ASTM Logo

Removed

Designation: E 260 - 96

Standard Practice for

Packed, Column Gas Chromatography 1
This standard is issued under the fixed designation E 260; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (.) indicates an editorial change since the last revision or reapproval.

1. Scope 3. Terminology
1.1 This practice is intended to serve as a general guide to 3.1 Terms and relations are defined in Practice E 355 and
the application of gas chromatography (GC) with packed references therein.
columns for the separation and analysis of vaporizable or 4. Summary of Practice
gaseous organic and inorganic ),mxtures and as a reference
for the writing and reporting 0f OC methods. 4.1 A block diagram of the basic apparatus needed for a
gas chromatographic system is as shown in Fig. 1. An inert,
NOTE I-This praCtice excludes any form of gas chromatography pressure or flow-controlled carrier gas flowing at a measured
associated with open tubular (capillary) columns. rate passes to the injection port tir gas saniple valve., A
sample is introduced into the injectiori 'port~ where it is
1.2 This standard does not purport to address all the safety vaporized, or if gaseous, .into a' gas sample valve, and then
concerns, if any, associated with its use. It is the responsi- swept into and through the column by the carrier gas.
bility of the user of this standard to establish appropriate Passage through the column separates the sample into its
safety and health practices and determine the applicability of components. The emuent from the column passes to a
regulatory limitations prior to use. Specific hazard statements detector where the response of sample, components is mea-
are given in Section i and 9.1.3. sured as they emerge from the column. The detector
electrical output is relative to the concentration of each
resolved component and is transmitted to a recorder, or
2. Referenced Documents electron1cdata processing system,dr 'both, to produce a
2.1 ASTM Standards: ,record of the separation, or chromatogram, from which
E 355 Practice for Gas Chromatography Telll'ls and detailed analysis can be obtained. The detector emuent must
Relationships2 be vented to a hood if the emuent coritains toxic substances.
E 516 Practice for Testing Thermal Conductivity Detec- 4.2 Gas chromatography is essentially a physical separa-
tors Used in Gas Chromatography2 tion technique. The separation is obtained when the sample
E 594 Practice for Testing Flame Ionization Detectors mixture in the vapor phase passes through a column
Used in Gas Chromatography2 containing a stationary phase possessing special adsorptive
E 697 Practice for Use of Electron Capture Detectors in properties. The degree of separation depends upon the
Gas Chromatography2 differences in the distribution of volatile compounds, organic
E 840 Practice for Using Flame Photometric Detectors in or inorganic, between a gaseous mobile phase and a selected
Gas Chromatography2 stationary phase that is contained in a tube or GC column.
E 1140 Practice for Testing Nitrogen/Phosphorus In gas-liquid chromatography (GLC), the stationary phase is
Thermionic Ionization Detectors for Use in Gas a nonvolatile liquid or gum coated as a thin film on a
Chromatography 2 finely-divided, inert support of a relatively large surface area,
2.2 eGA Publications: and the distribution is based on partition. The liquid phase
CGA P-1 Safe Handling of Compressed Gases in should not react with, and should have different partition
Containers3 coefficients for, the various components in the sample. In
CGA G-5.4 Standard for Hydrogen Piping Systems at gas-solid chromatography (GSC), the stationary phase is a
Consumer Locations3 finely divided solid adsorbent (see 4.4).
CGA P-9 The Inert Gases: Argon, Nitrogen and Helium 3 4.2.1 After separation in the analytical column, the com-
CGA V-7 Standard Method of Determining Cylinder ponents are detected, and the detector signal is related to the
Valve Outlet Connections for Industrial Gas Mixtures3 concentration of the volatile components. Tentative identifi-
CGA P-12 Safe Handling of Cryogenic Liquids3 cations can be made by comparison with the retention times
HB-3 Handbook of Compressed Gases3 of known standards under the same conditions, either on a
single column or preferably by injecting the sample onto two
columns of different selectivity. Ancillary techniques, such as
I This practice is under the jurisdiction of ASTM Committee E-19 on mass spectrometry or infrared spectrophotometry, are gener-
Chromatography and is the direct responsibility of Subcommittee E 19.03 on ally necessary for positive identification of components in
Methods and Specifications, Gas Chromatography. samples.
Current edition approved April 10, 1996. Published June 1996. Originally
published as E 260 - 65 T. Last previous edition E 260 - 96. 4.2.2 Prior to performing a GC analysis, the following
2 Annual Book of ASTM Standards, VoI14.02. parameters must be considered:
3 Available from Compressed Gas Association, Inc., 1725 Jefferson Davis 4.2.2.1 Sample preparation.
Highway, Arlington, VA 22202-4100.
4.2.2.2 Stationary phase and loading on support.
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E 260

RECORDER

I
I

'---+:
'---+-:-----{
I
I
I
·INTERFACE ,~,."""
DATA SYSTEM
COMPUTER
j)
I
GAS OVEN I
CHROMATOGRAPH :

---------------------~

FIG. 1 Block Diagram of a Basic Gas Chromatographic System

4.2.2.3 Column material required. very large number of possible compounds in existence and
4.2.2.4 Solid support and mesh size. the finite number of peaks that a chromatograph might
4.2.2.5 Column length and diameter. resolve. Additional characterization data may be provided by
4.2.2.6 Instrument and detector type that will be needed. ancillary techniques, such as spectrometry.
. 4.2.2.7 Injector, column oven, and detector temperatures
required for analysis. 5. Significance and Use
4.2.2.8 Injection techniques, such as flash volatilization,
on-column technique, purge and trap, pyrolysis, etc. 5.1 This practice describes a' procedure for packed-
4.2.2.9 Carrier gas and flow rate. column gas chromatography. It provides general comments,
4.2.2.10 Data handling and presentation. recommended techniques, and precautions. A recommended
.4.3 In gas-liquid chromatography, the degree of separa- form for reporting GC methods is given in Section 14.
tion possible between any two compounds (solutes), is
determined by the ratio of their partition coefficients and the 6. Apparatus
separation efficiency. The partition coefficient, K, is the ratio 6.1 Carrier Gas System-Common carrier gases are he-
of the solute concentration in the liquid phase to the solute lium and nitrogen. Paragraph 7.6 provides more details on
concentration in the vapor phase at equilibrium conditions. carrier gases. Means must be provided to measure and
The partition coefficient is affected by temperature and the control the flow rate of the carrier gas. Any flow or pressur~
chemical nature of the solute (sample) and solvent (sta- control and measurement combination may be used that will
tionary phase). give an accurately known and reproducible flow rate over the
4.4 Another mechanism for separation is gas-solid chro- desired range. .
matography. With this technique there is no liquid phase, 6.1.1 The main gas supply is regulated with a two-stage
only a porous polymer, molecular sieve, or solid adsorbent. regulator which .must have a stainless steel diaphragm.
Partition is accomplished by distribution between the gas Rubber or plastic diaphragms permit oxygen or water to
phase and the solid phase. . ' diffuse into the carrier gas. In addition, instruments will have
4.5 After the sample is resolved into individual compo- a flow controller between the pressure regulator and column
nents by the chromatographic column, the concentration or inlet to maintain a constant flow during temperature pro-
mass flow of each component in the carrier gas can be gramming. Copper or stainless steel carrier gas lines, not
measured by an appropriate detector which sends an elec~ plastic tubing, should be used to avoid diffusion of oxygen
trical signal to a recording potentiometer or other readout (air) into the carrier gas. When using the thermal conduc-
device. The curve obtained by plotting detector response tivity detector, variations in the flow will change retention
against time is referred to as a chromatogram. For flame and response. The' carrier gas line pressure must be higher
ionization and thermal conductivity detectors, either the than that required to maintain the column flow at the upper
peak areas or the peak heights are proportional to the temperature limit for the flow controller to operate properly.
concentration of the components in the s~l.mple within the A pressure of 40 to 60 psi is usually sufficient.
linear range of the detector system. However, response 6.2 Column Temperature Control-Precise column tem-
fractors are not necessarily the same for all compounds, and perature control is mandatory if reproducible analyses are to
linearity of detector response may depend on operating be obtained. Temperature control must be withinO.l°C if
conditions. (Testing of detector performance is discussed in retention times are to be compared with another instrument.
ASTM Standard Practices for the appropriate detector, see 6.2.1 Air Bath-The thermostated forced-air bath is gen-
2.1). erally accepted as the best practical method of temperature
4.6 Components in a mixture may be tentatively identi- regulation for most applications. Temperatures can be con-
fied by retention time. Ideally, each substance has a unique trolled by regulators or proportionally controlled heaters
retention time in the chromatogram for a specific set of using a thermocouple or platinum-resistance thermometer as
operating conditioJ1s. However, caution is required because a sensing element. The advantage of a forced-air bath is the
the GC separation may be incomplete and a single peak may speed of temperature equilibration. Air bath ovens are
represent more than one compound. This is especially true of readily adaptable to temperature programming and are
unknown mixtures and complex mixtures because of the capable of operating over a range of 35 to 450°C. This range

