emission ccll wall transmission factor cell and emr\slon monochrom~ltor,h = W'lrn', f i t -
(typically, 96%) as controlled (equation 15-17) by the t2L,,,(rn')L, , ~ n dh' = H'lrn', whele W' and / I ' arc the
emission cell wall absorptance (a:) and the csll wall eml\\lon nionoch~om,itoisllt w~dthmcl \ht licrght m d
rcflcctance (pi). 12' I \ the hc~ghtof the volume element v~cwcdIn the
The secondary absorption factor 1; is the product \ample ccll. Iilrnce the o v c ~ , ~throughput
ll 1s dcte~ni~ncct
of two factors, as shown in equation 15-18. The first by the Fin of the enllssron monochroni,rtor Collecting
exponential factor cquals the transmittance in the post- the tluorcvxnce w ~ t hn \ol~clmgle l a ~ g c rt h m 12,,,, rc-
absorption region (Tfr) betwccn the vicwcd face of V:' sults In a cm,dler volume element (I e , smaller h ) bcmg
and the cmission ccll wall. It can be calculated if the viewed 101 't glvcn s l ~ tw~dth Often h' i- bi ,lnd the
total spectral absorbance [A:"(hr)] ovcr the postfilter he~ghtof the excltatlon heam rather than If' controls
pathlength (h,,,) is known. The second hctor in cqu a t'Ion the u e of V IIowever, ~f the rnCrgn~llc,1t~on 14 1,ilge
15-18 accounts for the absorption of emitted photons enough th,tt h' < h, H ' controlc the lie~ghtot the volume
within i f : :and depends on the total solution absorbance elcmcnt v~ewcd,11I\ repl,~ccctby h' in equation5 15-16
(A:(h1)] at the cmission wavelengths across the width m d 15-20, dnd (I,, 14 cot r cspond~nglyreduced by the
of the exciting beam (w).This factor is more complex f x t o r lz'llz
becausc different photons travel different pathlengths For a tluorornctcr, T,,,,(X1) = T:T,(hl), where
within V::'depending on whcrc they are emitted. T,' IS the tranmlsslon t x t o r for all lmdg~ngcomponents
The spcctral radiant powcr observed by the PMT u\ed w ~ t hthe filter and T,(Xr) i \ the spectral trmsm1t-
is the product of B,, and the cmission optics throughput tancc of the fdtcr. The latter factor detcrm~ne\the
factor, Y(X1) (equation 15-19). From equation 15-20, wdvclcngth range o v a wh~chequ'ltlon 15-22 I \ inte-
Y(hf) is determined by the area of the viewcd face of grdtcd 'I he niagn~tudeot b I \ dcterm~ncdby some
V " (hb'"), the collection solid angle of the cmission q c r t u r e rn the c m ~ \ \ ~ obemin The product h'f2' 15
optics (fi'), and the cmission optics transmission factor u\ually much larger than that with a n ernl\\lon mono-
[T,,,,(X1)]. This last factor accounts for the efficiency chrom,itor. It 15 po\\lblc to collect fluorescence o v e ~'1
with which a given wavelength is transmitted between pathlength of about O 5 cm w ~ t han Fin of 1 to 2 and
the sample cell and the detector; it thus determines the to Intercept the complete Im,igc w ~ t hthe I'MT photo-
range of emission wavelengths passed to the detector cathode
(assumed to be a PMT). In a spect~ofluororneter,the v~ewedvolume elc-
Suhstitilting equations 15-16 and 15-20 into cqua- rnent IS normally only a \mdl Iract~onot the total vol-
tion 15-19 yields equation 15-21 for the lumincsccnce ume elcmcnt In whlch exnt,lt~onoccur\, ar ~llu\tratcd
spcctral radiant power incident on the PMT. The in- rn F~gurc15-14d As 5hown In Figure 15-14b, the viewcd
tegral of equation 15-21 ovcr the wavelength interval pathlength h m d V c m hc cubstant~dlyrnclen5ed by
passed by the cmission wavelength selector is the total u\lng 'I hor~zont~il c l ~ nr
t rmgemrnt
Sec. 15-2 1 Signal and Noise Expressions 455
Final Signal Expressions. By combining equa- tion. The solid angle of luminescence collected and the
tions 15-13 and 15-22, we obtain equation 15-23forethe emission pathlengths, w and b,,, vary for photons col-
photoluminescence signal. Equations 15-24 to 15-26 are lected from different regions in V'.
limiting cases of this general equation for a spectro- We have also ignored some other factors, such as
fluorometer in which it is assumed that all wavelength- stray radiation and elastic scattering by particles in the
dependent quantities are constant over the excitation sample solution. Reflection of excitation and emission
and emission bandpasses. It is further assumed in equa- radiation from the cell walls opposite the excitation and
tion 15-25 that the absorbances at the excitation and emission cell walls, respectively, will slightly increase
emission wavelengths are small such that f,, = f, = 1 the observed luminescence signal. For some molecules
and a,"f, = 2.303 A:. Finally, in equation 15-26 we at high concentration, reemission must be considered.
substitute ~ b * for
c A,", where c and e are the analyte Here analyte molecules in the emission beam absorb
concentration and molar absorptivity. It is further as- luminescence photons emitted by other analyte mole-
sumed that measurements are made at the maximum cules; a fraction of this absorbed radiation is reemitted
of an emission band that is Gaussian in shape. In this as luminescence.
case we can substitute [YLI(1.06AX')] for Y,,, where In the literature. the luminescence radiance is often
YL is the luminescence power yield and AX' is the half- calculated by dividing the total radiant power of lu-
width of the emission band. Note that the photolumi- minescence by 4nhbxq2 rather than 4nhbx (see equa-
nescence signal is directly proportional to the analyte tion 15-16). where q is the refractive index of the test
concentration. Because s' and b y are proportional to solution. The refractive index factor accounts for the
W ' , E L is proportional to (W')2. With 1: 1 imaging in fact that emission rays, not parallel to the normal to
the emission beam, b" = W' and 0' = a,,. the emission cell wall, are bent away from the normal
when passing from a higher refractive medium (the so-
Refinements of Signal Expressions lution) to a lower refractive medium (air). Because the
emission radiation is expanded into a larger solid angle
The equations in Table 15-1 tell us how instrumental after passing through the cell wall. the actual solid angle
variables affect the luminescence signal. They should of collection from V ' is less than CL' by q p 2 However,
be considered semiquantitative in terms of predicting the refractive index effect also changes the magnifica-
the absolute value of the luminescence signal. The equa- tion of the image focused on the entrance slit of the
tions could be refined to account for some assumptions monochromator. Thus the viewed volume element be-
that have been made. comes larger in proportion to q2; this counteracts the
We first note that the region of excitation and reduction in solid angle. It has been shown that under
viewing (V') is not a perfect parallelepiped. The ex- some experimental conditions. the efficiency of collec-
citation beam is not perfectly collimated such that b,, tion of fluorescence from a given fluorophore in sol-
and b* vary for different excitation rays. The irradiance vents of different refractive index does not vary signif-
of the excitation beam often varies across its cross sec- icantly.
Chap. 15 I Molecular Lurnitiescence Spectrometry
0.5
Log QS concentration, r*g/rnL Pathlength
FIGURE 15-15 Fluorescence calibration curve FIGURE 15-16 Relative excitation beam inten-
for quinine sulfate (QS). Curve a is the actual sity as a function of distance traversed through
calibration plot. The calibration is linear to within the cell for different sample transmittances: (a)
1% up to 1 pg mL-' because f,,, = f , = 1 and 80% T;(b) 65% T:(c) 50% T; (d) 25% T;(e)
a> 2.303 A:. Above 1 pg mL-': nonlinearity 10% T;(f) 1% T;(g) 0.1% T. The section be-
due to primary absorption effects is observed be- tween the two vertical lines labeled h and i rep-
cause f,, < 1 and a: = (1 - lo;*). Above about resents b*, the pathlength over which lumines-
100 kg mL-', the prefilter effect causes roll-over. cence is viewed. The difference in transmittance
In curve b, the signals have been corrected for between h and i is the fraction of the excitation
the attenuation due to primary absorption and radiation absorbed within V* as shown for curve
linearity is restored. Conditions: A,, = 365 nm: d. The pathlength from the front of the cell to h
A,, = 450 nm. (From M. C. Yappert, Ph.D. is the prefilter pathlength. Thus the transmittance
thesis; Oregon State University, 1985; with per- at point his the prefilter factor Cf,,) or the fraction
mission of author.) of the incident radiation that reaches the viewed
volume element.
Corrected Measurements
rected excitation spectrum is a plot of relative quanta
The equations developed help us understand the cor- per second per nanometer vs. A,,. If the quantum ef-
rection schemes described in Section 15-1. ficiency is independent of the excitation wavelength,
primary absorption effects are negligible, and the op-
Source Intensity Correction. The lumines- tical characteristics of the beam splitter are independent
cence signal is proportional to B,. In source intensity of the excitation wavelength or are compensated, the
correction, the instrument is configured to keep B, con- shape of the spectrum reflects the dependence of the
stant or to divide E L by a reference signal proportional molar absorptivity on wavelength; it has a shape iden-
to B,. tical to the absorption spectrum. If a thermal detector
is used as the reference detector, the reference signal
Corrected Excitation Spectra. During an ex- is proportional to energy rather than quanta. The ex-
citation scan, all emission variables are held constant. citation spectra can be normalized to the shape obtained
From Equation 15-26, B,, Top, T,, and E vary with with a quantum counter by multiplying the reference
excitation wavelength. With an instrument configured signal by A,,.
for corrected excitation spectra, EL is ratioed to a ref-
erence signal obtained from a reference detector (see Corrected Emission Spectra. Excitation vari-
Figure 15-9a). The reference signal is proportional to ables are held constant during an emission scan. From
B,,,T,, with a quantum counter and to BAT,,, with a equation 15-25, R(A1), Y,,, T,, and TAP are dependent
thermal detector. With a quantum counter, the cor- on the emission wavelength. For corrected emission
Chap. 75 I Molecular Luminescence Spectrometry
\pectr,i, the m\trument I\ conhgurcd to rnult~ply C, two different positions along the emission axis (i.c.,
by ,I factor tli,~t 15 propolt~onalto [R(A1)T:,,,I ' 01 different I?,,,).
[ K ( A 1 r,'),,]
)- 1 he con ecteci emltrmn spcctrum thu, 111-
d~cate\the true \h,ipe of the emlc\lon \pcctrurn ( I c ,
how Y,, or (11,~ depend\ upon A,,,,) IJ\u,dly. T i I \ ,IS-
wmed to he ~ndependcntot A,,,,
With laser excitation, it is possible t o approach satu-
A b s o r p t ~ o n Corrected Measurements. For ration conditions for fluorescence in which a significant
ahorptlon corrected ~nea\uremcnt\,the oh\ervcd lu- fraction of the analyte molecules is in the excited state.
mlnescence vgnal E ; 1s div~dedby a prim,n-y ,~h\orpt~on I t can be shown that thc fli~orescencesignal is related
correction factor (jl,) and a xcondnry ,~hsorptroncor- to the incident irradiancc (El,),,in photons c m 2 s ' by
rection factor (j,):
TABLE 75-2
Signal-to-noise expressions for molecular photoluminescence spectrometry
a limiting best value at the point that flicker noisc in mine\ccnce by calculating typ~calvalue\ 01 the cal~hrn-
the background signals becomes dominant. t ~ o ncurve dopc (tn) and the blank n o i w (A,,,) under
As the analytc concentration is increased, blank favorable c o n d ~ t ~ o nFrom
\. equation 15-26, the slope
noise becomes negligible and noise carried by the an- reterenccd to the PMT photocathode 1s h , ImGt or
alyte signal dominates. Analyte luminescence shot noise
can he limiting at modcrate conccntrations in which case
i , should be increased as for the blank noisc case or
the electronic bandwidth ( K ) should be reduced t o im-
prove precision. At yet higher analytc concentrations, In t h ~ \cqu'lt~on @,, i\ the rnc~dcnt r,idl'~nt power
flicker noisc in the analytc luminescence signal is usually ( = WHS1,xsT,,,,H,), m d we estirn,ite thdt
limiting. With high blank noisc, the signal-shot-noise- (T, I': T&,ll 06) = 0.22 W ~ t ha t y p d \pectrotluoro-
limiting region is often not observed (i.e., when the meter and c x c ~ t a t ~ owcivelcngtb
n and bandpas\, (I,,,
analytical signal-carricd noise becomes greater than the In the range 0 01 to 10 mW. FOI c,~lculatlonpurpo\es,
blank noise, the signal shot noise is negligihlc relative we w ~ l laswine the follow~ngt y p ~ c dvdlues tor @,, m d
to signal flicker noise). The sourcc flicker factor (t,) cell and cml\\lon par crmcter\:
with conventional sources is typically 0.003 to 0.03 with
a 1- to 10-s time constant or integration time. Double-
beam systems compensate for sourcc intensity fluctua-
tions and rcduce the source flicker factor.
At even higher analyte concentrations, where the
prefilter effect causes the analytc luminescerice signal
t o decrease with increasing concentration, analyte lu-
minescence shot noisc or even blank noise can agair~
become dominant. Because the background signals arc
also attenuated, dark current and amplifier-readout noisc nf = 0.004 (Fl4 system)
-
are relatively more significant. 4Tr
A common spectrofluoromcter specification is the
SIN for the Kaman band of common solvents under
specified conditions (e.g., for water, X,, = 350 nm. Note that (b lh)(s'IAX')(L2'14n) = 4 x 10 ' o~ o d y
X ,,,, = 397 nm, s = s' = 5 to 10 nm, T = 1 s). This about 0 004% ot the lum~nesccncein the cell I \ collected
specification is most useful for comparing cliffercnt by the emission monochr omator. Sub,t~tutrngthcse val-
spcctrofluorometers. The Kaman signal is proportional ues into cquat~on15-40 yields
to the same instrumental parameters as the Iumines-
cencc. Thus thc SIN of the Kaman hand is higher for
instruments with larger source radiances and excitation
and emission throughput factors. For good spectrom- The blank noisc is given by the denominator of
eters the SIN of the Raman band is in the range 30 to equation 15-38, If background signals arc low
100. The SIN of thc Raman band has little relevance to (i, < i,,). dark current shot noise is limiting and
Sluorometric measurements in which background lu-
minescencc noise limits the SIN.
The D L calculated above corresponds to an an- blank noise in samples can be much greater. Second,
alyte absorbance of 4 x 10-"n a 1-cm-pathlength cell. the calibration slope can be less due to quenching. Third,
In conventional absorption measurements. the noise the calibration curve can be nonlinear at subnanomolar
limited DL is reached when the analyte absorbance is concentrations due to contamination problems. as il-
about 10-" At this point, signal flicker noise limits our lustrated in Figure 15-17. Finally, the practical D L can
ability to distinguish two large signals (the sample and be limited by the variability of the uncompensated blank
reference signal) which differ by only about 0.01%. For component from sample to sample rather than noise.
photoluminescence measurements of appropriate mol-
ecules, the difference between the total signal and the
blank signal can still be distinguished even when the 15-3 MOLECULAR FLUORESCENCE
net analyte absorption is very small. SPECTROMETRY
Theoretically, the D L calculated above could be
lowered substantially. By cooling the PMT and using Molecular fluorescence spectrometry has become es-
photon counting, the dark current noise, and thus the tablished as a routine technique in many specialized
blank noise, can readily be reduced by an order of applications. For some species. molecular fluorescence
magnitude. The slope of the calibration curve could be spectrometry can yield lower DLs and greater selectiv-
increased two orders of magnitude with an emission ity than molecular absorption spectrometry and is the
filter (i.e., collection of a larger fraction of the total preferred technique when these characteristics are para-
luminescence emitted) and another two orders of mag- mount. However, it should be noted that the instru-
nitude by increasing the incident radiant power (i.e., mentation and theory are more complex.
laser excitation). Thus it would seem that femtomolar Although fluorescence can be observed from al-
or even attomolar DLs are possible. most any molecule with a sufficiently intense excitation
The best experimental DLs for efficient fluoro- beam, only a small fraction of molecules exhibit fluo-
phores such as fluorescein are about 0.1 pM. Such de- rescence characteristics (e.g., high +
), that are favor-
tection limits have been obtained with laser excitation able for analytical purposes. This makes fluorescence
with the analyte dissolved in prepurified solvents. Ex-
perimental DLs are much larger than the theoretical
DLs in the femtomolar or attomolar range predicted
by the discussion above because scattering or back-
ground luminescence noise becomes limiting when ex-
perimental variables are adjusted to maximize the cal-
ibration slope. For a given fluorophore, the DL becomes
independent of the values of instrumental variables (see
equation 15-40) that affect m when background flicker
noise is limiting (i.e., a,, x i, x cD,. s ' . a',
etc.).
To understand the magnitude of the background
signal. it is useful to define a blank equivalent concen-
tration (BEC). It is the concentration of the analyte that
yields a luminescence signal equal to the dark signal- LOG (CQS3 XIO), p p T r
corrected background signal (BEC = i,lm). For pur- FIGURE 15-17 Fluorescence calibration curve
ified solvents. and efficient fluorophores, the BEC is for quinine sulfate (QS). Here the fluorescence
rarely lower than about 10 pM. Under these conditions, signal is given in counts and the concentration in
the experimental D L is usually no better than about part per trillion (pptr) or pg mL-'. The large
0.1 pM. At this concentration, the analyte lumines- dynamic range of fluorescence is illustrated.
cence signal is at most 1% of the blank signal However, below about 100 pg mL- ', nonlinearity
(SIB = 0.01) and the RSD in the blank signal is at best is observed (the fluorescence signal is greater than
about 1 % . In complex samples, the BEC is much higher expected). This behavior is believed to be due to
due to luminescing concomitants. For example. the BEC contamination problems (e.g., desorption of flu-
orescent contaminants from glassware or the cell
for human serum diluted 1: 100 can be 1000 times greater walls). Under these conditions, the calibration
than that for water. slope at higher concentrations cannot be used to
The discussion above indicates that the D L for a evaluate the DL. [From J. D. Ingle, Jr., and R.
given analyte must be experimentally determined for a L. Wilson. Anal. Clzem., 48, 1641 (1976), copy-
given instrument. The value obtained for the analyte right 1976, with permission of the American
in a pure solvent must be used with caution. First. the Chemical Society.]
462 Chap. 75 I Molecular Luminescence Spectrometry
Ic\s ~1niver4,iIthan ahsorption tcchnrque\, but more s - potent~~tl d c c ~ e ~ In e )IN must he xceptablc Al-
~ rthe
lcct~ve kven rt many \pecie\ 111 a s'lmple tluorc\ce \ig- tern,~tlvely,,I shutter can be used \o thdt the w n p l c IS
n ~ l ~ c ~ n i t\elect~v~ty
ly, ts otten cnhancetl by nn appro- expo5ed to cxc~tdtionr ' ~ d ~ ' ~ tonly ~ o n dunng the mcas-
prr'ltc choice of e x c ~ h t ~ omn d cmrssion wnvclcngth\ urcment t m ~ c
LJlt~~n,~tely, the \clcct~v~ty15 lnn~tcdby the plcscncc of For qu,lnt~t,~t~ve work, ,I \pcctrolluorometer or
background \lgn,ll\ Flcctrrc,rl n o ~ s cIn the dark m d lluoronieter may be employccl Pluorornctcr\ ,ire lecc
backg~oundvgn,~l\u\ually control4 thc detect~onIrm~ts c x p c n ~ v em d can prov~dcbctter detect~on111nrt\it the
ach~cv,~blc. wurcc rAlCtntpowcr 14 re,i\on,lbly h ~ g hW ~ t hIrnc wurce
exc~tatlon,tlie n v d ' ~ b l eexet,lt~onwavelengths mdy not
wltdbly n1'1tch the abwrpt~onspcctruni of the m'llyte
Fol devclopmentnl w o k. ~ \pect~olluorometer\arc In-
Mmt tluoromctr~cdeternim,ltmn\ 'ire cnrricd out in 40- v , ~ l u ~ ~ hExcitat~on
le m d eml\\lon spcctr'l of 'I \t,lndard
lution with extcrn'~lst,~nd,t~dirat~on I hcrcforc, wlid ~ndic,ttethe wavelengths m d b,indpa\scs thc~tmaxlrnm
\nniple\ mu4t be d~ssolved In spcc1~111zed ,q>pI~cdion\ the signnl honl the tluoropliore or the type\ and char-
to be discu\\ed I,~terIn thi\ \ect~on,get\ or \old phcl\e actci~st~c\ ot hlters to u\c I n ,I fluoromctcr Spectr'i ol
fluo~csccnceine(~\uremcnt\,Ire ucclul the s,~mplecan be comparcd t o tho\e of a stmd,rrd to
In tn'lny .~ppl~c,tt~ons, m i n ~ m dsample treatment delmcatc the nature of the bxkground lluorc\cence
15 r c y ~ l ~ r cbecauw
d the natwe fluoresccnce p r o p e ~ t ~ c s and ccattering c~gnal\m d to d\\e\\ prlmary m d we-
ot the analytc 'ue u x d . Eor case\ In wh~chthe mCrlyte o n d x y absorption by concom~tmts W,ivelengths m d
14 not tluorc\ccnt or the fluorescence quantum efti- v,rlues 01 bandpa\\ can be cho\cn to optiniue the DL,
crency ~nadcqu'ltc,derivnt~~ntlon re,~ctionr'Ire u\ed SIB, or JIN to1 '1 part~cuI,~r type ol wrnple
to convert the m,~lytcInto ,I product w ~ t hgood fluo-
rocence ch,li c~ctcrlstlcsThe fluorecccnce \lgn;rl 01 the ackground Signals
product is re1,rted to the andyte conccntratmn. Most
conmonly, the malyt~calrcxtion I \ allowed to reach Because background signals and noise are critical, it is
cq~uhbr~urn b e f o ~ emeacurcrncnt\ 'ire mnde A\ w ~ t h important to understand their origin. Common sources of
rnolccul,~rabwrpt~on\pectromctry, tltratiom or kinet- background signals are listed in Tahle 15-3. The origin
~ c \ - b a dmcarurement\ c m be ccu-ried out by moni- and characteristics of the elastic scattering and Kaman
toring n fluorescent product or r c x t m t mvolvccl In 1' scattering signals are discussed in detail in Chapter 16.
\elect~vcrcdcllon
Such \elution condihons as ionlc ctrcngth, pH, Elastic Scattering and Reflection Signals.
v~sco\~ty, ,md temperature mu\t be caretully controlled, The wavelengths of the scattering signals can be prc-
,I\ they c m 'tffect the m o l x absorptivity, the q i ~ ~ u i t u n ~ dictcd as shown in Figure 15-IS. The elastic scattering
ylcld, and the t r x t ~ o not the malytc in the lluo~e\cent signal, as well as tlie reflcction signal, occurs over the
chcm~cdlform When ,I dcr~v,rt~~allon re'lctlon I \ e n - wavclc~~gth range passed by the excitation wavelength
ploycd, redgent conccntratlon\ ,ire nclju\ted to m a - selector. Ideally, these can be totally eliminated if the
r n m the fluore\cence slgn'd , ~ n dthe conver\lon cfti- excitation and emission bandpasses do not overlap. In
crency 01 the cin~ilyt~cal rcc~ction.The b,rckground reality some of thc scattered or reflected radiation is
fluorc\ccnce from ie,igcnt\, or contamrnCmt\111 the re- passed by the emission wavelength selector as a stray
,tgent\, inu\t be conwkred durlng optm17'1tion radiation component. Also, excitation radiation outside
In gcnc1'11, the rate 01 cxteinal conver\ion In-
c ~ e , ~ \ ewith
\ mcrcawlg temperature hecnusc the colll-
TABLE 75-3
\~oii~ll e \ , reduce\ 0, Tcmpel,lturc
l,lte ~ n c r c ~ ~ \thi\
Sources of background signals
coefticient\ ,tre typ~c~illy 1 to 2(/0 per "(', cllthough they
c m be much hlghcr Thu\ a tcmper;rturc-controlled 1 . Elastic scattering of cxcilation r ; ~ d i ; ~ t ~by
o nthe solution. sample
smiple cell holdel 14 recon~mendcdto en\ulc thLlt\tmd- cell. or optical component\
arcls and wrnple\ arc '1: the wmc tcrnper,~turc 2. Kcflcclion of excitation 1-atli:~tionhv the s;unplc cell walls. opti-
cal components, or other \urlnces within the sample compart-
I'l~otodccompo~itionor photochcni~c,rl rcactlon\ ment
rnvolv~ngthe cxcttcd st,ttc c a u x the Iluorc\cencc s~gn,~l 3. Ramall \cat:ering hy the solvcnt
to decre'lw dunng expowre to cxc~t,~tion radi&ion. The 4. Stray radiation tiuc to room light lcahs
degree of photoclcconipo~itlon 14 pr~portion~rl to the 5. Fluorcscencc from the solvent and conco~nilants(dissolvccl or
~ n c ~ d c nr,ld~,lnt
t powcr m d the cxpowrc tlme It pho- adsorbed on particles o r sample ccll walls)
0. Lu~nincsccnccIron] the \ample cell walls o r optic;~lcomponents
todcconilx)\itim 14 vgnrtlcmt, the ineldent r,ul~,lntpowc~ (lcnsc\, lillers)
c m be reclucetl (e.g , \m,lller exc~tation4ht wldth) I he
Sec. 15-3 i Molecular Fluorescence Spectrometry 463
electrostatic environment of the molecule and change excited fluorescence are considerably poorer than those
E or YhF. obtained with conventional spectrofluorometry.
Spectral analyte interference is caused by concom-
itants that absorb significant amounts of excitation or Linearity. The linear range of molecular fluo-
emission radiation. This can be compensated as dis- rescence can be four to six orders of magnitude for
cussed in Section 15-2 (equation 15-31). Presently, com- efficient fluorophores with low detection limits. As de-
mercial instruments do not provide on-line correction scribed in Section 15-2, nonlinearity begins to occur at
for absorption. However, the sample absorbance at the analyte concentrations where primary absorption be-
excitation and emission wavelength can be measured in comes significant (i.e., the absorbance over the prefilter
a spectrophotometer to ascertain if there is significant and viewed pathlength, b,, -t b*, is greater than 0.01).
primary or secondary absorption (i.e., A > 0.01 over Secondary or self-absorption is usually insignificant at
b,, + b* or A ' > 0.01 over u1 + b,,). emission wavelengths near the maximum of the emis-
Fluorescence measurements are less dependent sion band, but it can distort the emission spectrum on
than absorption measurements on uncompensated con- the short-wavelength side. Self-quenching (i.e., colli-
comitant absorption. Consider a case where the analyte sional deactivation of an excited analyte molecule by a
and concomitant absorbances are each 0.001. In ab- ground-state analyte molecule) or dimer formation can
sorption spectrometry, the absolute error would be 100% be significant at relatively high concentration (e.g.,
if the blank did not compensate for the concomitant > 1mM). Contamination can cause nonlinearity at very
absorption. The concomitant absorption would cause low concentrations (see Figure 15-17).
less than 0.1% error in fluorescence measurements. Even Other potential causes of nonlinearity are similar
with relatively high background absorption, the abso- to those in molecular absorption measurements (see
lute error caused by uncompensated background ab- Section 13-3). Chemical equilibria involving the analyte
sorbance is less with fluorescence measurements than can cause significant nonlinearity. Usually, one chem-
with absorption measurements. ical form is fluorescent. However, to achieve linearity
Blank fluorescence interferences are minimized the fraction of the analyte in the monitored chemical
by careful selection of the excitation and emission band- form must be independent of the total analyte or an-
pass and wavelength. In some cases, separation from alytical concentration.
fluorescent concomitants is mandatory. More spe- Changes in solution bulk properties with analyte
cialized techniques are sometimes used to enhance the concentration or instrumental factors (e.g., polychro-
SIB or DL or to permit the determination of several matic excitation radiation, stray radiation, variability in
fluorescent species in one sample. These include deriv- pathlengths of different excitation rays, multiple re-
ative, synchronous, and wavelength modulation tech- flections, or dichroism) can cause nonlinearity in ab-
niques (see Section 15-I), methods that sharpen the sorption measurements. These sources of nonlinearity
structure of the excitation and emissiori spectra (see are not usually significant in fluorescence measurements
later in this section), and lifetime discrimination (see because they only become important at high analyte
Section 15-6). concentrations, where primary absorption effects usu-
Analyte interference due to absorption. scatter- ally dominate.
ing, or quenching can often be eliminated by simple Measurements can be made in the nonlinear re-
dilution if the DL is adequate. Standard additions can gion if the shape of che calibration curve is established
also be useful in these cases. with a sufficient number of standards. Precision can be
Two-photon excited fluor.escence involves the si- poor in the region yielding maximum signal before roll-
multaneous absorption of two photons to reach an ex- over because the fluorescence signal changes only slightly
cited state whose energy is the sum of the energies of with concentration ( i . e . , the calibration slope ap-
the two photons. Because the transition probability is proaches zero). It is also possible to linearize the cali-
much less than that for one-photon absorption. laser bration curve with correction factors (see equation
sources are required (i.e., a two-photon cross section 15-31 and Figure 15-15).
is typically lop5' cm4 s molecule-I photon-'). Selec- With proper design, the characteristics of the PMT,
tivity can be enhanced because different selection rules signal processor. and readout device should not limit
apply for two-photon processes. Because the fluores- linearity. If photon counting signal processing is em-
cence occurs at wavelengths lower than the excitation ployed, nonlinearity due to pulse overlap can occur at
wavelength, Rayleigh and Raman scattering signals can high incident light levels.
be virtually eliminated. Also. primary absorption ef- Generally, it is preferred to make measurements
fects are usually minimal at the longer excitation wave- in the linear region of the calibration curve. This can
lengths. To date, the DL,s achieved with two-photon be accomplished by simple dilution or by changing the
Chap. 15 I Molecular Luminescence Spectrometry
cxcltcttion wavelength to one where the 'tbwrbance 15 or physically bound to specific sites in molecules such
l a \ . Nonlincar~cydue to pret~lteretlcct$ 15 rnlnlmi7eti as proteins. The magnitude of the observed fluores-
by u\lng m~crocellfor by movmg the cell to reducc the cence signal provides information about energy transfer
prclllter pathlcngth ( I c , recluc~ngI?,,,) processes, the distances between reactive sites, or the
polarity of the protein in the vicinity of the binding site.
The nonfluorcscent tag aminonaphthalene sulfonic acid
becomes highly tluoresccnt when bound to proteins. It
Fluorescence spectrometry is not considered u major can be ~ s c dto titrate the number of available binding
structural or qualitative tool for solution measurements. sites.
Often molccules with suhtlc structural differences have
similar spectral characteristics. Also, excitation and
emission hands are relatively broad in solution at room
tempcr;~ture.Libraries of tluorcscencc excitation and
emission spectra of many molecules are available. These The fluorcscence quantum cfficiency (yield) or powcr
reference spectra can he compared t o sample spcctra yield is also of fundamental importance. The ~nagnitudc
to suggest the possible presence or to confirm the ab- and variation in magnitude of these quantities with so-
sence of certain species. Only corrected spectra should lution conditions provide information about mecha-
be compared as the shapes of uncorrected spcctra arc nisms and rate constants of deactivation and quenching.
highly instrument dependent. Care must be taken that The determination of the absolute quantum cfficiency
spcctra are acquired at a low enough analytc concen- or power yicld is a challenging experimental problem.
trations that inner filter effects do not distort the spec- Mere we review briefly some of the techniques utilized.
tral characteristics. Depending on the specific technique, either the quan-
Fluorescence spectrometry has proved :I valuable tum yield or powcr yield is determined. The other quan-
forensic tool for oil spill identification. For example, tity can be calculated from the determined quantity, as
the source of an oil spill can be confirmed by cornpar- discussed in Section 12-6.
ing the emission spectrum of the environmental sample To determine the fluorescence quantum cfficiency
to the emission spectrum of the suspected source of oil. or powcr yield directly, it is necessary to measure the
This fingerprinting is made possible by the high degree total radiant power of fluorescence and the total radiant
of vibrational structure of polycyclic hydrocarbons in power absorbed by a givcn amount of fluorophorc. The
oil. ratio of these quantities is ( 1 1 ~ or Y F (see Table 12-7).
