C H A P T E R 2 1
Special Instrumental Methods
Strictly speaking, there is no such thing as a
non-instrumental method, since even ord-
inary volumetrie analysis depends ultimately
on use of an analytieal balance for standard-
ization. This chapter will be restricted to
explanations of the functions in analytical
chemistry of several instruments that are
"special" in that they are not apart of the
common equipment of analyticallaboratories.
These instruments differ in relative availability
in rough proportion to their cost. For in-
stance, flame photometers, polarimeters and
less expensive types of fluorometers are found
in quite a few food laboratories, while
mass spectrometers and nuclear magnetic
resonance devices are uncommon. In modern
times, absorption spectrophotometers for the
visual-uItraviolet range such as the Beckman
DU have co me into almost universal use, so
we feel there is no need to discuss these
Instruments for Qualitative and
Quantitative Analysis of Mixtures
Atomic Absorption
Spectrophotometers
It has been pointed out! that atomie
absorption spectrophotometry "is identieal
459
F. L. Hart et al., Modern Food Analysis
© Springer-Verlag New York Inc. 1971
instruments in this chapter.
The 15 instruments whose usefulness for
partieular types of analytieal problems we
shall attempt to explain can be divided
roughly into two categories: 1) those that are
primarily used for qualitative and quantita-
tive analysis of mixtures of compounds; and
2) those that are primarily used for identi-
fying single pure compounds. It should be
emphasized that this division is a very rough
one; for example, while the infrared spectro-
photometer is almost uniquely valuable in its
ability to identify an organic compound and
the groups that it contains, there are numerous
references to its use in quantitative analysis of
mixtures of several compounds (more often,
however, in the drug than in the food field).
Nevertheless, the classification is a useful
one, and we have employed it here.
in principle with solution absorption spectro-
photometry except that the sampie to be
examined is in the vapor phase, produced
and maintained by a flame." Atomic absorp-
tion instruments find their practical use,
460] Special Instrumental Methods
however, as do flame photometers, primarily
as cheaper substitutes for the emission spect-
trograph.
Introduction of the atomic absorption
spectrophotometer by Walsh in Australia in
1957 2 was rapidly followed by the develop-
ment of commercial instruments, and this
technique has become quite popular in the
years that have elapsed since that time. Like
the flame photometer, a solution of the
sam pie is sprayed into a flame where it is
heated to incandescence. It differs from the
flame photometer in this respect: in the
flame photometer, the emitted light of a
particular wavelength, which comes only
from the activated (ionized) atoms of an
element, is measured; in the atomic absorp-
tion spectrophotometer, the vaporized union-
ized atoms absorb some of the light emitted
from a hollow cathode lamp whose cathode
gives off light of the resonance frequency of a
particular element, and it is the amount of
this absorption that is measured. One of the
claims for the atomic absorption instrument
is that because only a minor proportion of
the vaporized atoms of an element is ionized,
atomic absorption spectrophotometry is more
sensitive than flame photometry. Claims for
superiority over emission spectrography are
also made on the basis that instrument read-
ings can be made direct1y proportional to
concentration. On the other hand, deter-
minations of many elements can be made
simultaneously with an emission spectro-
graph (and it is not even necessary to have
prior knowledge of what elements are pres-
ent), while the atomic absorption spectro-
photometer requires aseparate hollow cath-
ode lamp, and a separate determination, for
each element (or at most each pair of ele-
ments) estimated.
As a practical matter: 1) the flame photo-
meter (which is the cheapest of the three
instruments) finds its chief use, and is most
efficient, in the determination of the alkali
metals Na, K, and Li; 2) the atomic absorp-
tion spectrophotometer, because its cost is
one-fifth as much or even less, is within the
means of more laboratories than the emission
spectrograph, and in addition is a more
sensitive means of determining certain ele-
ments such as zinc; and 3) as noted above,
the emission spectrograph can determine the
widest range of elements in the shortest time,
without prior knowledge of what elements are
present. Under normal conditions only
metallic elements can be determined by any
of these instruments, and none determines
how the element is combined in the sampie.
Method 6-24 is an example of an atomic
absorption method.
Manufacturers of atomic absorption spec-
trophotometers include the following:
Aztec Instruments, Inc., 2 Railroad Place,
Westport Conn.