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can be extended down to -100·C by using cryogenic is relatively cool (below 50·C) to avoid contact of stationary
equipment. phase in a hot column with air (danger of oxidation). After
6.2.2 Other Devices-Liquid baths, drying ovens, incuba- the septum is changed, return the inlet temperature to that
tors, or vapor jacket enclosures are less stable, less conve- . whic!} was originally set. The inlet temperature should be the
nient means of providing a SOlloJ;ce, of heat t<) maintain or ,optimum for ,the particular analysis, as well as within the
raise the temperature of a chromatographic column. These recommended operating temperature of the septum. If the
devices are not recommended,fdfprecision chromatographic septum is 'pui;lctured too many times, it will leak air into the
applications. gas chromatdgfaphic system, even though it is under pres-
6.3 The Injection Port-The purpose of the injection port sure. Athjgh temperatures, above 150 to 200·C, air (oxygen)
is to introduce the sample into the gas chromatographic in the carrier gas from a septum l~ak will degrade the
column by instantaneous volatilization following injection stationary phase. An excessive septum leak will also produce
into the gas chromatographic system. Two sample inlet types a ch~nge in carrier gas flow rate (a change in retention time)
are in common use in gas chromatography:' the flash and losso£ sample (irreproducible peak heights) due to
vaporization and the on-column injection inlets. outflow from the leak. When installing the septum, do not
6.3;1 The temperature of the flash. vaporization ;i~et overtighten the retaining' nut." The septa will 'swell' at high
should be above the boiling points of the sample components temperature and extrude"out ofthe injection port. A snug fit
and is limited by the am.ount of septum bleed generated and at room temp.erature is' suffiCient It is important for septum
the temperature stability of sample components; It should be' life to make sure the injection ileedle is sharp With no bent
set at that temperature above which no improvement in peak tip. Fine emery cloth, or a'fine sharpening stone, can be used
shape occurs but should be determined bY.,the nature of the to sharpen the point. .
sample and the volume injected, not by tbe' tem.perature of
6.3.4.2 Ghost" peaks' may be obSerVed in temperature
the column. If the iDlet'temperature is too low, broad peak
programmed runs due t6 septum bleed. Septlim bleed IS due
with a slowly rising front edge' Will result froni slow vaporiZa-
tion of the sample .. If the temperattri-e is set (ar above what is to the thermal decomposition, 300·C or higher, of the
necessary to produce' fast vaporiiation, thetmal decomposi- septulIl tpat pr9duc~s primarily lower moletular weight
tion of the sample, decreased septum life, and ghost peaks cyclic dirrtethylsiloxailes. It contributes.to baseline response
due to septum bleed may be observed. Generally; a good and is frequently obserVed as eveJ11y spaced peaks in a
guide]jlle, ~s to maintaintp~:jnlet temperature 25 to 30~C tempe.rature" programmed run in. ~ffi.ch no sample sa.~ple has been
higher ~an the highest bojling point <?(anysa111ple compo- injected,. This ,situatipn '. can be.. dePlQnstrated 'by the disap-
nent. . ,. ., , i ' ., ;
pearance of ghost peaks afterplacmg' aluminum foil (pre-
, 6.3,7 A glass liner placed insiej,e the injection POrt·;'YW cleaned with solve:rit~ ~uch as methy,ene chloride 'or toluene)
eliminate samplecontacCwith hot metal inner walls of the "in;p.er Jace;;~rtl1e
over the "iAper ,/ace};~rtl1e septum or by turning off *e
inlet; which can c~t3.iYzethermal· d,ecomposiiions. Any temperature and making several blankruns~
injector temperature' blank runs~ Septurp.
debris left in the liner, especially from biological samples, bleed can, be decreased. by, using either air~ or water-cooled
can be a source of excessive ,sample adsorption. Ifa liner is septum r~taining nuts., by using a septum"flush head, or..py
used, tbe debris can e~~ily..be rerilOved by replaqitlg
replaqill,g the liner. using sP~~ialLlPgh-teinp~rature septa, which ~~ avaihtble
Deactivation ofthegl~.Il,s,li,ner by treatment with dimethy~- from a ngmber ,of gas chromatograplPc'supply;,);lOuses.
d,ichl,orpsilane m~ybe nec~s~~ry for some compounds. . 6.4 Detector Temperature Control-The detector temper-
6.3.3 With on-cqltllllP: injectioll technique, the l sample is ature should always be above that of the maximuD;l operating
deposited in the liqm<htate,qirectly on,tl,J,e:~olumn,packing. analytical temperature, to prevent the possibility of conden-
The sample must be smaU,enough t.o preclude, flooding of the sation of sample components or stationary phase bleed in the
column, with possible detrimental, effects,tq;peak shape and detector and connecting line .. Because there is usually some
column life. Ideally, ~he on-colQ,mn inlet)s a part. of the temperature. gradient, aCf(';)ss:!a detector, the temperature
temperature,
column, so its temperature m~y ,be contr,oUed as th~ column should be set at 30 ,t050·C above the maximum analysis
temperature is controlled. Inpr~ptice, 1:lec~use an on-column temperature toensure,that the 'entire detector is hot enough
inlet usually has a somewhat ;higher thermal ;m~ss than an to pteveilt condensation; Usually, it is neither necessary nor
equivalent sectqr of the rest ofthe;J;olumn"thejnletmust be desirable to \use ,an excessively high 'temperature' since this
heated somewhataMve the maximumana1ysis tenl,perature can 'result .in reduced" sensitivity; increased' noise level,
of the column. oven, The criteria ,of good 'peak shape and frequent need t6 dean the'detector., and thermal decomposi~
quantitation should be used to .determine the maximum iion (jfsample or 'stationary phase." ., '\
required terilperaturefor the ,inlet. One should conSider,the , 65 Measurement of Temperature~The ~choice of
temperature limit of the colun;m packing when heating the sertsirigelements used to measure temperature depends on
injection inlet and detector. With some samples, a nonheated the desired accuracy (control about aset point) aftd'precision
injection port is adequate, especially with temperature- of the measurements. Instrument read-outs should be veri-
programmed operation. fied periodically. Some common tempera:hii'e'lineasurement
6.3.4 Injection Port Septum;~' ,' devices are as fo116ws: ." ....,
63.4.1 The septum is a disc; usually made of ,silicone 6~5.1 'Standardized Mercury Thermometer:'
rubber; which .. seals, onelend of. the injection port. It is , ' , r
Range, D¢ ,AccUracy, DC
important to change ,the septuinfrequentlyafter two to three o to 100 . :l:O:OZ
dozen injections, or preferably at the end of the. working day, to
100 200 ±O.OS'
The best techriique, is to change the septum when the column 200 .to 400 ·±O.SO