The quantum yield can be evaluated with chemical ac-
ical actinometes is a device for
measuring the absolute radiant power absorbed by a
Fluorometry is uscd to study chemical equilibria and solution by determining the photolysis products of a
kinetics in much the same way as spectrophotometry photochemical reaction of known yield. For example,
(see Section 13-5). Thus common applications include a sufficiently concentrated solution of potassjum ferri-
determination o l equilibrium constants and reaction oxalate absorbs all radiation below 490 nm. The ab-
stoichiometry (e.g., mole ratio method, method of con- sorbed radiation reduces ferrioxalatc to ferrous oxalate
tinuous variations), rates of reactions (dcldt tdE,ldr), with near-unity conversion efficiency. The number of
rate constants, and reaction mechanisms. The enhanced moles of ferrous ion produced during a givcn exposure
selectivity makes it easier to monitor one species in- time is determined by titration or by another technique.
volved in the reaction. eactions can be studied at lower For quantum cfficiency determinations, the sam-
reactant concentrations hecausc of the improved de- ple cell is totally surrounded (except for small windows
tection limits. to transmit the excitation radiation) by a chemical ac-
The shape and spectral position of bands in the tinomctcr solution. This yields a measurcinent of the
excitation and emission spcctra of a molccule yicld in- total Sluorcscence radiant power emitted after corrcc-
formation abo~itits molecular structure and energy level tion for the window losses. The same actinomctcr is
structure. As we have already noted, the corrected ex- then uscd to determine thc incident radiant powcr.
citation spectrum is often equivalent to a conventioni~l Calorimetric methods provide a simple and ele-
absorption spectrum. However, the excitation spectrum gant method of indirectly determining the luminescence
can be obtained at much lower concentration if +,. is power yield. The efficiency with which the luminescent
favorable; this is advantageous if a compound has lin- species (x) converts absorbed radiation into heat is
itcd solubility. measured. 'This is compared to the ell'icicncy with which
Fluorescent probes or tags can be covalently linked a nonluminesccnt, photochemically inert reference spe-
Sec. 15-3 i Molecular Fluorescence Spectrometry 467
cies (r) converts absorbed radiation into heat: excitation or emission variables if different excitation
or emission wavelengths or different solvents (e.g., re-
fractive index effects) are used.
Quantitative Analysis
where H is a signal proportional to the heat produced
corrected for the solvent blank, 0, is the incident ra- Inorganic Species. Although many fluorome-
diant power, and a is the fraction of the incident ra- tric methods for metal ions have been proposed, few
diation absorbed. Traditionally. H is determined by of these methods are used routinely. Atomic spectro-
conventional calorimetry (e.g., a Dewar flask or a ther- metric techniques are more universal and in general
mostatted compartment with a temperature transducer provide better detection limits and selectivity.
to measure AT). The response H has also been meas- Only a few unchelated metal ions, primarily lan-
ured with photoacoustic and thermal lensing techniques thanides [Ce(III), Tb(III), Eu(III)] and actinides [UO:,
(see Chapter 17). Th(I)], exhibit appreciable fluorescence in solution. The
Other methods for the determination of +,utilize fluorescence, or actually phosphorescence in some cases,
a more standard spectrofluorometer setup. The fluo- is due to d-f and f-f electronic transitions, is rather
rescence signal measured with a spectrofluorometer is long-lived (7, = 1 ms), and often consists of several
only a relative signal. Moreover, only a fraction of the narrow emission bands. Detection limits for some ele-
total fluorescence emitted is collected and detected. By ments are as low as 1 ng mL-l.
completely characterizing all geometric and instrumen- Inorganic reagents can increase the luminescence
tal factors (see Table 15-I), calibrating the wavelength of metal ions. The quantum efficiency for uranyl ion,
selection-detection system, and making additional ab- for example, is greatly enhanced in concentrated phos-
solute measurements of the incident and transmitted phoric acid. In concentrated HCl and HBr. several non-
radiant power, we could eventually calculate the total rare earth elements [e.g., Tl(I), Sn(II), Pb(II), As(III)]
radiant power absorbed and fluoresced. Such absolute luminesce. The luminescence is often significantly in-
calibration is exceedingly difficult. Therefore, relative creased at very low temperatures (e.g., 77 K or lower).
methods based on some type of reference species are Rare earths can also be excited to luminescence by
usually employed. energy transfer from excited states of aromatic carbonyl
The Weber-Teale approach involves ratioing the compounds produced by photon excitation.
fluorescence flux seen by the detector to the photon The majority of inorganic fluorometric determi-
flux measured with a scatterer (e.g., glycogen) in the nations are based on the formation of fluorescent che-
sample cell. Under suitable conditions (e.g., the solu- lates with nonfluorescent ligands. The chelating re-
tions have identical absorbance). the scattering solution agents shown in Table 15-4, or their derivatives, are
is considered an ideal emitter (i.e.. a yield of unity). commonly used. Selectivity is often limited by the spec-
Complications arise due to polarization effects and dif- ificity of the chelation reaction. The best DLs are often
ferences in the spatial distribution of scattering. In this obtained with low atomic number, diamagnetic metal
and other methods, integrating spheres and quantum ions (Be2+, Mg2+, Ca2+, A13+), which are less likely
counters are often used to minimize these effects. to promote intersystem crossing. The fluorometric D L
The most popular indirect method for determining for A13+ with several reagents is in the range 0.1 to 1
quantum efficiency is the Parker-Rees method. Here ng mL-I, better than that achieved with most atomic
+F is evaluated by comparing the corrected emission spectrometric techniques.
spectra of the analyte (x) and a reference fluorescent The fluorometric determination of selenium in bi-
species (r) of known +:, ological materials with 3,3'-diaminobenzidine, of boron
with 4'-chloro-2-hydroxy-4-methyloxybenzophenone,
and of beryllium with morin (2',3,4',5,7-pentahydroxy-
flavone) are still popular methods in specialized appli-
cations.
Some proposed fluorometric determinations are
based on the inorganic ion acting as a reducing or ox-
where A is the absorbance at the excitation wavelength, idizing reagent or a quencher. These methods have gen-
cP, is the relative incident radiant power (photons s-I), erally proved to be less selective than more direct fluo-
and J E dh' is the integral of the corrected emission rometric techniques.
band (relative quanta per second). Additional factors As in spectrophotometry, reaction-rate methods
may be included to account for other differences in can yield added selectivity by discriminating against
Chap. 75 I Molecular Luminescence Spectrometry
(hero lor-111)
steady-state signal\ (uncompensated background fluo- or solid-state reactions. The fluorometric determination
rescence) or species that react at rates significantly of uranium fused in a NaF matrix has been widely used
different from the analyte. For example, All' can for decades. In a solid matrix, the absorption and emis-
be determined down to 0.1 ng mL by fluorometric sion bands become much narrower. With a tunable dye
monitoring of the rate of formation of its chelate with laser, it is possible to achieve high selectivity by exciting
2,4,2'-trihydroxyazobenzene-5'-sulfonicacid. Many ki- a specific absorption line of a given rare earth in a
netics-based determinations have been proposed ln wh~ch particular environment or crystallographic site. Detec-
the analyte metal ion acts as a cataly\t. tion limits for many lanthanide ions in CaF, are in the
Metal ions, part~cularlyrare earth ions, can al\o range I to I00 pg mLP1.
be fluorometrically determined if they are incorporated Nonf-luoresccnt ions can be determined by probe
into a host d i d matrix by precipitation, crystallization, ion techniques. Here the analyte is incorporated into a
Sec. 15-3 1 Molecular Fluorescence Spectrometry 46
solid matrix where it forms associates with the fluores- TABLE 75-5
cent rare earth ions. The associates experience a changed Types of organic compounds with native fluorescence
crystal field splitting and the spectral bands are shifted.
Class Examples
It is then possible to obtain single-site spectra of only
rare earth associates with the same detection limits as Aromatic amino Tyrosine, typtophan,
achieved for rare earth ions directly. Sample prepara- acids phenylalanine
tion can be tedious, and cryogenic facilities are usually Vitamins A . B, (riboflavin), B,
used to cool the sample for narrowing the bands. (pyridoxine), Biz. E. folic
acid
Flavins Flavin mononucleotide
Organic Species. Fluorometric methods of
(FMN). flavin
analysis are used routinely for the determination of many d~nucleotide(FAD)
types of organic molecules. They are employed exten- Reduced NADH, NADPH
sively in the clinical and biochemical area; however. pyridine
environmental applications are also quite important. nucleotides
Clinical analysts are often required to determine Cathecholamines Dopamine, norepineprine,
very low concentrations of analytes (often 1 kg L - l or epinephrine (adrenalin)
less) and to work with small samples (e.g., serum from Porphyrins Chlorophylls
a newborn). The detection limits and selectivity pro- Steroids Estrogen
vided by fluorescence are thus well suited for clinical Drugs Quinine, salicylates,
tetracyclines, barbiturates
assays. Before measurement, some form of sample
cleanup is usually required. Red blood cells are usually
removed from blood by coagulation and centrifugation be coupled to a product of the initial enzymatic reaction
before enzymatic assays. For other species, deprotein- to produce a fluorescent product.
ization of the blood serum or urine is also required to Most fluorometric enzymatic assays are carried
reduce background absorption and fluorescence from out in solution. However, special front-surface tech-
proteins and to release analytes that are bound to pro- niques have been developed to monitor reactions on
teins. Solvent extraction or simple column cleanup pro- solid surfaces. The reagents can be adsorbed or im-
cedures are used to separate the analyte from the ma- mobilized on the solid surface. A drop of sample is
jority of the sample constituents. Tissue samples are placed on the active surface.
usually homogenized in a blender, and then the ho- Fluorescence is used in environmental applica-
mogenate is filtered or extracted with a suitable solvent. tions for detecting certain aromatic hydrocarbons. The
Several biochemical compounds exhibit signifi- most extensive applications are for determining poly-
cant native fluorescence at room temperature as shown cyclic aromatic hydrocarbons (PAH) such as chrysene,
in Table 15-5. Thus their determination requires only perylene, pyrene, fluorene, and 1,2-benzofluorene. These
a separation from potential interferents. The residues compounds are released into the environment by fossil-
of highly fluorescent tryptophan account for about 90% fuel burning, and many are carcinogenic.
of the fluorescence from proteins.
Many selective derivative reactions have been de-
Automated n/leasurements. Automated batch
veloped for nonfluorescent organics or organic mole-
or continuous flow analyzers are available for fluoro-
cules with inadequate natural fluorescence properties.
metric determinations. They are similar to the systems
Some common reagents are listed in Table 15-6.
described in Section 6-6 except that the spectropho-
The detection limit of enzymatic assays is often
tometer or photometer is replaced with a spectro-
improved by using fluorometric instead of spectropho-
fluorometer or fluorometer with a flow cell. Their use
tometric monitoring. Substrates can be determined by
equilibrium-based or kinetics-based procedures; en-
zyme activity must be assayed by reaction-rate methods. TABLE 756
Often, the same chemistry used in spectrophotometric Common fluorometric reagents for organics
monitoring is employed in cases where the coenzymes
Analyte Reagent
NADH or NADPH are involved in the reaction (see
equations 13-59 to 13-61). The fluorescence quantum Estrogens H,SO,
efficiencies of the reduced coenzymes are about 2 % . Corticosteroids H,SO,
To improve the DL, the NADH formed can be coupled Amino acids o-Phthaldehyde (condensation)
Thiamine Fe(CN),3- or Hg" (oxidation to thiochrome)
to another reaction which produces a product that is Ascorbic acid Phenyldiamine (condensation)
highly fluorescent. Alternatively, other reactions can
470 Chap. 15 / Molecular Luminescence Spectrometry
Other techniques can be used to prepare samples against. The loss in absolute calibration sensitivity and
in inert matrices. In matrix isolation techniques, the increased background signals and noise often degrade
sample is vacuum co-deposited with N, or a Shpol'skii detection limits.
solvent on a cold finger at 10 to 15 K. Samples can also Because of the difficulties noted above, many
be dissolved and solidified in plastic matrices. companies market dedicated HPLC fluorescence de-
tectors specifically designed to illuminate and collect
Spectrofluorometric Titrations fluorescence radiation from microvolume cells. Fluo-
rometers often provide adequate selectivity and higher-
Fluorescence indicators are used in titrations where col- radiation throughput than monochromator-based de-
orimetric indicators lack sensitivity or are difficult to tectors. Specially designed emission selection systems
observe, as in highly turbid or colored solutions. Titra- as shown in Figure 15-21 are used to enhance the emis-
tions with visual endpoint detection are carried out in sion signal detected. Although spectrofluorometers may
a dark room or a viewing box with illumination by a yield poorer detection limits than fluorometers, the ex-
UV lamp. In instrumental fluorometric titrations, the citation and emission wavelengths can be changed at
titration vessel is located in the compartment of a fluo- appropriate times during the chromatographic run to
rometer or spectrofluorometer. Acid-base indicators are optimize conditions for specific compounds.
based on changes in bF for the acid and base forms of Researchers have designed specialized "window-
the indicator. In precipitation titrations, the difference less" flow cells to reduce background signals from silica
in 6, for the indicator in solution as opposed to ad- cell walls. In one design, eluate in drop form is illu-
sorbed on the precipitate is exploited. Redox titration minated as it flows out of the HPLC column. In another
indicators exhibit different fluorescence characteristics design, the eluate is confined in a sheath of solvent.
in different oxidation states. Indicators for complex for- Laser excitation is well suited for fluorometric
mation titrations, often called metallofluorescent indi-
cators, usually form fluorescent complexes with the un- Excitation
titrated metal ion. beam
4
Fluorescence Chromatographic Detection
('J)
EXCITATION
IR
LAMP
Micelle- and Cyclodextrin-Based Measure- tants depends on the chemical structure of the surfac-
ments. More recently, solution-based RTP tech- tants and the nature of the solvent, as shown in Figure
niques have been developed. Micellar solutions have 15-24.
been found to be particularly attractive for RTP meas- Phosphorescence from many organic molecules is
urements. Molecules having a nonpolar hydrophobic observed at room temperature in solutions containing
group attached to a polar hydrophilic group are known detergents (sodium lauryl sulfate is common) at con-
as surfactants or detergents. The nonpolar group is usu- centrations above the CMC. In a typical sample prep-
ally a hydrocarbon chain ("tail"), and the polar group aration procedure, a small amount of sample is mixed
("head") can be either an ionic or a neutral compound. with a micellar solution. An ultrasonic bath often aids
Above a certain concentration known as the critical mixing. The sample is usually spiked with salts con-
micelle concentration (CMC), these surfactants dynam- taining heavy atom ions [Tl(I). Pb(II)] to enhance +,
ically associate in solutions to form colloidal aggregates (external heavy atom effect). Next, the sample solution
called micelles. The type of aggregation of the surfac- is transferred to a container where it is deoxygenated
TABLE 75-7
Summary of some analytical features of different RTP methods
70 ernision
monochromator
From sxcitntion
monochromator
Sample
Fillcr
paper
Excitation
plane From excitatrm
mlrror monocl~romat(x
(2)
FIGURE, 15-25 Phosphor-oscope designs. In (a) the rotating can or cyl-
indcr design is shown. The hollow cylinder has one or more apertures
equally spaced o n the circumference. During part of the rotation cycle.
the excitation radiation passes through an aperturc and strikes the sample
cell while the emission r;ldiation is blucketl from r c x h i n g the emission
monochromator. As the can rotates, it reaches a position where the cx-
citation radiation is blockcd, but thc cmission radiation can pass through
an aperture and be detected. It is suitable for the right angle excitation
emission geometry. The Becquerel or rotating disk desigll (17) provides
the same type of out-of-pliusc excitation and viewing with an in-line ex-
c i t a t i o n - e n s s i i geometry. Thc cam-shapcd disks can be replaced by
circular disks with apertures. In both (a) and (b) the shalt is cirivcn by a
variable-speed motor and the satnple is in a Dewar flask for L I T meas-
urements. 'l'hc Dewar flask can he rcp1;iced by a normal cell lor miccllc-
based RTP tncasurements or by a suitable holder for solid substrate-hascd
R'I'P measut-cmcnts (see Figure 15-23>),A rotating mirror phosphoro-
scope is shown in (c). The filter paper- strip can be moved to allow viewing
by purging with nitrogen and applying vacuum to the 3. 'The altered microenvironment can provide fa-
solution. Finally, the sample solution is transferred to vorablc polarity and aciclibase equilibria lor en-
a sample cell for excitation and observation of the phos- hancccl phosphorescence quantum efficiency.
phoresccnce. 4. The miccllar solutions can improve the dc-
The incorporation of the analyie molecule into the tection limit lor hydrophobic species in iry ucous
core of a micellc imparts certain advantages: solution by increasing their solubility.
5. The proximity of interacting species (phos-
I.The structural conformation ol the micclle\
p h o r s and heavy atoms) is increased. and can
protects the phosphorescing triplet \tatc of
result in a more efficient spin-orbit coupling.
the analytes from external qucncher\.
2. The o r ~ e n t a t ~ o ncon4t1
~ i l a n t decreases v 1 1a-
~ Although d e o ~ y g c n ~ r t ~I \ oneeded,
n ~t c~inprovide
tional deact~vation. an advantage. The oxygenated wlutlon contnlnlng the
Sec. 75-41 Molecular Phosphorescence Spectrometry 477
analyte produces little or no phosphorescence because (i.e., that observed without a phosphoroscope) is given
of the efficient quenching by 0 , . Hence this solution by
can serve as a blank that accounts for scattering signals
and luminescence signals that are not dependent on 0,. T, exp( - tdhp)[ l - exp(t, /T,)]~
Cyclodextrins are doughnut-shaped polymerized CY = (15-48)
t, [ l - exp ( - t,/~,)]
sugar molecules. They form inclusion complexes with
molecules in solution. Phosphorescence and fluores- To maximize the phosphorescence flux from a given
cence are enhanced for molecules included in the cavity. phosphor, it is important to adjust the cycle time so
In contrast to micellar solutions. some phosphorescence that the decay of the phosphorescence is small during
is observed without deaeration. However, deoxygena- the delay and observation time. If T, >> t,, cw is inde-
tion does increase the phosphorescence signal. Heavy pendent of T and the cycle time and
atom-containing reagents (e.g.. 1,Zdibromoethane) are
added so that they are incorporated into the cavity with
the analyte to promote formation of the triplet state.
Selectivity can be greater than in micellar solutions, due
to the stringent size limits imposed by the inclusion The maximum rotation rate of rotating can and
process. disk phosphoroscopes is about 6000 rpm, which yields
a minimum t, of about 10 ms. Thus phosphoroscopes
are generally useful for T, > 1 ms.
Sensitized RTP Measurements. In sensitized
RTP techniques, a weakly or nonphosphorescent ana- Pulse Source, Gated Detection. Time resolu-
lyte (donor) is excited and transfers its triplet-state en- tion can also be achieved with pulse excitation and elec-
ergy to the triplet state of an acceptor with a good tronically gated detection. The sample is repetitively
phoshorescence quantum efficiency. The phosphores- excited with radiation from a pulsed or chopped light
cence signal from the acceptor is measured. Good ac- source to populate the triplet state. The resulting phos-
ceptors are bromonaphthalenes and diacetyl which ex- phorescence from a given excitation pulse is observed
hibit significant phosphorescence in solutions even at after a suitable delay time measured from the end of
room temperature. To obtain specificity, chromato- the excitation pulse. Often, a boxcar integrator is used
graphic separation normally precedes mixing of the do- to allow control of both the delay time and the period
nor and acceptor and detection. The donor can also be
determined from the quenching of its phosphorescence
by the acceptor.
The advantages and limitations of different RTP
methods are summarized in Table 15-7.
(a) Timing
Instrumentation
of oh\crv,lt~on,md to (iver'igc the pho\pho~c~ccnce \IS- r x h a t ~ o n Zhc Liter ctlcct 14 \m,lllcr hec<~uwot thc
n,il\ lrom mmy pul\e\ I n some c<i\e\, the I'M I I>I,I\ \lnAler cell p,~thlength\rn L I P ,lnd c\pec~,~lly In RTIJ
volt,lgc i\ tur ncd oft d u ~ n gthc exclt'itlon pulw to prc- w ~ t hf~ont-\urt,ice\ o l ~ d - \ ~ ~ l > \m
t ~c~dt \t ~e ~ r ~ ' n l ~ n t s
vent o v e ~ l o d r n gthe PMT w ~ t hthe \c,ittcred ~ ~ i c h , ~ t ~ o n For good phosphor\, the hc\t DL\ ,ire gener,llly
vgnd. In the r'rnge I to 10 nM wrth I 11' m d ~n~celle-ba\ctl
In e d y \tudlo, ,I contlnuou, Xc ,ire I'in~p,mcl '1 R I P Thc\e Dl,\ ,ire s ~ g n ~ t ~ c mWOI t l y\e th'in lot the
mcch,~nrc,~l chopper were employed More rcccntly, the best Iluo~ophorc\ Ilowevcr, DL\ tot many ,rn,rlyte\
preterred soulce 15 ,I pul\ed Xc ,lrc Li\h Imip wh~ch ,n e better wrth phmphol cxcnce t h m Iluo~cxcnce ~f
y~eldspulse w~dthsoi I to 10 l~ls7h14 c ~ l l opho\phors
~\ +,,- - +, tor the ,tn'ilytc Dctect~onllm~tsw ~ t hsol~d
w ~ t h-rl, value\ below 0 I In\ to be ~ne~isurcct and oftcr\ sub\tr,~teRTP ,ile typ~cdly1 to 10 ng, c ~ l t h o ~ ~v~ilucs gll
more convenrent control 01 the del'ty and ob\erv,it~on near 1 pg have been 'lchlevetl w ~ t h\ome compouncl\.
tmes. The SIN 14 better t h m w ~ t hchopped \ystcm\ For '1 2 5-pL \cimplc SIK, ,I DL oi 1 ng colre\pond\ to
bec'lu~ethe peak ~rracllance15 much h~ghert h m the 2 l~lMlor ,i compound wlth a molecul'~rwe~ghtol 200
average lrrdclmnce for cont~nuurnlamp\ Pul\ed l a v n The hnear range I \ typ~callytwo to tour order4 ot
have been ir\cd. m'ign~tude dcpcndlng on thc D L Nonl~ne,rt~ t yoccur\
dt hrgller conccntrntlon\ when the t r x t ~ o no l rath,tt~on
abso~bedIS n o longer proport~on,~l to the m,dyte con-
centr,lt~onor pref~ltercliect\ o c c u ~111 solut~onRTP
The methodology used in phosphorimetry is similar in
many respects to fluorometry except for sample prcp- Applications
aration and prcsent;ition. Most analyses arc based on
species with native phosphorescence; derivative reac- The fact that phosphorescence is rarer than Sluores-
tions are rarely used. Reagents containing heavy atoms cence restricts its applicability, but enhances selectivity.
are often employed to increase +,,.
These heavy-atom It has found use in situations where excellent (c.g.,
matrix modifiers must be uscd with care because the nanomolar) but not spectacular (e.g., picomolar), DLs
lifetime is also reduced ( i . ~ . .oc in equation 15-48 is are required and selectivity is paramount.
reduced for a given t,). Phosphorimetry has been uscd primarily for the
Wavelength and bandpass optimization, deriva- determination of organics in environmental applications
tive techniques, and synchronous scanning are em- (e.g., PAHs) and in the drug and pharmaceutical field
ployed to improve the SIN or the selectivity and to carry (e.g., theophylline, sulfonaniidcs). Thc rangc of ana-
out multicomponcnt determinations as in fluorometry. lytes is demonstrated by the list of data in Table 15-8.
Time discrimination is an added dimension ol' selectivity Solution RTP can be used for HPLC detection.
and SIN cnhancernent. The delay and observation timcs Phosphorimetr-ic determination of inorganic spe-
can be adjusted to discriminate against other phosphors cies is rare (other than for rare earths, as discussed in
with half-lil'es much shorter or greater than that of the Section 15-3). However, some metal ions can be de-
analyte. termined by forming phosphorescent complexes with
nonphosphorescent ligands.
ance Characteristics At this time, phosphorimetry is not used as rou-
tinely 21s fluorometry. 'Ihc development of KTP meth-
The measurement precision of phosphorescence for ods could change this situation. Much research is being
concentrations wcll above the D L can be 1 to 2%. How- conducted to improve RTP methods and to make them
ever, sampling irrcproducibility often limits sample-to- adaptable to flow systems and FIPLC detection.
sample precision to 5 to 105%.In low-temperature phos-
phoresccnce measurements, the SIN and DL, can be less
due to the smallcr V , the loss of radiation in passing
through many interfaces, and convection Slicker noise
(from radiation passing through liquid N, with bubbles Chcmilumincsccnce (CL) techniy ucs arc based on the
and refractive index changes). fact that in a few reactions, a significant Gaction of
The factors limiting accuracy arc much the same interniccliatcs or products are produced in excited elec-
as for fluorescence measurements. The primary blank tronic statcs. In other words, part of the chemical en-
interferencc is uncompensated background phospho- ergy released in the reaction is used to produce excited
rescence from concomitants. Analyte interference can species. The emission of photons from the excited mol-
be due to concomitants causing quenching or ahsorbing ecules is measured. Often, it is possible to arrange re-
of a significant fraction o f the primary and secondary action conditions so that the luminescence signal is re-
Sec. 75-5 1 Chemiluminescence 479
TABLE 75-8
Phosphorescence data for some organic compoundsa
Adenine WM 278 406 2.9 0.02 Pyridine EtOH 310 440 14 0.0001
RTP 290 470 - 4.1 Pyridoxine HC1 EtOH 251 425 - 0.008
6-Amino-6-methyl- WM 321 456 0.66 0.0002 Salicyclic acid EtOH 315 430 6.2 0.05
mercaptopurine RTP 320 470 - 0.7
Anthracene EtOH 300 462 - 0.05 Sulfamerazine EtOH 280 405 0.7 0.0001
EPA 240 380 2.1 0.10 Sulfamethazine EfOH 280 410 0.8 0.0001
Aspirin EtOH 310 430 5.3 0.007 Sulfanilamide EtOH 297 411 2.9 0.012
Benzoic acid EPA 240 400 2.4 0.005 RTP 267 426 - 3
Caffeine EtOH 285 440 2.0 0.2 Sulfapyridine EtOH 310 440 1.3 0.0001
Cocaine HC1 EtOH 240 400 2.7 0.01 Tryptophan EtOH 295 440 1.5 0.002
RTP 285 460 - 0.04 RTP 280 448 - 4
Codeine EtOH 270 505 0.3 0.01 Vanillin EtOH 332 519 - 0.1
DDT EtOH 270 420 0.2 0.007
Diacetylsulfanamide EtOH 280 405 1.3 0.001
- p~p~ - -
aFor LTP measurements the solvent is indicated (WM = water!methanol in 9:l ratio) and the D L is in k g mL-I. For RTP measurements on different support
materials the D L is in ng.
lated to the analyte concentration. Note that unlike For CL to occur, three general conditions must
photoluminescence measurements, no external radia- be met. First there must be sufficient energy to produce
tion source is required. an excited state. Thus the reaction must be sufficiently
exothermic such that
Principles
hc
Three common reaction sequences used to produce CL -AG r - (15-51)
he*
are the following:
.
The C1 ratl~antpowel In photon4 per second <II
t m e t [@, ,,(t)] i\ g~vcnby
lininesccnce signals since no external source is required.
The C l , signal must be distinguished from only dark
current noise in some cases. $11 other- cases, the CL
signal due to the analytc rnust I-)c distinguished from a
background CL signal observed with no analyte. This
background signal is often tieternmined by contaminants
where drz,(t)/dt is the rate of the reaction (i.e., the in the reagents and solvents. Selectivity and linearity
rate of clisappearance of the CL prccursor in rnoleculcs arc very dcpcndent on the rcaction and rcaction con-
a n 3s I) and V is the volume of the CL mixture viewcd. ditions chosen. As for photoluminescence measure-
Thus after the reactants are mixed, the CI, signal even- ments, absorption of emission radiation by the analyte,
tually decays with time as the reagents are consumed, products, or concomitants can cause nonlinearity or
as illustrated in Figure 15-27. Thc shape of the intcnsity spectral interference.
vs. time curve may also be clepcndent on +,,. if it changes in some C1, reactions ( ~ w a l l yin the gas phase),
with time. For example, products formed in the rcaction the analyte is thc CL precursor. For many solution C L
may inhibit the reaction pathway leading to excited spe- applications, the analyte is not the CL prccursor. Rather,
cies (change +), or may quench the luminescence the analyte is a necessary reagent for the C L reaction
(change (I),,). Although most CL reactions arc carried to occur or a species that affects the rate or efficiency
out in one phase (gas or solution), heterogeneous CI, of the GI, reaction (c.g., (I),,). For the latter case, the
reactions can be employed (e.g., a gas in contact with malyte can be an activator (c.g., catalyst), an inhibitor,
21 solution or solid, or a species in solution in contact or an acceptor. The increase or decrease in the CE
with iminoblized reagents). For such cases, the CL sig- signal caused hy the presence of the analyte is meas-
nal depends on both the reaction kinetics and the rate ured. In some c;~scswhere the analyte cannot be directly
involved in a C1, reaction. it is possible to use coupling
rcactions. Here the analytc reacts with suitable reagents
to produce a product which is a reactant in a CL rcaction.
Membrane
Reagents
Sample
Reagents -
Sample
Sample gas
Power
supply
Calibrated
FIGURE 15-29 Continuous flow CL
photometers. In (a) the sample and
reagents are combined at a tee and
sent to a flow cell for observation of
the CL. Mixing aids are often em-
ployed. The design in (b) is also used Signal processor
for solutions. In this case, the reagent and readout
Pressure
is forced through a membrane into gauge
the flowing stream of the sample. A
reaction chamber for CL determi- Pump
nation of gases is illustrated in (c). (c)
4 Chap. 75 1 Molecular Luminescence Spectrometry
truni can be collected by increrncnting the wavelength as shown below. This product has been identified as the
between each run. Multichannel detectors, such as in- luminescing species.
tensified diode arrays, have proven useful in this ap-
plication. Monochromators or spectrographs are not NH, NH,
used for quantitative measurements because of the loss I I I
in radiation throughput.
Because CL, signals are often very weak, the ef-
Iicient collection and co~~plitig of the CL radiation to
the detector arc critical. 'l'he sample cell is placed as
close to thc detector as possible. Additional mirrors,
lenses, and othcr dcviccs are used to increase the solid
angle of collection. Often, dark current noise limits the For the lucigenin reaction,
detection limit, in which case cooling the P M F and
photon counting signal processing cnhancc the DL and C'H ,
SIN.
Solution Chemiluminescence
there is still controversy as to whether the luminescing 1 ng m L 1 and as low as 10 pg m L P ' have been obtained
species is one of the products (N-methylacridone) or for several metals, including Fe(II), Cu(II), Co(II),
an intermediate. Chemiluminescence measurements in- Cr(III), and Cr(V1). Detection limits and specificity
volving peroxyoxalates employ sensitized CL. depend on the CL reagent. Although reaction condi-
tions can be somewhat optimized for a given metal ion,
additional steps are often necessary to enhance the se-
lectivity. The determination of Cr(II1) with luminol can
be made quite specific by determining the CL signal
immediately after adding EDTA to the sample. The
EDTA masks most metal ions, but the complexation
reaction with Cr(II1) takes hours. In other cases, rapid
separation schemes have been developed to isolate the
analyte from the major interfering species.
In the absence of H,O,, the luminol system is
much more selective. Thus oxidants such as hypo-
chlorite or ferricyanide can be determined selectively
in alkaline luminol solutions. With 0, as the oxidant,
Fe(I1) can be determined through its enhancement of
the luminol CL reaction. Under proper conditions,
Cr(V1) can be determined selectively in alkaline lophine
solutions with H,O, as the oxidant. The linear range
extends from 0.1 ng mLP1 to 100 mg mLP1.
The determination of hypochlorite can also be based
on the following simple CL reaction:
Chemiluminescence Indicators. The first an- Organic Determinations. Several types of or-
alytical use of solution CL involved CL indicators. As ganic species are determined by their effect on some of
for fluorescence indicators, they are useful for titrations the common CL reactions discussed above. Metal-con-
of colored or turbid solutions in which colorimetric end- taining compounds such as hemes and vitamin B12 en-
points are difficult to observe. Their use is based on hance the rate of the luminol CL reaction. Formalde-
the initiation of CL due to the change in solution com- hyde is determined by its enhancement of the CL reaction
position that occurs at the equivalence point. For ex- between gallic acid and H,O,. The lucigenin reaction
ample, in the titration of an acid with a base with added involves some reduction steps. This makes possible the
H 2 0 2 , CL from the indicators luminol. lucigenin, or CL determination of reducing species such as glucose,
lophine occurs only after the acid is completely titrated ascorbic acid, and uric acid in the absence of H,O,.
and the solution p H is high enough for the CL reactions Some organic analytes can be determined by sensitized
to proceed. Redox titrations can be carried out with CL with the peroxyoxalate reaction. The overall CL
luminol as the indicator and hypochlorite or hypobro- efficiency with acceptors such as rubene or some an-
mite as the titrant. Chemiluminescence appears at the thracenes can be greater than 20%. Postcolumn detec-
equivalence point when excess oxidant is present. An- tion schemes have been developed for the determina-
other CL indicator is siloxene, a polymeric silicon oxide tion of separated dansyl or fluorescamine derivatives of
(Si,H,O, units). It is used for redox titrations with an amino acids and steroids, porphyrins, and polycyclic
oxidant as the titrant in acidic solutions. hydrocarbons. Attomole DLs have been achieved in
some cases.