Instrumentation Laboratory, Inc., 113
HartweIl Ave., Lexington, Mass.
Jarrell-Ash., 590 Lincoln St., Waltham,
Mass.
Perkin-Elmer Corp., 702G Main Ave.,
Norwalk, Conn.
Emission Spectographs
The emission spectograph consists of a
power source to heat the sam pIe to incan-
descence, a device (either a prism or a grating)
to break up the spectrum of the emitted
light, and a means of recording the spectrum
(either a photographie plate or film or a
bank of photocells). Grating spectrographs
have largely replaced prism instruments.
Emission spectrographs find their chief use
in food analysis either in the quantitative
analysis of plant materials for metallic ele-
ments or in semi-quantitative tests for toxic
elements such as arsenic, lead, beryllium and
thallium. For such purposes the sampie is
usually ashed, taken up in dilute acid, and
applied to a crater in a carbon or graphite
electrode. An are or spark is then formed
between this electrode and another of the
same material, and the spectrum of the
 Instruments for Qualitative and Quantitative Analysis of Mixtures [461
burning sam pIe is recorded and measured.
In the metal industries, automatic recording
spectrographs employing photocells are used
to measure the intensity of the chosen line in
each element being determined, but such
devices are quite expensive, and in food
analysis the cheaper instruments in which the
spectrum is recorded on a photographic
plate or film are still in use. In photographic
recording, after development the film (or
plate) is placed in a densitometer which
first locates the lines corresponding to the
various elements present and then measures
their intensities. For semi-quantitative trace
element detection, a d-c arc of 5-30 ampere
current is the preferred means of excitation
because it is the most sensitive, but in quanti-
tative analysis the a-c spark is employed
because it yields more uniform spectra.
For a more extended discussion of the
details of emission spectography the reader is
referred to the artic1e by Scribner and
Margoshes in Chapter 64 of Part I, Volume 6
of Kolthoff, Elving and Sandell's Treatise on
Analytical Chemistry (New York-London-
Sydney: Interscience Publishers, 1965). Two
things should be noted here, however: I)
The densities of the spectrallines obtained in
emission spectography are not strictly pro-
portional to the concentrations of their
respective elements; and 2), because it is
nearly impossible to take two "shots" under
completely identical conditions, it is the
common practice in quantitative analysis to
correct for small variations in burning, etc.,
by adding a uniform amount of an element
such as cobalt (whieh is not presen~ in
measurable amount in the sam pIe) to both
sampIe and standards. The ratio of the den-
sity of the line of the element being deter-
mined to the density of a chosen cobalt line is
then what is calculated, and graphs are con-
structed relating these density ratios to con-
centrations of the elements in question.
Examples of emission spectrographic
methods especially designed for plant and
food analysis are given in Text Reference 3;
note also particularly Selected Reference W.
The method of the late Waddy Mathis was
worked out in the laboratory of one of the
authors of this book, and is still in use there.
Both of these methods (with slight modifi-
cation) have been adopted by the AOAC
(AOAC Methods 41.001-41.016).
Manufactures of emission spectrographs
inc1ude:
Applied Research Laboratories, Inc., 3717
Park Place, Glendale, Calif.
Baird-Atomic, 33 University Road, Cam-
bridge, Mass. and Jarrell-Ash Co., 590
Lincoln St., Waltham, Mass.
Spex Industries, Inc., 3880 Park Ave.,
Edison, N.J., is a source of spectrographic
supplies; graphite electrodes are available
from National Spectrographic Laboratories,
Inc., 6300 Euclid Ave., Cleveland, Ohio, and
Union Carbide Corp., Carbon Products Div.,
270 Park Ave., New York, N.Y. Extremely
high purity elements and their salts for
standardization, manufactured by Johnson-
Matthey Co., London, England, are available
from Jarrell-Ash Co.
Flame Photometers
Flame photometry is essentially emission
spectrography using as excitation source a
flame rather than the higher-temperature
eleetric are or spark. As a matter of fact,
Lundegardh wrote two books describing
results obtained with an ordinary spectro-
graph equipped with an acetylene-air flame
source. 4 However, the modern-day flame
photometer, besides being a much less ex-
pensive device, in our opinion is most useful
just when the temperature of its flame, e.g.,
propane-air, is not high enough to excite most
elements, and it consequently can determine
the alkali metals with minimum interference.