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6.5.2 Calibrated Platinum Resistance Thermometer: lower starting temperature and greater sensitivity because of
Range, °C Accuracy, °C sharper peaks for the higher boiling components.
-140 to 500 ±0.01 6.8.1 Column Heater and Temperature Programmer-It
is of utmost importance that the column temperature
6.5.3 Thermocouple (iron - constantan, or other). program be reproducible, and that the difference between the
6.6 Analytical Column: set (desired) temperature and the true average column
6.6.1 The analytical column is a length of tubing (glass, temperature be as small as possible. However, these require-
metal, or plastic) that is filled with a packing material. It is ments are difficult to achieve at high heating rates and with
discussed thoroughly in Section 8. columns of large diameter. The mass of the column and its
6.6.1.1 Column Characteristics-Specified by method. heater should be kept as small as possible. This will minimize
6.6.1.2 Carrier Gas-Specified by method. thermal lag and will give proportionately small variations
6.6.1.3 Sample Size-Variable within limits. around the set temperature at any time. Proportional tem-
6.6.1.4 Flow Rates of Carrier Gas and Detector Gases- perature controllers supply almost full power to the heater
Variable within limits. until the set point is very closely approached.
6.6.1.5 Column Temperature-Usually specified by 6.8.2 The recirculating air bath is the recommended
method, and method of heating in programmed temperature gas chroma-
6.6.1.6 Physical or Chemical Characteristic ofCompouizd tography (PTGC). The obvious advantages are extremely
Analyzed, or both. rapid heating (and cooling after an analysis is completed)
6.6.2 Detector Characteristics-Desirable detector charac- with very little temperature lag.
teristics should include the following: 6.8.3 Liquid baths may be used for very low heating rates.
6.6.2.1 Good stability (low noise level, minimum re- They are commonly contained in taped Dewar flasks.
sponse to changes in temperature and flow rate). 6.8.4 No matter what type of heating device is used,
6.6.2.2 Ruggedness and simplicity. accurate control of the temperature program is necessary.
6.6.2.3 Sensitivity to the components for which analysis is This is usually accomplished by appropriate electronic sys-
desired. Use either a selective detector for materials of tems that develop linear (or other) programming rates· as
.interest or one with a similar response for all components. desired.
6.6.2.4 Linearity of response versus sample concentration. 6.8.5 Detectors for programmed temperature gas chroma-
Wide linear range. tography should be relatively insensitive to minor tempera-
6.6.2.5 Rapid response to changes in column effluent ture and flow fluctuations and insensitive to stationary-phase
composition (small internal volume or flow-through design, bleed. These difficulties can be overcome by operating the
or both). detector at or near the upper temperature limit for the
6.6.2.6 Detectors, which are nondestructive and do not analysis and by using adequate flow controllers. If stationary-
contribute to band broadening maybe used in series with phase. bleeding is excessive during PTGC runs, a second
other detectors. conditioning procedure (Section 9) might improve the situa-
6.7 Types of Detectors-The detector is located at the tion, Alternately, a duplicate analysis column can be used on
outlet end of the chromatographic column and both senses ,the reference side of the detector. By equalizing substrate
and measures the amount of components that have emerged bleed on both sides of the detector, the baseline drift can be
from the column. The optimum detector should have high substantially compensated. However, this technique does not
sensitivity, low noise level, a wide linearity of response, a improve column life and is detrimental to detector linearity.
response to all compounds of interest, and yet be insensitive . If at all possible, operate the column within its recommended
to changes in flow and temperature. Selective detectors are temperature range.
characterized as having selective, or greatly enhanced re- 6.8.6 When using the, temperature programming tech-
sponse to certain components. Linearity is decreased for all nique, the resistance to carrier gas flow in the gas chromato.-
detectors by column bleed. As many as forty detection graphic column increases with increasing temperature. The
systems have been reported, yet only about ,a dozen are flow controllers needa positive pressure 000 psi to operate
commonly used. Table 1 shows some of the more commonly proPerly. By setting the second stage ofthe regulator to 40 to
used detectors. Of these, the thermal conductivity, the flame 60 psi, there will usually be sufficient excess pressure to
ionization, the electron capture, the nitrogen-phosphorus, maintain a constant gas flow through the column. Higher
and the flame photometric detectors are the most popular. pressures might be required to maintain flow when using
Nondestructive detectors should be vented to a hood to relatively long columns of 10ft, or longer, or packings finer
remove any toxic effluents from the workplace. The effluent than 120 mesh.
from destructive detectors may also be toxic. Details on
detectors can be found in the applicable methods in Practices 7. Hazards
E 516,E 594, E 697, E 840, and E 1140. 7.1 Gas Handling Safety-The safe handling of com-
6.8 Programmed Temperaiure Operation-The apparatus pressed gases and cryogenic liquids for use in chromatog-
used in programmed temperature gas chromatography dif- raphy is the responsibility of every laboratory. The Com-
fers in some respects from that normally used for isothermal pressed Gas Association, a member group of specialty and
work. Basically, the column temperature is varied with time bulk gas suppliers, publishes the following guidlines to assist
(program rate) to enhance speed of separations. The advan- the laboratory chemist to establish a safe work environment:
tages of using programmed temperature operation. include CGA P-1, CGA G-5.4, CGA P-9, CGA V-7, CGA P-12, and
better resolution of lower boiling components because of HB-3.

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E260
TABLE 1 App.ljcl[lbility of Commonly Used Gas Chromatographic Detect,ors ,
A Applicability Range of Minimal Detectable
Detector
,(Type ofGompound) Amou~ts (grams)

TnermalConductivity All Hi 6 to 10-7

Flame 10l"!i,zation Qrganic , , '" 10-12
Electron Capture Halogenated and Oxygenated 11)""'12 to 10-15
Flame Photometric Sulfur, Pho$phorus 10-11
, , 10-12 (even lower for phosphorus)
Alkali Flame Nitrogen, Phosphorus
A Further information can be found in Practices E 516,E 594 and E 697. '

8. ' Materials exposed by removal' of liquid phase, thus' shortening column

8.1 Stationary Phases-The stationary phases (paqi- life. Bleeding also can ~xpoS'e bare support surface that can
tioning agents) that have been successfully used for specific adsorb molecUles being analyzed and reduce column effi-
separations are found most quickly by a literature search. ciericy. In'extremecases, phase bleeding will result in fouling
M~y phases are listed in ASTM publicatiqns AMD-25A
the detector and connecting lines. The observed maxirilUm
andAMD-25A-51.4 The most desirable stationary phases do temperature will depend upon many experimental variables,
not volatilize (blee9) slginficantly"frQm the solid supports at such as type of liquid phase column, conditi01:llng, phase-
temperatures required to elute the sample. loading level, columritemperature, serisiii~ty setting of the
8.1.1 The polarity of the stationary phases is currently detector, and purity of the carrier gas. In programmed
best characterized by McReynolds Constants. 5 The higher temperature runs, the column can sometimes be 'operated for
the McReynolds Constant, the more polar the phase. short periods about 25°C above maximum temperature.
Rohrschneidet constants can also be used to measure the However, column bleed should be minimized for quantita-
PQlarity of stationary phases. 6 tive results since it decreases the linear range of all detectors.
8.2 Active Solids:
8.1.2 The effects of using polar and nonpolar stationary
8.2.1 Molecular Sieves-The sYnthetic zeolite molecular
phases are summarized as follows:
sieve sorbents separate molecules by size and structural
8.1.2.1 Nonpolar stationary phases separate compounds
shape. Isomers with a more round shape, as branched versus
primarily by order of relative volatility or boiling point. straight chain molecules, diffuse ih and out of the zeolite
8.1.2:2,PQlar stationary phases separate compounds;by structure more easily than isomers with the long chain
order of both relative volatility and -relative polarity. With structures. Separations are affected 'by the differences in
polar phases, nonpolar compounds will elute before polar times reqtiiied for molectiles of different sizes tafind.their
compounds of the same boiling point. way into and out of the sieve-like structure of the adsorbent.
8;1.2.3 Polarity alone is insufficient to describe the sepa- Molecuiat sieves are·most useful for separating H 2 , O2 , N 2 ,
ration power of a columri; One must consider the ~he overall CO, and CH4 • Carbcin molecUlar sieves are also availa:ble,
selectivity of a column towards a se't of analytes'."
anaiytes'." This
selectiVity is a summation of the effects of dispersive interac-
°
and can be used to separate 2 , N 2 , CO, CO2 , H 2 0; and C1
to C4 hydrocarbons.
tions, acid-base interactions and the dipole interactions 8.2.2' Porous Polymers:
offered by the various pendent groups ·in the stationary 8.2.2.1 ,One tYPe of porous pblymer used in gas chroma-
phase."",. tography is' available in the form of microporous cross-
8.1.3 The stationary philses used in gas chromatographic linked, p0lymer beads produced, by copolymerizing styrene
cohinuis havebothmiriimum and maximum temperature and divinylbenzene or more polar copolymers, or both.
limits. The chromatographer must be aware of the limits for These materials are generally used as received without
the phase being used. Below the minimum temperature, the coating with any liquid phase. They provide symmetrical
phase will 'behave as either a Viscous liquid or a solid. Less peaks for polar" hydrogen-bonding compounds such, as
efficient separation will De observed, arid the chromato- water, alcohols, free acids, amines, ammonia, hydrogen
graphic, results Will qe exhibited as broader peaks in the gas
graphic g~s sulfide, etc., iand organic compounds up to molecular
chromatogram due 'to poor mass transfer' of components in m weights icorresponding to ,about 170. -:,
the stationary phase.' ' . > , 8.2.2:2 Ai::tother 'porous polymer is 'PQly(2,6--dipheIiyl-p-
8.1.3.1 Above the maximum temperature limit, the pnase phenylene"oxide)~This material is usefuVfortheanruysis of
will begi)1 to bleed off the colu~n at an accelerated rate, and arirines, alcohols, and. hydrogen-bonding compounds. It, is
the observed results will include "a driftirig baseline or also used, as an, adsorbent for trapping trace organic com-
excessive spiking on the baseline. Under these conditions, pounds in water and air. c " -

the liquid phase will decompose or volatilize, and thus be 8.2.3 Silica .Gel, Alumina; and Carbon-Among the ac-
removed from the column. This situation will eventually tive solid adsorbentsare silj,ca gel;· alumina, and activated
lead to decreased retention times with broader peaks re- carbon. They are useful· for IQw-boiling hydrocarbons.
resolutio:Q. of very close peaks. Peak taili~g
sulting in poorer resolution 8.2.4 Solid adsorbents mQdified' by' low concentrations of
will ,also, be observed as the uncoated surface becomes liquid phases may retain theadvantageouspmperties <;>f
both. Some solid adsorbents can. Qe modified by the addition
of surface activating compounds."such",as wetting agents,
4 ~as Chromatographic Data Compilation. ASTM, 1981. silver nitrate, and the metal salts offatty acids.
S McReynolds, W.O., Journal ofChromatojp:aphy Science, V018, 1970, p. 685.
'6 Supina, W. R., and Rose, L. P., Journal of Chromatography, Vo18, 1970, p.
8.3 Diatomaceous Earth Supports-The most popular gas
214. chromatographic supports are those prepared, from diato-