Inorganic Determinations. Most of the CL re- Clinical assays of substrates and enzymes can be
agents identified in Figure 15-30 have been used for performed by coupling to CL reactions. These assays
determination of various inorganic species, primarily often involve an initial oxidase enzymes-catalyzed re-
transition metal ions. In most cases, the metal ions en- action in which H,O, is a product. The amount of H,O,
hance the rate of the reaction and the observed CL produced is monitored with the luminol CL or the per-
signal with H 2 0 , as the oxidant. Detection limits below oxyoxalate CL reaction. Often, it is necessary to run
Chap. 15 I Molecular Luminescence Spectrometry
the c n ~ y m ~ i t and
l c incl~c~rtor
r c x t ~ o m\equentially be- ,~ctionto other rcactlon\ ~nvolvmgthc formatmn ot
cauw the optmum pH for thc two mrction, d ~ l t c r \ FMNH,. Mo\t applicat~onsinvolve coupling to reac-
In most 01 the , ~ p p l ~ c n t ~ oabove,
n\ the CL pre- tion\ that form NADH or NADPH; the\e can be cou-
cur\m i \ d rcClgent More rcccntly it h,~sbccn \hewn pled to the bacterial B1, reaction with the lollowmg
th& wrne analyte\, I n partlcul,n orgmics with poly- rcactlon.
.
phenolic g r o u p (e.g humic acid, t m n ~ n ) u, n be dc-
termmcd from the CL produced hy t h a r o x ~ d ~ i t ~wo~nt h
permangunnte
-
polymers. This CL is often enhanced by oxygen or hcat- bascd on the following CI, reactions:
ing. In sorne cases, the intensity o f CL can he related
to the condition or age of the product. NO + O, NO, + 0,
-
Bacterial BL is bascd on the following reaction:
FMNII, + 0, + R-CIPO
luc~tcr,tri
FMN
The luminescence spectrum corresponds to the fluo-
+ RCOOH + lI, + hv rescence spectrum of the parent compound. Excited
species can also be produced by oxidation of hydro-
where FMNH, is the reduced form of tlavin mononu- carbon anion radicals generated by oxidants generated
cleotide (FMN) and RCHO is a long-chain aldehyde. at the electrock. Formation of cxcimers that luminesce
Clinical assays are based on coupling this indicator re- is also possible.
Sec. 75-6 1 Lifetime and Polarization Measurements 485
To date, analytical applications of electrochem- pensate for the residual baseline signal. For example,
iluminescence are rare. A linear relationship between in the Guggenheim method, the natural logarithm of the
the luminescence signal and analyte concentration has difference in the luminescence signal between pairs of
been demonstrated for some molecules. data points along the decay curve, separated by a con-
stant time interval, is plotted vs. time. The lifetime is
obtained from the negative reciprocal of the slope.
5-6 LIFETIME AND POLARIZATION For multicomponent mixtures. the decay curve
MEASUREMENTS can be fit to a weighted sum of exponential terms to
obtain the lifetime of each component. This approach
In this final section we discuss the instrumentation and is also used for fitting a nonexponential decay from one
applications of luminescence lifetime and polarization species even though the lifetime calculated may have
measurements. little physical meaning.
We will now discuss some of the primary ap-
Lifetime Measurements proaches used to obtain lifetime data. They are based
on time resolution. phase resolution. and correlation
The luminescence lifetime is an important characteristic techniques.
of a molecule and its environment. Lifetime measure-
ments provide information that is useful in fundamental Pulse Excitation Method. In the pulse excita-
studies of energy transfer and quenching. In analytical tion method, the analyte is excited with a short-duration
applications, the luminescence lifetime is an additional pulse of radiation. and the decay of the resultant lu-
parameter that can be used to enhance selectivity or minescence signal after the excitation pulse is moni-
permit multicomponent determinations. tored. Thus the method involves time resolution. Either
Several experimental approaches are used to ob- a pulsed source is used or the radiation from a contin-
tain lifetime data. The primary goal of any of these uous source is gated with an external modulator. Data
approaches is to obtain data representing the time de- from many excitation pulses are summed for signal-
pendence of the decay of the luminescence. For a single averaging purposes. If the excitation pulse width (t,) is
component sample in which the excited state decays by much shorter than the luminescence lifetime ( T ~ ) the
,
first-order processes, the luminescence time-decay pro- true decay of the luminescence is observed as shown in
file is described by Figure 15-31a. The log slope method is illustrated in
Figure 15-31b.
Phosphorescence lifetime measurements are rea-
sonably straightforward because phosphorescence life-
where EO,is the initial luminescence signal and 7, is the times are relatively long. The pulsed Xe arc lamps used
luminescence lifetime. Thus, from E L ( t ) vs. time data. in the pulse source-gated detection technique usually
the lifetime is the time for the luminescence signal to provide a sufficiently short pulse. With mechanical shut-
decay to lle of its initial value. It can be determined by ter arrangements (phosphoroscopes), the excitation and
fitting the decay data to equation 15-55. observation steps are completely separated, and the
Before the widespread availability of laboratory
computers, lifetimes were commonly determined by
graphical procedures. For example. by taking the nat-
ural logarithm of both sides of equation 15-55, we obtain
first part of'the dccay curve is not observed (sec Figure critical that the shape ol thc excitation profile be re-
15-26), producible. With tieconvolution procedures it is pos-
Fluorescence lifetime measurements arc more dif- sihlc to measure a I-ns lifetime with a 2-11s excitation
ficult because the lifetimes arc usually five lo seven pulse width.
orders of magnitude smaller than phosphorescence lifc- Detection systems with subnanosecond response
times. To observe the true decay of the fl~~oresccncc times arc difficult to construct. Spccinl fast-response
signal, the cxcitation pulse width should be on the order PMTs are employed. The typc of dynode biasing and
of nanosccoucls or even smaller. Moreover. the re- wiring are critical for a fast response. The dccay curve
s p n x time of the detector, signal processor, and rcad- could be directly displayed o n u oscilloscope or digitized
out device must be sufficiently smi~llto prevent ciistor- (e.g., transient recorder) for later display. Because these
tion of the truc decay curvc. devices do not provide time resolution of I ns, it is morc
Most available pulsed light sources such as nitro- common to use a pulse sampling method (also called
gen or deuterium flash lamps or pulsed lascrs yield pulse the stroboscopic ~iicthod)in which the fluorescence sig-
widths of several nanoseconcts' duration. Only modc- nal over a small time wintlow is sampled and storcd
locked lascrs provide picosecond-width pulses. With na- after a controlled delay period relative to the cxcitation
nosecond excitation pulse widths, the decay curvc rep- pulse. The signal from different points of the dccay
resents a composite or convolution of the source and curvc are measured by varying the delay time. 'I'h~rsthc
tluorescencc decay as illustrated in Figure 15-32, and timc-resolved dccay is reconstructed from the data. The
the following equation applies: sampling can be accomplished with a boxcar integrator
or a sampling oscilloscope. Alternatively, the PMI' bias
voltage can be pulsed to a high value for a short period
(typically, 0.2 to 1 ns) to increase the I'MT gain at some
point along the fluorescc~~ce decay curvc. The signal
where E,.(tl) is a reference signal related to the time resulti~lgfroni thc gain pulsc is digitized and storcd.
profile of the excitation pulse, E;. ( t ) is the measured
decay curve, and t' is a dummy integration variable Time-Correlated Single-Photon Method. 'lhe
(t > t'). The truc fluorescence dccay curve is extracted time-correlated single-photon (TCSI') method is the most
froni thc experimental decay curvc by deconvolution widely used time-resolution method Sor lifctimc meas-
methods. This requires that the convolution integral be urements. As shown in Figure 15-33, the detection sys-
solved for E,.(t). Thus it is necessary to know the ex- tem measures the time between the excitation p ~ ~ landse
citation pulsc profile, which is experinlentally obtained the arrival of the first fluorescence photon. The time
by measuring the signal from a scattering solution. It is of photon emission is random but weighted according
to the decay kinetics. By measuring the emission time
for a large number of excitation pulses, the fluorescence
dccay curve is reconstructed. To obtain the true dccay
curvc, it is necessary to arrange conditions (c.g., source
irradiance) such that the probability ol' observing morc
than a single luminescence photon per excitation pulsc
is very small. In practice an average count rate of 0.01
to 0.05 photon per excitation pulse is used. This tech-
nique provides a resolution oS about 0.2 ns. Typically,
103to 106counts are accumulatecl in the peak channel.
With a low repetition rate and pulscd light source, scv-
era1 hours are rcquircd to accumulate the data.
Other L~fetlme Measurement Methods. 111the follow thc excitation source. The Fourier transform of
correlation fluorometer appioach, the \~gnnl l ~ o ma the powcr spectrum obtained is the impulse response
pulsed mode-locked law-, detected by a reference dc- function or the fluoresccrxx decay curve. The roll-off
tector, and the t l u o ~ e ~ c c n cvgnal
e ,tre rnult~pl~cdwith of the power spcctrum at higher frequencies is Loren-
a microwave mixer The le\ultnnt \ign,~li\ time dvcr- tzian if the fluorescence decay is exponential. Thus the
aged with a low-pa\s t~ltei A plot ot the corlel,~t~on lifetime can be obtainccl from the reciprocal of the Lor-
functmn vs delay tune yields the tluore\ccr~cetlec,~y cntzian half-width without resorting to Fourier trans-
curve 'The delay t m e tor the ~clerenccslgncll 1s vnr~ctl Sormation. Note that the powcr spectruin of a scattering
optically by changmg the di,tatlce ot the reference dc- solution will be flat if the source is motlulated equally at
tector f ~ o mthe law-. Correldtron techmques based on a11 frcquencics. Thus white noise in the frequency do-
opt~calmlxlrig or excltcd-\t&c populatmn mxmg have main correspollds to a infinitely narrow p~dsein the time
alw been employed domain (i.e., the Fourier transform ot 3 pulse is flat).
'I'imc-rewlut~onmethod,, \uch as the pulse wurce Experimentally, this approach has hcen irnple-
method, mvolvc me~lsuringthe llnpuisc re\pon\e f~ulnc- mcnted by using a CW lascr as the source. 'l'hc laser
twn of the tluore\cencc \peclcs In the tlme domnin In mode noise results in a lascr output that is aniplitucle
the phdse modulatmn method, the trcquency re\ponse modulated at discrete frcqucncies scparatcd by an amount
function of the fluorophore 1s monitored In the trc- equal to the laser's mode spacing. For a typical 1-m
quency donidin. Because the wurce I \ n~odul'~tcd ,it one laser, the nodes oscillate at Srequcncics from 150
frequency, the frequency lespon5e 'tt one frequency 15 up to as high as 4.5 GHz with a mode spacing of 150
measured at a given time MfIz. The powcr spectrum of a scattering signal and
The complete frequency rcsponw lunction e m he the fluorcscencc signal elicited is measured with a mi-
obtained by excitlng with a wurce th'tt IS modulated '11 crowave analyzer. 'Thc ratio of these two spectra is taken
all frcquencm m d nionltor~ngthe frequency rc4pon\c to producc :I normalized power spectrum that accounts
of the fluorophorc with ,I \pectrum m l l y ~ e rThe 'lm- for the diSSerent aniplitudcs of the laser modes. Life-
pl~tudcof thc fluoropholc u g n d I\ mcnu&ed '11 hg11 times arc extracted from this power spectrum as ciis-
frequcnc~esbec,luse the excited-\tate popu1,rtion cmnot cussed above.
Sec. 15-6 I Lifetime and Polarization Measurements 489
modulation method\. In the latter case, the phase dlf- time. Thus the anisotropy of a mixture of antibody,
ferencc between the p o l a r u a t ~ o ncomponents I5 m e w tagged antigen, and analyte antigen decreases with in-
ured and ult~matelyrelated to r . creasing analyte concentration because more tagged an-
tigen is displaced.
Applications. Fluorescence polarization is used I'olarizers have been used to reduce the elastic
in many fundamental studies o f thc hydrodynamic prop- scattering signal. Only the parallel polarization com-
erties, size, conformation, and shape o f n~acromolc- ponent of the excitation beam is scattered. Thus a po-
culcs. It has also been used t o quantify protein dcna- larizer in the excitation o r emission hcam that is ori-
turation, protein-ligand association reactions, and ented to pass only the perpcncticular polarized component
rotational rates of proteins. T h e time-dependent decay can significantly reduce the viewed scattering signal.
of anisotropy provides additional information about t h e Fluorescence-detectcd circular dichroism has also
diffusive motions of a fluorophore. gained popularity. H e r e the sample is excited with cir-
T h e primary analytical use of polarization meas- cularly polarized light. 'The corrected excitation spec-
urements is in fluorescence homogeneous irnmunoas- trum yields the same information as a conventional ab-
says. T h e anisotropy of a tagged antigen increases sub- sorption-detected circular dichroism measurement.
stantially when it is b o ~ ~ ntod a high-molecular-weight FIowever, measurements can be made at lower con-
antibody because of the increased rotational correlation centrations if +, is significant.
15-1. Calculate the peak wavelength of the liaman band Describe the instrurnent;tl differences between mo-
of water with cxc~tat~on at 365 nrn. lecular fluorcscencc and molecular phosphorescence
15-2. Discuss the major reasons why molecular phospho- spectrometry.
rescence spectrometry has not heen widely applied Contrast chcmiluminesccnce and photolurninescencc
as an analytical tool. Will thc development of room- tccliniques in terms of the factcm that control the
temperature phosphorcscencc methods hc likely to selectivity.
change this situation drastically'? Why or why not'? Saturated fluorescence can be advantageous since the
15-3. A method for obtaining molecular fluorcscencc re- fluorescence signal beconies independent of quench-
sults that arc independent of quenching has been pro- ers and prefilter effects. Consider an analyte niole-
posed [ G .M. Hieftje arid 6. R. Flaugcn, Annl. Clli~n. culc with a quantum efficiency of 0.10, a molar ab-
Acru, 123, 255 (1981)l. The fluorescence signal B, is sorptivity of 1.0 x 104 L mol c n i ' , and a
normally related to the concentration of analyte by fluorescence rate constant of 1.0 x 10" s The ~ I I -
E , = kcf),.c,where k is a constant and cb, is the quan- alytc concentration in the samplc cell is 1.0 pM.
tum yield ofiluoresccnce. Because the quantum yield (a) Calculate the source irradiance incident on the
of fluorescence is the ratio of the observed lifetime sample required to reach 50% of saturation.
of fluorescence T , to the radiative lifetime (I-,),, the (b) Ifow docs this irracliance compare to that pro-
proposed method measures both the fluorcscencc sig- vided by a conventional arc lamp-excitation
nal E, and the fluorescence lifetime I-,. Show math- monochromator configuration (= 10 mW cm '
ematically that the ratio E , / ~ , i sindcpe~idcntof yuan- over 10 rinl centered at 300 nm)?
turn yield. (c) Assume that 25% (0.50 x 50%) of the analyte
15-4. Describe the major experimental differences between molecules are excited at a given time. Calculate
the following typcs of molecular fl~~orescence spectra. the total number of fluorescence photons cmittcd
Tell which monochromator(s) are scanned (emission per second in a viewed volume clement of 1.0 x
01-excitation), which arc kept at a constmt wave- I0 ' cm'.
length, if any, and the relationship of the spectrum In Section 15-2, equations were prcsentcd to cstimatc
obtained to the molecular absorption spectrum, if the detection limit in photoluminescence mc;tsure-
any. nicnts that are limited by dark current shot noise.
(a) The excitation spcctruni. Assume that the slope of the calibration curve for the
(h) Thc emission spectrum. analyte referenced to the photocathode is given by
(c) The sync1mm)us fluorescence spectrum. equation 15-41 and that the molar ;tbsorptivity i s
(d) The cxcitatio~ilcrllissionmatrix spcctrum. 1 .O x 10' L r n o l ' cm ;lnd tlic power yield is 0.50.
15-5. Discuss why a molcc~~lar fluorcscencc calibration plot The blank equivalent concentration (BEX) is I .O nM.
becomes nonlinear at high concentrations. C';ilculatcthe following assuming that the noise band-
Chap. 15 1 References 491
width constant (K) is 1.0 x lo-'" and the source 0.10 pM-' cm-I and the fluorescence signal for 0.01
flicker factor is 0.0050. ILMof the analyte is 1.0 mV.
(a) The detection limit when limited by background
(a) Calculate the primary absorption correction fac-
luminescence shot noise. How does this detection
tor and the observed fluorescence signal at ana-
limit compare to that calculated with a typical
lyte concentrations of 0.1, l , and 10 pM.
value of dark current shot noise?
(b) What is the BEC for which background lumi- (b) Calculate the error in the measurement of an
analyte concentration of 0.10 pM caused by sec-
nescence shot and flicker noise are equal?
ondary absorption by concomitants if the ab-
In a pulse excitation experiment, the fluorescence sorbance at the emission wavelength over 1 cm
signal decays to one-tenth of its initial value in 10 ns. is 0.10 and 1.0.
Calculate the fluorescence lifetime. Calculate the
SIB for measurements at 2 and 10 ns after the exci- Discuss the use of derivative and wavelength mod-
tation pulse. Assume that the SIB is 1.0 at zero time ulation techniques for photoluminescence measure-
and that the background luminesence has a lifetime ments.
of 1.0 ns. Calculate the fraction of the total phosphorescence
Contrast molecular absorption and fluorescence signal observed with a phosphoroscope for analytes
measurements with respect to selectivity and detec- with phosphorescence lifetimes of 10 and 100 rns with
tion limits. a delay time of 2 ms and exposure and observation
In a fluorescence polarization irnmunoassay, why does times of 3 ms.
the anisotropy decrease at the analyte concentration Phase modulation measurements are made with a
increases? source modulation frequency of 30 MHz. The analyte
Fluorescence measurements are made in a l-cm-path- fluorescence signal is phase shifted 60" relative to a
length cell under conditions that b,, = b,, = 0.40 cm scattering solution. Calculate the fluorescence life-
and b" = 0.10 cm. The absorbance of the analyte is time and the demodulation factor ( m ) .
REFERENCES
The following references deal with molecular luminescence E. L. Wehry, ed., Modern Fluorescence Spectroscopy,
spectroscopy. Plenum Press, New York, vol. 1, 1976: vol. 2, 1976; vol.
3; 1981; vol. 4,1981. This series includes many excellent
G. G. Guilbault, Practical Fluorescence: Theory, Meth-
review chapters on recent advances in techniques and
ods, and Techniques, Marcel Dekker, New York, 1973.
applications of molecular luminescence in such areas as
S. G. Schulman, ed., Molecular Luminescence Spec- modulation, synchronous, and derivative techniques, laser
troscopy: Methods and Applications, Part I ; Wiley, New excitation, chemiluminescence~clinical and biochemical
York, 1985. applications, reaction-rate methods, fluoroimmunoas-
J. D. Winefordner. S. G. Schulman, and T. C. O'Haver, says, array detectors, probe ion techniques, and low-
Luminescence Spectrometry in Analytical Chemistry, temperature measurements.
Wiley-Interscience, New York, 1972. E. J. Bowen, ed., Luminescence in Chemistry. Van Nos-
R. S. Becker, Theory and Interpretation of Fluorescence trand, London, 1968.
and Phosphorescence. Wiley-Interscience, New York, D. M. Hercules, ed., Fluorescence and Phosphorescence
1969. Analysis, Wiley-Interscience. New York. 1965.
W. R. Seitz. "Luminescence Spectrometry (Fluorimetry A . J. Pesce. C. J. Rosen, and T. L. Pasby, Fluorescence
and Phosphorimetry)," in Treatise on Analytical Chem- Spectroscopy: A n Introduction for Biology and Medi-
istry, 2nd ed.. I . M , Kolthoff and P. J. Elving, eds., pt. cine, Marcel Dekker, New York. 1971.
I., vol. 7, Wiley-Interscience, New York. 1981, pp. 161-
204. S. G. Schulman, Fluorescence and Phosphorescence
Spectroscopy: Physiochemical Principles and Practice,
C. A. Parker, Photoluminescence of Solutions, Elsevier,
Pergamon Press. Oxford, 1977.
Amsterdam, 1968.
S. Udenfriend. Fluorescence Assay of Biology and Med- C. E. White and R. J. Argauer, Fluorescence Analysis:
icine, Academic Press, New York. vol. I, 1962; vol. 11, A Practicalilpproach, Mercel Dekker. New York. 1970.
1969. P. A. St. John. "Fluorometric Methods for Traces of
J. R. Lakowicz, Principles of Fluorescence Spectroscopy, Elements," chap. VII-B in Trace Analysis. J . D .
Plenum Press, New York. 1983. Winefordner. ed.. Wiley-Interscience, New York. 1976.
J . N . Miller, Standards in Fluorescence Spectrometry. B. L. Van Duuren and T. L. Chan, '-Fluorescence Spec-
Chapman & Hall, London. 1981. trometry;" chap. 7 in Spectrochemical Methods of Anal-
2 Chap. 75 1 Molecular Luminescence Spectrometry
vsis, J . D. Winefordner, ed., Wilcy-Intersciencc, New 33. 1). 13. Paul, "licccnt Analytical Developrncnt llsing
York, 1971. (:hcmilurninesce~~cc in Solution," 'Iiilnntu,25, 337 (1978).
18. T . C. O'Haver. "The Development of Lumincscencc 34. E. Schrani and 1'. E:..Stanley, cds.. I'rocwdirrg o/'ilrc
Spectrometry as an Analytical Tool." .I. ( ' l i r n ~Etluc., Irllc~rrrationalSyrnpriunl on Analytic~i~I Applic(ltiorrs o f
5.5, 423 (1 978). Hiol~unitzrsccncerlntl C'lrc~rrrilult7inc~,r(~c~11c~c~,
State Printing
19. F. Van Creel, E. Voigtman, m d .I. D. Wincfordner, and Publishing, Westlakc, Calif., 1979.
"General Intensity Expression in Molccul:u- Imnines- 35. M. Cormicr, I>. M. Hercules. and J. Lcc. eds., Biollc
cence Spectrometry ," Appl. Spectrosc., .?8, 228 (1 986). nlirlcwmcp and C'i~c~ir~ilr~tnitic~.sc~(~~/(~c~,
P I C I ~ Press,
L I ~ New
20. I. M. Warner, G. Fatonay, antl M. 1'. Ihomas. "Mul- York, 197 I .
tidimensional Lumincsccncc Measurements," A M / . 36. W. R . Seitz and M. P. Ncary. "C'hemilumiriesce~iccand
C'hc~nt.,57, 403A ( 1985). Biolurnincsccnce." Ancll. ('hcwz., 46, l SXA (1974).
21. K. D. Mielenz, cd., "Measurcmcnt o f Photolumincs- 37. M. A. Deluca and W. D. McElroy. eds., Hioltrr~~irrc~s-
cencc," vol. 3 in Opticd Rrcrliution Meczs~~rernrnis, F. wtzct, (itzd C,'hci~~il~~~nincscc~~~cc~Acadeniic
Press, New York.
Grum and C. J . Bartleson. eds.. Academic Press, New 1981.
York, 1982. This reference includes exccllcnt chapters 38. M. A . Dcluca, ed., "Iliolunlinescc~~cemci Chernilu-
on correction of cxcitation and emission spectra and minesccncc," in Method,s of Enzyrnolo,yv. vol. 57. Ac-
quantum yield me;rsurcmcnts. ademic Press, New York, 1978.
22. "Molecular Fluol-csccnce, Phosphorcsccnce, and Chcm- 39. A . K. Campbell, M. 13. Fiallett, and 1. Wccks, "Chcm-
ilumincscence Spectrometry," Anal. C'hem. This review iluminesccnce as a n An;~lyticalTool in Cell Biology and
articlc appears once every two years in the Fundamental Medicine," in M(4rocl.so/'BioclrcwlicdAn(~lv.si,s, D . Glick,
Reviews issue of Anrdylicul C'lrrvnistry. Applications as e d . , vol. 31, Wiley-lntcrscience. Ncw York, 1985. p.
well as recent advances in instrumentation and chemistry 317.
arc covered. 40. L. J . Kricka antl G.I I . C;. Thorpc, "Chemiluminescent
The following references deal with phase-resolved and lifc- and Bioluminescent Methods in Analytical Chemistry,"
time measurements. Analyst, 108, 3 274 (1983).
41. L. J. Kricka, P. E . Stanley, <;. H. G.Thorpe, and J .
23. L. McGown and I;. Bright, "Phase-Kcsolved Fluorcs-
P. Whitehead, eds., Anrllyticul App1ii.alion.r. o f Biolu-
cence in Chemical Analysis," CRC' Crii. RPV. Anal.
rninescrt~ceurrd C'hetnilri~ninr.sc~c~t~ee, Acadcmic Press,
C'lwrn., IS, 245 (1987).
London, 1984.
24. J. N. Demas. Exiled St[& Lzfktinze Meuswwmxts, Ac-
ademic Press, New York, 1983. 'The following refercnccs deal with fluorescence IIPLC de-
25. G. M. Hieftjc and G . R. Flaugcn, "Correlation-Based tection.
Approaches t o Time-Resolved Fluorimetry," Anal.
C'hrm., 53, 755A (1981). 42. D . C. Shelly and I . M. Warner. Detectors
26. T. D. Harris and F. E. Lytle, "Analytical Applications in High-Performance Liquid Chromatography." chap. 3
of Laser Absorption and Emission Spectroscopy," chap. in Liquid CI~r~wr(lfogrupi~y I)etrctor.s, 7'. M . Vickery,
7 in Ultrasansitive Laser Spec,/roscopy,D . S. Kligcr, ed.. c d . , Marcel Dekker, New York, 1983.
Acadcmic Press, New York, 1983. 43. A. T. Rhys Williams, Fluorcwcwc,e L)ciec,iiorr in Liq~lid
27. D . V. O'Connor and D. Phillips, Titne-Corrclalcd Single Chron~togrccphy,I'erkin-Elmer, Bucl<inghamshirc.
Photon Counting, Academic Press, London, 1979. England. 1080.
Thc following references dcal specifically with phospliorin-
etry. The following refercnccs deal the absorption-correctccl lu-
minescence measurements.
28. T . Vo-Dinh, Room Ternpcrutwc, Phosplzorirnctry ,for
C'hemicul Annlysis, Wilcy-lntcrscience, New York, 1984. 44. J . F. Holland, I<. 6. Teets, P. M. Kelly, and A . Timnick,
29. M. Zander, Pho.sphorirnc~try,Academic Press, New York, "Correction of Right-Angle Fluorescence Mcarure-
1968. ments for the Absorption of Excitation tiadintion." Arrrcl.
30. Ii. J . Hurtubise, "Phosphorimetry." Anal. Chrrn., 55. Cllrnl. , 49, 706 ( 1977).
669A (1983). 45. D. R . Christman, S. R. Crouch, and A . Timnick, "Au-
tomated Instrument Sot- Absorption-Correctcci Molec-
The following references deal with chcmiluminescencc.
ular Fluorescence Measurements hy the Cell Shift
31. W. R. Seitz, "Cherniluminesce~iccand Riolun~incsccnce Method," Anal. Chcrn.. 53, 276 (108 I).
Analysis: Fimdamcntals and Biochemical Appliczi- 46. K. Adamsons, J . E . Sell, J . I;. Holland, and A . Timnick,
tions," C'RC' Crit. Rev. Anal. Chetn., I.?, 1 (1981). "Cell Rotation for Absorption Correctcci Measurements
32. V. Isacsson and G. Wettermark, "Chernilu~nincscc~lce in Molecular Fluorescence," Am. I,ab., /(,(I 1). I6 ( 1984).
in Analytical Chemistry," A n d . Cl?inz. Acts, 68, 339 This article includes a review of thc different techniques
( 1974). used for ;thsorption correction.
Chap. 15 I References 493
Other specific applications of molecular luminescence are dis- 51. E. M. Chait and R. C. Ebersole, "Clinical Analysis-
cussed in the following references. A Perspective on Chromatographic and Immunoassay
Technology," Anal. Chem., 53, 682A (1981).
F. D. Snell, Photometric and Fluorometric Methods of
Analysis: Metals, Wiley-Interscience, New York, 1981. 52. E . Soini and I. Hemmila, "Fluorimmunoassay: Present
F. D. Snell, Photometric and Fluorometric Methods of Status and Key Problems," Clin. Chem., 25. 353 (1979).
Analysis: Non-metals, Wiley-Interscience, New York, 53. I. M. Warner and L. B . McGown. "Recent Advances
1981. in Multicomponent Fluorescence," CRC Crit. Rev. Anal.
P. A . St. John, W. J. McCarthy, and J. D. Winefordner, Chem., 13. 155 (1982).
"Applications of Signal-to-Noise Theory in Molecular 54. C. G. delima, "The Shopol'skii Effect as an Analytical
Spectrometry," Anal. Chem., 38, 1828 (1966). Tool," C R C Crit. Rev. Anal. Chem.. 16. 177 (1986).
F . E. Lytle, "Solution Luminescence of Metal Com- 55. W. R. Seitz, "Fluorescence Derivatization," C R C Crit.
plexes," Appl. Spectrosc., 24, 314 (1970). Rev. Anal. Chem.. 8 , 367 (1980).
In prcvious chapters we have considcrcd the absorp- the incident heam. The Kaman effcct can he treated in
lion, cniission, or luminescence of electromagnetic ra- most aspects with classical theories. Wc discuss the the-
diation by atoms and molecules. When radiation inter- ory here qualitatively in order to develop selection rules
acts with matter, however, several additional processes for Rarnan transitions. Thc instrumentation used t o ob-
can occur (recall Figure 1-1). The radiation can, for serve Raman hands is ncxt described. Also considered
cxamplc, be refracted, reflected, or scattered. While are resonance Kaman methods, nonlinear Raman
the first two processes do not find many direct analytical methods, such as coherent anti-Stokes Raman, and var-
applications, the scattering of clcctromngnetic radiation ious applications o f Raman spectroscopy.
forms the basis for several analytical techniques, in- The ncxt section considers the analytically useful
cluding Raman spectroscopy. molecular scattcring methods known as turhidimetry
In this chapter the principlcs o f radiation scattcr- and nephclometry. The chaplcr concludes with a de-
ing are first considered in order to put the Rarnan eflcct scription of various laser light scattcring methods and
and various other scattering phenomena into a11 overall includes a discussion of remote sensing with laser sources.
perspective. Many scattering processes irivolvc n o changc
in the energy of the scattered beam compared to the
incident heam. On the surface, these processes appear
rnerely t o rantloniize the direction o f an incident beam SCATTERING
of photons. In actuality, however, the dctails of the
various scattering processes arc quite complex: only a The general principlcs 01 radiation scattering were in-
q~ialitativediscussion o f the theory of radiation scat- troduced in Section 2-3. We concern ourselves here with
tering is givcn hcrc. two general classes of scattcring phenomena: elastic
Following this general introduction to radiation scattcring, in which the scattcrcd radiation is of the
scattcring, Raman spectroscopy is considcrcd. The Ila- same frequency as the incident radiation, and inelastic
man effcct is a scattcring pl~eno~ncnon that does involve scattering. in which the scattered radiation is of a dif-
a net cncrgy change hetween thc scattered beam and ferent frequency.
Sec. 16-1 1 Principles of Radiation Scattering 495
TABLE 76- 7 tion has oscillating electric and magnetic fields that can
Scattering classes interact strongly with the electron clouds of the analyte
molecules. If these molecules are polarizable, charges
Type of Refractive index Size are induced that oscillate at the frequency of the field
scattering requirement" requirementb
as shown schematically for a single molecule in Figure
Rayleigh - I)! << 1 d, < 0.05 A 16-1.
Debye ( % - 11) = 0.1 0.05 A < d, < A The oscillating dipole induced in the analyte mol-
Mie (% - 1) >> 0 d, > A ecule produces a field of its own which also oscillates
--
dq,, Relative refractive index, q,/q,: q,. refractive index of scatterer: q,,. at frequency v. This field acts as a source of radiation
refractive index of medium. itself so that radiation of the same frequency as the
bd,, Major dimension of scatterer
incident beam is sent in all directions from the particle.