It is possible, by using higher-temperature
flames (hydrogen-oxygen at 2900 0 K or
cyanogen-oxygen at 4800 0 K s), and separating
462] Special Instrumental Methods
the emission lines with a monochromator, to
adapt the flame photometer to the deter-
mination of numerous other elements, but
only at the price of loss in simplicity and
freedom from interference, and increase in
cost.
An example of the determination of sodium
and potassium by flame photometry is
Method 1-17.
Relatively inexpensive flame photometers
that yield accurate results for Na and K
(particularly when using a lithium internal
standard) are available from:
Advanced Instruments, Inc., 45 Kenneth
St., Newton Highlands, Mass.
Coleman Instruments Corp., 42 West
Madison St., Maywood, Ill. and
Jarrell-Ash Co. (See above).
Fluorometers
Disregarding the cause of fluorescence, the
visually observed phenomenon is that when
a substance is illuminated with light of one
wavelength, it emits light of another wave-
length; if the emitted light persists after the
illumination ceases, the substance is said to
phosphoresce. These phenomena have been
explained as being due to the release of energy
in the form of light by an excited moleeule
when it reverts to the ground state. A co m-
plete explanation would require a knowledge
of quantum mechanics, but an understanding
of the theory of fluorescence is unnecessary
for its use as an analytical too1.
In principle, a fluoro meter is a device in
wh ich a solution of the substance being
analyzed is illuminated by an ultraviolet
light source of fixed wavelength, and the
intensity of the (usually visible) light emitted
by the irradiated solution is measured. The
incident radiation of the source must, of
course, be separated from the light emitted
by the sampie ; this can most readily be done
by measuring the light from the sam pie at a
right angle to the be am of light from the
iIluminating ultraviolet lamp. For accurate
analysis it is necessary to ensure, by the use
either of filters or of prisms or gratings, that
both the incident and fluorescent light are
monochromatic.
Fluorometric methods are valuable where
they are applicable (i.e., when the compound
being determined is fluorescent or can be
converted to a fluorescent derivative) because
they are usually highly specific and about 10
times as sensitive as absorption methods; the
calibration curves are also usually linear
over a 50-fold concentration range. There
are pitfalls, however; for instance, other
compounds present in a solution of a sam pie
can quench the fluorescence of the fluores-
cing ingredient, and a practical method must
be so designed as to avoid such interferences.
The better grade fluorometers use grat-
ings or prisms and a recorder of some sort to
separate and record the fluorescent spectrum
of the substance being measured; when
this is done, a fluorescence spectrum is
obtained, frequently a mirror image of the
absorption spectrum, wh ich serves to charac-
terize the substance in addition to deter-
mining how much of it is present. 6
Examples of fluorometric methods of food
analysis are those of Watkinson for selenium, 7
McFariand for diethylstilbestrol in meat,8 and
Roberts for cholesterol in egg noodles. 9
Fluorometers of the quality needed for
such methods may be obtained from:
American Instrument Co., Inc., 8030
Georgia Ave., Silver Spring, Md.
Baird-Atomic, 33 University Road, Ca m-
bridge, Mass. and
Farrand Optical Co., Inc., 535-A South
Fifth Ave., Mt. Vernon, N.Y.
Gas Chromatographs
It can be questioned whether gas chroma-
tography is an "instrumental" method of
analysis in the same sense as are the deter-
minations made by means of the other instru-
 Instruments for Qualitative and Quantitative Analysis of Mixtures
ments described in this chapter. It is true that
commercial gas chromatographs carry various
dials and controls on their faces and display
their results on x-y recorders, but all that
these controls do is adjust temperatures and
gas pressures, maintain a detecting device, and
record when that device indicates that an
ingredient of a sam pie has come through a
column and how much there was of it. In
contrast to the other instruments we will
describe, the gas chromatograph analyzes a
sampie not by detecting its ingredients
through their characteristic intramolecular
properties but by physically separating them.