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E 260

maceous earth, also called diatomaceous silica or kieselguhr. character of a silanized diatomaceous earth, even coating of
The two main types are white and pink in color. The white the most polar liquid phases is difficult to achieve. 'Acid-
supports are recommended over the pink supports because washed, silanized grades of white diatomaceous earths are
of their more inert surface. The former are, however, very recommended as supports for the polar liquid phases, such as
friable and must be handled very carefully when preparing polyesters and silicones of high cyano-group content.
packings and loading into gas chromatographic columns. 8.3.5 If the column is 6 ft (2 m), or less, use particle size of
Before using these supports, check the manufacturer's litera- 100 to 120 mesh (125 to 149 J.Lm) for highest efficiency under
ture for comments on their use. isothermal conditions. If the column is longer than 6 ft, use
8.3.1 The white-colored supports are produced by calcina- 80 to 100 mesh (149 to 177 J.Lm) particles. If temperature
tion of diatomaceous earth with sodium carbonate as a flux. programming is used, 80 to 100 mesh particles should be
In this process, the diatomaceous earth fuses, due to forma- used to lessen resistance to carrier gas flow.
tion of a sodium silicate glass. The product is white in color 8.3.6 Further information concerning the liquid phase
due to conversion of iron oxide into a colorless complex of loading is given in 9.3.
sodium iron silicate. These white materials are used to 804 Halocarbon Supports-The two types of halocarbon
prepare the more inert gas chromatographic supports. How- supports are those prepared from poly(tetrafluoroethylene)
ever, they are fragile and subject to abrasion from excessive and poly(chlorotrifluoroethylene) ... These supports are rela-
handling in the course of sieving, packing, or shipping. tively inert and are nonpolar. They eliminate peak tailing
Abrasion will produce finer particles, or fines, which will observed in the analysis of organic compounds capable of
decrease column efficiency. hydrogen bonding, such as water, alcohols, amines, etc. They
8.3.2 The pink-colored supports are prepared by crushing are the preferred supports in the analysis of corrosive halogen
diatomaceous earth firebrick that has been calcined with a compounds such as HF, BCI3, UF6, COCI2 , F2 , and HCI.
clay binder. The metal impurities remaining form complex 8.4.1 Poly(tetrafluoroethylene) supports require special
oxides that contribute to the pink color of the support. These handling procedures. When used as received, they are soft
pink supports are denser than the white supports because of and tend to form gums upon handling. They can also build
the greater destruction of the diatomite structure during up a static charge and spray out of the column during the
calcination. They are harder and less friable than the white packing operation. These problems can be. virtually elimi-
supports and are capable of holding larger amounts of liquid nated by cooling the support to O°C before coating with
phase (up to 30 %) without becoming too sticky to flow liquid phase and by avoiding the use of glass vessels. Rinsing
freely. Their surface is generally more adsorptive than white poly(tetrafluoroethylene) with methanol and drying before
supports. For this reason, they are not recommended for use use is another way to eliminate the static-charge problem:
in the gas chromatographic analysis of polar compounds. 804.2 Supports prepared, from poly(chlorotrifluoro-
However, pink supports provide excellent efficiencies for the ethylene) are structurally harder and are much easier tp
analysis of hydrocarbons and organic compounds of low handle and to pack into a column. .
polarity. 8.5 Tubing Materials-Tubing materials should be
8.3.3 Chemical Treatment of Diatomaceous Earth Sup- chosen on the basis of the following criteria:
ports-Neither the pink nor the white materials give gener- 8.5.1 They should be nonreactive with the stationary
ally acceptable analysis of more polar compounds without phase, sample solvent, and carrier gas.
further treatment. With these compounds, severe peak 8.5.2 They should possess physical properties to withstand
tailing is often observed, especially with the dense pink temperature and pressure of operating conditions, and
supports. This tailing is due to the presence of adsorptive and 8.5.3 They can be shaped to fit in'the column oven of the
catalytic centers on all diatomaceous earth supports. The chromatograph.
adsorptive sites are attributed to metal oxides (Fe, AI) and 8.504 Satisfactory materials include glass, nickel, stainless
surface silanol groups, -SiOH, on the support surface. The steel, and glass-lined stainless steel. Glass is the material of
latter are capable of forming hydrogen bonds with polar choice, unless conditions prohibit its use. Nickel tubing is
compounds. more inert than stainless steel in most applications. Less
8.3.3.1 Metal impurities are removed by washing with frequently used column materials are poly(tetrafluoro-
hydrochloric acid, which leaches out iron and aluminum and ethylene), aluminum, and copper.
renders the surface both less adsorptive and less catalytically 8.6 Carrier Gas-The use of an impure carrier gas will
active. However, even with acid washing, the pink supports produce problems in gas chromatography. Trace water and
are still more adsorptive toward polar compounds than the oxygen can cause decomposition of the liquid phase coated
white-type supports. Acid washing is sometimes followed by on the support. The common carrier gases, helium and
base washing, which seems to remove only minor amounts nitrogen, should contain less than 5 ppm water and less than
of metal impurities, but is a good pretreatment for supports 1 to 2 ppm oxygen by volume. An oxygen adsorption trap
that are to be used for the analysis of basic compounds. can be used to remove trace oxygen, while trace amounts of
8.3.3.2 Neither acid or base washing is effective in re- water and hydrocarbons with molecular weights higher than
ducing peak tailing due to hydrogen bonding with the surface methane, can be trapped on a molecular sieve trap. Place the
silanol groups, -SiOH. These groups are most effectively molecular sieve drier nearest the gas supply. Calcium sulfate
masked by treatment with c1imethyldichlorosilane. has been used in drying tubes, but cannot dry carrier gas to
8.3.4' Acid-washed silanized grades of white diatomaceous the same level as molecular sieve.
earths are recommended as supports for nonpolar and 8.6.1 For some applications, hydrogen may be the pre-
medium polarity liquid phases. Because of the hydrophobic ferred carriet gas. However, additional safety precautions are

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required due to hydrogen's explosive nature. this compressed air usually. contains an,oil aerosol from the
8.6.2 Air (oxygen) can leak into the. ,gas chromatographic pump.
system through loose fittip.gs or "a septun;t, that has been NotE 3-Mostchromatographicsupply houses provide metal tubing
punctured too many times, even though the carrier gas is that has been washed with solvents and is .ready for use.
under a pressure of 40 to 60 psL,J<.eep all fitting~ on the gas
9.1.3 An alternative procedure is recommendedfof,tllckel
delivery lines tight, and check them at periodic intervals.
tubing and can be used to clean stainless steel tubing. Rinse
Change tl1e septum in the injection port frt'jquently. Plastic
the nickel tubing With ethyl acetate, methanol, and distilled
tubing should never be used for carrier gas; hydrogen fuel
water. Then fill the tube with 20 volume % nitric acid and let
(for flO), or maJce-up gas lines due to the possibility of
it stand for 10 min. (CAUTION: Work in a hood and wear
oxygen or moistu,re diffusing through the tubing wall ..
safety equipment when using nitric acid.) Next, nnse the
8.6.3 Each cylinder of carrier gas has its own impurity
tube with distilled water to neutrality and then rinse with
level. Occasional tanks contain large. amounts of impurities
methanol and acetone. Finally, dry the column by blowing
which might overcome a low-capacity oxygen adsorption
nitrogen ot helium through it.
trap .and destroy a gas chromatographic .column at ,high
9.1.4 The column length is generally 3 to'6 ft (1 to 2 m).
temperatUre.. A new tank or a fresh oxygen adsorption unit,
or both will improve this situation. " Shorter columns can be used to decrease the time of analysis
or to separate high boiling compounds. Longer columns are
8.6.4 Always change the ta]1k when the pressure is less
used to improve resolution, but have longer analysis times;
than 200 psi. As the total pre!lSw;e in the cylinder decreases,
(Columns longer than 20 ft "'(6.1 m) require excessive
there is an inciease in the partial pressure of the water and
pressures to maintain the proper carrier gas flow.) A compro-
other impurities. adsorbed on the Imler walls of the, gas
mise is usually made between analysis time and resolution.
cylinder. As a result, the last amounts, of gas delivered from
As a general rule, an increase or decrease of column length
the gas cylinder contain high levels of impunties.
bya factor of3 to 41isnecessaty to see a significant change in
8.6.5 Carrier Gas for Insiruments with:thermal Conduc-
peak separation.
tivity Detectors-A major·factor in sensitivity is the differ-
9.1.5 The diameter ofthe column can be lis in. (3.2 mm)
ence in thermal conductivity of the compound being ana-
or' 1/4 in. (6.4 mm) outside diameter. The lis-in. column has
lyzed and the thermal conductivity of the carrier gas. Helium
less sample capacity, but greater efficiency, and is the most
(thermal conductivity =~33.60 cal/cm-s-°C) is usually the
common type. Glass columns are generally 2 mm or 4 mm
carrier gas of choice. '
inside diameter. Some analysts have found that 3/16 in. (4,8
8.6.6 Carrier Gas for Instruments wit/z Flame Ionization
mm outside diameter) metal columns are the ideal combina-
Detectors-The most: commo:i1ly used carrier gases are
tion between ,the capacity of 1/4 in. (6.4 mm outside
nitrogen or helium. A maximum impurity l~vel of 0.05
diameter) columns and the efficiency ,of lis in. (3.2 mm)
volume % does not generally ,interfere with most applica-
outside diameter columns.
tions. Hydrogen is less commonly used inihe US but is more
9.2 Choice of Diatomaceous Earth Support for Packed
popular in Europe because of availability and relatively low
cost. ' Columns-See 8.3.
,9;3 Phase Loading on. Diatomaceous Supports~For pre-
NOTE 2-Ifhydrogen is used, special precautions must be taken due parative work and analysis of substances boiling below room
to its explosive nature, to ensure that the system is free from leaks and temperature, use 1:5 wt % loadings. for white-type supports
that the effiuent is properly vented. and 30 wt % for pink~type !lUpports; For general work, uSe
8.6.7 Carrier Gas for Instrunients.with Electron-Capture loadings of the range,of3 to 15 wt %. For highest efficiency,
Detectors-Usetsshould 'follow the manufacturers' recom- shortest retention'times, and the. least amount of bleed
mendations for the choice of carrier gas. Some common ones during high-temperature operation, use 3 wt. % loadings.
are mtrogen or 95 % argon/5 % methane. When using a The lower phase loadings have lower sample capacity and
tritium source in the detector, do not use hydrogen as the elute components ;:more;rapidly and at lower temperatgr,es.
carrier gas. Hydrogen will replace ~tium in the source. Always check, the manufacturers' literature for suggested
phase loadings for a particular support. For some applica-
tions (especially head,space analysis) loadings as low as 0.2
9. Preparation of Packed Gas Chromatographic Columns wt. % are used which ,res].l1t in ~ery narrow peaks and short
9.1 Preparation of the Tubing Material: analysis times. High,. p4ase loadings tend to produce less
9.1.1 Glass columns should be cleaned and deactivated, reactive packings.
first by rinsing with 30 mL acetone and then 30 mL toluene. 9.4 Preparation of the Gas Chromatographic Packing-
Next, fill the. column with 10 volume % solution of The following. procedures describe the cqati,ng of a, solid
dimethyldichlorosilane in tolpene. AllQwth~ solution to support with stationary phase; ~he following four methods
stand in the column for 30 min. Finally, rin.se the column are comII).only used to prepare gas chromatograp!:ric
withanbydrous toluene and then anhydrous methanol to cap packings; (a) Filtrati9n or Solution Coating Method, (b)
unreact.ed DMDGS CL groups. Dry the column by passing a Rotating Evaporator Method (c) Evaporative Method; and
stream of dry nitrogen through it Cap, both ends of the (d) Vacuum Evaporative Method. Wh.en preparing packings
coIu.:rpn until such time .that.it can be, packed. with .l()adings in the r~nge of less than, ~ wt %, the Filtration
9.1.2 Metal columns showd be cleaned thoroughly before or Solution Coating Method is recommended. This method
packing by rinsing with methanol,· acetone" an<i chloroform. is preferred because it' P!ovides minimum handling ". of the
The column should be dried bypassing nitrogen or dry air friable white-type supports. For loadings of more than'. 5
through it Do not blow house air through the colum,n since wt. %, other methods can be used. The Rotating Evapor~ior