From a quantum mechanical viewpoint, elastic
Elastic Scattering scattering can be considered to be the removal of energy
from the incident beam and the subsequent reemission
In Section 2-3 elastic scattering was classified into three of that energy. We can view scattering in terms of tran-
types: Rayleigh scattering, Debye scattering and Mie sitions between energy levels. A molecule in the ground
scattering. It should be noted that this classification is state absorbs an incident photon and reaches an inter-
not universally followed. Debye scattering is often re- mediate level called a virtual level because it does not
ferred to as Rayleigh-Debye scattering and is some- correspond to a real energy level of the atom or mol-
times omitted as a class. The type of scattering observed ecule. Despite this, there is a finite transition proba-
depends on the refractive index of the particle relative bility connecting the ground state to a virtual state. The
to the surrounding medium and the dimensions of the closer the virtual state is to a real state, the higher the
particle relative to the wavelength of the incident ra- probability of absorption. The molecule returns to
diation. Approximate criteria for the types of scattering the ground state emitting a photon of the same energy
are listed in Table 16-1. that was absorbed. Elastic scattering and resonance flu-
Radiation scattering, like reflection and refrac- orescence are similar processes, except that the level
tion, has its origins in the induced secondary emission reached upon absorption is virtual in the scattering case
of particles in the path of an incident beam of radiation. and real in the resonance fluorescence case. Scattering
Scattering occurs only if the particles have dimensions occurs essentially instantaneously, while the upper level
on the order of or smaller than the incident beam wave- in fluorescence has a measurable lifetime.
length and they are randomly distributed in a medium Let us now examine in more detail the types of
of refractive index different from their own. In other elastic scattering.
cases only reflection and refraction are observed.
From a classical viewpoint, let us first consider the Rayleigh Scattering. Rayleigh scattering is
sample to be a collection of spherical molecules with characteristic of scattering from such particles as atoms
polarizable electron clouds illuminated with electro- and molecules where the dimensions of the scatterer
magnetic radiation of frequency v. The incident radia- are much smaller than the incident beam wavelength
. Particle
(uncharged)
Dipole polarity
varies with field
(typical molecular dimension5 are on the order of na- ing. Figure 16-3a illustrates the interference that can
nometers). In the case of Rayleigh scattering. we can occur in large-particle scattering. As a result the inten-
consider the scatterer to be a point source of secondary sity distribution pattern shifts to one showing predom-
emission. inantly forward scattering as depicted by the center scat-
Lord Rayleigh was the first to investigate the de- tering envelope in Figure 16-3b. Note that the envelope
pendence of the scattered radiation intensity on the is similar to Rayleigh scattering in the forward direction,
incident wavelength. For a single particle, a syrnmet- but quite different in the reverse direction.
rical intensity distribution of the type shown in Figure Mie scattering occurs from large particles of rel-
16-2 is obtained with unpolari~edincident radiation. atively high refractive index. Here, not only are there
The irradiance of the scattered radiation at angle 0. fixed phase relationships between waves scattered from
(E,,),. is given by different parts of the same particle. but there are also
distortions of the electric field of the incident and scat-
tered radiation. The result is the complex scattering
envelope shown in the inner part of Figure 16-3b. Note
again the similarity to Rayleigh scattering in the forward
where a' is the polarizability of the particle in m3, X is direction and the complicated angular dependence in
the wavelength of the incident radiation, 0 is the angle the reverse direction.
between the incident and scattered ray, E, is the inci-
dent beam irradiance, and d is the distance from the Scattering from Liquids and Solutions. Pure
center of scattering to the detector. The polarizability liquids and solutions of small molecules are generally
is a measure of how efficiently a given incident fre- poor light scatterers. Because they contain a fairly reg-
quency induces a dipole in the particle. Because the ular array of molecules, lateral destructive interference
polarizability varies roughly with the volume of the scat- occurs; this virtually ensures that most of the incident
tering particle, equation 16-1 predicts that the scattered radiation is refracted or reflected at the surface. Very
radiation intensity increases with increasing particle size. weak scattering can occur, however, because of inhom-
Thus, in a sample that contains particles of various sizes, ogeneities (localized density and concentration varia-
the larger particles tend to contribute most heavily to tions) produced by thermal forces. These lead to some
the scattering. volume elements with more particles than the average
Equation 16-1 also predicts that the scattered ra- and some with less. Each volume element that differs
diation intensity varies inversely with the fourth power from the average can act briefly as a local scattering
of the wavelength (directly with v4). This dependence center of polarizability Aa'. where Aa' = a;,,,, - a:,.
on wavelength is responsible for such natural phenom- For such a case we can replace (a')2 in equation 16-1
ena as the blue color of the sky and ihe red color of with Since the latter is very small, scattering is
the sun at sunset. The sky appears blue because short expected to be quite weak.
wavelengths of sunlight are efficiently scattered by small If the particles in solution are large molecules,
dust particles and water vapor in the atmosphere. The such as polymers or sols, the scattered radiation can be
red color of the sun as seen through haze, smoke, or primarily the result of concentration fluctuations which
fog results from the efficient transniission of the longer increase with increasing concentration. With molecules
wavelengths of sunlight relative to the shorter wave- that have dimension> between approximately 1.5 and
lengths which are effectively scattered. Note also from 40 nm, the scattering is still of the Rayleigh type.
Figure 16-2 that with unpolarized incident radiation.
the scattered radlation at 90" is linearly polarized.
Inelastic Scatterin
Large-Particle Scattering. The comprehensive
theory of large-particle (Debye and Wlie) scattering was Two types of inelastic scattering can be distinguished.
formulated by Mie. In Debye scattering we can no longer In Raman scattering, relatively large frequency shifts
consider the particles to be point sources. Instead. cen- occur that are independent of scattering angle. Raman
ters of scattering are found in various areas of the par- scattering is caused by rotational and vibrational tran-
ticle. When such particles scatter radiation. the scat- sitions in molecules. In addition, scattering can occur
tering centers are far enough apart that some interference with a relatively small frequency shift that varies with
is likely between the rays emitted from one area of the the scattering angle. This type of scattering, caused by
particle and those from another. This leads to an in- thermal fluctuations in the medium, is known as Bril-
tensity distribution that is quite different from the fairly louin scattering. Because Brillouin scattering requires
uniform distribution characteristic of Rayleigh scatter- a high-resolution spectrometer (e.g.. Fabry-Perot in-
Chap. 16 I Molecular Scattering Methods
(t))
FIGURE 16-3 Scattel-ingfrom large particles. In (a) the interference that
occurs in the scattering from ;I large particlc is illustrated. Note that the
rays emitted in a backward direction are highly susceptible to dcstructive
interference because of the large path differences possible. Thus scattered
radiation originating at ccnters X and Y and observed at point I) is likely
to show dcstructive interference effects hecause there is a large path
differencc between the route S X I ) and the routc S Y D . In the forward
direction, there is a much smaller path difference to detector position I)'.
Hence rays traveling the route S X D ' and those arriving via S Y D ' arc
more likely to constructively interfere. In (b) the distributions of scattered
radiation intensity in the plane of propagation are shown for thc three
scattering types. I n Debyc scattering, the highest intensity is observed in
the forward scattering direction hecause of dcstructivc interference in the
backward dircction. Mic scattering is much more complex and diffraction
effects appear in the angular distribution diagram. Notc that the scattering
anglc 0 is taken as the angle betwccn the incident and scattcrcd rays. Thus
B is 0" for forward scattering and 180" for scattcring in thc backward
direction.
terferorneter) to observe and has kw analytical appli- 5a1d t o be ,I Warnan-active rnodc The rewltmg change5
cations, it is discussed only briefly in Section 16-4. In the anplitucle ot the {cattercd r a d ~ a t ~ o;ire
n ~llu\-
111discussing Rayleigh scattering. it was assumed trnted rn F~gurc1 6 4 . For ,I monochroinat~cmcrdent
that the polarizability a' of the scattering molecule re- b e m ~ the
, c~~nplrtude-~nodulated wavclorm 14 \hewn ~n
mains constant. A changing polarizability gives rise to Sect~~jil 10-2 to havc three lrcquency component\. One
a scattered radiation intensity that changes correspond- component c o ~ ~ e \ p o n tto
t \ the R,~yle~ghscattered ra-
ingly. A moleculc can undergo a change in polarizability di,it~onand occur\ at the trcquenq o f the monochro-
during one of its norinal modes o f vihration. A normal rndt~cwurce, the rcm'lining two component\ colic-
mode that involves such u change in polarizability is ,pond to R a m m 5catteled r a d ~ a t ~ omn d occur \hlttcd
Sec. 16-2 / Raman Spectroscopy 499
-
make a more comprehensive comparison between Ra-
frequency man and IR spectroscopy.
Time
cidation, multicomponent qualitative ;~nalysis,and obtain an expression for the induced dipole moment
quantitative determinations of minor and trace constit- k,,,:
uents.
Although Kaman scattering can be used to obtain PIl1= @I&, COs ( 2 n v J )
information about rotational, electronic, and vibra-
tional transitions, we will be concerned primarily with
its use in vibrational spectroscopy.
+ E,,,r,,, ($1 C
cos (Znv,,r ) cos (2nv,,i) (1 6-71
for the integral in equation 14-3 to be nonzero. that is. on the order of cm2 for a good Raman scatterer.
for the transition to be allowed. there must be a change Fundamentally, the strength of a Raman line depends
in dipole moment during the vibration. For Raman spec- on the square of the polarizability derivative (a(v,,) is
troscopy, on the other hand, there must be a change in proportional to [(da/dr),J2). In actual practice. the in-
polarizability during the vibration. An easy way to de- tensities measured depend on averages of the polariz-
termine whether a transition is allowed or not in IR ability derivatives for isotropic systems (e.g., liquids
and Raman spectroscopy is to use group theory and the and gases). For a sample such as a single crystal. the
symmetry properties of the states and the operator (di- radiant power of Raman-scattered radiation depends
pole moment or polarizability) involved. For integrals experimentally on the specific crystal orientation.
such as those in equations 14-3 and 16-10 to be nonzero, Raman intensities also are directly proportional
they must be totally symmetric. That is, the product of to the source irradiance E,,, as shown in equation 16-
the ground state, the operator and the excited state 11. For this reason and for several others discussed
symmetry must be totally symmetric (i.e., totally in- later. laser sources have replaced conventional sources
variant under all symmetry operations). This leads to in modern spectrometers. The radiant power of Raman-
the conclusion that allowed IR transitions generally in- scattered radiation is usually directly proportional to
volve unsymmetrical vibrations. By contrast, Raman the concentration of the active species. Because of this,
active vibrations include those that are themselves to- quantitative analysis with Raman spectrometry has be-
tally symmetric. come quite popular.
The differences between Raman and IR activity The dependence of the detected Raman scattering
arise from the different symmetry properties of the di- radiant power on instrumental parameters such as source
pole moment and the polarizability operators. For mol- irradiance, the emission collection efficiency, and con-
ecules with a center of symmetry, these differences lead centration is quite similar to that observed in molecular
to the conclusion that there are no IR active transitions luminescence spectrometry (see Table 15-1). The major
in common with Raman active transitions, the mutual difference is that the Raman cross section replaces the
exclusion principle. For example, the symmetric stretch- &YLterm in the luminescence expression. Thus many
ing mode of CO, is IR inactive because there is no of the techniques used in luminescence ypectrometry
dipole moment change during the vibration (recall Fig- for maximizing the analytical signal and for optimizing
ure 14-1). On the other hand, the polarizability varies the signal-to-noise ratio (see Chapter 15) are also ap-
during the vibration. which leads to Raman activity. plicable with Raman spectrometry.
For the asymmetric stretch of CO,, the dipole moment A typical Raman spectrum is shown in Figure 16-
changes during the vibration. However, as the polar- 6. It is common in Raman spectrometry to calibrate the
izability of one of the C-0 bonds increases as it length-
ens. that of the other decreases, and overall. there is
no change. Thus the asymmetric stretching vibration of
GO, is Raman inactive.
For noncentrosymmetric molecules, there are many
cases in which the mutual exclusion principle still holds.
However. many other vibrational modes may be both
Raman and IR active. One interesting case is that of
C, symmetry, in which there is no symmetry. Here all Anti-Stokes lines
vibrations are both Raman and IR active. In cases where
vibrations are Raman and IR active. the intensities ob-
served for the same vibration may be quite different.
intensity increases as (v,J4, A r f and Kr- lasers can capacity dumping accessory (see Section 4-3). By fre-
provide a significant improvement in Raman intensity quency doubling the output, tunable UV radiation can
over the He-Ne laser. In addition, the most commonly be produced. The short (e.g.. picosecond) pulses also
used detectors have higher responsivities in the blue or allow time-resolution techniques to be employed to re-
green than in the red. ject fluorescence.
The excitation wavelength for Raman spectrom-
etry must be chosen carefully. Photodecomposition of
samples can be a severe problem with short-wavelength Sample Cells and Cell Configurations. Raman
excitation. In addition, fluorescence can also occur with spectra can be readily obtained with gas, liquid, and
some samples. Colored samples can absorb either the solid samples. Since the exciting and scattered radiation
incident radiation or the Raman-scattered radiation. is in the visible (occasionally ultraviolet) region, cell
Hence a compromise is often made in selecting the materials, windows, and optical materials can be made
excitation wavelength. The best wavelength would pro- of glass or quartz. With laser sources, many different
duce high Raman intensities, but minimal photodecom- sample cells and configurations can be used.
position, fluorescence, and, except for resonance Ra- The most common cells for liquids are capillary
man spectrometry (discussed later), minimal absorption. tubes (typically, 1.5 mm 0.d.) in which pure liquids or
Tunable lasers are also used for exciting Raman solutions are contained. Two widely used sample cell
spectra. Often these are CW dye lasers pumped by configurations for micro samples are shown in Figure
powerful A r t or Kr+ lasers. These sources allow op- 16-8. Cell configurations have also been described for
timization of the excitation wavelength. In addition, macro samples. Flow-through cells, while not com-
they are highly useful for resonance Raman spectros- monly used for Raman spectra, have been described
copy (see later). A potentially important source is the for examining liquid samples without realignment.
synchronously pumped dye laser. This is produced by Another popular cell design, shown in Figure
a mode-locked Ar' laser pumping a dye laser with a 16-9, is the rotating cell, originally developed for ob-
Mirror
11
Wavelength
selector
Mi
I \ capillary
I I
F?
From laser
Lells
sible to obtain time-resolved Raman spectra for studies With photon counting we will assume that amplifier-
of the dynamics of molecular processes. readout and excess dark current noise are negligible.
Microcomputers and microprocessors are playing We can thus write an expression similar to equation
increasingly important roles in Raman instruments. Their 5-61 for the SIN.
major use is in signal averaging. Here, an automated
spectrometer can use long integration times and photon S Y ~ R
N - [ ( n , + n , + n, + n,,) + [:(nR + nF + n1,)2]112
-
counting for SIN enhancement. With time-resolved Ra-
man spectrometry of repetitive processes, computer av-
eraging of multiple spectra can be used. In addition,
recent monochromators have been operated under
computer control. Often these use the simple sine bar. where n, is the number of fluorescence counts from the
linear wavelength drive (see Figure 3-46) rather than analyte, concomitants and the cell in time t , n,, is the
the more complicated cosecant drive; the computer then number of counts due to elastic scattering, n, is the
converts wavelength to the Raman shift in wavenum- number of dark counts, and [,
is the source flicker
bers. Computers have also been used to perform many factor.
other functions, including digital filtering, smoothing, Equation 16-13 shows that for weak Raman sig-
calculation of depolarization ratios, line-shape analysis, nals. the SIN can be improved by increasing n,, or by
spectral matching, and wavelength calibration. reducing n,, n,, or n,,. The Raman signal can be in-
creased by using a higher source irradiance or a higher
Signal-to-Noise Ratio Considerations. The excitation frequency and by improving the collection
signal-to-noise ratio (SIN) expressions in Raman spec- efficiency. The dark signal can be reduced by cooling
trometry are quite similar to those in molecular lumi- the PMT. The elastic scattering signal can be reduced
nescence spectrometry (see Table 15-2).The major dif- by using a monochromator with better stray radiation
ferences are that the scattering signal is the analytical rejection. Reducing the fluorescence background is often
signal in Raman spectrometry, and fluorescence from one of the more difficult tasks in Raman spectrometry.
any source is a background signal. Sometimes changing to a different excitation frequency
Since photon counting is widely used in Raman will reduce fluorescence. It may also be necessary to
spectrometry, we will write the SIN expression in terms clean up the sample or to purify the solvent if the flu-
of the number of Raman counts n, observed in time t . orescence is from concomitants and not sufficiently re-
Single-channel
display
Mult~channel
display
enters the interferometer. Multistage interference fil- tectors respond very rapidly, Raman spec-
ters and very high rejection ratio filters called Chevron trometry can be used to study short-lived or
filters have been used to achieve rejection ratios in the transient species and to follow the kinetics of
range lo-' to 10P1('. In addition, it has been proposed rapid reactions.
to carry out FT-Raman measurements with excitation 5. A single Raman spectrometer can cover the
in the near-IR region. In this region, the fluorescence entire range of vibrational frequencies, whereas
stimulated is minimal and photodecomposition of the even with FTIR systems, changes in detectors
sample can be largely avoided. Although spectra are or beam splitters must be made to cover this
weaker because of the wavelength dependence of scat- range. With conventional IR spectrophotom-
tering and detectors are much less sensitive, very high eters. two or more instruments must be used
average-power lasers can be used. With solid-state de- over this range.
tectors rather than photomultipliers. the multiplex ad- 6. Raman spectra are usually much simpler than
vantage comes back into play. and it has been calculated IR spectra because overtone and combination
that these detectors should outperform the PMT at bands are not very intense. Overlapping bands
moderate signal levels. In any case, theoretical analysis are thus much less common.
has suggested and experimental results have demon- 7. Totally symmetric vibrations can be observed
strated that FT-Raman measurements in the near-IR with Raman spectrometry, whereas they are
region provide SIN values in the range 10' to lo", which not with IR spectrometry.
is in the same range as ordinary Raman measurements. Polarization measurements add an extra di-
8.
In addition to the freedom from fluorescence and
mension to the information obtained by Ra-
photochemistry, FT-Raman benefits from using the same man spectrometry. This aids in band assign-
interferometer and computer system that is used for
ments and structure determinations.
FTIR measurements. We should see further improve-
9. Raman intensities are directly proportional to
ments in instrumentation and many new applications of
this method. concentration and to the laser power.
The resonance Raman effect can substantially im- state. Note the simplicity and the sharpness of the Ra-
prove the selectivity of Raman measurements on com- man spectrum. The combination of Raman and IR spec-
plex molecules. Because an electronic transition is trometry is extremely powerful for structure elucida-
somewhat localized in one part of a complex molecule, tion.
bands of the chromophore can be observed with little Raman spectrometry is also applied in measure-
interference from other Raman bands of the molecule. ments of physical properties, such as hydrogen-bond
strengths, acid dissociation constants, and phase tran-
Applications sitions. In addition, Raman spectrometry is becoming
quite useful in organic chemistry for the study of short-
Raman spectrometry is widely used in the chemical sci- lived species, such as reaction intermediates or excited
ences. We consider here only a few typical applications states.
of the Raman technique in qualitative and quantitative
analyses. Inorganic Applications. Raman spectrometry
is an essential technique in inorganic structural analysis.
Organic Applications. Raman spectrometry has It is widely used in conjunction with IR data for the
been applied to organic systems since the 1930s. Be- vibrational analysis of inorganic solids. Such studies al-
cause of its complementary nature, Raman spectrom- low the assignment of internal modes of the ions and
etry is often used in conjunction with IR to give qual- crystal lattice modes.
itative and structural information. With transition metal complexes, the vibrational
Group frequencies regions are used to identify energies of many metal-ligand bonds are in the region
functional groups just as they are in IR spectrophotom- 100 to 700 cm-l, a region that is difficult to study by
etry. Raman scattering is particularly useful for such IR spectrophotometry. Such metal-ligand vibrations
groups as -C-S-, --S-S-, -C-C-. -N=N-, are frequently Raman active and readily observed by
and -C=C-; IR spectrophotometry is well suited to Raman spectrometry.
identify such groups as -0-H, C = O , P = O , -NO,, The ease of obtaining Raman spectra in aqueous
and S=O. Correlation charts and spectral compilations solutions has also contributed to the popularity of Ra-
are now available for functional group and compound man measurements on inorganic compounds. Highly
identification. colored samples can be studied with the rotating cell
Raman and IR spectra are both used for structural technique described earlier. Resonance enhancement
studies of organic compounds. Figure 16-13 compares is becoming increasingly useful in inorganic applica-
the IR and Raman spectra of cystine in the crystalline tions. Figure 16-14 shows the resonance Raman spec-
trum of Cs,Mo,Cl,. By bringing the exciting line into actual b~ologicdproce\\es Normnlly, concentr,itlom
coincidence with thc lowest allowed electronic transi- on the order ot 0.1 M ,rle rcqu~red Rcson,mce R , m m
tion, the overtone progression of tlie totally symmetric spectrometry, howeve1 , ccrn caddy detect moleculcc 'lt
Mo-Mo vibration is observed. o n W ~ t hrewnmcc Kc]-
the 1 0 p 5 M c o n c e n t ~ ~ i t ~lcvel
man, only th,~tpnrt o f the molecule clo\ely '~ssoc~ated
Biological Applications. Rarnan spectrometry w ~ t hthe clectronlc trnns~t~on I \ enhanced Thu4 ~ n t e r -
has recently become an important technique for ob- ,tct~onsmvolving ,I chromopho~icgroup m,ly bc jtudied
taining structural information on biological systems. The In I'lrgc molecules such ,I\ protcmc For cxample in
ability to use small sample volumes and aqueous so- hemoglobm, ~t is po\\~bleto sclcctively cnhmce vibra-
lutions is a major factor. In acldition, tlic ease with t i o n ~dcwc~atcdw ~ t hthe hcmc group (chroniophorc)
which information can be obtained o n samples such as by appropr~atec h o m of e x c ~ t ~ r t ~trequcncy
on
lipids and membranes has also contributed to the in-
creasing use of Raman techniques. Raman mcasure- Surface-Enhanced Raman Scattering. Con-
ments arc also quite sensitive to conformational changes, ventmnal R'~m,tnscattering m d rewnmce K,nnan tcch-
which arc of great importance in biological systenis. nlquec have been applied to ctudres 01 x i w r p t ~ o non
Since many molecules of biological interest are also surfaces Surlacc-enhmccd 1L1rnm \c<~tter~ng (SEKS)
tluorcsccnt, a major problem has hccn to separate tlie 1s '1 recent tcchn~cluethat allow\ very s n ~ t l mwunts
l of
Kanian scattering from the fluorescencc background. ,idcorhC~to to he invest~g,ltecl Or~g~nally, 'In cnhance-
Conventional Raman spectrornctry has bccn used rncnt ot == 10'' w,rs tound 111 the R,~in,in spectrum of
to monitor conformational changes of peptidcs and pro- pyr~dlneo n ,I s~lverelectrode, 'tfter ,In electrochcrn~c,~l
teins in aqueous solutions. Nucleic acids and polynu- rcdox cyclc had occurred. h h m c c m e n t s wcre Lrter
clcotides have also been studied. For cxamplc, the ion- ob\erved for ,i v,~riety01 ad\(~-b,rtc\on 4evcral metJlic
ization equilibria of the bases and ol the phosphate sul t x e \
groups in nuclcic acids have been invcstigatccl with the M m y e x p a ~ m e n t a det,lil\
l ot the SkRS phenon-
aid of Raman spectrometry. The secondary structure cnon lic~vebeen elucidalcd ,tnd \cve~,rl'ittempt\ h'ivc
of RNA has been studied, and the binding of nictal ions been n ~ d to c develop ,t theor etlcd model to expl'nn
to various sites on nuclcic acids has been monitored. the enhancement One rnodel 1nvolve4 the ~ o u p l ~ notg
Raman spectrometry has been used to measure the rates a molecular d~polew ~ t h1t4 ilndgc In thc mctd, wh~ch
of hydrolysis of several nucleic acids. l e d \ to an ~ n c r e ~ l wpolar~z,rhil~ty.
d Other model\ h'lvc
Because of its poor detection limits, convcntional ~nvolvcda rewnmcc R m i m ettcct ot w m c hmd St111
Kaman spcctromctry is of limited utility i n studying o t h e ~ sIiwc ~nvokedthe roughncsc of the surface At
Sec. 16-2 1 Raman Spectroscopy 511
present the theoretical explanation for the SERS effect urement practical. The nonlinear Raman techniques de-
is incomplete. However, SERS is being applied to sev- scribed below have begun to open up many new chemical
eral problems involving surface adsorption and catal- applications including several in analytical chemistry.
ysis. Energy-level diagrams illustrating the nonlinear
Raman processes are shown in Figure 16-15. The or-
Nonlinear Raman Scattering dinary or spontaneous Raman process was illustrated
in Figure 16-5. The techniques considered here are
Ordinary or spontaneous Raman scattering is a linear stimulated Raman scattering (SRS). the hyper-Raman
process that originates from induced sample polariza- effect, inverse Raman scattering (IRS), stimulated Ra-
tion P that is dependent on the first power of the electric man gain (SRG), coherent anti-Stokes Raman spec-
field strength of the incident beam. Nonlinear Raman troscopy (CARS), and coherent Stokes Raman spec-
effects can be observed at high laser irradiances where troscopy (CSRS, pronounced scissors).
the electric field strength exceeds about 10" m-'.
These effects depend on the polarization induced by Stimulated Raman Scattering. Stimulated Ra-
second- and higher-order terms in the electric field man scattering is a nonlinear technique in which the
strength. Nonlinear Raman effects are usually treated spontaneous Raman emission is amplified by factors as
theoretically by introducing the phenomenological non- large as 1013.The energy-level diagram for SRS is iden-
linear susceptibilities as was done in Section 4-3 when tical to that for ordinary Raman scattering, as shown
treating nonlinear optics. When higher-order terms are in Figure 16-1Sa. In SRS, however, the laser irradiance
included, the polarization can be described by equation is such that a high population is created in the virtual
4-20. Although nonlinear effects were observed in the state. Interaction with the laser field causes stimulated
early days of laser-Raman spectrometry, the introduc- Raman scattering. The scattered beam is coherent and
tion of the dye laser was necessary to make their meas- coincident with the direction of the incident beam.
can write the steps involved as "reactions": available, the nonlinear Karnan techniques, particularly
CARS, are certain to become more widely used.
v, = v, +v, incident pump photon
v, = v, - v,, incident probe photon
v, + v 2 = v, sum frequency Turbidimetry and nephelometry are methods that meas-
ure the concentrations of particulate matter in a sus-
v, - v, = v, difference frequency
pension. Both are based on the elastic scattering of
radiation. Turbidimetric methods measure the decrease
Note that the overall reaction is 2v, = v, + v, and the that occurs in the transmitted radiation as a result of
new CARS photon appears at v, = 2v, - v,. particle scattering, while nephelometric methods meas-
We are now prepared to understand some of the ure the radiant power of the scattered radiation itself.
features of GARS. Since two photons at v, and one Turbidimetric methods are preferred when the
photon at v, are required to produce the CARS photon concentration of suspended particles is high. Here it is
at v,, the CARS signal should vary with the square of simple to detect changes in the transmitted radiant power,
the pump laser power and linearly with the probe laser whereas when the amount of scattering is high, nephe-
power. The CARS signal should also vary with the square lometry suffers from interferences and nonlinearities.
of the sample concentration. This is because the number On the other hand, when the concentration of scatterers
of vibrations created in the first step will depend linearly is low and the radiant power of scattering is small, ne-
on concentration. Similarly, the number of vibrations phelometric methods are preferred.
mixing with the pump laser will depend linearly on con- The scattering of radiation by suspended particles
centration, giving the overall process a square depend- is often called the Tyndall effect. In the UV and visible
ence. Even in the absence of the resonance condition, regions the scattering particles are usually of colloidal
photons are generated at v,. Thus there is a continuous size, from 1 nm to 1 pm in diameter. The scattering
background level associated with CARS; the signal is can be Rayleigh. Debye, or Mie, depending on the size
greatly enhanced when resonance occurs. of the particles.
The CSRS technique involves a process similar to We consider here briefly the instrumentation, the-
CARS, as can be seen in Figure 16-15f. Here Stokes ory, and a few applications of turbidimetry and nephe-
radiation is generated by the mixing of a pump fre- lometry.
quency with an anti-Stokes-shifted probe frequency. The
CSRS output occurs at v, = 2v, - v,. CARS is by far Instrumentation and Theory
the most popular of the two techniques primarily. be-
cause the frequency generated is in the anti-Stokes re- The instrumentation used in turbidimetry and nephe-
gion, where it can be spectrally distinguished from flu- lometry is normally quite simple, as illustrated in Figure
orescence. 16-16. In both methods an incident beam passes through
Like IRS, CARS is a coherent process. The co- the sample cell and strikes a radiation transducer. An
herence, directionality, and spectral nature of CARS interference filter is often used, either before or after
allow the CARS output to be readily separated from the sample cell, so that the detected beam is in the blue
incoherent background such as fluorescence emission. region of the spectrum, where scattering intensity is
The major limitation to CARS is the nonresonant back- high. In turbidimetry, the transducer is placed at an
ground mentioned above. In conventional CARS this angle of 0"with respect lo the incident beam, whereas
background has limited the limits of detection to the in nephelometry a 90" geometry is used.
1% level in solution. Resonance enhancement has been Turbidimetric measurements are often made with
used to improve the detection limits (DLs) in CARS. simple filter photometers or modifications of these.
When the pump frequency approaches the frequency Nephelometric measurements. on the other hand, can
of an allowed electronic transition, a greatly enhanced be made with fluorescence instruments. In some cases
CARS output is obtained. With resonance enhance- more elegant light-scattering photometers, such as those
ment, DLs of M have been reported. described in Section 16-4, are employed. These can
At present CARS is still a new and relatively ex- detect the scattered radiation as a function of angle.
pensive technique. It has been shown to be useful in The sample cells are often rectangular absorption (tur-
combustion diagnostics, in studies of strongly fluores- bidimetry) or fluorescence (nephelometry) cells.
cent biological compounds, and in HPLC detection. As When an incident beam of radiation is scattered
less expensive and more routinely useful lasers become by suspended particles, the radiant power appearing at
Chap. 16 I Molecular Scattering Methods
any angle is a function of the concentration of particles, p r e p i ~ ea c,il~br~rt~on curve of - log(4),/(D,)v\ c, where
their size and shape, their rel'ractive indices relative to (I), rs the rad~antpower tr,ln\mittcd w ~ t hthe snmple
the medium, and the wavelength of the incident ratlia- (\tantl,~rd)In the cell and (I), 1s that wlth the rcterence
tion. Although the comprehensive Mie theory of ra- (usudly the \olvent) In the cell The ~c\ultlngwolking
diation scattering is applicable to turbidimetry and curve is then ~ s c dto o b t a ~ nthe conccntr;ltlon\ ol un-
ncpl-lclometry, it is seldom applied. Instead, thesc tech- known\ There 15, ol courje, a 1nutu~11 ~ n t c ~ f c r e n cbe-
e
niques arc usually based on empirical relationships bc- tween ordmiry ah\orption 'ind turbdiinetiy Scnttenng
tween the scattered or transmitted radiant powcr and from turbrd \ o l u t ~ o n \c,lu\es (tn mterlercnce In n b \ o ~ p -
the conccnlrations of the particles. tlon me,l\i~rements, while h o r b ~ n g\peclcs ~ntertere
Let us consiclcr a parallel, monochromatic, inci- in tur b i d l r n e t ~ ~mcnwrcmcnt\
c
dent beam of radiant power (I,,, passing through a SO- In nephcloinetry, the sc'ittercd r d l a n t power [I),,
lulion containing suspended particles. The attenuation I\ ernp~r~c~llly related to the c o n c e n t r ~ t ~ oofn \uspended
by scattering in a dilute suspension Sdlows the rela- part~clesby the cquntlon
tionship
= a),,k,, c (16-16)
perature, concentration of reagents. order of mixing, instruments, called light-scattering photometers, are
ionic strength, and time before measurement must be available for such measurements. Many of these are
carefully reproduced to obtain precise and accurate re- similar in design to the photometer shown in Figure
sults. 16-17.