The heart of the gas chromatograph is a
tube of glass or metal, of one of two types:
either it is of appreciable diameter (about i"
o.d.) and fi11ed with an inert base coated
with an adsorbing medium, or it is a capillary
whose inner surface bears a coating of adsor-
bent. A portion of the sam pie being analy-
zed-usually in the form of a solution con-
taining only milli grams or less of the ingred-
ients being determined--is injected onto the
heated column, and a stream of inert gas
(nitrogen, argon or helium) is passed through
the tube and onto the detecting device. The
gas carries the sam pie ingredients through the
tube at rates that vary inversely with the
tenacity with wh ich they are held by the
adsorbent the tube contains; in effect a sort
of distillation takes place, and the efficiency
of a tube packing is sometimes caIculated in
terms of so many theoretical plates. As each
compound reaches the detector at the end
of the tube, it causes the detector to activate a
recorder. This recorder plots a height vs time
curve showing a peak for each time interval
that a compound leaves the tube; the height
of each peak (or more exactly, the area
enc10sed by it) is proportional to the quantity
of the compound present in the sam pIe
aliquot.
The various gas chromatographs differ in
control features such as their ability to make
rapid temperatllre changes to permit pro ces-
sing a sam pie in increasing temperature
[463
stages as the more volatile ingredients are
removed. The more important developments
in gas chromatography, however, have
involved the discovery of new detectors that
either were more sensitive than previous
ones, or had the desirable qllality of reacting
toward only certain types of compounds. The
first and most general type of detector was
the thermal condllctivity gauge, wh ich was
basically an electrically heated fine wire
mounted axially in a narrow tube; as the
current of gas flowing through the gas
chromatograph tube carried a sampIe ingred-
ient over this wire, its temperature (and
consequently its resistance) changed, and a
Wheatstone bridge device converted this
change to a small e.m.f. that activated the
recorder. A later device was the flame tem-
perature detector; this consisted of a hydro-
gen flame impinging on a thermocouple. As
an organic compound emerged from the
column, its heat of combustion was added to
that of the hydrogen, causing an increase in
flame temperature and change in e.mJ. of
the thermocouple that was translated into a
peak on the recorder.
Nearly a11 of the later more sensitive detec-
tors, as weil as those possessing specificity
for particular types of compounds, have been
devices involving ionization. One exception
is Coulson's microcoulometric detector, in
which the effluent compounds are burned
and then titrated for their chlorine or sulfur
contents. This device has been employed
almost exclllsively for the detection and
determination of organic chlorine or sulfur
pesticide residues in foods; for this purpose
it has the advantage that non-pesticide
residues carried through the column do not
affect the recorded curve.
Ionization detectors, in general, possess
higher sensitivity than thermal conductivity
devices. They also differ in that they do not
respond to all compounds. There are several
types, but one that was and still is being
extensively used in pesticide residue analysis
is the electron capture detector (Fig. 21-1):10
464] Special Instrumental Methods
Teflon tube sensitive to most organic phosphorus com-
pounds it is much less satisfactory for the
estimation of residues of organie phosphorus

-
pesticides that contain no chlorine atoms.
Inlet Outlet This deficiency was corrected with the
discovery by Giuffrida l l that a detector
composed of a hydrogen flame impinging on
a platinum coil coated with an alkali metal
salt had a sensitivity for phosphorus co m-
gauze
I cm
source pounds up to 10,000 times that of a regular
flame ionization detector. If the alkali salt
Fig. 21-1. The electron capture detector (after used were ahalide, the response to halogen
Lovelock). compounds was actually depressed. The
original coating material was sodium sulfate,
This deviee consists of achamber con- but potassium chloride was eventually adop-
taining two electrodes and a radioactive ted as the salt giving the best results.
ionizing source. A potential is applied across As was pointed out in Chapter 16 (Method
the electrodes just sufficient to collect all ions 16-11, Note 5), "gas chromatographic
and electrons. As the carrier gas passes methods identify aseparated compound only
through the chamber it is ionized by the beta to the extent that they show that, with a par-
partieies from the radioactive source, but ticular column and a particular form of detec-
when a sampie ingredient which is electron- tion, the compound gives a peak on achart at
capturing emerges from the chromatograph the same place as does the compound it is
tube and enters the ionization chamber, assumed to be." In order to make more
electron capture occurs to give negative ions certain identification of gas chromatographie
which are rapidly removed from the system by fractions, various authors have worked out
recombination. The ion current is thereby means of transferring these fractions to infra-
reduced, and this drop in current is translated red spectrophotometers or mass spectrometers,
into a peak on the recorder curve by suitable particularly the latter. This subject is discussed
devices. 10 The fact that this detector is below under "Mass Spectrometers".
exceedingly sensitive to halogen-containing For a more detailed yet concise discussion
compounds explains its appeal to the of gas chromatography the reader is referred
pesticide residue analyst. to Selected Reference U-l.