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Method is recommended, but should only be used if a TABLE 2 Solvent for Liquid Phases
rotating evaporator is available, which turns very slowly at Uquid Phase
TypeA Solvent
20 to 30 rev per min.
Dimethyl Silicone Toluene
NOTE 4-A 5 wt. % loading of stationary phase consists of 5 g Phenylmethyl Silicone Ethyi Acetate
stationary phase added to 95 g of support. Cyanopropylphenyl Silicone Methylene Chloride
Trifluoropropyl Silicone Ethyl Acetate
9.4.1 Filtration or Solution Coating Method-Prepare Polyethylene Glycol Methylene Chloride
Cyanopropyl Silicone Methylene Chloride
100 mL of a solution of the desired phase in a vacuum filter O!her Uquid Phases Use solvent as recommended by supply house.
flask. Use a suitable high boiling solvent (boiling point more
than 60°C). The actual loading of the liquid phase on the
support will depend upon both the viscosity of the phase 1/4-in. thick. A borosilicate glass baking dish makes a suitable
solution and the density and mesh size of the support. container.
9.4.1.1 Add 20 g of support to the filter flask. Reduce the 9.4.2.5 The critical stage occurs when excess solvent has
pressure in the filter flask for a few minutes with a water evaporated, but the bed is still quite damp with a slight excess
aspirator, then release the vacuum. Repeat this procedure for of solvent. Break up the damp bed by gently raking it with a
several cycles in order to remove air bubbles from the pores spatula. As the solvent evaporates from the surface of a static
of the support particles. Be prepared to release the vacuum if bed of support, it leaves a higher concentration of phase at
the slurry foams excessively. the bed surface. Therefore, the bed must be broken up
9.4.1.2 Allow the slurry to stand for several minutes. Pour frequently during the final stages of solvent evaporation to
the slurry into a coarse-frit sintered-glass filter funnel, and prevent formation of an unevenly coated support.
allow the solvent to drain freely until the support settles. 9.4.2.6 Continue to air-dry the material in the hood until
9.4.1.3 Apply vacuum cautiously and stop instantly when the last traces of solvent are gone. Avoid excessive handling
the solvent stops dripping. Dump the support into a flat of the particles to prevent formation of fines due to abrasion,
borosilicate glass dish, and allow it to dry. Do not scrape the especially in the case of the white-type supports.
particles out of the funnel, since this might crush the 9.4.3 Rotating Evaporator Method-Prepare the slurry of
particles. Do not resieve before use. support and phase as described in 9.4.2.1 to 9.4.2.3, except in
9.4.1.4 The actual phase loading will depend upon the an indented, round-bottom flask. Connect the flask to a
viscosity of the phase solution and both the density and rotating evaporator. Rotate the flask very slowly (less than 20
particle size of the support. For example, a 2 % solution of to 30 revolutions per minute) and evaporate the solvent
dimethyl silicone gum liquid phase will give a 3.8 wt % under reduced pressure (water aspirator). Very slow rotation
loading on white-type supports. A 10 wt % solution of a less is necessary to prevent the particles from abrading against
viscous liquid phase will give a 5.5 wt % loading on each other. Use of a heat lamp increases the evaporation rate.
white-type supports and 7.5 wt % on pink-type supports. This method is not recommended for fluorocarbon supports.
Loadings obtained with other phases on other supports are 9.4.4 Vacuum Evaporative Method-Prepare a slurry of
best determined by experimentation. support and phase in a filtration flask of sufficient capacity.
9.4.1.5 The best way to determine the percent loading is (Suitable solvents are given in Table 2.) Attach the flask to a
to extract it from the support by extraction in a Soxhlet vacuum source (water aspirator) and apply vacuum briefly;
apparatus and determine the weight loss. Alternatively, (Be prepared to release the vacuum if the slurry foams
measure the volume of solution recovered and calculate the excessively.) Repeat this procedure several times in order to
volume of solution held up by the support. Calculate the remove the air bubbles from the pores in the support.
approximate percent loading on the support by assuming 9.4.4.1 Apply the vacuum fora longer period, and swirl
that the concentration of the solution does not change. the contents of the flask occasionally until all the solvent is
9.4.2 Evaporative Method: almost evaporated. This is the critical stage.
9.4.2.1 Weigh out the desired amounts of support and 9.4.4.2 Now shake the contents of the flask by gently
phase. Use a larger amount than that required to account for bumping the flask on a wood or plastic board. This will break
attrition, spills, etc. Dissolve the liquid phase in a chemically up the bed of packing: Do not allow the solvent to evaporate
inert, low-boiling solvent contained in a filtration flask (see from the surface of the support bed. Otherwise, the solvent
Table 2). (Most catalogs of gas chromatography equipment will evaporate and leave a higher concentration of phase at
suppliers contain lists of suitable solvents.) the bed surface.
9.4.2.2 Gradually add the support to the solution with 9.4.4.3 Continue to apply vacuum until the packing is a
gentle swirling or agitation but with no mechanical stirring. freely flowing powder, then transfer it to a tray for air-drying
(Suggested solvents are given in Table 2.) The amount of in a hood. .
solution should be just enough to wet the solid support and 9.4.5 Fluidized Drying Thhnique-This technique has
form a slurry with little excess solvent. been used to produce efficient, uniformly coated packings.
9.4.2.3 Evacuate the flask briefly several times to remove During the drying stages of methods 9.4.1 to 9.4.4, when the
air bubbles from the pores of the support. Be prepared to packing has reached the consistency of a wet sand, add it to a
release the vacuum if the slurry foams excessively. fluidizer. Then dry the packing by passing a flow of inert,
9.4.2.4 Transfer the slurry to a large flat borosilicate glass warmed gas (nitrogen or helium) through the bed of packing.
dish, and slowly evaporate the solvent in a hood with no 9.5 Packing the Gas Chromatographic Column-The pur-
further handling. The dish must be of a size that the packing pose in packing a gas chromatographic column is to fill the
is spread on the bottom in a thin layer, no more than about column. with packing as completely as possible, leaving no