In recent years laser sources have begun to replace
Applications conventional sources in such photometers. There are
several reasons why CW lasers are nearly ideal for ra-
Turbidimetric or nephelometric measurements are often diation scattering measurements. First, the spatial co-
used to estimate the amount of suspended matter in herence. directionality, and low divergence of the laser
liquids. The determination of the clarity of water is one beam reduce some of the optical constraints placed on
example. These methods are also applied to determine instruments with conventional sources. This permits
the effectiveness of water treatment processes. scattering to be measured at very low angles. Second,
The concentrations of various species in solution the laser allows a high output power to be transmitted
can also be determined by turbidimetry or nephelom- in a beam of low cross-sectional area: this permits meas-
etry. Here precipitating agents are added to the urements to be made on small sample volumes. The
sample to form the suspended particles. Often a surface- outputs of many laser beams are polarized, which is
active agent, such as gelatin, is added to prevent co- used to advantage in many radiation scattering meas-
agulation beyond a certain size. urements.
A few of the species that have been determined In this section we first discuss the principles of
by turbidimetry or nephelometry are given in Table 16- molecular weight determinations by scattering meth-
3. The most widely used method is that employed for ods. Then, low-angle laser light-scattering techniques
the determination of sulfate. With nephelometry, sul- are described. These methods are not only applied to
fate concentrations as low as a few kg mL-' have been determine the molecular weights of isolated materials,
obtained with relative standard deviations on the order but they are also used as molecular weight-sensitive
of 5 % . detectors for size-exclusion chromatography. Infor-
Turbidimetric methods have also been applied to mation regarding molecular geometry is also provided.
determine endpoints in precipitation titrations. With Laser sources are also used in Brillouin scattering meas-
titrations. the instrument employed can be as simple as urements and in quasi-elastic (dynamic) light-scattering
a white light source and a photocell placed on opposite methods. The information obtainable from these tech-
sides of the titration vessel with an appropriate readout niques is briefly discussed in this section.
device. Turbidimetric detection has been employed for
titrations of fluoride with calcium, of bromide with sil- Molecular Weight Determinations
ver. and of sulfate with barium.
To see how radiation scattering can be applied to mo-
lecular weight determinations, it is useful to consider
16-4 LASER SCATTERING METHODS again the equation that relates the intensity of Rayleigh
scattered radiation to polarizability and scattering angle
Radiation scattering methods have been widely used (equation 16-1). Let us consider the sample to be a
for many years in the determination of molecular weights, collection of isotropic polarizable particles (e.g.. mol-
particles sizes, and particles shapes. Several commercial ecules). The polarizability a' is directly proportional to
TABLE 76-3
Turbidimetric and nephelometric methods
Ag AgCl NaCl T, N
As As KHZPO, T
Au Au SnClz T
Ca CaC20, H?CZO, T
CI - ~ g c l AgNO, T. N
K K,NaCo(NO,), NajCo(NOz), T
SO,'- BaSO, BaC1, T. N
Se Se SnCl? T
Te Te NaH,PO, T
aT, Turb~dimetr!, N , nephelometr~
Chap. 76 I Molecular Scattering Methods
GURE 16-17 L ~ g h t - s c a t t e r m g
photomctcr. In (a), collimated sadla-
tlon from a mercury source and in-
terference filter is polarized and passes
through the sample cell. 'I'hc scat-
tered radiation passes through a sec-
ond p l a r i z c r and is detected by a
p h o t o r n ~ ~ l t i p l i ctube
r (PMT). 'l'hc
PMT is mounted on a turntable, which
can be positioned to receive scattcrcd
radiation at any angle from 0" to 180".
l'hc semioctagonal cell (b) has faces
that allow measurements at 4S0,90•‹,
and 135" to the incident beam. Often
the walls not used by the incident or
detected beams are blackened to ab-
sorb radiation. The polarizers allow
the use of polarized light or the meas-
urement of dcpolarization ratios, as
discussed in the text.
molecular weight [a'=- Mq,,jdq/dc)/(2nN)], whele M ratio K,, is defined by the cquation
1s the molccu1,tr we~ght,N I \ Avogadlo's number, rlois
the ~elractivemdex ot the pure solvent, m d ciqldc i \
the rchactive mdex chmge ot the solution with con-
ccntrat~on Hence the Intcnslty of RCryleighscattering
from a m g l e p,irtrcic I \ proportlon'il to the square of
the molecul,~rwe~ght
bxperimentally, one d o o not dctcrm~ncthe scat-
t c m g f ~ o m,i single part~clc,ol course, but r,ithcr the This is often writtcn simply as
sc,ittering per unit volumc Fol 1' sxnplc of conccnlr,l-
tlon ( (g empi) with NclM pdrlt~clcsper imlt volume, R, - KcM ( I 6-19)
the scattered ~adiatlonlntcncrty per unrt volumc is ob-
tamed from equation 16-1 by substltutmg the value of where K = 2 ~ r ~ r ~ ~ ( U ' q / d ~with
)~(N ~ ) cm2 g *
theX units
a' given rn the prcvlous paragraph and rnultiplymg by ~nol.Actually, it is something of a misnomer to call K ,
the number ot part~clcspel unit volurnc, NtlM. For a ratio since it has units of cm ', but this is standard
unpolarized incldcnt r a d ~ a t ~ othe n , scattered irradiance practice.
pel u n ~ volumc,
t ( E , , ) ,can be wrlttcn In theory, then, measurcments of the Rayleigh
ratio should allow determination of the nlolecular weight
M. In practice, however, the equations above hold only
for pure Kayleigh scattering (i.e., small particles) and
for ideal solu'tions. For nonideal solutions, local fluc-
tuations in concentration must be taken into account.
This is usually accomplished by a virial expansion in-
Note again that B is taken as 0" fol torward c'rttering volving virial coefficients A, and A,. In dilute solutions
and 180" for scattermg back toward the wurce. only the second virial coefficient A, is used. Adding
Sec. 16-4 1 Laser Scattering Methods 517
I{ Y
="-
T
'I S,,12h
Soon 'itlcr the I,rsc~ was developed, ~t w,lr wldcly rec- where \,, I t the photornultrpl~er~caclout\ignd f o ~tlic
ogni7cd to be ,I u,elul source In light-,c<ittcrmg Inves- sc,rttcrcd r c i t l ~ c ~ t ~.\,,
on 1s,the PMT q n d for the ~ n c ~ d c n l
tlg'lt~ons HOWCVCI, 111 e,~rly~n,trumcnt\ the convcn- r , ~ d ~ , ~ t ~I o1snthe
, tr,rn~rl~~tt,rncc ot the lnudent ,ittell-
tlond Inel cur y source w,t\ repl,~ccclw ~ t h'I He-Nc 1,1tc1 u'ltor, 11 14 thc \ohd angle through wli~chthe \cdtcrcd
mct the unrque characterist~csol the 1,ner r,idutlon were ~ a d ~ , t t i o14ncollected, m d h thc length ot the x a t t c r ~ n g
not eniploycd In the mid-1970t 1,lser I ~ g h t - x i ~ t t c r ~ n g volume p'lr,rllel to the ~ n c ~ d e n b et a n . Note that IE ,tnd
photomctcls were developed w ~ t ho p t ~ c d\ystc~n\de- h ale gcomctilc,rl constmt\ th,rt can be ~ndcpcnrlently
s~gnecla-ound the l,l,cr obt,uncd Also, r c'in be m e ~ \ u r c doptlc,~lly Hence
l c I ~ g h t - s c ' ~ t t c ~( E
The 11rtt l o ~ - ~ u n gInser ~ nAgL L S ) d b s o l ~ ~v,tlue\
tc ot R,, a e obtd~ncdw ~ t h o st,rntl,~rcl\
~~t
photonictc~.developed by C h r o m , ~ t ~ xInc , . w~14dc- Although the LALLS method g~c,itly ~ n i p l r l ~ c ~
t~gneclto take nclvmt,~geot the exccllcnt ~pCrt~,rl co- the dcterni~n,ltionof molecul,~rwelgIit4, u e mcl ,h,ipe
herence of L r w 1,ldlatlon 'rnd to p c ~ r m tsc'lttcrlng rn- informit~on,ir c lo\t 1hew ~ l er only o1~t~1maI~le t l i ~O L I ~ ~
ten4rtres to be mc~isurcd,it <rnglc\ below 2" A\ \hewn mc,isurements ol scattering vcl\u\ L~rlgle dncl conccn-
111 Frgurc 1(>-19, ,I low output p o w c ~(3 niW) I lc-Ne tration
I ' i w 14 utcd ,is the source The small sc,itterrng volume I'he LALLS method h,l\ ,rlw bccn ,~ppl~ccl to w e -
~eclucctthe p r o l ~ b ~ l ot ~ t y'I lorclgn p ~ r t ~ c suchl c a\ ,i exclusion chrorn,ltogr,lphy A c o ~ n m c ~ cLAY,LS ~ , ~ l tie-
dust p ' i ~ t ~ c l1~c1ng
e plewnt w ~ h the n ~~r,idr,~tccl reglon tcctor 14 clva~lnblefrom C'hroni,~t~.i., Inc The detector
m d I \ ,ldv~nt,rgcouswhen thc 'mount of nv~ul,blema- is u\~l~rlly u\cd 111 4er1es wrtli ,I conventional, concen-
terul 1 4 Imrtecl I~rntli,rtronw ~ t hthe He-Nc I;lscr (613 trcltlon-scnt~t~ve cletecto~,such 1' s a d~tfercntlal~etr,lc-
Sample Field
I Lcns ston
ilensi ly /'
f~ltcr~ Annulus
the ~ n c ~ d c nbenm
t 1s pcrmtted to pass t h ~ o u g hthe smlple to the PMT
B c c a u x the incident iri-acli;mce is on the ordcr of 10" timcs the irrdiance
of t l ; ~scattered beam, a series of noutral density filters i s inserted during
the mcasuremcnt o f the signal .S,, (see equation 16-23).
Sec. 16-4 1 Laser Scattering Methods 519
tive index detector. The concentration detector re- volved (- 5 x 1014 Hz with a He-Ne laser) to fre-
sponse is used to evaluate the concentration dependence quencies near 0 Hz that can be readily measured. In
of the LALLS signal. A typical application of this sys- most QLS photometers, a photomultiplier tube (PMT)
tem is to determine the molecular weight distribution is used as a nonlinear mixer since its output is propor-
of a polymer sample. tional to the square of the electric field falling on its
photosensitive surface.
Quasi-Elastic Light Scattering Let us consider the QLS photometer illustrated
in Figure 16-20. The sample is irradiated with laser
The determination of particle sizes has become increas- radiation. The scattered radiation at some angle 0 is
ingly important in recent years. One of the promising incident on a PMT. The PMT output can be processed
new techniques for particle-size analysis in solution is by photon counting techniques as shown or as a pho-
quasi-elastic light scattering (QLS). also known as dy- tocurrent. Photon counting is preferred because of its
namic light scattering and photon correlation spectrom- digital processing compatibility. Particle-size informa-
etry. The QLS technique can obtain particle-size infor- tion is obtained from an autocorrelation analysis of the
mation in a few minutes with particles ranging from less processed signal as described below.
than 50 A to about 2 pm in diameter. In order to see how the PMT acts as a mixer, let
The QLS technique involves the measurement of us assume that the scattered radiation contains two fre-
the Doppler broadening of the Rayleigh-scattered light quencies w, and w,. The electric field E can be written
as a result of Brownian motion (translational diffusion) as
of the particle. The name quasi-elastic distinguishes the
technique from the elastic light-scattering techniques E = El sin w,t + E, sin w,t (16-24)
discussed previously and from inelastic light-scattering
methods, such as Raman spectrometry. The PMT output signal S(o) is proportional to the square
In quasi-elastic light scattering the thermal motion of the electric field and can be written
of the particles gives rise to time fluctuations in the
scattering intensity and a broadening of the Rayleigh
line. The Rayleigh line has a Lorentzian shape. In ma-
cromolecular solutions, concentration fluctuations are + E,E, [cos (w, - w,)t - cos (w, + w,)t])
usually dominant. Under these conditions the width of
the central line is directly proportional to the transla-
tional diffusion coefficient D,. Conventional spectrom-
eters and even interferometers cannot be used to meas- where A is a proportionality constant. The PMT cannot
ure the small line widths (1 Hz to 1 MHz). respond directly to frequencies w,, w,, or the sum term
To measure the Doppler widths. the QLS pho- since these are greater than 1014 Hz for visible laser
tometer uses optical mixing or light-beating techniques. sources. The PMT can respond, however, to the dif-
These techniques translate the optical frequencies in- ference frequency term (w, - w,) which can be as small
Sample
cell Lens
n Laser
Rayleigh
line \
Anti-Stokes
\
where v, is the frequency of the incident radiation, v, nents become broader, and their position shifts toward
is the velocity of sound in the medium, c is the speed the central line. Brillouin scattering is useful for study-
of light, 7 is the refractive index of the medium, and 0 ing the behavior of liquids near the critical temperature.
is the scattering angle. Equation 16-27 predicts that When the temperature of the liquid approaches the
there should be Brillouin lines appearing on either side critical temperature, the intensity of the central line
of the unshifted Rayleigh line. The frequency shifts are increases rapidly, while the Brillouin components get
generally on the order of 0.05 A. Because of the small much weaker and disappear in the background. The
frequency shifts and low intensities, Brillouin scattering width of the central component is used to measure the
was not observed experimentally until the 1930s. Figure thermal conductivity near the critical temperature. Bril-
16-22 shows Brillouin spectra of CCl, taken at various louin scattering has also been noted in viscous liquids
scattering angles. and has provided experimental evidence for the solid-
The frequency shifts observed in Brillouin scat- like behavior of such liquids toward hypersonic waves.
tering require high-resolution spectrometers to meas- In addition, hypersonic velocities have been obtained
ure. High-dispersion grating spectrometers were once
used, but in recent years the scanning Fabry-Perot in-
terferometer has become the wavelength-selection de-
vice of choice. A typical setup for observing Brillouin
scattering is shown in Figure 16-23.
The frequency shifts of the Brillouin lines have
been used to obtain hypersonic velocities [the frequen-
cies of sound waves are often classified as hypersonic
(>500 MHz), ultrasonic (20 kHz to 500 MHz), audible
(20 Hz to 20 kHz), and infrasonic (<20 Hz)] in many
7' Lens
To vacuum
pump
A
bbl PMT
o n amorphous and crystalline solicl5 with Rrillouin detection limits (DLs), they suffer from quenching cf-
scattering. fects with thc more abundant atmospheric species. The
atomic fluorescence (AF) method is best suited for
monitoring trace constituents, such as sodium, in the
uppcr atmosphere. Molecular tluorcsccncc is also highly
Laser-based techniques are being rapidly developed for uscful. It finds major applications for dctccting oil spills
rernotc-sensing applications. I x x r s have been uscd to and monitoring water cluality from the air.
detcr~ninethe optical characteristics of the atmosphere, Because the cross section for Mie scattcring is
to probe the upper atmosphere, and to monitor nt- much largcr than that for Kaylcigh scattering (ciuc to
mospheric pollutants. I n addition, laser sources have the much larger particles detected in Mie scattering),
been mounted in aircraft and pointed downward to study Mie scattering often dominates in the atmosphere. 'I'hus
the hydrosphere. Many of the laser-based methods uscd quite low concentrations of atmospheric particles can
in such studies are based on radiation scattcring. In this bc detected with Mic scattering. Rarnan scattering is
section we consider briefly some of the techniques that also employed in atmospheric studies when molec~rlar
have proven most useful in remote sensing. Many of specificity is desired. Raman back-scattering techniques
these are called laser radar or lidar methods. The prin- are highly useful in air pollution studies, because most
ciples of lidar systems are first introduced and thcn a of the molecules of interest arc Rarnan active. Although
few of the most important applicaiions are discussed. the cross sections are small for Raman scattcring, the
resonance Raman technique can he employed to im-
Laser Radar prove detection limits. Rarnan scattering is highly useful
for remote monitoring of effluent plumes from indus-
Following the introduction of lasers in the 1960s, it was trial operations. Here the concentrations of the pollu-
soon realized that these sources allow optical rneasure- tants of interest can be in the tens to hundreds of
rnents to be made at sites remote from the probed area. kg m L p l .
Indeed, studies of the atmosphere with lascr sources Differential absorption lidar or DIAL can achieve
date from 1963 when the first laser radar system was low limits of detection and excellent spatial resolution.
developed to study aerosol distributions. In the years In the DIAL technique, two lascr beams arc used, and
that followed, enormous aclvances have been made in the differential attenuation is obtained from the back-
the develop~nentof lidnr systems (lidur stands for light scattered signals. One lascr is tuned to a molecular
detection und ranging). transition while the other is slightly detuned. The ratio
The term "lidar" is actually a generic term that of the two signals gives the differential absorption. The
docs not imply a particular detection process. For ex- large Mie scattering cross section is used to advantage
ample, lidar systems can use Rayleigh scattcring, Mie in this technique to provide both spatial resolution and
scattcring, Raman scattering, fluorescence, and differ- a strong return beam. 'I'he DIAL technique is most
ential absorption. Typically. such systems are con- advantageous for long-range monitoring of specific mo-
structed inside astronomical d o n e s or placed in shelters lecular constituents.
on the tops of buildings. They have also been placed
in mobile vans and in aircraft. It is anticipated that the
Space Shuttlc or a space station will bc widely used for
global monitoring with lidar in the future.
A typical large-scale lidar system uses a pulsed Although the use of lasers in remote sensing is still in
laser source and a telescope receivcr at the same lo- its infancy, major advances in o ~ r rknowledge havc al-
cation. The laser transmitter and the telescope receiver ready been made. For example, it is now common to
can be collinear or offset. In the ofl'set geometry the obtain information o n the particulate and aerosol con-
laser beam enters the field of view of the tclcscope tent of the atmosphere using lidar. Particle sizcs can
optics only beyond some preestablished distance. The readily be determined from Mic scattcring. Multiwavc-
offset geometry avoids detector saturation from the near- length lidar has been employed to obtain aerosol s i x
field back-scattered radiation. With collinear gcorne- distributions.
tries near-field back-scatter is overcome hy either gating Atomic fluorescence measurements havc shown
the PMT detector or by using a fast optical shutter. that there is a "sodium layer" in the uppcr atmosphcre.
The various scattering ~ncthocisand diflcrential Several groups have investigated the spatial and t e n -
absorption arc most widely uscd for ground-hascd re- poral behavior of this layer during metcor showers. For
mote sensing. Although fluorescence methods havc low example, a fourfold increase in the sodium concentra-
Chap. 76 I Problems 523
tion was observed during one shower. It was concluded Laser-induced fluorescence has been used for a
that meteor ablation was probably the source of sodium. variety of high-altitude environmental studies. Flu-
Raman back-scattering measurements have been orescent dyes have been dissolved in water and used to
used to measure the oxygenlnitrogen balance in the study water movement. Oil pipelines have been mon-
atmosphere. Similarly, measurements of the CO,IN, itored for leakage and oil spills have been detected and
ratio and long-terms drifts in this and the O,/N, ratio classified. In some remote studies of water quality. the
are being made. Spatial profiles of nitrogen in the at- Raman band of water has been used as a built-in ref-
mosphere have been obtained with Raman back-scat- erence for fluorescence measurements to compensate
tering and the temperature profile calculated from the for laser fluctuations and for changes in some of the
molecular concentration profile and the pressure. The variables inherent in quantitative fluorescence meas-
DIAL technique, with the multiwavelength CO, laser, urements.
has been used to measure the vertical profiles of at- From this brief discussion it is apparent that re-
mospheric ozone and atmospheric water vapor. mote sensing is a rapidly growing field. Improvements
Differential absorption and Raman back-scatter- in laser and detector technology, particularly with in-
ing are being increasingly used in pollution studies. For frared detectors. are making atmospheric monitoring
example, the DIAL technique was used to obtain the studies routine. Already laser remote sensors have helped
KO, distribution over a chemical factory: the Raman make major contributions to our understanding of the
technique was used to measure S O , in the plume of a atmosphere and our immediate environment.
power plant.
PROBLEMS
A molecule has a Raman shift of 859 cm-' when length were changed to 365.0 nm? Assume purely
excited by an argon ion laser at 514.5 nm. What are Rayleigh scattering.
the wavelengths in nm of the Stokes and anti-Stokes For isotropic scattering the Raleigh ratio and the tur-
Raman lines? Which of these is more intense, and bidity are related by R, = 3 ~ ( 1+ cos2 0)/167i. For
why? the solution described in problem; 16-6, find the Ray-
A tunable dye laser has approximately the same ir- leigh ratio at a 60' angle of observation.
radiance at 400 nm as at 500 nm. It is used in a The absorbance of a solution is measured and found
Rayleigh scattering experiment. Compare the irra- to be 0.18. Upon removing the 1.00-cm cuvette from
diance of Rayleigh scattering at 90" at the two laser the spectrophotometer. the solution was found to be
wavelengths. slightly turbid. The turbidity was then measured and
A typical Raman spectrum of an organic compound found to be 0.014. What was the true absorbance of
covers Raman shifts ranging from 0 to 3500 cm-l. the solution?
For excitation by an argon ion laser at 488.0 nm. what Contrast normal IR spectrometry and Raman spec-
is the equivalent wavelength range? Recalculate for trometry with respect to applications, factors limiting
the 514.5-nm argon ion laser line. What types of pho- the SIN, and instrumentation.
tomultiplier tubes would be most suitable for use with A Raman shifter with H2 gas was used to produce
these two sources to acquire Raman spectra (see Fig- new wavelengths from a fixed wavelength KrF laser.
ure 4-23). The output wavelength of the excimer laser is 249 nm
Assume that the ratio of anti-Stokes to Stokes Raman and the fundamental vibrational frequency of HZis
intensities is governed entirely by the Boltzmann dis- 4400 cm-I. Find the wavelengths of the five Stokes
tribution. For temperatures of 25•‹C and 35•‹C. find and the two anti-Stokes lines produced at the output
the ratio of intensities for the CC1, lines at 218 cm-I of the Raman shifter.
and 790 cm-I. In a CARS experiment, a laser pump beam at 500
From the Stokes Br~lloulncomponent of CC1, ~51th nm was incident on a cell containing COZ gas. The
l v = 1887 MHz (see F ~ g u r e16-22). find the hyper- symmetric stretch of C 0 2 is Raman active with a
sonic veloclty z*, if = 1 46 vibrational frequency of 1340 cm-I. What should be
Calculate the percentage decrease in radiant power the approximate wavelength of the probe beam? At
when a 546.1-nm radiation beam passes through a 3- what wavelength will the CARS output appear?
cm-thick layer of a polymer solution with a turbidity In a low-angle light-scattering experiment to deter-
T of 0.01 cm - I. What would the percentage decrease mine the molecular weight of a polymer. values of
in radiant power be for the same solution if the wave- the Rayleigh ratio were obtained at various concen-
Chap. 16 / Molecular Scattering Methods
Thc references txlow arc general references o n light scattcr- M. D Lcvcnson, Iiliroduct~orl/o N o r ~ l ~ n c Ltrtrrw Sprc -
ing, including cluasiclastic and low-angle laser light scattering. troscopy, Academ~cb e \ , , New York, 1982 Include\ an
M. Kerker, 7'hc Scurtcring of Light unri Other Eleclro- cxccllcnt chapter o n nonhnenr Rnmm techn1clue5
nlugnetic Rudi~rtion,Academic Press, New York. 1969. G.L. Eescly, C'ohrrcrit Reurluur .Yp.iroscoyv, I'crgarnon
A classic book on light scattering. Press, Elmsfo~-d,N.Y.. 1981. A very good discussion of
B. J . I3crnc and R . Pecora, Dynainic Light Scuttcrirlg nonlinear Icaman methods.
with Applicatioizs to ('hernktty, Biology and Phjsic:~, B. S. Hudson, "Coherent Anti-Stokes Jiamm Scattering
Wiley-lntcrscicnce, New York. 1976. An outstanding Spectroscopy ," in Mrw Applic~ltiotlsof' I.LI.SC~,S1 0 C'11c~ir~-
book on dynamic light scattering. istry, G.M . I licftje. etl., American C'hcmical Society,
G. Ostcr, "Light Scattering," in 'I'cchniqcrc~sof C'hcw- Washington, U.C., 1978.
istry, vol. I , pt. IIIA, A. Wcisshcrgcr and R . W. Ros- J. J . Valentini, "Cohel-ent Anti-Stokes Raman Spcc-
sitel-. e&., Wiley-Interscience, New York, 1972. troscopy," in Speclromrtric Tc,clrrliquc~.s,vol. 1V, G . A .
B. Chu. Lu.sc,r Light Sccrfterirlg, Academic Press, New Vanasse, ed., Academic Press, New York. 198.5.
York, 1974. Highly mathematical. hut a very useful book. I:. S. Yeung, "Spectroscopy by Inverse Raman Scatter-
Contairis a wealth of information. ing," in New App1icntiorl.s of Lasers to Clrrrni.stry, G. M.
N. C . Ford, "'l'hcory and Practice of Correlation Spcc- I lieftje. ed., American <:hemical Society. Washington,
troscopy," in Mc~a.surcrncnt.sof Si~sprntlrctI'articles hy U.C., 1978.
Quasi-~lustic.Light Scallcrirrg, R . ti. Dahneke, ed., Wilcy, W. F I . W o o d r ~ ~ and
f t ' S. Farclui~xson,'"l'irne-Resolved
New York, 1983. I<csonance Raman Spectroscopy (I'R')and Related Vi-
dicon Raman Spectrography: Vibration Spectra in Na-
M. L. McConncll, "Particle Size Uetcrmination by
Quasielastic Light Scattering," A n d . Chml., 5.1, 1007A noseconds." in Ncw App1ic~utior1.s of'Luwrs to C,'hcrrlistr~,
(1981). G . M. Ilieftje, ed., American Chemical Society, Wash-
ington, D.C., 1978.
A . T . Young. "Rayleigh Scattering," Phys. Today. 42
(Jan. 1982). J . C. Wright, "Nonlinear Optics,"iin 1~u.srr.sin C'hc~rrricul
Analy.si.s, G.M. Hieftjc, J. C . Travis. arid F. E. Lytle,
W. Kaye, "Low-Angle L'tw L g h t Scattcrmg," Arlul
cds., 1 lumana Press, Clifton. N . J . . 1981. Includes a good
Chcnl., 45, 221A (1973).
discussion of nonlinear Iiamini ~nethocts.
The following are references that deal specifically with Kaman A . Anderson, ed.. Thc. Rutnun I</;li,c/, vol. 1 ., Marcel
spectrometry, including in many cases resonance Rarnan and Dekkcr, New York, 1971. Includes a chapter on Bril-
nonlinear Iiaman methods. louin scattering and onc on the stimulated liiiman effect.
D. A . L m g , R a m m Sprcrro.sc~opy,McGraw-Hill, New M. D. Morris :und D. J. Wallan. "Rcson;~ncc 13am:un
York, 1977. A modern and useful tcxt on Raman spec- Spectroscopy---C'LI~I-~II~ Applications and Prospects."
troscopy. Inclucles theory. instrumentation. and appli- Arrul. C'hc~m.,51, 182.4 (1979).
cations. S. A . B o r m m , "Nonlinear R a m m Spectroscopy,'' Arlnl.
J. C'. <;rassclli. M. K. Snavely, and B. .I. Hulkin, C'hc~nz- Ch(v~z..54. 102 1 A ( 1082). A very good introductory
icul Applicatiorls of Rnrnrrrl Sprctroscopy , Wilcy , New review of the various noinlinear technique\.
York, 1981. A small and quite readable hook on appli- B. S. Hudson, "l<csonance Raman in the Far-Ultravi-
cations of Raman spectrometry t o inorganic, organic. olet Region." Sprc/roscapy, l ( 1 ) (1980).
and biological systcms. I'. I lirschfield and 13. Chase, "FI-fiarnan Spectroscopy:
N. R . Colthup, I > . H. Daly, and S. E. Wihcrlcy, Intro- Development and Justification," Appl. Spc~trose..,411,
duction to Iu~/kurcrlunrl Rrnrrlun ,Sprc.tro.w)p~2nd cd. , 133 (1980).
Academic Press. New York, 1974. Incl~ldessurvey o f B. Wopenka and J. I>. I h t c r i s , "Limitations to Ouam-
theory, instrumentation. and ;~pplications. titative Analysis of I;luicl Inclusions in (ieological Sam-
M. C. Tohin, Laser Rulnutl Sp(,ctro.sc.opy. Wilcy-lnter- ples by l,;lscr liaman Microprohe Spectroscopy," Appl.
science, New Yol-k. I97l. S p c ( ~ f r m c 40,
. . 144 (1986).