Until very recently the most common radio- Gas chromatographic methods described
active source was tritium adsorbed on in this book are Methods 2-13, 11-9A,
titanium. This source always had the dis- 13-17, 16-11, and 16-28.
advantage that as the temperature of the There is a rather large number of manu-
system rose above 200°, tritium began to facturers of gas chromatographs in the United
evaporate. To avoid this restrietion on States, inc1uding:
usable temperature and its associated hazard,
sources composed of nickel 63 have come on Barber-Colman Co., Industrial Instruments
the market,u Div., 1300 Rock St., Rockford, 111.;
Because of its high sensitivity and relative Beckman Instruments, luc., 2500 Harbor
specificity for halogen compounds, the Boulevard, Fullerton Calif.;
electron capture detector is nearly ideal for F & M Scientific Division of Hewlett-
the determination of residues of chlorinated Packard Co., P.O. Box 245, Avondale, Pa.;
organic pestieides, but because it is not very larrell-Ash Co.;
 Instruments for Qualitative and Quantitative Analysis of Mixtures [465
Packard Instrument Co., Inc. 2200
Warrenville Road, Donners Grove, Ill.;
Perkin-Elmer Corp.; and
Varian Aerograph, 2700 Mitchell Drive,
Walnut Creek, Calif.
Manufacturers of gas chromatographie
supplies inc1ude:
Analabs, Inc., 80 Republic Drive, North
Haven, Conn.;
Applied Science Laboratories, State
College, Pa.; and
Pierce Chemical Co., P.O. Box 117,
Rockford, 111.
Neutron Activation Analysis
Apparatus
Neutron activation analysis depends in
principle on the use of a neutron source to
convert a nonradioactive isotope to a
radioactive isotope, followed by determina-
tion of the concentration of that element by
measurements of its radioactivity. An example
is the determination of nitrogen (and there-
fore protein) by the N 14(n, 2n) N 13 reaction.
This method of analysis is often very
sensitive; it can be both rapid and specific,
and requires little previous sam pIe prepa-
ration. The chief obstac1e to its more frequent
use has been the initial cost of the apparatus
and the shielding for the neutron source.
Manufacturers supplying apparatus of this
type include:
General Dynamics Corp., General Atomic
Division, P.O. Box 608, San Diego, Calif.;
Kaman Nuc1ear, 1700 Garden of the Gods
Road, Colorado Springs, Colo.
Polarimeters
The polarimeter is of course an instrument
for measuring the optical rotation of a solu-
tion. Polarimeters calibrated to read directly
in terms of per cent sucrose when the rotation
of a solution of 26 grams of sam pIe in 100 ml
is measured at 20° in a 200 mm tube, using
sodium D light, are called saccharimeters.
While these instruments may be employed
in the determination of other optically active
compounds than carbohydrates, they find
little use in food analysis other than for the
estimation of sugars, as in Method 16-6,
"Determination" (A). Among the few ex-
ceptions are AOAC Method 20.062 for
laevo-malic acid (which depends on enhance-
ment of rotation of the acid by uranium
salts) and Method 9-14 for lemon and
orange oils in extracts.
A discussion of the principles of optical
rotation may be found in Text Reference 13.
American manufacturers of better-grade
polarimeters are:
Cary Instruments, 2724 South Peck Road,
Monrovia, Calif.
O. C. Rudolph Sons, Inc., P.O. Box 446,
Caldwell, N.J.; and
Rudolph Instruments Engineering Co.,
Inc., 61 Stevens Ave., Little Falls, N.J.