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empty spaces. Two variations are noted in 9.5.3 and 9.5.4 (a water aspirator. (If a water aspirator is used"a 500-mL filter
pressure-fill proc~dure and a vacuum fill procedure). flask, or the device shoWn in Fig. 2B, shQuld be placed in the
9.5.1 It is preferable to coil the cQlumn before packing to line between the pump and the column.) Do not turn Qn the
prevent crushing of the support particles. Metal columns can vacuum yet.
be coiled after loading to ;meetequipl;lleni 'requiren;uints. 9.5.4;3"Add 1 t6 2 mL of packing 'to the funnel, and tap
Bends in the packed region1l).ust never be made with radii the column gently to settle the packing. A pencil or a wooden
less than those specified in 9.5.2; to avoid crushing the spatula hanQIe can be used. Alternatively, the column can ,be
packing in the cQlum.n. stroked with a plastic saw. The use of an ele~tric vibratQr is
9.5.2 Right-angle 'bends are often necessary to make not recommended. Excessive vibrati():Q.. ~WW c~llse the parti-
connections to injection and detection systems, and must be cles to abrade against each other, prod*i~g fines and newly
made before packing the columnsincesometubingdeforma- fractured surface~ that are not co~ted with stationary phase.
tion will occur, which will crush some of the solid support. 9.5.4,4 Turn on the vacuum source.C<intinueto add the
Bends for such purposes should be within 4 in. (10 cm) of the packing in small increments with tapping' until'the column is
column ends. For coiled. columns, minimum' diameter full. Finally,. apply pressure to the head'hf t4e column to
mandrels should be as' follows: for lis in. (3.2 mm) OD pack it a little tighter. However, take care to make sure the
column use al l/:z-in.· (38-mm) mandrel; for 1/4.in. (6.4 mm) pressure jsequalized slowly, because p:icklngwill be blo~
OD column use a 2-in: (51-mm) mandrel. These configura- out of the column if the p!,"essu:(e is released too suddenly.
tions do not preclude the use ofU- or W-shaped columns. If 9.5.4.5 Next, tap 'out enough pac;king to crj:ate a lis in. (3
a U-or'W-shaped column is to be .used,the miliimum 180° mpl) void space at the injector port end of the column. Plug
bend diameter must be at least that given for the above this end. with a, silariized glass wool plug. Do not .pack the
mandrel sizes. 7L plug tpo tight,lY. This. will either impede the carrier gas flow
9.5.3,;Pfessure,Fill Procedure~To'each end of the column or crush the packing particles. .
to be"filled, fit a nut;:a ,back ferrule, and a suitable front 9.5.4.6 Higher' efficiencies are always observed if the
ferrule. Place a small plug of. silanized glass wool into the column is packed for on-column injection. In this techriique,
detector end of the column, arid cap thecolll,nin by screwing the column is p~cked so that there is space at the injection
in Ii metal cap With a 1/16-in; vent hole drilled into it. When port end Qf the column, which is then piaced inside the
analyzing trace.- acidic compounds,asorganic acids and injectiQn port. This void space should. be of such a length
phenofs, adsofptioncan be decreased by using phosphoric that the injection needle just reaches, or slightly penetrates,
acid-treated glass wool to plug the column ends. Wear safety only the glass wool plug, not the packing, when ,the column is
glasses when pressure-packing columns.-" " insta,lled. Thus the sampJe is injected almost directly onto the
9.5.3.1 Attach the end of,the empty column to ali column.. .
apparatus similar to .that shown in,. Fig·.'.2A; Add to the , 9.6 :Conditionirzg of Packed GC Columns:
reservoir sufficient packing ,niaterial to fill the -column·, .plus 9.6.1 The pU1]Jose of·· the conditiQning process is to
about 30 %. Attach the ,upper .end of the reservoir to a remove. extraneous material (solvent and adsorbed material)
nitrogen, supply line controlled· to' provide' approximately 40 from the column before analytical usage. Since the column is
psi. Check thaVall connections are tightened, place a safety heated, the liquid pha~e' also redistributes itself Qver the
shield'in front of theisetup, and, apply AO"psi to the system. support surface to provide a more even coating.,
, ·9.5.3.2 As theistationaryphase startS.to fill the column, 9.6.2 Install the column int.o the gas chromatograph atthe
gently tap thecohimn with:a.'wood rod: (handle of spatula or injection side. only. Do not connect the column to the
screwdriver) or an electrical vibrator set at a very low detectQr during the conditioning st~ge. Any column bleed
vibration level. Continue tapping until the 'packing shows no might foul the detector and the connection lines between the
voids ;and ,the level of packing in the reservoir· remains column' and detector. Turn o.n the normal analytical carrier
constant. gas flow and flush air out of the column at ambient
9.5.33 Shut off the nitrogen supply and wait for the temperature for JO nUn. .
pressure ." to 'dissipate.. Disconnect the,: ;column from, the 9.6.3 Heat the..·column at a.,rate of 2°C/min to. the
reservoir.. Do not discbnnect the column while it is under conditiomng temperature, The latter temperature shQuld be
pressure. Have';,a. Cleanbeaker ..available to collect excess at least 25~C higher'than the analytical temperature but 25°C
packing material that will fall from the opened reservoir. Tap lower than the· maximum. Qperating. temper;lture 'recom-
out about lis in. (3 mm) of column packing, and replace it mended for the \liquid phase. Maintain this temperature
with a silanized glass wool plug. Affix a" metal column' tag overnight with .carrier gas flow.,
engraved ,with 'Ii "descaption of the stationary phase, loading, 9.6.4 The next day cool the column and connect it to the
support, and the assigned column number. detector. Detectors operated in: very sensitive modes, partic-
9.504 Va(Juum~FillPrQcedute: . ularly the,.electron .capture detector, might require two or
9.5.4.1 Chimp thebohlmnc.;so -that the detector 'and more days of conditioning at the higher temperatures before
injector 'ends point upwatd:Plug the detector. end of' the a satisfactory baseline is obtained.. (Othersources for baseline
colunm with a 1/4~iit plug of silaruzed glass. wool. Use drift 'and. noise ...are <impurities in the Garrier gas,. a dirty
phosphoric~a:cid treafed glass wO'ol when analyzing for trace detectQr, air .leaks ,in the.gas-line fittings, insufficient carrier
organic acids arid phenols. .. , ." gas pressure,. a .much-punctured septum, chemical decompo-
9~5.4.2 Attach Ii 'small funnel to the ,injection 'port end of sition of the phase (due to presence Qf traces.of acid or base
the' column. Attach the detector end of the, column to a on the support, in the phase, or on the inner Golumn walls,
vacutim· source, 'either a vacuum pump (preferably) or a and incQrrect fuel gas ratios to the flame ionization detector.)

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Vacuum

FIG. 2A Vacuum Fill Apparatus

functional groups other than phenyl or methyl.

~
+ Nitrogen at
40 psig ., 9.6.6 Many of the liquid phases are commercial-grade
material, and conditioning might require several days before
\ the noise level is low enough to provide :usable baselines at
high sensitivity. The use of gas chromatographic-grade
pha.ses is recomm.ended since they have been carefully
purified and long periods of conditioning are usually not
necessary.

10. Sample Injection Procedures

10.1 Injection Technique-Useful analyses are obtained \
only by injecting representative samples into the gas I
. ,;'"
chromatograph. Since chromatographic samples are small
the best practices and procedures must be followed. '
10.1.1 If elution is to be otherwise, the sample injection
must be almost instantaneous in order to introduce the
sample as a plug. Avoid unnecessary sample dilution and
inadvertent trapping. .'
10.2 Sample Size-Sample sizes used for analysis with 2-
to 5-mm ID packed columns are in the range of0.1 to 10 JlL
multi-component liquids. Gas samples usually range from 10
JlL to 2 mL.
10.2.1 There is frequently a reduction in retention times
with sample size when the column is overloaded; Therefore
when identifying by comparison with a known sample, th6
same amount of each component must be used in an
FIG. 28 Pressure Fill Apparatus amount that does not overload the column. The sample size
that overloads the column is that size which decreases the
efficiency (Section 11) by 10 % compared to a smaller
, 9.6.5 There is a special "no-flow" conditioning procedure
s~mple s~e. Sample overload is sometimes shown by peaks
,with certain silicone phases, as methyl and
which can be used .with
With slopmg fronts and backs that are almost perpendicular
methylphenyl silicones With or without low vinyl content. It
to the baseline. (Another reason for this peak shape is
has been reported to improve analysis for drug compounds.
insufficient vaporization of the sample. Either the injection
The procedure starts by conditioning the colurpn for 112 h as
port or the column temperatures; or both, are too low.)
described in 9.6.2. Turn off the carrier gas flow, cap the free
end of the column with a metal cap, and heat at 31O·C for
10.3 Sample Injection Devices-Sam,ples may be intro-
duced by syringes, automatic sample injectors using syringes,
1.5 h with no carrier gas flow. Cool the column to 100·C.
or sample valves. (Thert~ are also devices that. that, introduce
Uncap the oven, turn on the carrier gas flow, and continue
capsules containing sample into the injection port.) For
the regular conditioning procedures listed in paragraph 8.6.3.
rigorous quantitative work, any sample introduction device
.NOTE •5--:-This no flow conditioning procedure may damage or should be flushed and filled at least three times with the
destroy non-silicone containing stationary phase or silicone containing sample immediately before use. .' .'