There are a number of spectrochemical techniques that photoacoustic and thermal lensing spectrometry which
do not readily fit into the usual emission, absorption, are related to each other. Then the atomic laser ioni-
photoluminescence, and scattering categories. Among zation methods are presented. In the final section sev-
these are photoacoustic spectrometry, thermal lensing eral of the new laser-based methods are described.
spectrometry, laser-enhanced ionization spectrometry
and several new laser-based methods. In this, the final
chapter, these newer spectrochemical methods are de- 17-1 PHOTOACOUSTlC SPECTROMETRY
scribed. Many of these have great potential in trace
analysis. In spectrophotometry, one measures the source radiant
We have chosen to title this chapter "Spectro- power transmitted by the sample. The absorbed source
chemical Techniques on the Horizon" because the radiation produces excited states which deactivate by
methods discussed are relatively new. It can be argued radiative processes. A signal proportional to the frac-
that photoacoustic spectrometry is now a well-estab- tion of the absorbed radiant power that is emitted as
lished method. However, it can also be argued that we photons is monitored in photoluminescence spectrom-
have only scratched the surface in applying photo- etry. In photoacoustic spectrometry (PAS), the signal
acoustic spectrometry to analytical problems. Because detected is related to the other fraction, the fraction of
of this and also because commercial developments in the absorbed radiation that is converted eventually to
the photoacoustic area are very recent, we have chosen heat. If the excitation source is modulated, the periodic
to include photoacoustic methods in this chapter. heating and cooling of the sample results in pressure
The remaining methods discussed here have not waves at the source modulation frequency; these are
been applied routinely in analytical chemistry. Many detected with an acoustic transducer such as a micro-
are techniques that depend on laser sources. These have phone or piezoelectric device.
the potential to become standard tools of the analytical The photoacoustic (PA) effect, also termed the
chemist in the future, but they await further experi- optoacoustic or acousto-optic effect, was first reported
mental, theoretical, and economic developments before by Alexander Graham Bell in 1880. Until the mid-1970s,
they can become routine tools. We begin by discussing there was little interest in PAS, but in the last decade
Chap. 17 1 Spectrochemical Techniques on the Horizon
there havc bccn r,rpid development\ 111 the t h c o ~y, 111- pcns'lte or correct tor the vari,~t~on ol the ~ n c ~ d e r,i-
nt
\truinentat~on,,md .~pplicat~on\. drmt power w ~ t h~ ' ~ v e l e n g t,md h t ~ m c A \drnplc ot
c,~rbonb l x k 14 oltcn u\ed ,i\ ,I reference bcc,~u\e~t
,~bsorbse \ w l t ~ a l l y,111 the ~ n c ~ d e nr dt a t r o n The ret-
ercncc vgn,~lder~vcd14 p o p o ~ t ~ o n . rto l the ~ n c ~ d c n t
A general block diagram of a pliotoacoustic spectrom- tpectral r x l i m t power m d clet1ne4 the m,ixmurn PA
ctcr is shown in Figure 17- 1 . In commercial instruments \~gn,rlo b t ~ ~ ~ In ~ b tlnglc-bcCrm
l e in\trument\, thc s'm-
for the LJV-visible-N1K region, sourcc radiation over a ple 'tnd rclcrcnce \pectra arc rne'~\ured ,rt \ep,rr,ltc t r m o
selected wavelength band is usually provided by a 300 mtt \tored A t t e ~tl&l ,icqul,ltion, .I n o ~ m i l ~ ~spcc- ed
to 1000-W Xe arc lamp and a monochromator. The t ~ u m14 produced by rdtioing the smlplc PA \ign,rl , ~ t
excitation radiation is n~odulateclwith a mechanical ro- e x h w,ivelength to the corrc\pond~ngreference PA
tating choppcr. L,ascr sources have also bccn employed s~gnal In double-beam m\trumcnts, the cxc~tationhem1
in place of the conventional so~irce-rnonochr(>111:itc)r m s p l ~ tinto two beams which are wnultancou\ly 11ic1-
combination, A modulator is still required with u CW dent on the t a t and reference \,lmplc\ The rc\ulting
laser, whereas with pulsccl lasers, n o external modulator \,rmplc and rete~enccPA \ignci14 ,ile \ep,~r,ltcd, de-
is ncccssary. The photoacoustic sample compartment is tected, and proce\\ed m d then r , ~ t ~ o etod p i o v ~ d co n l ~ n e
a sealed chamber with appropriate windows I'or trans- ~ ~ t ~ao wdvclength
n o ~ n ~ a l ~ ~durmg ii \ c m In wrnc In-
mission of the excitation radiation. It contains thc sanv \truinentt, '1 sm'ill fract~onol thc exnt,itlon beam 1s
plc and also houses the aco~lstictransducer and pream- d~vertedto ;1 rctcrence therm'rl clctcctor to prov~dcn
plifier. For gas samples, thc s ~ n p l cfills the chamber reterence \~gnaltor n o ~ m A ~ ; l t r o Inn ,111 c~r\e\,the nor-
and is in direct contact with the transducer. For con- m'1117ed PA spectrum c m be rcl,~tcdto the ,~h\orption
clensed samples, the gas-coupling method is conlrnonly \pcctrum ot the samplc in much the \amc way '15 the
employed. The heat generated by absorption of 1-adia- cxcitatlon \pectrum 1s In photolummetccncc spectrom-
tion in the samplc is transferred to thc surface of thc etry
sample to create pressure waves t h ~ arc t carried to the The design ,ind rn,~tcri,il\ of conttructlon of the
transducer by a coupling gas. Direct coupling tech- photoacou\t~ccell m d the ch,~r,lcteri\t~c\ of the xoustlc
niques have also bccn employcd. particularly for liq- tr,m\ducer arc clit~c,rl Ihc dctectwn lnnit ot photo-
uids. In this case, the transducer (usually piezoclectric) ci~ou\ticmenturement\ 15 dctcrmlncd by the r&o ot the
is inserted into the samplc without the intervention of blank iiol\e to the le\pon\e obt;lincd lor I' given miount
a gas medium. 'l'ypically, the PA signal is further proc- of rdd~,rtionabwrbed by the \ample The hlmk i m \ e
csscd with a lock-in amplifier. 'l'hc phase of the PA is that oh\erved III the a b w i c e o f ,in ,~b\orbcror w ~ t h
signal lags behind the rcfcrcnce signal that tracks the a nonabsor bing 4amplc , ~ n dinclude4 , ~ d d ~ t nolsc ~ v c com-
nmdulatcd incident radiation. Thus the phase of the ponents ~ndependentof the wurce ,ind dependent on
lock-in amplifier is adjusted to maximize the PA signal. the tourcc ~nten\ity Source-~ndependentnoltc ~ncludc\
'1'0 obtain a PA spectrum, it is necessary to com- electr~c~il nolw from the m ~ c r o p h o n e - c ~ ~ i icombl- tler
nation, noise due to Brownian motion of molecules, sound (344 m s I ) ] . For modulation frequencies around
and noise generated by mechanical vibrations. The last 50 Hz. the appropriate dimension is several meters.
type of noise is minimized by insulating the cell from A gas-piston model is often evoked to describe
external acoustic waves and incorporating acoustic baf- the generation of the PA signal from solids. Radiation
fles within the sample compartment. Source-dependent is absorbed by species in the upper layers of a solid.
background signals and noise arise from the acoustic Nonradiative deactivation produces heat. The heat dif-
waves due to radiation absorbed by the cell windows fuses to the surface and is transferred to the gas mol-
and walls. Because the detector does not directly re- ecules near the surface. With source modulation, the
spond to scattered or reflected radiation. these optical gas molecules next to the surface act like a piston as
signals are much less of a problem in PAS than in pho- they are periodically heated and cooled by the thermal
toluminescence spectrometry. However, scattered or waves generated in the sample. Except for very weakly
reflected radiation can eventually strike the cell walls absorbing samples, the contribution of vibrations due
or windows, be absorbed, and generate a PA signal. to expansion and contraction of the solid to the total
Under many conditions, the PA signal is inversely PA signal is minor in contrast to gas and liquid samples.
proportional to the distance of the sample from the A solid sample can be characterized by its thick-
transducer. Thus cell volumes are commonly less than ness ( b ) ,its optical absorption length ( b o p )and
, its ther-
1 cm3. If the cell volume is too small, the acoustic waves mal diffusion length (b,,). The optical absorption length
produced by the sample suffer significant dissipation to is usually defined as l l k ( h ) , where k(X) is the spectral
the cell walls before they reach the transducer. absorption coefficient in cm-l at the wavelength of the
The PA signal is proportional to llf", where f is incident radiation. As the radiation passes through the
the modulation frequency and n is typically in the range sample, its intensity decreases exponentially with pen-
0.5 to 1.5. In practice, modulation frequencies from 10 etration depth according to Beer's law. Thus the optical
to 1000 Hz are used. Although very low frequencies absorption pathlength is the penetration depth where
produce the largest magnitude acoustic wave, the SIN the intensity has decreased to lie of its initial value.
is better at moderate frequencies because of the llf Most of the radiation is absorbed in the first few optical
character of the source-independent noise. The fre- absorption lengths. The amplitude of the thermal wave
quency response of most microphones is also quite poor generated by the radiation absorbed in a particular sub-
at low frequencies. layer of the solid decays exponentially as it propagates
to the surface with a thermal absorption coefficient (a,,)
in cm-'. The thermal diffusion length is defined as
Principles lla,, and thus represents the distance over which the
amplitude of the thermal wave decays to lie of its initial
The theory for the generation of photoacoustical signals value. Most of the detected acoustic signal arises from
is quite complex and highly dependent on the nature absorption occurring in the first few thermal diffusion
of the sample. We will summarize the results of math- lengths.
ematical treatments in a semiquantitative fashion. For a thermally thick ( b >> b,,) and optically thick
For gas samples and liquid samples with direct ( b >> bop) sample, the magnitude of the PA wave
coupling. a fraction of the radiation absorbed is con- generated in the coupling gas, and thus the measured
verted into an acoustic energy that can be detected. PA signal, is related to the optical and thermal diffusion
Thus the radiant power absorbed (Q,) and the PA sig- lengths as follows:
nal ( E p A )are proportional to @,(I - e-k(A)"), where
@, is the incident radiant power, k(X) is the absorption
coefficient in cm-', and b is the sample pathlength.
Usually, PAS is used with weakly absorbing gases or
liquids in which case the PA signal is proportional to
The P A signal is also proportional to the incident source
@,k(X)b. If one species is responsible for the absorp-
radiant power, the ambient pressure of the coupling
tion, k(X) and E p Aare proportional to the analyte con-
gas, and the square root of the thermal diffusivity of
centration.
the coupling gas.
For gas samples, multiple-pass or resonance cells
For a sample that is more thermally thick than
are sometimes used to imprwe detectability. In the
optically thick (b,, << b,,), equation 17-1 reduces to
latter case, one or more cell dimensions are adjusted
to be equal to one-half the wavelength of the acoustic
wave to create a standing wave and amplification for
that signal [XI2 = v,l(2f), where v , is the velocity of
Chap. 77 I Spectrochemica1 Techniques on the Horizon
trometry. In general, PAS is more useful for weakly profile described by a Gaussian function. When a weakly
reflecting samples. In the IR region, PAS provides an absorbing liquid is placed in the laser beam, the liquid
alternative to attenuated total reflectance (ATR) and is heated as a result of optical absorption. A temper-
Fourier transform infrared (FTIR) spectroscopy. Be- ature profile is established in the liquid that matches
cause of the low radiance of conventional IR sources, the intensity profile of the laser. As a result of heating,
Fourier transform instruments have been developed in the liquid expands, which leads to a refractive index
which a photoacoustic cell is used as the detector; a gradient in the sample. Most liquids have a positive
SIN advantage is realized as with conventional thermal coefficient of thermal expansion and a negative tem-
detectors. Recently, there has been interest in a new perature coefficient of the index of refraction, leading
coherent Raman technique called photoacoustic Raman to a diverging lens. In certain cases, however, the tem-
spectroscopy (PARS). In PARS, a given energy state perature coefficient of the index of refraction is positive,
is selectively populated by the stimulated Raman effect resulting in a converging lens.
(see Section 16-2) induced by two incident laser beams. Although the effect was originally thought to be
The frequency difference of the two beams is adjusted a nuisance, thermal lensing has since been shown to be
to equal the frequency of a Raman-active transition. a valuable spectroscopic tool. For example. the thermal
Relaxation of the excited mo!ecules generates an acous- lensing effect can be used to measure extremely small
tic wave which is detected by a microphone. At present absorptions in ostensibly "transparent" liquids. Ther-
PARS has only been applied to obtaining vibration- mal lensing has also been used to increase our under-
rotation spectra of gases and gaseous mixtures. The standing of nonradiative processes in luminescence
PARS signal has been shown to vary linearly with gas spectrometry, of vibrational-translational relaxation, of
concentrations, and detection limits for gases have been thermal diffusion in gases, and of photolytic reactions.
estimated to be in the ppm range. In this section we first describe the instrumenta-
The use of PAS for qualitative analysis of solids tion for observing the thermal lens effect and discuss a
in now established. Its use as a quantitative tool requires model of thermal lens formation. Analytical applica-
further assessment. Accuracy is limited by absorption tions of thermal lensing spectrometry are then pre-
of radiation by concomitants in the sample and differ- sented, and several closely related techniques are con-
ences among the bulk properties of different types of sidered briefly.
samples. Even if the analyte is the only absorbing spe-
cies at the analysis wavelength, the PA signal for a given instrumentation
amount of analyte is affected by the matrix-dependent
thermal diffusivity, thermal diffusion pathlength, and A simple single-beam thermal lensing spectrometer is
scattering properties. illustrated in Figure 17-2. A shutter defines the start of
a thermal lensing experiment. A computer data acqui-
sition system is shown. With a computer system the
17-2 THERMAL LENSING SPECTR beginning of each experiment is defined by a start pulse
generated by the computer. This pulse causes the shut-
The thermal lens effect, also known as thermal bloom- ter to open, which allows the laser beam to strike the
ing, was accidentally discovered in the mid-1960s when sample and lens formation to begin. The photodiode
a Raman scattering cell filled with an organic liquid was current, proportional to the beam center intensity, is
placed inside the cavity of a He-Ne laser in order to converted to voltage prior to analog-to-digital conver-
increase the scattering signal. When the laser was turned sion. Typical thermal lensing transients are shown in
on, large power transients were noted in the laser output Figure 17-3 as a function of the number of transients
that would last for seconds. These transients were ob- averaged.
served to buildup and decay with time and were very Several different instrumental arrangements have
sensitive to cell positioning. Upon placing liquid sam- been used for thermal lensing spectrometry. It is com-
ples in the cavity, the laser beam spot size was observed mon to use a chopper or modulator instead of the shut-
to increase as if a diverging lens were present in the ter shown in Figure 17-2. Here lock-in detection is used
cavity. It was later shown that the thermal lens effect to monitor the result of lens formation. A two-laser
could be observed with the liquid sample placed outside system has been used in which an intense dye laser beam
the laser cavity, and this is the configuration most often causes lens formation and a weak fixed-wavelength probe
used today. beam allows the lens to be monitored. This arrangement
The thermal lensing effect is relatively easy to in which one laser acts as a stimulus and a second probes
understand qualitatively. The fundamental (TEM,,,) the response is often called a pumplprobe geometry. In
mode of a laser gives a beam with a spatial intensity- addition, a differential system has been used to cancel
Chap. 17 I Spectrochernical Techniques on the Horizon
any lens formed by the solvent itself. Mere two cells are positioned correctly, the lcns formed by the solvent,
are used and they are symmetrically positioned around if it is not too strong, is effectively canceled.
the focus of the lens used to define the beam. Solvent
alone is placed in one cell and the sample solution in
the other. A diverging lens, placed after the beam focus,
causes an increase in beam divergence. If the same type To model the formation of the thermal lens, several
of lcns is placed before the beam focus, it causes a assunlptions must he made. We will assume that the
decreased convergence before the focus and a de- electric field of the laser beam is described by a Gaus-
creased divergence after the focus. Thus if the two cells sian function (TEM,,,,modc) and that the beam prop-
agates along the z axis. As the Gaussian beam passes
through an absorbing medium, a time-dependent tem-
perature increase develops in the medium. For a tem-
perature rise AT, the refractive index distribution as a
function of radial distance r and time t is given by
f(t) of the lens can be obtained: Enhancement factors per milliwatt of laser power are
8.93 and 7.02 for CCl, and benzene, respectively, while
methanol (3.06) and water (0.21) show much lower en-
hancements.
The parabolic model above gives good qualitative
where K is the thermal conductivity of the medium (W agreement to experimental data. This model has been
cm-' K-l), T,is the characteristic time constant of the modified, however, to take into account lens aberra-
lens, w is the beam waist (cm). b is the pathlength (cm). tions. This more extensive aberrant lens model better
k(A) is the absorption coefficient (cm-I), and @ is the fits the experimental data in the case of a relatively
laser beam incident radiant power (W). Note from strong lens and better predicts the time dependence of
equation 17-5 that at t = 0 (at the beginning of the lens formation. In the aberrant lens model, the irra-
experiment), dqIdT = 0 and there is no lens [f(O) = x]. diance as a function of time is given by
The lens strength builds up over time (usually milli-
seconds to seconds depending on K and o ) to its steady-
state focal length f(x) given by
amount of absorption is small, the relative change in M should be detectable. Indeed, Cu(I1)-EDTA has been
irradiance is given by determined at ultra-trace levels with a 4-mW He-Ne
laser. It was predicted that 10-I, M would be detectable
with current 1-mW lasers.
Thermal lensing has also been applied to several
practical analytical problems. For example, phosphate
was determined in seawater as the reduced hetero-
polymolybdate (see Section 13-5) by thermal lensing
where AEIE = [E(O) - E(x)]/E(x), A is the solution spectrometry. Detection limits in the pg mL-I range
absorbance, and K is the thermal lensing enhancement were claimed. Many of the standard colorimetric pro-
factor encompassing all the terms in brackets. cedures appear to be applicable to thermal lensing
Note from above that K represents the factor by methods.
which the thermal lensing signal is enhanced over the Matrix interferences have not often been reported
normal absorption signal. In normal absorption spec- with the thermal lensing technique. Much of the initial
trometry the source radiant power does not affect the work was, understandably. carried out with ultrapure
magnitude of the signal obtained since a ratio meas- solvents containing the analyte at ultratrace levels. As
urement is made. However, in thermal lensing, the the method is applied to practical situations, interfer-
magnitude of the signal is directly proportional to the ences will undoubtedly appear. It was recently found,
source radiant power; the enhancement can be quite for example, that the presence of NaCl strongly affects
large when lasers are used as sources. Because of the the thermal lensing signal of the reduced antimony1
dependence of K on thermal conductivity and refractive molybdophosphate complex. A 28% enhancement in
index gradient, different solvents give different en- the thermal lensing signal was found in a matrix equiv-
hancement factors. Most nonpolar solvents have larger alent to that of seawater over an aqueous dilute solu-
enhancement factors than polar solvents because the tion. Solvent extractions. followed by thermal lensing
long-range dipolar interactions in polar solvents allows measurements, did not solve the matrix problem be-
efficient heat conduction away from the heat source. cause of ionic strength effects on the extraction effi-
Chap. 17 I Spectrochernical Techniques on the Horizon
cuit containing a pair of biased collection electrodes as can be achieved. However. the noise in the background
shown in Figure 17-5. Detection of the ions produced ionization signal can influence the SIN. Laser ionization
has several advantages over detection of the photons methods are also susceptible to certain interferences
emitted by fluorescence processes. These advantages that are not present in optical detection methods. Con-
are responsible for the extremely low detection limits comitants in the sample that affect the laser-induced
achieved for certain elements. First, biased electrodes ionization rate cause an interference. Continuous ion-
can collect essentially all the ions produced. By con- ization in the flame can produce charge sheaths at the
trast, only a fraction (i.e.. the fractional solid angle electrodes. Concomitants that influence these charge
N 4 r ) of the emitted photons are collected in fluores- sheaths can also cause interferences. In addition, laser
cence techniques. By detecting ions, the quantum ef- ionization methods have not been very successful with
ficiency losses in photon detectors are avoided and there elements of high ionization potential. The best detec-
are no spectral interferences in the detection process; tion limits have been achieved with elements of low-to-
there can still be spectral interferences in the excitation moderate ionization potential.
process as well as nonspectral interferences (e.g., va-
porization interferences). Unlike resonance AF, scat-
Laser-Enhanced Ionization Methods
tered laser light is not detected with ionization methods.
Ion detection also does not suffer from self-absorption
Enhancement of the ionization of metal vapors in flames
losses.
irradiated by intense laser beams was first observed in
The laser ionization methods do have certain dis- 1976 by scientists at the National Bureau of Standards
advantages, however, when compared to techniques that and by van Dijk and Alkemade in the Netherlands. These
use optical detection. First, the continuous background initial studies prompted many other workers to begin re-
ionization from the analyte. the atomizer, or from con- search in the area, and an international conference on
comitants must be discriminated against. This is usually laser ionization methods was held in 1986. Research stud-
done by using a high-pass filter and a gated integrator ies have been aimed at understanding the fundamentals
to process the ionization signals. as shown in Figure of the ionization process, improving the ion collection and
17-5. The integrator is turned on only when the laser detection systems, extending the range of elements de-
pulse irradiates the flame. Since the pulsed lasers used tected, and applying LEI in trace analysis.
have relatively low duty cycles, excellent discrimination
--i!I! Boxcar
integrator
young that many new developments should occur with Cs is introduced into the counting gas. A pulsed dye
further research. laser tuned to 455 nm excites the 62Sl,,-72P,,, transi-
tion. The 455-nm photons are also capable of photoion-
Resonance Ionization Methods ization of atoms in the 7p level. The proportional counter
signal is linearly related to the number of ions produced.
Resonance ionization differs from LEI in that laser ra- By this experiment, Oak Ridge scientists were able to
diation is used to photoionize the atom (or molecule) conclude that at the lowest signal level measured, a
in the former instead of relying on collisions from an single Cs atom in 1019 atoms of the counting gas was
excited state. Resonance ionization spectroscopy (RIS) ionized.
originated in the mid-1970s at the Oak Ridge National Because of its youth, the RIS technique has only
Laboratory. Today there are several different forms of been applied at present to a few problems. In chemical
RIS. The original RIS method uses laser photoioniza- physics, it has been used to study the diffusion of se-
tion and a proportional counter for electron detection. lected atoms among many other atoms and molecules.
A RIS ionization source has been combined with a mass In chemical kinetics, RIS has been applied to the study
spectrometer to provide excellent detectability with ex- of reactions of highly reactive atoms such as alkali halide
tremely high selectivity. The RIS process has also been atoms. Here, the ability to detect extremely low con-
used in conjunction with flame atomizers where it is centrations is a distinct advantage because corrosion of
more commonly called dual laser or direct laser ioni- the apparatus and the possibility of side reactions can
zation (DLI) . be minimized.
A proportional counter RIS detector is illustrated In analytical chemistry, RIS has been receiving
in Figure 17-6. Proportional counters are capable of increased attention because of its single-atom detection
counting single electrons. They are usually filled with capabilities. At the National Bureau of Standards (NBS)
90% Ar and 10% CH,. If charged particles (ions and an apparatus has been constructed for laser ablation
electrons) are formed in the counter, they are accel- and RIS with a proportional counter as detector. Here
erated by the nonlinear electric field of a small-diameter one laser is used to ablate atoms from a solid, while a
wire held at a high voltage. As the charged particle second laser is used to photoionize. With this system,
moves it collides with atoms of the counting gas pro- NBS scientists were able to detect 5 x lo1' Na atoms
ducing more charges and thus amplification. In the ex- per cm3 of electronics-purity Si. The detection limit was
periment shown in Figure 17-6, a low concentration of such that less than 1 Na atom per integrated-circuit
device could be detected in principle. The combination
of RIS and mass spectrometry is also of considerable
interest. One possible use of a RISIMS instrument is
for isotopic selectivity. Here the total selectivity is the
Proportional
counter / product of the selectivities of photionization and mass
separation. An alternative approach is laser-induced
fluorescence of the ions selected by a conventional mass
spectrometer.
The DL1 method has also been receiving attention
for flame-based determinations. The DL1 technique is
closely related experimentally to LEI in that the de-
Cesium
metal tection system employs a biased probe or a pair of biased
probes. The major difference is that in DL1 either a
second laser is used to photoionize excited atoms by
schemes 4 and 5 in Figure 17-4 or direct photoionization
by scheme 6 is employed. Thus the ionization scheme
is closely related to that of RIS. Schemes 4 and 5 are
most commonly employed. Here the first laser is used
FIGURE 17-6 Proportional counter detection to produce a large population of excited atoms (satu-
system for RIS. A small amount of Cs is intro-
ration is desirable). while the second laser promotes
duced into the counter with the counting gas (Ar
and CH,). A laser beam causes photoionization
those excited atoms into the ionization continuum. In
of the Cs. Acceleration of the charged particles DL1 the rate constant (k,)P1 for photoionization from
by the field of a small wire produces additional excited level j is given by
charges and thus amplification. The signal is proc-
essed by counting electronics (not shown).
Chap. 17 I Spectrochernical Techniques on the Horizon
whel e (J-/"
1s the crw4 xctlon tor photo~onv,~tion l~ om
level j and A,, 1s the ~ncrdentphoton ~rr,rtllanceof the
1on17ingbedm (4 cm ,) In 'iny DL1 experiment in In summary, the laser ionization methods are relatively
a flwne, the coll~\~on,il lonmtlon gwcn by equatlon new analytical tools. As such they have not yet been
17-10 cannot be eliminated, 40 that the total rate ot applied to a variety of practical problems as have such
ion17,1t1onIn D I J , cln,,,,ldt, 14 given by established techniques as AA and AE. The lascr ion-
ization process competes with atomic fluorescence. Hon-
ization and fluorescence are in fact somewhat exclusive
processes. Much research remains to be done t o develop
the quantum mechanical basis of the methods and to
establish various performance characteristics. Since they
We arc assuming here that any ground-state thermal all depend on laser sources, the future of these tcch-
ionization is negligible or has hecn discriminated against niqucs is linked to future developments in pulsed lasers.
by a high-pass filter and gated electronics. However, already the ionization methods can achieve
Although photoionization and collisional ioniza- outstanding detection limits for certain elements. They
tion compete in DL1, conditions can be chosen in some have great potential as Suture routine analytical tools.
cases so that photoionization will dominate. For this to
happen, the overshoot energy of the ionizing laser beam
(energy in cxcess of that needed to reach the ionization
continuum) should be srnall. Photoionization cross scc-
tions decrease with increasing overshoot cnergy. In ad-
dition, the energy defect (E,,,,, - E,) shoi~ldbe large
only slightly exceeds that necessary for oscillation, small olecular Multiphoton Ionization
changes in the absorption losses in the cavity can cause
very large changes in the laser output. Unfortunately, Laser ionization methods for atoms were described ear-
there is also an increase in laser instability when the lier in this chapter. Lasers can also be used to ionize
gain is near threshold, which makes this method of molecules via multiple photon processes. Multiphoton
enhancement difficult to use in practice. ionization (MPI) was first reported in the mid-1970s and
With multimode dye lasers an additional enhance- has since been widely used in molecular spectroscopy.
ment effect can be observed. If selective modes are The MPI process involves a multiphoton excita-
attenuated by absorption by molecules inside the cavity, tion to an intermediate vibronic state followed by a
the gain for the other modes increases. By using a fre- multiphoton excitation to the ionization continuum
quency-selective detector, the power loss in the atten- through a dense manifold of autoionizing states. Early
uated modes can be measured and used to obtain the experiments were carried out in the gas phase in cells
amount of sample absorption. containing collection electrodes. Here only the total ion
Intracavity absorption has been used to make current (electrons, positive ions, and negative ions) could
quantitative measurements of several molecular spe- be obtained without any specific means to identify the
cies. By placing a gas cell containing I, inside the cavity detected species. In more recent work a mass spec-
of a CW dye laser an enhancement of 10' over a single- trometer has been used to identify the ionic fragments
pass external cell was observed. An I, vapor pressure obtained. In addition, the MPI technique has been ex-
of 1.5 x torr could be detected. Free radicals, tended to solutions with simple collection electrodes
such as O H , NH,, and HCO, have also been quanti- used to monitor the total charge produced by the in-
tatively determined by intracavity absorption. cident laser beam.
Vacuum chamber
1
I beam
molecular beam in (b) produces a
pencil-shaped beam of atoms or mol-
(a)
ecules. The sample, a metal or a salt,
is placed in an oven and heated to
the desired temperature. A small hole Molecular
beam
in the side of the oven allows the beam Velocity-selection
of molecules to effuse into a high-
vacuum region. The pinhole provides
collimation of the beam. Often a ve-
locity selector is used to allow only
molecules in a particular velocity range
to pass. This can be a pair of slits Collimating
pinhole
displaced relative to each other by a
certain angle and separated by a cer-
tain distance. Only molecules with a Collimated
particular velocity pass both slits. (b) beam
Chap. 17 1 Spectrochemicai Techniques on the Horizon
A block diagram of a laser multiphoton ionization zation source with a time-of-l'iight mass spectrometer
molecular beam mass spcctrometer is shown in Figure with both p ~ ~ l s eand
d continuous molecu1;rr bcam in-
17-7a. An apparatus for producing a collimated rnolcc- troduction systems.
ular bcam is illustrated in Figure 17-7b. With the MPl Multiphoton ionization has also been actively
mass spcctrometer system either the total ion current studied in solution. Hcre various pirlscd lasers (e.g., an
or the ion current corresponding t o a selected fragment N, laser, an N, pumped dye laser or an excimer lascr)
can be monitored. Figure 17-Xa shows the MPI spec- have been used t o provide the excitatiolllionization en-
trum of benzene in the total ion current mode, while ergy. The current produced as a result of irradiation hy
Figure 17-8b shows the spectrum obtained while mon- the pulsed laser beam is measured with two electrodes;
itoring C + , the most abundant ion. By tuning the dye a boxcar integrator is used to acquire the photoioni-
laser t o various wavelengths and recording the mass zation signal. Multiphoton ionization signals have been
spectrum of the positive ions, a two-dimensional vi- obtained for polycyclic aromatic hydrocarbons and scv-
broniclmass spectrum is obtained. That is, mass spectra era1 drugs of analytical importance. The multiphoton
can be measured corresponding to different vibrational ionization method shows considerable promise as an
excitation bands. HPLC detector. Indeed, a windowless flow cell has heen
Although this technique is quite new, it has ob- designed that allows simultaneous detection by molec-
vious potential in analytical chemistry. 'l'racc analysis ular tluorescence, the photoacoustic effect and pho-
of atmospheric pollutants is one potential application. toionization methods. These techniques were choscn
The selectivity is such that analyses of isomeric mixtures because they monitor all the major deactivation path-
may be feasible without GC separations. Recent vari- ways for excited molecules.
ations in technique have included using the laser ioni-
earn Spectros-
copy. One q q r o . ~ hfor rcducmg the Doppler width
FIGURE 17-8 Multiphoton ionization spectra. 15 to mtroduce the w m p k as a well-collimated molec-
Curvc a shows the MPI spectrum of benzene. This ular beam (scc F~gure17-7b) In wch '1 beam the ve-
was obtained while scanning the laser wavelength loclty component perpendlcul'u to the hcnm ax15 can
and monitoring the total positive ion current. For be ,I factor ol20O or more lower t h m thnt in the beLlm
curve b the most abundant ion C ' was selcctcci directmn To obtain ' l b x q t ~ o ns p e c t ~ , ~,t ,monochro-
with the mass spcctrometer and monitored while m,it~claser beam I \ crowxl pcrpendicul,u to the mo-
the lascr was scanned. (With permission from
lecul,ir bean1 and tuned over the ~ihsolpr~on prot~le
D. A. Lichtin, L. Zandee, and R. B. Bernstein,
"Potential Analytical Aspects of Laser Multipho-
Smce the velocity component 14 5mdl in the d ~ r c c t ~ o n
ton Ionization Mass Spectrometry," in Lasers iiz ot the h e r beam, CL reduct~onin the Doppler w~dth15
Cheinical Analysis, G . M. Elieftje, J . C:. Travis, ach~eved,In m m y case\ to the polnt where the natural
and F. E. Lytle, eds., FIurnana Prcss, Clifton, hnewidth become5 the lmltlng t x t o r
N.J., 1981.) l o t k c full ndvant;lgc ot the high retolut~on'if-
Sec. 77-4/ Miscellaneous Laser-Based Techniques 539
forded by reducing the Doppler width, a very narrow Consider a gas-phase collection of free atoms that
laser bandwidth must be used. With dye lasers, this can absorb different frequencies of laser radiation be-
usually involves using one or more Fabry-Perot etalons cause of the Doppler effect. Atoms moving away from
in the laser cavity, employing bandwidth narrowing out- the source absorb lower frequencies than those moving
side the cavity or switching to a single-mode laser. Be- toward the source. The absorption lines of such atoms
cause of very small absorption pathlengths and rela- are said to be inhomogeneously broadened because dif-
tively low molecular densities, the absorption spectrum ferent atoms are responsible for different parts of the
is often obtained by measuring the total fluorescence absorption line. If any frequency within the absorption
from the laser-excited levels (fluorescence excitation line could excite every atom with equal probability, the
spectrum). As an example of the improved resolution line would be homogeneously broadened. Atomic and
that can be obtained, Figure 17-9 shows excitation spec- molecular beams crossed with a laser beam can produce
tra of NO, taken in an ordinary gas cell and in a col- homogeneously broadened lines because the Doppler
limated molecular beam. . . .. previously.
width is so small, as discussed
Let us now consider a two-laser experiment on a
Saturation Spectroscopy, Saturation phenom- collection of gas-phase atoms. We will use an intense
ena have been studied since the early days of powerful monochromatic pump laser beam to saturate the atomic
laser sources. We have previously discussed saturation population and a weak tunable probe beam to monitor
in Sections 11-3 and 15-2 in conjunction with atomic the saturation effect. If the pump beam is of very narrow
and molecular fluorescence. Selective saturation of mo- bandwidth, only those atoms that have the appropriate
lecular transitions can be used to produce Doppler-free Doppler velocities will absorb the laser radiation. Hence
spectra. we can cause saturation of only those atoms within a
I!:
I
/
((II
A v represents
frequency shift
frorn514.5 nrn
REFERENCES
The references below are general references on recently de- 5. G. M. Hieftje, "Approaching the Limit in Atomic Spec-
veloped laser spectrometric methods. trochemical Analysis," J. Chem. Educ., 59, 900 (1982).
Part of seven articles taken from a symposium "Ap-
M. D . Levenson, Introduction to Nonlinear Laser Spec- proaching the Limits of Chemical Analysis" at the Spring
troscopy, Academic Press. New York, 1982. Includes 1982 National American Chemical Society Meeting.
chapters on saturation spectroscopy and multiphoton ab- Covers a variety of laser techniques for single-atom de-
sorption and a useful catalog of nonlinear phenomena. tection.
D . S. Kliger, ed., Ultrasensitive Laser Spectroscopy, Ac- 6. C. Th. J. Alkemade, "Single Atom Detection," Appl.
ademic Press, New York, 1983. Covers several topics Spectrosc., 35. 1 (1981). Covers theory of single-atom
relevant here, including thermal lensing, laser ioniza- detection and possible techniques to achieve it.
tion, photoacoustic spectrometry, two-photon spectros- 7. E. H . Piepmeier. ed., Analytical Applications of Lasers,
copy, and laser intercavity-enhanced spectrometry. Wiley, New York, 1986.
N. Omenetto, Analytical Laser Spectroscopy, Wiley, New
York, 1979. An excellent book dealing with laser prin- The following references are devoted wholly or in part to
ciples and the applications of lasers in analytical chem- photoacoustic spectrometry.
istry. 8. J. F. McCelland, "Photoacoustic Spectroscopy," Anal.
G. M. Hieftje, ed., New Applications of Lasers to Chem- Chem., 55, 8 9 A (1983).
istry, American Chemical Society, Washington, D . C . , 9. D . Betteridge and P. J. Meylor, "Analytical Aspects of
1978. Contains excellent chapters on laser applications Photoacoustic Spectroscopy," CRC Crit. Rev. Anal.
written by experts in the field. Chem., 1 4 , 267 (1984).
42 Chap. 17 1 Spectrochemical Techniques o n the Horizon
10. A . Roscncwaig. "Photoacoustic Spectroscopy: A New K . ('. Lin, P. M . Hunt, < i . E. 1,croi. and S. R . Crouch.
'l'ool for Investigation of Solids," Arrnl. ('hc~n1.. 47,502A " t k r g y Consideration in Dual I x c r Ioni/ation I'roc-
( 1975). csses in Flarnes," A n d . ('irc7trr.. 5.7, 2382 (1983).