Polarographs: Amperometric
Titrations
The technique of polarography and the
name itself were originated by Jaroslav
Heyrovsky at the Charles University in
Prague about 1929. 14 Polarography is a
process of recording the variations in current
that take place as a constantly increasing
voltage is applied to a system composed of
a dropping mercury electrode and a reference
electrode immersed in a solution of the
sampIe being analyzed. The mercury electrode
consists of a drop of the metal hanging for a
few seconds at the surface of a glass capillary
from which mercury regularly drops out.
Figure 21-2 is a schematic diagram of the
polarograph; Figure 21-3 shows the type
of curve obtained by plotting current vs
voltage.
These polarograph curves are completely
reproducible, and if experimental conditions
are kept constant they depend only on the
466] Special Instrumental Methods
composition of the electrolyzed solution. In
the presence of substances which undergo
reduction or oxidation at the surface of the
dropping electrode, or substances that
catalytically affect the electrode process,
or those that form stable compounds with
mercury, an increase in cathodic or anodic
current is observed. The current rises in a
given potential range, and this increase is
Current
meosuring
device
Mercury
reservoir
Nitrogen
inlet
Electrolysis
vessel
Fig. 21-2. Schematic Diagram of Polarograph.
followed by a region of potentials in which
the current has a limiting value. The S-shaped
portion of the current-voltage curve is called
a polarographic wave. The shapes and
positions of these waves provide information
on both the quantitative and qualitative
composition of the electrolyzed solution. The
difference between the limiting current and
the current before the wave rise, which is
Potentiometer
slide
contoct
Storage
bottery
Potentiometer
Reference electrode
 Instruments for Qualitative and Quantitative Analysis of Mixtures [467
Limiting
...e: current
~::J
U
Voltoge, E -
Fig. 21-3. Current-Voltage Relation in Polaro-
graphy.
called the wave height, depends on the
concentration of the electroactive substance
responsible for the wave, while the half-wave
potential, i.e., the potential at that point on
the curve at which the current reaches half
its limiting value, is independent of the
concentration but is a function of the nature
of the electroactive substance and conse-
quently can be used to identify it quali-
tativelyY
For a more detailed discussion of the
principle and practice of polarography, the
reader is referred to one of the special texts
such as Selected Reference 1-2. This tech-
nique has been used chiefly to analyze
sampies for different metallic elements that
can be made to yield specific polarographic
waves under the proper conditions; the
titrations are carried out in a stream of
nitrogen to eliminate the oxidation-reduction
effects of atmospheric oxygen. The method is
however also used to determine organic
compounds that yield half-wave potentials.
An example of the use of polarography in
food analysis is the method of Christian
et al. for traces of selenium. 16
Originally polarographie curves were
plotted manually, but because this was
laborious, Heyrovsky and Shikata introduced
an instrument in which the movements of a
galvanometer lamp were recorded photo-
graphieally. This was succeeded by various
instruments that plotted curves on achart
through an electrically-controlled pen. De-
rivative polarography has also been employed.
A method of analysis related to polaro-
graphy is amperometry. Without going into
detail, in amperometric titrations the current
whieh passes through the titration cell
between an indicator electrode and a de-
polarized reference electrode, at a suitable
applied e.mJ., is measured as a function of
the volume of the titrating solution. The end
point of an amperometrie titration is the
point of intersection of two lines giving the
change in current before and after the
equivalence pointP Amperometry is still
not a common technique, but it did find a
certain use in testing for chlorinated organic
pesticide residues in the days before gas
chromatographie analyses became common:
Extracts of the sam pIes were burned, and
chloride was determined in solutions of the
ash by amperometrie titration with silver
nitrate. t8
Manufacturers of polarographic apparatus
inc1ude:
Fisher Scientific Co., 711 Forbes Ave.,
Pittsburg, Pa.;
Leeds & Northrup Co., 4901 Stenton Ave.,
Philadelphia, Pa.;
Melabs Scientific Instruments Operation,
3300 Hillview Ave., Stamford Industrial
Park, Palo Alto, Calif.;
E. H. Sargent & Co., 4647 West Foster Ave.,
Chicago, IIl.