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10.3.1 The most common method for liquids is the use of release the sample, which is then swept into the gas chro-
a syringe injection technique through a self-sealing septum. matographic column by the carrier gas.
In the usual lO-I.LL syringe, there is approximately 0.8 I.LL of 10.4 Sample Container,-Care must be taken in the design
dead volume in the syringe needle. This volume is additional and construction of sample containers so that none of the
to the volume in the syringe barrel and can be measured by components of interest are in any way changed or removed
withdrawing the entire sample volume into the syringe from the sample by reaction, diffusion, or adsorption.
barrel. Stopcock grease and material desorbed from plastic cap
10.3.1.1 First, pump the sample back and forth vigorously liners are frequent sample contaminants.
in the syringe to wet the plunger and to remove air bubbles.
Then withdraw the sample back into the syringe so that the 11. Evaluatibn of Column Performance
entire volume can be read on the volume marks of the 11.1 Make, a test mixture which contains a normal
syringe barrel. When preparing the syringe for injection, aliphatic hydrocarbon and the compound being analyzed, or
never leave the sample solution in the needle. This technique one similar in structure. The aliphatic hydrocarbon peak
will minimize sample boiling out of the needle when it is should appear in the chromatogram near the second compo-
inserted into the hot injection port. nent in the test mixture. (Suggested aliphatic hydrocarbon
10.3.1.2 In an alternative procedure, called the solvent- mixtures are shown in Table 3.) The peaks of both compo-
flush technique, load the syringe in the following order: nents should be about the same size. The analytical condi-
solvent, air, sample solution, and air, with only airJemaini1l:& : tions should be chosen so that both chromatographic peaks
inside the needle. When the sample is injected, the solvent IS will appear about 4 to 6 times the retention time (distance) of
the last to leave the syringe, and it rinses out sample residue the solvent peak. (If a selective detector is being used, choose
in the needle. a compound that will give a response to that detector.)
10.3.1.3 Wipe the· syringe needle off before injection. 11.2 The different methods of determining column effi-
Insert the' n.eedle carefully through the GC septum, inject the ciency are shown in Equations 1, 2, and 3. Fig. 3 shows the
sample at once, and withdraw the needle in one continuous measurements made on a chromatographic peak which are
motion.··.· ' used to determine column ~fficiency.
10.3:1.4' Often tne tip of the needle becomes bent,
N = 16, (tRlwb)2 (1)
forming a fish hook that will tear the septum on subsequent
injections. This can be detected by brushing the end of the N = 5.54 (tRlwh)2 (2)
needle gently over the end of a finger. A few strokes on a H= LIN (3)
<;harpening stone will remove the fish ,hook where:
'j j

10.3.1.5 Syringes should be cl~~I1-ed witl:t"a solvent that N = number of theoretical plates,
will remove all traces of contamination. Con~ult the manu- tR = retention time, or distance, measured in mm,
facturer's literature for further information. Many chromato- Wb = width of the peak at base, measure4 in mm, (Deter-
graphic supplywndors sell suitable cleaning solutions and mined by extrapolating, as shown hi Figs. 3a and 3b.)
kits. Liqtrld ,sapiplevalves, in both autom~ted and manual Wh = width of the peak at one-half the peak height, h, all
versions, ar~a:I~o available. measured in mm, (See Note 6.)
10.3.1.6 Gas. samples are most conveniently injected H = height equivalent to a theoretical plate, HETP, and
using gas-tight syringes. These devices are q~te satisfactory L = length of the column in millimetres, or in centimetres.
for survey: ,work because the sample size caJ? easily be NOTE 6-The peak width may be measured to the nearest 0.1 mm by
HQwever, the syringe need1~ .is more likely to clog
verified. Hqwever, a magnifying loupe fitted with a scale graduated in 0.1 mm increments.
with pieces of septum material when,.gas samples are injected The peak to be so measured should be at least 10 mm wide; this is
than when liquids are injected. If no chromatographic peaks arranged by choosing an adequately large chart s~ed.
are observed, test'the syringe by injecting ::rir.intoa 1,iquid. If 11.3 Equation 1 is often used. Howe.ver, it involves an
no bubble~are
nQ bubbles are seen, uP-9lqg
up-clog the needle with a. wire or· by extrapolation of the baseline, shown in Fig. 3, which can be
filiing
filling thesyr.inge without' t\1e needle al;ld, forcing solvent
the sypnge without in error. The use of Eq 2 is preferred because the term, Wh
th,rough the needle, However" repeatable results in gas can be determined directly from the chromatogram without
analysis will only be obtained ,using gas sample valves that extrapolation. However,' the. width of ~he recorder pen line
have a fixed sample loop. The latter valves can easily be can be variable which can lead to difficulties in determining
automated. the true value of the peak width. Use a pen that writes with a
10.3.1.7/\. ~ealed, friable, or puncturable ampule col;l- 'sbarp:.ihln line. The peak Width shql.dd. be measuredfrqm
taining a Weighed a~()"!lnt"of sample maY,be placed, in the the ieading edge o( one line. to th~ leapi.:p.g edge of the <#lier
injection ,chamber. The ampule js p,hysicallY broken to lil)e,. as showl;1 in Fig. 3a.'.For a~ci1fa~Ylpetermine ,tpe, pe,il,k
Width twice '(see Fig. 3a),. and .l:J.se tll~. averag~. v~ue.:to
TABLE 3 n-Alkanes Used for Column EValuation at,200°C calculate "N'. To further miiIiinizeerrors in detehhiningJhe
, . '1.'/- '
. .. "J;.j :' J; '~'
1 m 10 % Dimethyl silicon'eC18,'C20/C;:~
, -"3 m10 % Dimethyl silicone "C14".G1il. G18
,1 m 10 % PhenylmethYlsilicone.C:!o. C22 • C24 TABLE 4G~~eral qiltirnum V~IJE!~ ~!Carrier~asFlow Rates.
3 m10 % Phenylrnethyi siliconeC16 • C;8. C20 General Optimum 9anier. Gall,F'ow, :."; ,.
1 rri 10'% Cyan'opro'pyl silicone C24 • Cli6. C28 Columri Dj'mensions
' .
~':,

3 m 10 %Cyanopropyl silicone Cllo• C22 • C24 Helium" Nitrogen ')'

1 m.10 % Polyetl)ylene ~Iycol C24 • C26 • C28 Vo In. 00 or 2 mm 10 ,. 30mL/min 20 mL/mln
3 m 1(:>% Polyethylene glycOl C18 • C20.G22 • 1ft! In. 00 or 4 mr'rl'lD 60mL/min 40 mqmin "

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I II
I t' R
I ..
I
I
I
(3a) I
I I

I
I
I-
a:
!/COMPONENT
: PEAK OF AN UNRETAINED

~
tn

-----'-----,,'----------------'-=_..: - - "" - ..;:...-

----l~~TIME :l~
. wb
.. t,.
FIG. 38 Use o.f the Chromatogram to Calculate Column Efficiency

TABLE 5 Typical Efficiencies and Ratings of Gas

Chromatographic Columns
Plates/ft Plates/metre HETP Rating
500 1640 0.6 mm Excellent
(3b) 400-500 1300-1640 0.60-0.76 mm Good
300-400 980-1300 0.76-1.0 mm Fair
300 980 1.0 Poor