I 1. A . Roscncwaig, l'lro~onc~o~r.s~ic.scmd ~ ' ~ O ~ ~ M C O L.Sp(,(.-
ISI~C K. (-I. Lin, P. M. II~unt,and S . R. C:rouch, "Flarnc Tcm-
troscq~y,Wilcy, New York, 1980. pcrature Dctcrnmination by Uual I , a w Ionization," C'lrrrr~
P11v.s.Lcti., 90, 111 (1982).
The Sollowing references discuss phototherm;d melhods, such G. S. I lurst. "l<csonnncc Ionization Spcctroscopy." Anid.
as thermal lensing spectromcti-y. Clrcrn., 53, 1448A (1981).
J<. L. Swofford, "Analytical Aspects of7'herrn;~lLensing .I. C. Travis, G. C'. Turk, and R. B. Grccn. "Laser-
Spectroscopy," in Lnsrrs in Cllrrrric~rrlArrn1ysi.s. G.M. Enhanccci Ionization Spectrometry, Arrul. ('hrrn., 54,
I lieftje, J. C. Travis, and F. E. Lytlc, eds., I l u ~ n a n a 1007A (1 982).
1'1-ess, Clifton, N.J., 1981. M. H . Nayfch, "Laser Detection of Single Atoms," Anz.
C'. M. Phillips, S. R. Crouch. and <;. E. Leroi, "Matrix Sci., 67, 204, March-April, (1979).
Effects in Thermal Lensing Spectromctry: Dctcrmina- G. S. Hur-st, M. H . Nayfeh, and .I. 1'. Young, "One-
tion of Phosphate in Saline Solutions," Atrul. Chrrrz.. Atom Detection Using Resonance Ionization Spcctros-
58, 1710 (1986). copy ," Phys. Rcv. A , 15, 2283 ( 1977).
M . D. Marl-is and K. Peck, "Photothcrrnal Effects in J. C . Travis, "Limits to Sensitivity in Laser Enhanced
Chemical Analysis," A n d . C'hcwl.. 58, 811A ( 1986). Ionization," J. Chcw~.E ~ L I c50,. , 909 (1982). Part of the
N. J. Dovichi and J . M. [Iarris, "Laser-Induced Thermal Spsing 1982 AC'S symposium on "Aplxoaching the Litn-
Lens Effect for Calorimetric l'race Analysis." Atznl. its o f Chemical Analysis."
C h ~ r n .51,
, 728 (1979). The follow~ngrcfcrences dcal w ~ t hspcc~al~zed
methods o f
mtercst In t h ~ schapter.
The 1efcrcnce5 below dcal w ~ t h1'1scr tonlzatlon rnethocls
2 D. A. Lichtin, L. Zandee, and I<. U . Bernstein, "Po-
16. J . C'. Travis and J. K. DeVoc, "The Optogalvanic Ef- tential Analytical Aspects of 1,ascr Multiphoton loni-
fect," in Lusers in Chrrr~icalAnulysis, G. M. Hicftje. zation Mass Spectrometry," in Lmcrs in C'hewzicnl A n d -
J . C. Travis, and F. F,. Lytle, eds., tlumana I'rcss. ysk, G. M . Hieftjc, J . C. Travis, and F. E . Lytlc, cds.,
Clifton, N. J., 1981. Ifurnma Press, Clifton, N. J.. 1981.
17. S. L. Chin and 1'. Latnbropoulos, Mul/iphotorz 1orrizr~- 25. I<. E. Smallcy, "Mass-Selective I.ascr Photoionization."
lion o f A t o m , Academic Press, Toronto, 1084. J. Chcm. E ; ~ L K . .59, 934 (1982). Also based on a pres-
18. J . C . 'Travis, Ci. C. 'Turk, and R. B. Green, "Laser- entation at the Spring 1982 ACS mccting.
Enhanced Ionization for Trace Metal Analysis," in New 26. J . M. Haycs a n d G. J . Small, "Supcrsonic Jets, Kota-
Applicution.~?/' L u s ~ r sto Chunistry , G . M. Hieftjc, ett., ."
tional Cooling and Analytical Chc~nistry A n d . Clrrtn..
American Chemical Society , Washington, D.C., 1978. 55, 565A (1983).
In most analytical determinations, practical consider- tions in variables or conditions that affect the magnitude
ations limit the number of measurements of a given of the measured quantity. The precision is a measure
sample and the number of samples in a total population of the magnitude of the random error. Random error
that we can analyze. Because of this and the uncer- causes measurement results to vary in magnitude and
tainties associated with measuring the magnitude of a sign. Random error can be reduced in some cases by
quantity of a given sample, it is important to report better control of the variables that cause the fluctuation.
some estimate of the accuracy and precision of results. For example, random fluctuations in temperature from
The accuracy indicates how close the measured mag- sample to sample may cause the signal measured for a
nitude of a quantity is to the true magnitude of the given analyte concentration to vary. Better control of
quantity in the population, whereas the precision in- the temperature will reduce the random error due to
dicates the reproducibility in the magnitude of the meas- temperature fluctuations. Also the effect of random
ured quantity or the variability of that magnitude of the error can be reduced by measuring many samples from
quantity among samples in the population. Accuracy is a population. This is not true for systematic errors.
affected by both systematic and random errors. System- Statistics provide us with a formalized strategy to
atic errors, also known as determinate errors, result evaluate the uncertainty in measurements due to ran-
from factors which cause the magnitude of the measured dom errors and to determine if systematic errors are
quantity to be biased too high or too low by either a also present. In this appendix, statistical concepts for
fixed fraction or an absolute amount. They are caused evaluating errors are presented and their application is
by factors affecting the measurement which are consis- demonstrated. The references for this appendix should
tently the same in or for a given population. For ex- be consulted for a more detailed discussion.
ample. a concomitant present in all samples in a pop-
ulation may cause the measured analyte concentration A-I STATISTICAL QUANTITIES
to be too high. Theoretically, a systematic error can be
eliminated, estimated, or corrected if the cause or factor Typically, we measure a given quantity or variable in
responsible for the error is known. a given population n times. This could be n measure-
Random error is caused by uncontrolled fluctua- ments on a given sample or one measurement for each
App. A 1 Statistical Concepts
The second form of equation A-2 is useful for calcu- and is called the z statistic or the standard normal dc-
lation purposes and is the equation preprogrammcd into viatc. Note that the mean has been normalized to zero
scientific calculators. The experimental mean is a meas- and the SD is normalized to 1.
ure of the central tendency of thc measurements or If we are concerned with the distribution of the
probability distribution and is an estimate of the pop- means rather than of indiviclual measurements (e.g.,
ulation or true mcan, p, the mcan for an infinite numbcr the distribution of X calculated from n individual mcas-
of mcasure~ncntsor for all rncmbers in the popul a I']on. uremcnts of X), then A' is replaced by X in equation
Standardized normal
probability distribution. 'The shaded
at-ca reprcscnts the probability that
i > z ,. P is thc probability.
Sec. A- l 1 Statistical Quantities 545
If this is compared to the z htatistic, we note that the r~ctuc~l(r due to r x d o m error, t mu\t be greater t h m z
only difference is that cr has been replaced by thc ex- (or the expermental m e m nlu\t be lurcher from the
perimental standard deviation, s . As for thc z statistic, popul,~tionmean) to obt'iin the \dmc level of conldencc
the difference between the expcrinlcntal and population a h i t the d~tferenccbetween the mean\ 1s due only to
mean has been norlnalizetl to the SD. The protxibility random errol Notc In Tcible A-2, thnt t,, 1s q u ~ t clarge
distribution for the t statistic is similal- t o that for the z for smlll n (e g , 2 or 3) due to the high proh,rtxl~tyol
statistic, but it is broader and the breadth of the dis- error In the o t ~ m ' t t eot 5 , but as n ~ n c r e < ~ \for
e \ ,I glven
tribution varies with 11. Probability tables (see Tahlc A- vdue of a , t,, dccren\e\ and lor r l = x,r, = z,,
2) indicate the probability ['(/ :-(,,) = tx that 1 is equal Uwdly. the m e m m d SD arc c\tlrnc~tedtrom the
to or greater than a specified value o f i denoted r,,, which sdme n me,t\urerncnts However, ~i they arc not, ~t 1s
is always positive. Notc in the table that the value of Important to r e A ~ cthat n In the d e f m t ~ o n sof the z
t,, varies with the number of mcasurcments rl used and t statlstlcs (equation\ A-5 dnd A-7) I \ the number
to calculate s or the number o f degrees of freedom of measurement\ u\ed to obt'un the m e m , while tor
v defined as 11 - 1. As for the z statistic, P(r < 1,") is the r \tat~stlc,v 1s determined by the number of me<r\-
(I - a ) , P(l r 1 < t,,) is (1 200, and here ['(I / I-2, t,) urement\ uwd to o b t m the SD. Oltcn, t ~ i n " or ~
is 2a, and P(0 < t < l,,) is (0.5 - tx). ,inLL,wh~ch,Ire denoted 6 dnd C, 1s dcimcd as the SD
For a given a , t,, is greater t h m z,,hecause r is a of the m e m , w h k cr or J 14 the SD oi an ~nd~vrdual
more conservative statistic which takes into account the result The SD of the mean repre\ents the SD ol a
uncertainty in deterniining s. Ilcnce to include the chance probability distr~but~on oi the medm rdther than of in-
that the calculated nlagnitucle of .u is greater than the d~vldualrn~~tsurements
TABLE A-2
Critical values of t
Sec. A- 7 / Statistical Quantities 547
z < z,, for cw = 0.OJ25. This predicts a significant dif- TABLE 61-3
ference at the 98.8% confidence level which is too lib- Propagation of random uncertainties for common
eral a confidence level. Similarly. for the two-tailed test, functions
the interval for 95%) confidence would be smaller by a
factor of (1.9612.776) = 0.71 sincc z,, = I .96 for oc =
0.025. Again this is not warranted due to the uncertainty
in the estimate of the SD.
The t and z statistics can be used only if the normal
distribution applies. In this case, test statistics are also
availablc to compare the experimental variance to the
pop~~lation variancc (chi-square test) and to conipare
two expcrimental standard deviations (F-test). Other
test statistics are available for different distributions
and, in addition, we can test the normalcy of data and
evaluate if other test statistics must be employed. Some- R log,,, A (0.4343) In A
3. - =
times the data can be made to follow the normal dis-
tribution by mathematical transformation. The log nor-
mal distribution is an example where the logarithm of
values follows the normal distribution. This distribution
applics somctinies to the concentration distribution of
trace clcmcnts in environmental samples where nega- A - c7
5 R = ------
tive concentrations are not possible. H - C'
1 I - R
where the variables A , B , and C' are a4sumed to be FIGURE A-2 Graphical interpretation of prop-
independent. agation of uncertainty mathematics. The partial
If the infinitesimal differentials are replaced by derivative of the function R with respect to a
small finite errors, the latter equation becomes variable A is the slope of the function with rcspect
to A at a particular value of A . This indicates how
a change in A causes a change in R. Thus if we
reflect the uncertainty in A , AA, through this
slope, we can deterrninc the uncertainty in R , AR,
caused by AA. To obtain an accurate value of
for systematic errors wherc AR, AA, AB, and AC are AR, it is necessary that the derivative be constant
estimates of the systematic crrors in R , A , B, and C, over AA.
App. A 1 References 549
F o r random errors, equation A-11 becomes A-13 it is assumed that errors o r standard deviations
are much smaller than m e a n values (e.g., AA << A ,
s, << A). If the variables a r e nonindependent o r cor-
-
- / ( E ) ~ ;+ (E)2si+ ( R ) +~. . ~
. ~ related, t h e n cross products which represent co-
SR
dA dB ac variances must b e included.
(A-13) Formulae for random errors are shown in Table
A-3 for several common functions. T h e graphical inter-
where s,, s,, s,, and s, are the standard deviations in pretation of propagation of uncertainty mathematics is
R. A , B , and C , respectively. I n equations A-12 and presented in Figure A-2.
REFERENCES
W. Mendenhall, Introduction to Probability and Statis- D. L. Massart, A. Dijkstra, and L. Kaufman, Evaluation
tics, 5th ed., Duxbury Press, North Scituate, Mass., 1979. and Optimization of Laboratory Methods and Analytical
Precision Measurement and Calibration-Statistical Cotz- Procedures, Elsevier, Amsterdam, 1978.
cepts and Procedures, NBS Special Publication 300, vol. J. C. Miller and J. N. Miller, Statistics for Analytical
1, National Bureau of Standards, Washington, D.C., Chemistry, Halsted Press, New York, 1984.
February 1969. R. Caulcutt and R . Boddy, Statistics for Analytical
M. G. Natrella. Experimental Statistics, NBS Handbook Chemists, Chapman & Hall, New York, 1983.
91, National Bureau of Standards, Washington, D.C., W. J. Dixon and F. J. Massey, Jr., Introduction to Sta-
August 1963. tisticalAnalysis, 3rd ed., McGraw-Hill, New York. 1969.
P. R. Bevington, Data Reduction and Error Analysis for 0 . L. Davies, ed., Statistical Methods in Research and
the Physical Sciences, McGraw-Hill, New York, 1969. Production, 4th ed., Hafner Press, New York, 1972.
P. Moritz, "The Application of Mathematical Statistics E. B. Wilson, Jr., A n Introduction to Scientific Research,
in Analytical Chemistry," in Comprehensive Analytical McGraw-Hill, New York, 1952.
Chemistry, G. Svehla, ed., vol. 11, chap. 1, Elsevier, C. A . Bennett and N. L. Franklin, Statistical Analysis
Amsterdam, 1981. in Chemistry and the Chemical Industry, Wiley, New
H. D . Young, Statistical Treatment of Experimental Data, York, 1954.
McGraw-Hill, New York, 1962. H. A. Laitinen and W. E. Harris, Chemical Analysis,
J. H. Zar, Biostatistical Analysis, 2nd ed.. Prentice-Hall, 2nd ed., chaps. 26 and 27, McGraw-Hill, New York,
Englewood Cliffs, N.J., 1984. 1975.
A . Savitzky and M. J. E. Golay, "Smoothing and Dif- W. J. Youden, Statistical Methods for Chemists, Wiley,
ferentiation of Data by Simplified Least Squares Pro- New York, 1951.
cedures," Anal. Chem. : 36, 1627 (1964). G . E . P. Box, W. G. Hunter, and J. S. Hunter, Statistics
C. G. Enke and T. A. Nieman, "Signal-to-Noise Ratio for Experimenters, Wiley, New York, 1978.
Enhancement by Least-Squares Polynominal Smooth- N. R. Draper and H. Smith, Applied Regression Anal-
ing," Anal. Chem., 48, 705A (1976). ysis, 2nd ed., Wiley, New York, 1981.
R. J. Harris, A Primer of Multivariate Statistics, 2nd ed., S. L. Meyer, Data Analysis for Scientists and Engineers,
Academic Press, Orlando, Fla., 1985. Wiley, New York, 1975.
Many different materials and coatings arc i~scclin op-
tical components and systems. I n this appendix a few
of the most important properties of materials are pre-
sented. Additional informatiol~can he found in tlund-
book o,f Optics, W. C . Driscoll and W. Vaughan, eds,
(McGraw-IIill, New York, 197S), and in catalogs and
guides available from optics houses such as Mcllcs Griot,
Oriel, Ealing, Ncwport, and many others.
TABLE B- 7
Refractive indices of optical materials at 20•‹C
propriate thicknesses. The MgF, coating also protects optical materials. The transmittance T of the glass (ne-
the glass from chemical deterioration. Multilayer glecting surface reflection losses) is given by T = e c k b ,
broadband coatings can give reflectances of less than where k is the absorption coefficient (cm-l) and b is
0.6% over much of the visible region. Multilayer coat- the thickness of the glass (cm). Figure B-2 shows k vs.
ings are also available to minimize the reflectance at a X for several materials.
particular wavelength; these can reduce the reflective Note that glass, quartz, and fused silica all absorb
loss to less than 0.25% at the chosen wavelength. in the IR region at wavelengths longer than 2 p m Hence
for IR spectrometry, halide salts (NaC1, NaBr, AgC1,
KBr) are used for optical components. The IR trans-
B-2 TRANSMlSSlON CHARACTERISTICS mitting regions for several IR optical materials are listed
in Table B-3. The water solubility of these materials is
Absorption of the incident radiation by the optical ma- also given.
terial can be a significant problem in spectrochemical
analysis. The absorption characteristics often dictate the
type of material suitable for use in lenses, windows, B-3 REFLECTANCE OF MIRROR
cuvettes, and other components. The transmission char- MATERIALS
acteristics of several typical UV and IR transmitting
glasses are shown in Figure B-1. The reflectance properties of typical mirror coatings are
Often, absorption coefficient data are given for illustrated in Figure B-3. Mirrors are usually front-sur-
TABLE B-2
Reflectances for several interfaces
Spectral Spectral
Interface reflectance Interface reflectance
Transmittance vs.
wavclcngth for scvcl-al IJV and I[<
transmitting glasscs. Ultrasil is a typc
of fused quartz and suprasil is fused
silica. (Reprinted with permission
from Handbook oJ' Optics, W. Dris-
coll and W. Vaughan, eds., McGraw-
Hill, New York, 1078.)
TABLE B-3
Properties of infrared window materials
Bare A1
Wavelength, nm
REFERENCES
1. W. L. Wolfe, "Properties of Optical Materials," in Hand- 4. Electro-Optics Handbook, Technical Series EOH-11, RCA
book of Optics, W . G . Driscoll and W. Vaughan. eds., Corp., Lancaster, Pa., 1974.
McGraw-Hill, New York, 1978. 5. J. Wilson and J. F. B. Hawkes, Optoelectronics: A n In-
2 . J. A. Dobrowolski, "Coatings and Filters," in Handbook troduction, Prentice-Hall, Englewood Cliffs, N.J., 1983.
of Optics. W. G. Driscoll and W. Vaughan, eds., McGraw- In addition, the catalogs from many of the optics companies
Hill, New York. 1978. (Oriel, Melles Griot, Corion, Ealing. Rolyn, Newport, Phys-
3. S. Musikant, Optical Materials: An Introduction to Selec- itec, etc.) often contain a wealth of information about optical
tion and Application, Marcel Dekker. New York, 1984. materials.
This appendix gives the transmittance characteristics of monly employed for rejection of stray radiation, for
various optical filters. Figure C-1 shows thc transmit- order sorting, and for many other applications that do
tance curves for 12 different colored-glass bandpass fil- not require thc narrow bandpass characteristics of in-
ters for the UV-visible region. Such filters are corn- tcrfcrence filtcrs. The transmission curves for a set of
02
5- 0 1 E C-1 Transmittances of
Corning glass filters. The numerical
: designations arc the product nunl-
+ bers for spccific filtcrs. [With per-
0 5R 0 62 0 66 0 70 0 74 0 78
Wavelength,pm
mission from K. S. Gibson, J. Opt.
@ Soc. Am., 2.5, 131 (1935).]
Sec. C / Characteristics of Optical Filters 555
Wavelength. nm
square passband tops instead of Gaussian or Lorentzian FIGURE C-4 Transmittance curves for a set of
shapes. Alternating layers of high (ZnS) and low (cry- multiple-cavity interference filters. (Courtesy of
olite. Na,AIF,) refractive index materials are deposited Melles Griot.)
556 App. C / Characteristics of Optical Filters
atomic lines are often stock itcms. Continuously vari- thc bandpass shape. Filters with narrower bandpass have
able interference filters are also available in which the less tuning range.
ccntral wavelength changes with position along the length Although filters for the UV-visible region are most
of the filter. One commercial filter (Oriel Corp.) iso- common, IR interference filters are also readily avail-
lates wavelengths from 400 to 700 nm along a 60-mm able. One manufacturer (Oriel Corp.) produces 1K in-
length. Circular wedge interference filters are also avail- terference filters in the rangc I .8 to 5.5 pm with central
able. wavelengths every 0.1 pn. The bandwidths (FWHM
Interference filters can be angle tuned over a small values) normally vary from 0.1 to 0.15 pm. Special
wavelength rangc as described in Scction 3-5. Filters filters with ccntral wavclengths as long as 20 pin can
with 5- to 10-nm bandpass valucs can be tuned over be purchased. Short- and long-wavelcngth cutoff filters
about 10 nm ( 4 0 " rotation) without drastic changes in arc also available for the IR region.
REFERENCES
Many of the general optics references for Chapter 3 are also 3. Optics Guide 3 , Melles Griot, Irvine. Calif. Morc than a
relevant here. catalog-almost an optics textbook.
4. B. Hecht and A. Zajac, Optics, Addison-Wcsley, 12ead-
ing, Mass., 1974. Good discussions on the principles and
1. Optics and Filters, Oriel Corporation, Stamford, Conn., construction of optical filters.
vol. 111 of Oriel catalog. 5. J. A. Dobrowolski, "Coatings and Filtcrs," in Harrdboolc
2. Opticul Filters und Coatings, Curion, Corp., Holliston, ofOptics. W . G. Driscoll and W. Vaughan, eds., McGraw-
Mass. Hill, New York, 1978.
APPE I
Photomultiplier
Tube Specific
Manufacturers of photomultiplier tubes (PMTs) such ample, some of the figures from the specification sheet
as RCA, Hamamatsu, and EM1 provide detailed spec- for an RCA 1P28 PMT are reproduced here (Figures
ification sheets for these devices which indicate their D-1 and D-2). This tube or its equivalent is used in
characteristics and operating considerations. As an ex- many spectrometers.
55 App. D I Photomultiplier Tube Specifications
Cb)
FIGURE D - l Spectral I-esponse, sensitivity, and current amplificxtion
characteristics. In (a) plots of quantum efficicncy [ K ( X ) ] and absolute
sensitivity [thc cathodic responsivity, /((A)] vs. wavelength are shown.
From these data, thc photocathodic current 01-pulse rate at a given wave-
length for a given incident I-ndiant power can be calculated. A plot of
relative sensitivity is the cathodic rcsponsivity normalized to 100% for thc
wavclength of m;lxirnum rcsponse [i.e., R(X)I(R(A,,,)].111 (h) the depend-
ence of the typical current amplific:~tion (the gain rrz) on the I'MT bias
voltage (6,) is illustrated. Note thc linear log-log relationship bctwccn tn
and E,,. The sensitivity [anodic responsivity, m K ( A ) ] is also shown as a
function of E,,. 'The rnaxi~n~um, typical. and minimuru plots illustrate tha!
the anodic rcsponsivity can vary about two orders of ~nagnitudeamong
tubes of the same type. This hehavio~-is due primarily to a vari;~tionin
gain ;lmong tubes. 'l'llus thc figure can be uccd to csti~natcnz within an
order of magnitude. Note that photometric units (e.g., lumens) arc often
used instead of more convenicnt radiometric units (e.g., watts). (Courtesy
ol R('A Ncw Products Division, I,ancaster, I'enn.)
e
Sec. D 1 Photomultiplier Tube Specifications 559
FIGURE D-2 Dark current, EADCI, and EN1 characteristics. In (a) the
dependence of the anodic dark current on the bias voltage (E,) is shown.
A t higher bias voltages, the log-log plot is reasonably linear because the
dark current is primarily due to cathodic thermal emission and propor-
tional to the gain (i,, = mi,,). Also shown are plots of the equivalent
anode dark current input (EADCI), the incident radiant power necessary
to produce a photoanodic current equal to the anodic dark current. A t
about 900 V, the ratio of gain to dark current is optimal. The effect of
temperature on the equivalent noise input (ENI) is illustrated in (b). The
EN1 is the incident radiant power that produces a photoanodic current
equal to the anodic dark current rms noise (i.e., an SIN of unity if meas-
urements are dark-current noise limited). As the temperature is reduced,
the dark current and dark current noise decrease exponentially. (Courtesy
of RCA New Products Division, Lancaster, Penn.)
Analytical methods usually require some sample trcat-
ment prior to the final, instrumental measurement. For
the majority of the spectrochemical methods discussed
in this book, the analytical sample must be in thc form e brought into solution by
of a solution. Some samplcs are obtained initially as osition with fluxes.
solutions (e.g., blood scrum, other biological fluids, samples, some alloys, and metallic oxides, as well as
samples from lakes, rivers, oceans, and wastewaters). purc solids to prepare standards, are commonly dis-
Solid samples, however, must normally be converted solved in mineral acids such as HGl, HNO,, H,PO,,
into solution samples in order to mix, aliquot, dilute. HF, flCI04, and H,SO,. Heating is often required and
separate, or otherwise treat and present to the spec- the type of acid or- combination of acids used depends
trometer. Sample clcanup methods are used, if neces- on thc composition of the sample (e.g., HF is priinarily
sary, for separation of the analyte, removal of primary used for decomposing silicatc rocks and minerals). In
interferences, or preconccntration of the analyte. This any acid dissolution method, potential contamination
appendix briefly reviews the most common sample dis- of the sample or standard by impurities in the acid must
solution and separation methods employed with spcc- hc considcred, particularly for trace determinations.
trochemical methods. The exact preparation method to Many mineral samplcs and materials of geological
be used depcnds highly on the nature of the initial sam- interest are solubilized by forming a sol~tblefused salt
ple, the likelihood of potential interferences being prcs- by heating a mixture of an appropriate tlux material
ent, and the characteristics of the specific spcctrochem- and the sample at high temperatures in special inert
ical methods to be employed. The refcrences for this crucibles (e.g., carbon or platinum). Common tlux ma-
appendix should be consulted for specific details. 111 terials are Na2C0, (m.p. X5I0C), Na,O, (dccomposes
addition, a good deal of information can be found in at high temperature), K,S,O, (m. p. 300•‹C),and LiBO,
thc "cookbooks" published by instrument manufactur- (m.p. 840•‹C). The use of fusion techniques for disso-
ers and in various applications-oriented monographs. lution is subject to contamination from the high con-
Sec. E l References 561
centration of the flux material or from the crucible and solution (i.e., lower surface tension and increased vapor
volatilization losses at the high temperatures employed. pressure), which improves the nebulization and desol-
Analysis of organic materials, including biological vation efficiencies and, in some cases, increases in flame
solids, for trace metals usually requires oxidative treat- temperature or excitation efficiency due to chemilu-
ment to convert the carbon and hydrogen to CO, and minescence. Ammonium pyrrolidine dithiocarbamate
H,. Oxidative decompositions are wet-ashing proce- (APDC) is widely used to extract many transition met-
dures, also called digestions, and dry-ashing methods. als simultaneously in conjunction with methyl isobutyl
In wet-ashing methods, the sample is treated with ox- ketone (MIBK) as the solvent.
idative mineral acids such as HNO,, HClO,, and H,SO, Cation, anion, and chelating (e.g., Chelex 100)
or mixtures of these acids at elevated temperatures. ion-exchange resins are often used in separating and
Precipitation and volatilization of analytes, the poten- preconcentrating trace elements. A large volume of the
tial of explosions, and contamination by impurities in sample solution is passed through the resin column and
the acids must be considered. Dry ashing is a very sim- certain ions are selectively retained depending on the
ple procedure, but prone to losses of some trace ele- type of resin. For a cation or chelating exchange resins,
ments (e.g., Hg) by volatilization. In dry ashing the a small volume of a strong acid is then used to elute
organic matter is heated in steps up to = 500•‹C in a the retained metals. Large concentration factors can be
muffle furnace in the presence of atmospheric oxygen achieved because of the high capacity of many resins.
until carbonaceous material has been oxidized to CO,, In certain situations undesired major constituents
after which the sample can be dissolved in dilute acid. of a sample can be separated by precipitation methods.
The use of microwave ovens for dissolution and ashing For example, proteins are often removed from biolog-
procedures is rapidly increasing. ical fluids prior to spectrochemical analysis by precip-
itation with trichloroacetic acid or occasionally sodium
tungstate and centrifugation. Deproteinization can pre-
vent clogging and fouling of nebulizers in plasma and
E-2 SEPARATION flame spectroscopy; the protein-free solution more nearly
AND PRECONCENTRATION matches the viscosity of aqueous standards.
Precipitation reactions can also be used to sepa-
A wide variety of separation techniques, including sol- rate the species of interest by forming an insoluble com-
vent extraction, ion exchange, and precipitation, have pound. The precipitate is then redissolved and ana-
been applied prior to spectrochemical determinations lyzed. For example, calcium and magnesium can be
for preconcentration purposes and for gross separations determined by precipitation with 8-hydroxyquinoline
from major matrix constituents. Solvent extraction followed by determination with atomic spectrometry on
techniques are commonly used to extract the metals of the redissolved precipitate. In some cases, trace metals
interest into an organic solvent with the use of a com- are coprecipitated in the precipitate of a metal added
plexing agent. If the volume of the organic solvent is in excess.
less than that of the aqueous sample and the extraction Many other separation and preconcentration
efficiency is near unity, preconcentration results. methods have been employed with spectrochemical
In flame atomic spectrometry, the organic phase methods. Simple evaporation is an effective but slow
is often directly introduced into the flame and can pro- method for preconcentration of aqueous solutions of
vide significant signal enhancements particularly with low salt content. Dialysis has been used for protein
spray chamber nebulizers. Such enhancements can re- removal particularly in continuous flow determinations
sult from alteration of the physical properties of the of various analytes in biological fluids.
REFERENCES
1. R. Bock, A Handbook of Decomposition Methods in A n - Techniques in Inorganic Analysis. Iliffe Books, London,
alytical Chemistry, translated and revised by I. L. Marr, 1968.
International Textbook, London. 1979. 4. T. C. O'Haver, "Chemical Aspects of Elemental Anal-
2 . T. T. Gorsuch, The Destruction of Organic Matter, Per- ysis," chap. I11 in Trace Analysis: Spectroscopic Methods
gamon Press, Oxford, 1970. for Elements, J . D.Winefordner, ed., Wiley-Interscience,
3. J. Dolezal, P. Povondra, and Z. Sulcek. Decomposition New York, 1976.
App. E I Sample Preparation Methods
The particles (electrons, nuclei) in a molecule or atom for a transition between states i and j, where (J, is the
can be described by the time-independent Schrodinger electric dipole moment operator (IJ, = er), e is the charge,
equation r is the distance between the charges, and d~ indicates
H* = E* the integration over all space. The square of the tran-
'F-l) sition moment or R2 in J cm3 is called the transition
probability.
where $ is the wavefunction. H is the Hamiltonian op-
erator, and E is the energy of a state characterized by We will concern ourselves primarily with electric
+. The product of )I and its complex conjugate $A rep-
dipole transitions involving the first-order interaction
of the oscillating electric vector of the radiation field
resents a probability distribution for the position of a
with the electric dipole moment of an atom or molecule.
particle in an atom or molecule in the state 4.The
Radiative transitions can also arise due to the interac-
Hamiltonian operator is composed of potential energy
tion of the radiation field with electric quadrupole mo-
and kinetic energy operators which can be determined
ments or magnetic dipole moments. These second-order
for any atom or molecule. For atoms or molecules with
effects are extremely weak compared to first-order elec-
only one electron, equation F-1 can be solved to obtain
tric dipole transitions.
analytical functions for the wavefunctions and energy
levels. For most atoms and molecules, such an exact Only transitions between certain energy levels are
probable as expressed by selection rules. Equation F-2
solution is not possible. In this case, approximation
methods (e.g., linear variation methods, perturbation is used to establish selection rules. Often, the symmetry
theory) and numerical methods are used to provide of wavefunctions can be used to show that R is zero for
certain transitions called forbidden transitions. If R is
estimates of the wavefunctions and the energy-level
structure. finite, the transition is an allowed transition. The mag-
nitude of R2 indicates the probability of the transition
The wavefunction can also be used to calculate
and hence the intensity of the transition. Transitions
the transition moment R as shown in the equation
that are dipole forbidden may occur because they are
electric quadrupole or magnetic dipole allowed; these
(F-2) are usually weak in intensity.