Raman Spectroscopes
The phenomenon of the Raman shift
interference effect was discovered inde-
pendently in 1928 by Raman and Krishnan
in India and Landsberg and Madelstam in
Russia. 19 When a beam of monochromatic
light is passed through a tube containing a
468] Special Instrumental Methods
liquid sam pie, and some of the light that is
scattered in a direction perpendicular to the
incident beam is passed into a spectrograph,
the resulting spectrogram shows (in addition
to the expected strong line at a position
corresponding to the wavelength of the
incident light) a pattern of relatively weak
lines or narrow bands on the red (long-
wavelength) side of the incident line, and
possibly a very few weak lines on the blue
side as weIl. These lines are the Raman
spectrum of the sampie ; the lines on the
long-wavelength side are called the Stokes
Raman fines, and those on the short-
wavelength side, the anti-Stokes fines.
If the wavelength of the incident light is
changed to larger or sm aller values, the
pattern of Raman lines (i.e., their relative
positions to each other) is unaltered, but the
group as a whole is shifted to longer or
shorter wavelengths. The difference in fre-
quency between aRaman line and the incident
light is independent of the frequency of the
incident light, and the set of frequency
differences or shifts, which is the Raman
spectrum, is characteristic of the molecules
in the sampie. These shifts are commonly
reported as wave-numbers.
Because the Raman spectrum of a com-
pound is unique for that compound, and the
spectrum of a mixture whose components
do not interact is a superposition of the
Raman spectra of the components, Raman
spectroscopy may be used for the identifi-
cation of both single compounds and
mixtures of as many as six different
substances. Since the intensity of each line
is proportional to the concentration of the
substance giving that line, quantitative
analysis is also possible.
The advantages of Raman spectroscopy
inc1ude: 1) the Raman spectrum of a com-
pound is relatively simple; 2) water may
be used as a solvent because the Raman
spectrum of water is diffuse and weak; and
3) the relationship between line intensity
and concentration is linear. Some disad-
vantages are: 1) analyses are practically
limited to c1ear, colorless (non-absorbing)
liquids; 2) the sam pie must be completely
non-fluorescent; and 3) the sam pie volume
required is larger than for infrared spectro-
scopy (5-15 ml is normally used). The
geatest disadvantage, however, has been the
difficulty in obtaining a light source that was
sufficiently intense and monochromatic to
yield Raman lines that were bright and
sharp enough. Until recently almost all
instruments utilized the Hg4358A line, but it
is now generally expected that the illumina-
tion difficulty will be solved by the use of
laser beams; in fact one instrument with a
laser source, the Cary, is already on the
market.
Manufacturers of Raman apparatus
inc1ude:
Cary Instruments;
Perkin-Elmer Corp.;
Spex Industries, Inc., 3880 Park Ave.,
Edison, N.J.
X-Ray Fluorescence
Spectrographs
There is a difference of opinion over the
name for the type of analysis to which we shall
refer here. Some authorities prefer the name
X-ray emission spectrography on the grounds
that this designation prevents confusion with
ordinary fluorometry as discussed above, and
"has a familiar ring to analytical chemists,
accustomed as they are to emission spectro-
graphy involving radiant energy in the visible
and in the ultraviolet."20 Nevertheless, while
such terminology may appear to prevent
confusion on the part of technicians knowing
little beyond, and interested only in, the
practical application of instruments as
analytical tools, it (perhaps deliberately) has
the effect of completely confusing the
distinction between emission and fluorescence
phenomena. We have therefore headed this
section "X-Ray Fluorescence Spectrographs."
 Instruments for Qualitative and Quantitative Analysis of Mixtures [469

When a be am of electrons impinges on a a bent mica crystal) that serves as a diffraction

metal target, X-rays characteristic of the
particular metal are expelled. This is X-ray
emission. To use it for analytical purposes
would require, however, construction either
of separate X-ray tubes for each analysis, or
of a single tube so designed that targets could
be changed. Because ofthe physical difficulties
in this method of analysis, it has had no
practical use, which probably explains in
part why some authors in their description of
X-ray spectroscopy leave uncIear the dis-
tinction between emission and fluoroscopic
effects. 21
The X-ray fluorescence spectrometer, on
the other hand, is a practical device that has
extensive use and may be the instrument of
choice for so me analyses. lt works in principle
as folIows: an intense beam from an X-ray
tube impinges on the sampie being analyzed,
producing fluorescent X-rays by secondary
emission. These rays are dispersed by a
crystal of known lattice constant (frequently
grating. The diffracted fluorescent X-rays are
detected by a Geiger counter attached to a
goniometer, and their intensities are auto-
matically recorded as functions of the
goniometer angles, which are proportional
to the wavelengths or frequencies of the
X-rays. Figure 21-4, which was obtained from
a chrome-nickel plating on a silver-copper
base, i1I ustrates the type of spectrum 0 bserved.
Each element is identified by its position on
the abscissa, while the height of the ordinate
peak is proportional to the quantity present
in the sample. 21
X-ray fluorescence analysis has an ad-
vantage over emission spectrography in that
X-ray spectra never consist of more than four
lines, while the transition elements yield
many atomic emission lines. In trace metal
analysis particularly this plethora of lines can
make it difficult to separate a line of the
element sought from the lines of a more
abundant element such as iron. Another

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Fig. 21-4. X-ray fluorescence spectrum of chrome-nickel plating on silver~opper brass. [From O'Connor,
R. T.: J. Assoc. Ojfic. Anal. Chemists, 51 :244 (1968).]
470] Special Instrumental Methods
advantage is that X-ray fluorescence analysis
is not restricted to the determination of
metallic elements, as emission spectrography
is in practice. Its disadvantages are that it is
not capable of extreme sensitivity, and
cannot be used to determine elements lighter
than sodium. 21 Tuchscheerer has applied
X-ray f1uorescence analysis to the de-
termination of K, P, Cu, Fe, Zn and Mn in
foods. 22
Manufacturers of this type of apparatus
Instruments Primarily for Identification of
Single Compounds
Infrared Spectrophotometers
Despite the fact that they are expensive,
infrared spectrophotometers are rapidlyon
their way to becoming part of the normal
equipment of all modern analytical labora-
tories. These instruments plot the absorption
curves of sam pies in the infrared in essentially
the same manner that instruments such as
the Beckman DU and the Cary do in the
visible and ultraviolet regions ofthe spectrum.
The source of light is different, being a
Globar (a bonded Sie rod) or a Nernst
glower (a mixture of Zn, Th and Yoxides);
the dispersing element of choice is rapidly
becoming a grating, but if it is a prism, it
must be made of a salt such as NaCI or KBr
rather than of quartz; and the detector is
usually a thermocouple. These differences in
the components of the infrared instrument
from those of the visible-ultraviolet apparatus
are necessary because the lamps used as
sources of visible and ultraviolet light are
poor sources of infrared, and glass and even
quartz are essentially opaque to the infra-
red.
We shall not enter into a theoretical
discussion of why the infrared absorption
curve of a substance teIls so much more about
that substance than do the visible and ultra-
include:
Baird-Automic; General Electric Co., X-
Ray Dept., 4855 Electric Ave., Milwaukee,
Wis.; Instrumentation Division, Engis Equip-
me nt Co., 8035 Austin Ave., Morton Grove,
Ill. and
Norelco Philips Electronics Instrument
Div., Philips Electronics and Pharmaceutical
Industries Corp., 750 South Fulton Ave.,
Mt. Vernon, N.Y.
violet spectra. For our purposes, it is sufficient
to note that the infrared curve of a compound
is a fingerprint that uniquely identifies it,
°
and further, that a specific group within a
molecule, e.g., CH 3 , C = 0, P = or C = C,
absorbs at nearly the same wavelength
regardless of the compound in which it is
found. Therefore, if an analyst is confronted
with a compound of whose structure he is
uncertain, he can run its infrared spectrum
and compare it with the spectrum of a
reference substance he suspects it to be; ifthe
two prove to be the same, identification is
certain. Ifthe curve for the unknown does not
match that ofthe postulated compound, it can
still be identified if a match can be found with
any ofthe thousands ofspectra recorded in the
literat ure. And if even this search fails, the
unknown will usually show some charac-
teristic group absorption peaks that will help
to point the way toward elucidation of its
structure. It should be noted, however, that
infrared spectrometry does not distinguish
between optical isomers.
Infrared spectrophotometry can also be
used for the quantitative determination of a
single compound or a mixture of several
compounds when each of the substances
being determined possesses a sharp peak that
is not shared with any other substance that is