4 App. F I Atomic and Molecular Transitions
can be shown to be
3 2 R 2 ~ 3 ~ o r n ,-v 32R2n'q,m,c
,,
fr, = - (F-10)
3e *hg, 3e 2hh,,g,
64n4vl,,K2 - 6 4 d R 2
A,, =
3hc'g, 3hAl,,g, (F-6)
(F-14)
where m, IS the mass of an electron, e IS the charge In
coulombs, and the factor g, is added to account for the
dcge~lcracyok the lower stdte brom this equdtmn and In the literature, the oscillator strength is often tabu-
equations F-3 m d F-6, the cl,~s\icalhnsteln coctf1c1ent5 lated as a weighted value, the "gf" factor. It is related
Sec. F-2 1 Oscillator Strengths 565
to the oscillator strengths for emission and absorption at a given frequency v or the decrease in spectral ir-
by radiance dE, (in W cm-= H z r l ) is given by
Oscillator strength is a convenient way to express tran- where S, is the profile of the line or band in Hz-I
sition strength because it is dimensionless and has a normalized so that J S, dv = 1. With the relationship
nominal range from zero to unity (or a small integer if E, = U,c, equation F-18 becomes
there is degeneracy). Typically, f,,for an electronic
transition from the ground state to the first excited state E, nihvS, db
(f,,), is on the order of unity for a strong spectral line dE, = - Bij - (F- 19)
C
or band. Experimental values of the oscillator strength
can be greater than unity although this is not predicted If equation 3-10 is written in terms of spectral
by the simple model used. irradiance, the decrease in spectral irradiance over
pathlength db is given by
J,S,,
rz, eL
k ( v ) = ---
4~,,rn,(7
TABLE F- 7
Relationships between quantities indicating absorption strength
Absorption cross
section (cn~'),'
Oscillator strength"
v can be considered constant over the absorption band F-4 UNIT CONSIDERATIONS
and equation F-29 reduces to
The units in the equations presented in the appendix
are complex and can be confusing because they vary
among literature sources. This is particularly true for
expressions involving the Einstein B coefficients. It must
be remembered that the product of radiation field in-
where v, is the frequency of maximum absorption. This tensity and the B coefficient must have units of s.-l.
approximation is only good for very narrow lines such Throughout this appendix, we have assumed that the
as observed in electronic atomic spectra or some pure radiation field intensity is expressed in terms of a spec-
rotational spectra, but is often quoted in the literature tral energy density U, in J Hzp1 so that B has
for broad molecular electronic or vibrational absorption units of J-I cm3 Hz s-l. This is true even in expressions
bands. that contain X rather than v such as the second form of
By substituting the expression for the classical equation F-5 (CIA substituted for v). If the spectral en-
transition probability (equation F-7) into equation ergy density is expressed in J cmP3 nm-l, the B coef-
F-30, we obtain an expression for the classical absorp- ficient has units of J P 1 cm3 nm s-l. To convert the
tion coefficient: expressions given into the form appropriate for B coef-
ficients in nm, we use the relationship dX = dv h2/c and
substitute B,c/X2 for B,, where the subscript indicates
the type of units. If the spectral energy density is ex-
pressed in photons Hz-', the B coefficient has
units of cm3 Hz s-l. In this case, the conversion involves
This now allows us to calculate the oscillator strength substituting B,llzv for Be. In some literature, the radia-
from the experimentally determined integrated absorp- tion field intensity is given in terms of spectral irradiance
tion coefficient [h, = K(exp)/E(classical)] or in W ~ m Hz-l, - ~ in which case B has the units
W-I cm2 Hz s-l. To convert to expressions with B
4eo~(exp)rn,c coefficients in these units, BEc is substituted for B,,
f..= where the subscript E denotes irradiance units and U
(I e2ni
denotes energy density units.
Another potential point of confusion has to do
In molecular spectrometry, it is more common to with the units of the charge of an electron (e). Through-
deal with the absorption cross section and particularly, out this book we express e in SI units of coulombs (i.e.,
the molar absorptivity. Using the relationship between e = 1.6 x 10-l9 C). Some authors express e in terms
the absorption coefficient and the absorption cross sec- of electrostatic units (esu), in which case e = 4.80 x
tion or molar absorptivity (equation F-17) and the last 10-lo esu. Expressions containing e2 in SI units can be
few equations, we can derive similar expressions that converted to those in esu (or Gaussian) units by sub-
relate ~ ( v )~, ( v )and
, the integrated molar absorptivity stituting 4ne,e2(esu) for e2(C). Note that h e o =
[Z = J e(v) dv] to the transition probability, Einstein 1.113 x 10-lo C2 N - I m-2 or 1.113 x 10-l9 C2 dyne-'
coefficients, and the oscillator strength. These are sum- cm-2 and that e2(C)/4.rreo= 2.56 x kg m3 s-' or
marized in Table F-1. 2.56 x g cm3 s-'.
lm lumen
L liter
m meter
in01 mole
M moles per liter
N newton
atnn atmosphere P poise (for viscosity)
A ampere rad radian (plane angle)
A angstrom S second
cal calorie sr steradian (solid angle)
cd candela v volt
C coulomb W watt
"C degree4 Celsius (1 ohm
dyn dyne
erg erg
eV electron volt
F fa1 ad g giga 10''
g gram M mega 10"
G gauss k kilo 10'
H henry c centi 1W2
Hz h e r t ~(cycle4 per second) m rnilli 10- '
.I pule micro JO-('
K degrees Kelvin n nano 1OF')
'Note that unit abbreviations arc uppercasc when the unit is narncd
p pico
alter a pcrson. Thc symbol for liter is capitalized to distinguish it t remto 10-'5
from the nurnhcr one. a atto 10-l"
Sec. 5 1 Common Abbreviations 569
=
rewltant total magnctlc quantum number
rewltant orbital magnetic quantum number
NEP noise equivalent power = resultant spin magnetic quantum number
NIR near infrared = prmcipal quantum number
NPS noise power spectrum = electron spin quantum number
op amp operational amplifier = resultant or total \pin quantum number
OPL optical pathlcngth
PA photoacoustic BLECULAR
PAS photoacoustic spectroscopy
PI9 photodiode .I = (rotational) angulai momentum quantum
PMT photomultiplier tube number
PT vacuum phototube S = resultant or total spin quantum number
Q quenchcr u = vibrational quantum number
QLS quasi-elastic light scattering h = quantum number for axial component of the
rms root mean square orbital angular momentum
KIA radioimmunoassay A = total axial electronic angular momentum
RIS resonance ionization spcctroscopy quantum number
RSD relative standard deviation R = total ax131 angular momentum quantum
RTP room-temperature phosphorescence number
Symbols
The symbols listed below are used in two or more chap- B = background signal
ters or several times in at least one chapter. IUPAC B or B = rotational constant B in J or B in
recommendations are used where possible. By necessity cm-'
some symbols have more than one meaning which can B, and B,, = Einstein coefficient for stimulated
be determined from the context. Subscripts and super- absorption between states i and j or
scripts are used with many symbols to designate specific stimulated emission between states j
values. Their common meanings and conventions for and i, cm3 J-I s-l Hz
their use are presented after the lists of Roman and Bi = spectral radiance, W sr-l
Greek symbols. nm-I
m ) = cumulative radiance, W sr-I
B? = spectral radiance for blackbody
ROMAN SYMBOLS radiator, W sr-I nm-I
c = speed of light, m s-I
= absorptivity, L g-I cm-' c = concentration (usually of analyte or
= damping constant or a-parameter for the absorbing species)
Voigt profile, dimensionless C = capacitance, F
= thermal diffusion coefficient, cm- l C = correlation coefficient, dimensionless
= absorbance, dimensionless CF = coefficient of finesse, dimensionless
= area, cm2 c(7) = correlation function
= spectral absorbance. dimensionless d = distance (between a source or
= integral or total absorption, m or Hz scatterer and a detector), spacing
= Einstein coefficient for spontaneous between grating grooves, or
emission between states j and i, s s l thickness of dielectric spacing in an
= pathlength of sample cell or length interference filter, cm
of base of prism, cm = modulation depth, dimensionless
= radiance or line radiance, = major dimension of a scatterer, cm
W sr-' = detectivity, W p l
Symbols
dimensionless v = -
Analytical sample, 3 source modulation. 273, 280, 282 signal and readout exprcssions. 19-20,
Analytic;~lsensitivity. 171-72 transtluccrs lor. 274. 294-95 25, 244-45
Analytical signal. 24 wavclength modulation, 295-96 signal-to-noise exprcssions and
in absorption rncasuremcnts, 25-27 Atomic absorption spcctrophotometry, optimization, 245-47
in emission measurcmcnts. 24--25 273-306 (Sea rrlso Flame atomic wavelength identilication, 252, 260-70
in luminescence measurcmcnts, 25 itbsorplion spectrc)photometry; Atomic fluoresccncc, 21. 219-20, 307-23
Analyzer (Src, Automated analy~crs; Electrothermal atomic absorption lifetimes, 220
Polarizcrs) s~~ectro~hotomctry) powcr yield of, 220-21, 314-15, 117
Angular dispersion, 63, 00-69 absorptancc, 217-19 quantuln eflicicncy ol, 220
Angular morncntun~,202-6. 320, 330 (Sw accuracy, 298-99 types of transitions, 307-0
iilso O u a n t u ~ nnumber\) ;ipplications, 303-4 Atomic fluoresccncc spectrometers, 309-13
Anhanmonicity constant, 327 background ahsorption correction, atomizers, 311- 12
Anharmonic oscillatnr, 327-28 288-94 Mock diagram. 309
Anisotropic crystals, 40, 43, (14, 106 characteristic concentrations, 299-301 optimiration, 318- 19
Anisotropy (of fluoresccncc), 348-49. dctection limits, 299-301 ritdiation sources for. 310- 1l
489-90 dependence o n conccntrntion, 217- 19, types of, 312- 13
Antireflection coatings, 53, 550-51 222 Atomic fluorcscence ycctromctry, 307-24
n-parameter, 214, 217 interferences, 199-202 analytical applications, 321. 522-23
Apcrture stop. 53, 443 lincxity, 296-97 background signals, 318- 19
Arc and spark atomic emission modulation signals, 282 curves of growth, 210-22
s ~ x c t r o n ~ e t c r207-08
s, performance charactcristics, 296-301 dctcction limits, 320-21
Arc and spark atomic emission precision, 298 intcrfcrences, 199-202
spcctrometcry, 257-72 sclcction of expcrimcntal conditions, 297. linearity, 320
applications, 270-71 30 1 localized diagnostics, 32 1-22
tletcction limits. 269 signal cxpressions, 217-1") 280-83 perlorrnance characteristics, 320-21
methodology, 269-70 signal-to-noise ratio exprcssions and primary absorption cffccts, 315
performance characteristics. 268-09 optimization. 283-86 resonance detection. 322
Arc emission (excitation) sources, 191. stray radiation. 297 quenching, 202
258-61 Atomic cmission, 1S-20. 214- 17 saturation, 316-18
ac and intermittent, 200-61 Atomic cmission (cxcitation) sources: secondary absorption effects, 315-16
anode and cathode fall, 258 arc dischargcs, 258-61 self-absorption, 314- 16
as atomizers for other emissio~isources, direct-current plasma\. 238-39 signal expression, 3 13- 18
191, 261 exploding thin films, 367 signa-to-noisc ratios, 3 18- 19
controlled atmosphere dc. 260 flnmcs, 227-33 A t o n m ionization, 196-98. 532-36
clcctrodcs, 258-60 hollow cathode discharge. 266-67 Atomic number density, 196-97, 245, 275,
free-burning dc, 258-60 idcal charactcristics of, 226-27, 233, 236, 282-83, 315- 17
sclcctive volatilization. 259 261 Atomic orbitals (AO), 202, 329-30
spectrochemical buffer. 260 inductively coupled plasma, 233-37 Atomic spcctra. 7, 205-7, 307-9
Arc lamps. "3-95, 410, 429. 441-38, 478 laser-induced breakdown. 266 Atomic spectroscopy. 2, 189-324, 532-36
Arc lines, 258 lascl- microprobe, 265-60 (See ulso specific atomic
Arc wander: low-lxcssure dischargcs (Grirnm glow spectrometric techniques)
in arc lamps. 441. 447-48 tlischarge), 266-67 analytc interferences, 199-202
in dc arcs, 260 microwave plasmas. 237-38 blank intcrferenccs, 198-99
Aspiration (Sw Nchulizers) spark discharge, 2 6 - 6 5 curves of growth, 197-98
Astigmatism. 56-57, 75 theta pinch discharge, 267 ideal systcrn, 220-27
Atomic absorpt~on,217-19, 273-306 Atomic emission spcctromcters, 239-44, sample introduction and atomization,
Atomic absorption spcctrophotonicters, 267-68 189-98
287-96 background correction, 242-43. 249-50 Atomic transitions, 7, 209-10
automated sequential measurcrncnts, 295 computcn control, 244. 250 Atomization, 189-92, 275
background absorption corrcction, entrance optics, 240 Atorniration efficiency, 196-97, 232-33
288-94 filter-based, 240, 254 Atomization proccsscs, 101-92
block diagrxn. 274 modulation techniques. 244 Atomizers or atomization deviccs, 14, 189.
burncrs, 274-75 monochromators for, 240-41, 240 190-97
continuum source bascd, 295-96 rnultielemcnt type. 241-43 flanies, 227-33, 274-75, 31 1
curve correction. 287 photographic detection, 267-68 free aton1 number density, 196-97
data presentation, 287 scquent~almeasurements, 240-41 clcctrotherrnal, 275-78, 31 1- 12
direct concentration readout, 287 single channel, 240 electrical dischargcs, 258-67 [ S w i d ~ o
double-beam, 288 transducer and electronic components, Atomic cmission (excitation)
electrothermal atomizers, 275-78. 302-3 243-44 sources]
flame types, 274-75, 301-2 wnvclcngth selection, 239-40 ideal charactcristics for AAS, 274
ideal atomize~-s,274 Atomic cmiss~onspectrometry, 19-21), ideal characteristics lor AES, 226-27
microcomputcr use, 287. 295 225-56 ( S w ulso Arc and spark idcal charactcristics for AFS, 3 11
monochromator, criter~afor, 274 atomic cmission spcctrometr!~; Flame laser microprobc, 191
rnultielemcnt, 294-96 atomic emission spectrometry; o r plasmas, 233-39, 31 1
peak mode, 287 Plasma atomic emission samplc introduction into, 189-95
radiation sources lor, 273-74. 288, spcclrometry) temperature of, 227
2% - 97 applications, 253-55. 270-71 Attenuated total rcflcctancc (ATR),
with resonance detectors, 322 curves of gl-owth, 214- 17. 222 432-34
samplc introduction methods. 278-80, interferences, 198-202. 249-50, 200 Aufbau order, 203
288 principles, 10-20 Autocorrclatmn iunct~on.158
\ample motlulation, 278 scll-absorption and reversal, 215-17. A u t o c o ~ ~ e l , r t ~tcchmqucs
on lor hght
single-beam, 273-74 221. 247. 261, 293, 310, 314-16 scattcnng me,~wremcnts,520
Index 579
continuous-wave (CW), 99, 515, 518, Littrow mounting, 75-76 Metal vapor lamps, 310
521, 526, 529-30, 537 Lock-in amplifier, 125, 526 Metastable states, 206
diode, 104-5, 416-17 Lorentz broadening, 211 Micelles, 474-77
dye, 102-4, 320, 441, 503, 529-30, Lorentzian orofile. 210-11. 520 Michelson interferometer, 81-83, 413-14
532-33, 537-39 Low-angle iaser light scatte'ring (LALLS), Microarc vaporizer, 261
excimer, 102-3 518-19 Microchannel plate, 116
frequency doubling of, 106 Low-pass filter, 124, 139-41, 158 Microcomputer instrumentation, 15, 182,
gain of, 94 LS coupling, 203-5 244 (See also under specific
gas, 101-2 ~uminescence,5 (See also Chemilumin- spectrometers)
helium-neon, 101, 414, 502, 518 escence; Molecular fluorescence; Microphotometer or densitometer, 114-15,
krypton ion, 502-3 Phosphorescence; Photolumin- 268, 270
metal vapor, 102 escence) Microscope:
mode locked, 100, 486 decay of, 342, 485-89 interference, 83
mode structure of, 100 infrared, 434 IR, 420
Nd:YAG, 101 polarization, 348-49 Raman, 505-6
nitrogen, 102, 441 sensitized, 344, 467, 479 Microwave-induced plasma [See Plasma
operational modes of, 99-100 signal expressions, 21, 25, 131-32, emission (excitation) sources]
population inversion in, 98-99 450-53 Microwave vaporizer, 261
principles of operation, 98-99 types, 21, 428 Mid-infrared region, 404
pulsed, 99, 320, 441, 486, 522-23, 526, Luminescence lifetimes, 342-43, 485-89 Mie scattering, 22, 497-98, 5~
532, 533 Luminescence power yield, 340-41 Mirrors, 48-51
Q-switched, 99- 100 Luminescence quantum efficiency, 339-41, beam splitting, 57-58
ruby, 100-101 344-48 Cassegrain, 312
saturation of transitions with, 316-18, dependence on environment, 347-48 dichroic, 58-59, 63
458, 539-41 dependence on structure, 344-47 formula for, 48-49
semiconductor, 104-5 determination, 466-67 interferometer. 80. 412-15
solid state, 100-101 expressions for, 340-41 laser, 98-99
spin-flip Raman, 417 Luminescence spectra, 338-51, 444-46 reflectance properties, 57, 551-52
synchronously pumped, 104 Luminescence spectrometers (See Modulation and modulation techniques,
use in AFS, 310-11, 320-21 Photoluminescence spectrometers;
use in FTIR spectrometers, 414 Chemiluminescence: Phosohor- in AAS, 273, 280, 282
use in ionization techniques, 532-38 escence spectrometers; Atomic advantages, 156-57
use in I R spectrophotometry, 414, 417 fluorescence spectrometers; or in AES, 244, 246
use in molecular fluorescence Molecular fluorescence spectrom- in AFS, 312
spectrometry, 441 eters) amplitude, 119
use in PAS, 526 Luminescence spectrometry or demodulation, 119, 125, 157-58
use in Raman spectrometry, 502-3, 508, spectroscopy, 21 (See also frequency, 119
511-13 Chemiluminescence; Phosphor- in molecular luminescence, 444-46
use in remote sensing, 522-23 escence spectrometry; Atomic phase, 486-88
use in scattering methods, 515-22 fluorescence spectrometry; or sample, 120, 246, 278
use in thermal lensing, 529-30 Molecular fluorescence spec- for selectivity, 180-81
Laser scattering methods, 515-22 [See also trometry) for signal-to-noise ratio enhancement,
Low-angle laser light scattering principles, 21, 338-51, 438-90 156-58
(LALLS) or Brillouin scattering] signal-to noise expressions, 146-50, source, 119-20, 180, 280, 312
Lateral diffusion interference, 201, 299 318-19, 459-60 in UV-visible spectrophotometry, 360-63
Lenses, 51-53, 410, 453 types, 21, 438 wavelength, 120-21, 181, 246, 295-96,
Lensmakers' formula, 51 L'vov platform, 278 312r445-46
Lens stop (aperture), 53 Modulators, 44-47
Lidar (laser radar), 522-23 Molar absorptivity, 35, 565-66
Lifetime broadening, 209-10 determination of, 385
Lifetimes: integrated value of, 337-38, 566
of excited atoms, 209-11 Molar ratio method, 386
of excited molecules, 342-43, 485-87 Mach-Zender interferometer, 83 Molecular band emission, 199
measurement of, 485-89 Macromolecules, scattering by, 495, 497, Molecular beams, 538-39
radiative or natural, 89, 210, 342 515-20 Molecular fluorescence. 339
Light (See Radiation) Magneto-optic modulators, 45-46 delayed, 339
Light emitting diodes (LED), 96, 417 Magnification, 50, 52, 54 lifetimes. 342-43. 349, 486-88
Light scattering photometry, 515-18 Malus's law, 42
molecular weight determination, 515-18 Masking agent, 177 prompt, 335
Rayleigh ratio, 516-18 Matrix, 3 quantum efficiency, 340-41, 344-48,
Zimm plot, 517 Matrix effect or error, 5, 165-67 439. 458
Limiting aperture, 53-54, 70 Matrix isolation, 471 quenching of. 343-44
Limit of detection (See Detection limit) Matrix match method, 177, 302 sensitized, 344
Linear birefringence, 42 Matrix modifiers, 177, 303 soecies exhibiting fluorescence, 467-69
Linear dichroism, 42 Maxwell's law, 211-12 spectra, 444-46,"463
Linear dispersion, 69 Mean, arithmetic, 135, 168-70, 173-75, Molecular fluorescence spectrometers:
Linear polarization, 35, 39-40, 42 544-48 block diagram, 439
Linear wavelength drive, 68-69, 505 Measurement, 13-15, 24-27 computerizaton, 443-44
Linear wavenumber drive, 69, 505 Meker burner, 231 corrected, 446-49, 457-58
Line profiles (See Spectral line profiles and Mercury arc lamps, 94-95, 410, 429, 441 double-beam type, 447-48
widths) Mercury vapor (See Cold vapor mercury filter type (fluorometers), 440
Line spectra, 27 technique) geometry, 443, 452
Index
Raman shifters. 106. 512 Resolution. 71 -72 (See also elastic; 494-97
Raman spectra. 501-2 Monochromators: Interferometers) principles, 494-99
Random errors. 168-70. 543-49 Resolving power. 72-73. 79-80 (See also Scattering. as interference, 199, 318-19.
calibration curve uncertainty. 170 Monochromators; Interferometers) 462. 465, 477
causes of. 168 Resonance detector, 322 Scattering factor. 517
statistical statements. 169-70. 543-49 Resonance fluorescence, 219-20. 307-8. Scattering methods, 21-22. 494-524 (See
Rapid scanning monochromators and 495 also Elastic scattering: Laser
spectrometers, 23. 129 Resonance ionization, 535-36 scattering methods; Low-angle laser
Rate constants: Resonance line. 7, 216-17, 219-20. light scattering; Nephelometry;
for transitions in atoms, 209-11 252-53 Quasi-elastic or dynamic light
for transitions in molecules. 341-44. 458 Resonance Raman spectroscopy, 508-9 scattering: Raman scattering;
Rayleigh criterion. 72 Resoonse function. 8 Turbidity)
Rayleigh ratio. 516-18 Reverse biasing of pn junction. 113-14. Scattering signal. 25
Rayleigh scattering. 21. 463, 495-99. 115-17 Schrodinger equation. 326, 563
516- 17 Rhombs (rhomboids); 44 Secondary absorption effects and
Reaction-rate methods, 181-82. 391-94. Rigid rotor model. 326 correction, 315-16, 454-58, 449, 465
469 rms noise, 136 Secondary combustion zone, 228
Readout: Rotating sector method, 268 Secondary emission. in PMT; 110-11. 143
in AAS. 274. 288-94 Rotational constant. 326 Selection rules. 7, 563
in AES, 240-44 Rotational correlation time, 348-49 for atomic transitions, 205-7
in AFS, 309, 312-13 Rotational diffusion, 348-49. 489 for molecular electronic transitions.
in molecular fluorescence spectrometry. Rotational overlap integral. 332 332-34, 335-36
443-44 Rotational relaxation time. 348-49 for Raman scattering. 500-501
in UV-visible spectrophotometry. 360-64 Rotational transitions and spectra, 7. for rotational transitions. 326
Readout devices. 14-15. 128 326-27 for vibrational transitions, 328, 333, 405
Readout expressions. 21, 25-27, 131-32 Rotation-vibrational spectra. 328-29 Selective volatilization. 259-60
(See also under specific techniques) Rowland circle mount, 76-78. 241 Selectivity, 10, 180-82
Readout system, 117-18. 128 Ruby laser, 100-101 Self-absorption, 215-17, 221, 247, 261,
Reciprocal linear dispersion. 68-72 293. 310, 314-16, 456
Reduced mass, 211, 327, 407 Self-reversal, 216. 261. 293. 296. 310
Reduced pressure discharges, 266-67 Semiconductor lasers, 104-5
Reference signal. 26-27. 132. 151-53 Sensitivity, 10, 171-72
Reflectance. 31-34 Sensitized chemiluminescence. 479
for common mirror materials. 551-53 Sagittal focus. 56-57. 75 Sensitized luminescence. 307-9. 344
of interferometer mirrors, 79-80 Sagnac interferometer. 83-84 Separation techniques. to minimize
Reflectance spectrometry: Saha equation. 196. 259 interferences, 176, 561
A T R in the I R region. 432-34 Sample acquisition, 168 Shot noise. 141-44
I R region. 429. 432-34 Sample and hold circuit. 127 Shpol'skii effect; 470
UV-visible region. 398 Sample cells: Signal:
Reflection, 4. 31-34 for fluorescence spectrometry. 442, ac. 137
diffuse. 32-33 463 - 64 analog, 118
multiple. 380 for I R spectrophotometry. 417-20. 429 background, 24
specular (law). 31-32 for phosphorescence spectrometry. bandlimited. 137
total internal, 33-34, 432-34 473-74 blank. 24-25
Reflection gratings, 64-67 radiation losses. 365-68, 372 dark. 24-27. 107-8, 131-32
Reflection losses, 33 (See also Fresnel for Raman spectrometry. 503-4 dc. 137
equation) for UV-visible and NIR digital. 118
with anti-reflection coating. 550-51 spectrophotometry. 279-80. 357-60. expressions. 143-44
at common optical interfaces. 550-52 427 frequency content of. 137-38
in molecular fluorescence spectrometry. Sample containers, 6. 24 mean of, 135
450. 452, 454 Sample dissolution, 560-61 reference, 26-27. 132
in UV-visible spectrophotometry. Sample introduction systems. 14, 189-98, rms, 136
365-66, 372, 380 278-80 (See also Sample cells; total (spectrochemical). 25
Refraction, 4. 31-34, 51 Nebulizers: Automated analyzers) Signal averaging (See Signal-to-noise ratio
law of (Snell's law). 32 for atomic spectrometry. 189-98. 227, enhancement)
Refractive index. 31-33, 40-41. 43. 62-64. 231-32. 234-35. 238-39. 261-63. Signal conditioning and circuits, 121-27
80-81, 520-21, 530-31 278-80 Signal processing. 14, 117-28 (See also
changes in UV-visible for molecular spectrometry. 396-98 Signal conditioning and circuits)
spectrophotometry. 373 Sample preparation. 168. 417-20. 560-62 with analog electronics, 118
of common optical materials. 550-51 Sample presentation. 13-14 analog-to-digital conversion. 118.
effects in luminescence spectrometry, 455 Sample types. 3 123-27, 243-44
extraordinary and ordinary, 43. 550-51 Saturated fluorescence: computer-based. 127. 287. 360. 443-44
of fiber optics. 59-60 in atomic spectrometry. 316-18 demodulation. 119, 125-26
Refractor plate, 121. 240. 242-43 in molecular spectrometry, 458 digital, 127-28
Relative aperture. 54 Saturation broadening, 212 digital-to-analog conversion, 127
Relative standard deviation. 8. 138. 249. Saturation method. 176-77 filtering (See Electronic filters)
269. 544 Saturation spectroscopy. 539-41 mathematical operations, 123-24
of absorbance, 150-54 Scattering. 4. 19. 21-22. 43 (See also modulation, 25. 119-21, 156-58, 282,
Remote sensing: Scattering methods; Brillouin 312, 360-63. 444 (See also
with fiber optics, 472-73 scattering; Rayleigh scattering; or Modulation and modulation
with lasers; 522-23 Raman scattering) techniques)
588 Index
Stimulated Raman gain (SRG), 511-12 evanescent wave, 433 scanning, 357, 362-63
Stimulated Raman scattering- (SRS), penetration depth, 433 single-beam type, 353-57, 360. 362-63.
511-12 Transducer (See Radiation transducers) 382-83
Stoichiometry, determination of, 386-87, Transfer functions: source modulation, 369
466 amplitude, 138-39 sources, 354
Stray radiation, 74 transducer, 127 specifications. 383
in AAS, 297-98 Transient recorder, 127 spectral bandpass selection, 375-77,
in AES. 249 Transition cross section, 35 381-83
in IR spectrophotometry, 423-24 Transition moments and probability, 89, temperature control. 358-59
in molecular fluorescence spectrometry, 332. 338, 40.5: 500-501, 563-67 wavelength selection, 375-76, 381-83
442 Transitions (See Spectral transitions: UV-visible spectrophotometry, 352-403
in Raman spectroscopy, 504 Selection rules) applications. 386-99
in UV-visible spectrophotometry, 357, Transmission, 4 automated measurements, 398
376-79 Transmittance, 20, 26-27, 31, 35 chromatrographic detection, 396-98
Student t statistic, 169-70. 545-48 for atomic absorption. 280-82 difference soectra. 394
Superposition. principle of, 35 of common optical materials, 551-53 dual-wavelength measurements, 394.
Surface enhanced Raman spectroscopy, internal, 367 396
510-11 for molecular absorption. 352. 360-61, molecular weight determination, 387
Synchronous demodulation. 125 366 multicomponent analysis, 390
Synchronous spectra, 444-46 Triboluminescence, 21 stoichiometry, 386-87
Systematic or determinate errors, 165-68, Triplet state, 203, 338-39, 342-47 reaction-rate methods. 391-94
543 t-statistic, 169, 174-75, 545-48 titrations, 390-91
due to calibration, 167-68 Tuned amplifier, 124-25 detection limits, 384
due to matrix (interferences), 165-67 Tungsten lamp. 90, 92-94, 354, 397, 410, interferences, 384-85
due to sample acquisition. preparation, 427 methodology. 381-85
and measurement, 168 Turbidimetry, 19, 22. 513-15 nonlinearity, causes of, 372-81
Symbols, 571-77 Turbidity, 514 chemical equilibria. 372-73
Turbulent flames! 232 circular dichroism. 380
Two-photon absorption, 465 fluorescence, 380
Two-photon excited fluorescence. 308-9, multiple reflections, 380
465 polychromatic radiation, 373-77
Two-photon ionization. 532! 535-38 refractive index changes, 373
Tangential focus, 56-57. 75 Tyndall effect, 513 stray radiation. 376-79
Tantalum boat, 279 variable pathlength. 379-80
Temperature: nonzero intercept, 372
in atomizers, 196-97, 227, 229, 235, performance characteristics, 383-86
238-39, 259, 264, 275-78 reflective losses, 365-67. 372, 386
control in spectrophotometers, 358-59 signal readout expressions, 365-67
effect on luminescence characteristics. Ultrasonic nebulizers. 193-94, 231 signal-to-noise ratio expressions. and
347, 462, 470, 473-74 Ultraviolet (UV) region, 2 optimization, 367-72. 376-79
Term symbols: Uncertainty in measurement, 8: 165-70, stray radiation, 376-79
for atoms, 203-5 543-49 wavelength calibration. 382-83
for molecules; 330-31 propagation of, 8, 548-49
Thermal absorption coefficient, 527 statistical statements, 169-70, 543-48
Thermal conductivity, 531 Units, tables of, 567-68
Thermal diffusion length, 527-28 UV-visible spectrophotometers, 352-65
Thermal equilibrium. 19, 87 block diagrams, 353
Thermal gradient lamp, 97 chromatographic detectors, 359-60 Vacuum phototube, 109-110, 360
Thermal lens effect (thermal blooming). computer-based, 360. 362-65 Vaporization (See volatilization)
529 concentration readout, 364 Variance, 136
Thermal lensing spectrometry, 529-32 derivative mode, 364, 394-95 Velocity. of species in atomizers. 211-12
Thermistor. 108-9 detectors, 360 Vibrational coupling, 408
Thermocouple. 108, 410-14 difference spectra, 394 Vibrational energy levels: 327
Thermoluminescence, 21 diode array, 354, 357-58, 360, 367, 397 Vibrational overlap interval, 332
Thermopile, 108 direct absorbance readout, 360-61; 369 Vibrational relaxation. 338-39
Theta pinch discharge, 267 display options, 363-64 Vibrational spectra, 7, 327-29
Thin lens approximation, 51 double-beam type, 353-57, 362-63, half-widths, 328
Throughput factors, 73-74, 425-26 382-83 normal modes, 328, 405-7
Time constant, 140 dual-wavelength mode, 364, 369. P, Q, and R branches. 328-29
Time-correlated single photon method, 394-96 Vibrational transitions, 7! 327-29
486-87 filter-based (photometers), 352-55. anharmonic oscillator model, 328
Time discrimination (resolution), 23, 180- 364-65 harmonic oscillator model, 327-28
81, 264-65, 450-51, 488-89, 508 fixed-wavelength, 361-62 Morse potential well for, 328
Time-division multiplexing, 294, 312-13 near-IR measurements, 360, 427-28 overtones, 328, 405-6. 408
Time domain, 14 rapid scanning. 357 potential energy, 328
Time-domain filtering. 159-60 ratio-recording readout, 363 selection rules, 328. 333. 405
Titrations, 182. 390-91. 471, 483, 515 readout scales, 360-63 Video fluorometer, 444-45
T-number, 54 reference signal adjustment, 360-61 Vidicon tube. 116, 443
Total absorption (See Integral absorption) reflectance attachments, 398 Vignetting, 53
Total consumption burner, 193, 232 sample cells, 358-60, 379. 427 Virtual level, 308. 495
Total internal reflection, 33-34, 432-34 sample compartment, 357-58 Voigt integral and profile, 213-15
critical angle, 33-34 scale expansion, 371-72 Volatilization efficiency, 196-97
590 Index