FIG. 3b Shows the Procedure to Measure the Peak Width, at wh ,

12. Use of the Gas Chromatographic Packed Column
to Account for the Thickness of the Pen. Use the Average of the Two 12.1 Certain precautions and preventive maintenance are
Determinations, w' h and w' I h' shown here necessary to obtain the best column performance. Some of
these points have been made before and will be referred to in
peak width; use a recorder chart speed that will give a this section. Further precautions will also be discussed.
minimum peak width of 4 to 5 cm. The peak height should 12.2 Carrier Gas Purity-Trace, or adventitious, oxygen
be about 40 to 80% full-scale. chart deflection. This can be and water in·· the carrier gas can produce degradation of
achieved by either changing the detector sensitivity, or liquid phases on the support. Purification of the carrier gas
changing the sample size. However, be sure that the sample and necessary precautions are discussed in 8.6.
size does not exceed the capacity of the column. 12.3 The Injection Port-The injection port is often a
11.4 The .optimum efficiency occurs at the optimum . source of trouble. The temperature can be either too low or
carrier gas flow which can be determined by plotting the too high. See 6.3. Another problem is a leaking septum. See
efficiency of the hydrocarbon peak versus carrier gas flow. 6.3.4.
General optimum values of the carrier gas flow rates are 12.4 Column Care:
'shown in Table 4, but should be determined for the . 12.4.1 Never heat a gas chromatographic column with air
particular column. in it.. When any column is first placed in a gas
chromatograph,. flush any air (oxygen) out of it by flushing
11.5 Calculate the efficiency of the second component.
with carrier gas at normal flow rates for 15 to 30 min. at
The results should be about the same for both components.
ambient temperatures. Then heat the column to the desired
If the efficiency of the second component is about 25 % operating temperature. Always cool the column to room
lower than that of the hydrocarbon, the column might not be temperature before removing it from the gas chromatograph.
the best choice for analyzing the compound(s) of interest. A If the column is to be used again, cap the ends with metal
strong indication of nonsuitability is shown by compara- caps to prevent diffusion of air (oxygen) into the column
tively greater tailing of the. nonhydrocarbon component. during storage. Contact of the stationary phase with oxygen
Tailing can indicate strong interaction with the column .when hot .or for prolonged periods of time at room temper-
packing (phase or support, or both), column tubing, or . ature cause degradation of the stationary phase. This is
adsorption in the injection port or in lines leading from the particularly the case with polyglycols· and polyester type
column to the detector. phases and to a lesser extent cyanosilicones. Other phases are
11.6 Typical efficiencies are shown in Table 5. Not all gas affected to a varying degree.
chromatographic packings are capable of excellent efficien- 12.4.2 After long periods of use, column performance
cies. For example, porous polymers, Teflon supports, and may degrade as shown by peak broadening, tailing, or
some liquid phases, such as trifluoropropyl silicone can give gradual merging of adjacent peaks. Often the problem lies in
efficiencies less than 500 plates per foot. the front end of the column. The injection port temperature

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might have been too high and destroyed the initial section of means of con;tparing the results obtained from different
the liquid phase on the column' packing. Residues or instruments (see Practice E 355).
decomposition products might have built up on the glass 13.4 Absolute compound identification or characteriza-
wool plug. These problems can be remedied by repacking the tion, must be m.ade with ancillary techniques such as mass or
first few inches of a glass column, or cutting off the first few infrared spectrometry, nuclear magnetic resonance, chemical
inches of a metal column. Use fresh silanized glass wool to analysis of the effluent, or spot tests for functional groups.
close the end of the column in both cases. 13.4.1 The samples for the analyses in 13.1 through 13.4
may be obtained by trapping components as they emerge
13. Methods of Qualitative Analysis from the chromatograph. A trap, glass capillary, or V-tube, is
13.1 Identification of compounds by gas chromatography cooled with ice or dry ice, and placed in the effluent stream
alone cannot be absolute, and the results must be considered of the column. Several collections may be required to obtain
with care. Elution of a compound is dependent upon carrier a sufficiently large sample.
gas flow rate, column temperature, support size, amount and 13.4.2 The· -collection of effluent is easiest with
type of liquid phase, column dimensions, instrument dead nondestructive detectors, see 6.7. In the case of destructive
volume, and column pressure drop. 'These parameters must detectors, a 'split is made for the collection just before the
be stable to obtain reproduciple results. The recommended detector.
format for a gas chromatographic method is given in Section 13.4.3 Apparatus is also available so that the effluent from
15. the gas chromatographic column can be analyzed directly by
13.2 Tentative identification ofa compound can be made mass spectrometers or infrared spectrophotometers.
by comparing its adjusted retention time against those of
known standards using exactly the same chromatographic 14. Methods of Quantitative Analysis
parameters, c"
14.1 Gas chromatography can be used to determine
13.2.1 The retention time is the time interval measured quantitatively the composition of complex samples. There
from the point of injection ,to maximum peak height of the are several factors that must be considered before the sample
sample. Adjusted retention time, t R, is derived by subtracting is analyzed; The recommended format for gas chromato-
the time required for art unabsorbed gas, like air, or methane, graphic metbods is given in Section 15.
tM , to traverse the column (also called the gas holdup time) 14.1.1 The Chemistry of the Sample-The chemistry of
from the retention time, tR. the sample; if known, allows a chromatographer to select
NOTE 7-0n some'solid adsorbent columns, such lis molecular more accurately a column compatible with the sample, and
si~v~s, there is no·nonadsorbedtcomponent. " to anticipate' 'potential iriterferences 'from reaction by-
tR = tR - tM (4) products.
13.2.2 Retention times are affected by. all chromato- 14.1.2 The Choice of a Detector-A detector must be
graphic parameters. As it result, direct comparison of reten- chosenwith,the needed selectivity and sensitivity. If compo-
tion times of the same components ort different instruments nents' will be analyzed at low levels" an electrolytic conduc-
or between laboratories should be done with caution. Use of tivity electron: capture, nitrogen phosphorus, 1Dicro-
relative retention time is an easy praotical techtlique for "coulQinetric, iQniza;tion; or flame photo.metric detector
providing elution data;. The retentionofa component. is !JhoUId ,be selected. The ,<ietec:tor ;may, be limited to Jhes,e
expressed relative to the retention ofa known reference lower concentrations and not app)icable to high concen:tr:a-
standllrd. The reference standard should possess structural or tions. . ' :
chemical similarity to the compounds being analyzed. 14.1.3 Initial Separation ofComponen~s..,.,-:Next, a column
13.3 The retention of a given weight of compound is must be chosen that will resolve the components of interest
usually independent of its concentration 'if the compound in the sample withln a reasonable a;mouQt..of·ti,me. First, a
does not overload the column producing skewed peaks. The rou&p-. separatioll &hould b~ achievy,c;l'witIl.known "standards.
retention' of the compound is also independe'nt of other Next, actual samples should be analyzed to ,determine 'if
substanceS present if there is no appreciable overlap with tl},ere"ar~ 3,J?-Y interferepc~s. A,secQl1cl cqlqmIl,;of,a,n anCillary
another compound. Substances that exhibit positive, ,or 't¢chniq~e(GC/m.as& jpectmme~rY,' GCjjn{rareQ' spectmm-
Langmuir-type, skewing (tailing) durlng,elution will produce '~tt;y, etc.,; shoUld,pe" useei'to veiify' .that ~a4(fitionaI compo-
a decrease in retentionas',the.concentrationjncreases; 'while nents ar~ not .elriting~tp the component of interest. Each
negative, 'or anti-Langmuir-type fronting will produce an new sample adds. the PbssibilitY. of an interference eluting
increase in retention time With increased concentration; , with the componeni .of interest; "therefore this should be
13.3.1 The logarithm of the retention time of members· of checked often. Ir-an i~teiference is detected,"the chromatog-
a homologous series. run isothernially is usually a linear rapher must change the' method to re'Iiiove it. The several
function of" the number of carbon latbms of a molecule. bpti6ns for dOlJi~ this are' as follows:' :; ,
Vsingthis characteristic, two or
three reference c'Ompounds 14.1.3.1 Select aCdfilinti statibriary phase with a greater
can provide sufficient information to prepare a plot of the selectivity for either' the iinterferenCe or the compohent of
logarithm of the retention time versus caroon number, and interest. . .... .
they can identify other members of the. 'series: Retention 14.1.3.2 Choose a different type of deteotorthat would
time, adjusted or not; is of little value in comparing the detect the component of: interest 'but nofthe interference.
results. from various instruments. The. use of Kovats reten- Exam:ple& would' be water 'not being detected by a flame-
tion index, based'onthe relative retention of,a·compound to ·ionization detector, or hydrocarbons not detected by an
the retention of normal paraffins; provides a morel'eliable electrolytic conductivitY or electron capture detector.

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14.1.3.3 Consider other types of chromatographic separa- 14.1.6.1 All manufacturers supply an integral electrom-
tion such as capillary gas chromatography for more efficient eter to allow the small electrical current changes to be
separation of volatile compounds, liquid chromatography for coupled to recorders/integrators/computers. The preferred
separation of non-volatile compounds, or another appro- system will incorporate one of the newer integrators or
priate separation technique. computers that converts an electrical signal into clearly
14.1.4 Detector Sensitivity and Linearity-Once the chro- defined peak area counts in units such as microvolt-seconds.
matographic separation has been optimized, the detector can These data can then be readily used to calculate the linear
be optimized and calibrated. Gas flows should be adjusted to range.
the optimum levels to get peak sensitivity at the concentra- 14.1.6.2 Another method uses peak height measurements.
tion range of the components of interest. The detector must This method yields data that are very dependent on column
also be clean and leak-tight. (See the manufacturer's manual performance and, therefore, not recommended.
for suggested procedures.) 14.1.6.3 Regardless of which method is used to calculate
14.1.4.1 The linearity of the detector over the desired linear range, peak height is the only acceptable method for
concentration range of the component(s) of interest is determining minimum detectability.
determined using prepared standards. This step will deter- 14.1.7 Calibration-It is essential to calibrate the mea-
mine what the response is to increasing amounts of compo- suring system to ensure that the nominal specifications are
nent. The peak area or height should be plotted versus the acceptable and particularly to verify the range over which the
concentration for about five concentrations near the ex- output of the device, whether peak area or peak height; is
pected sample concentration. There should be a linear linear with respect to input signal. Failure to perform this
correlation. Nonlinearity may be caused by reactivity, calibration may introduce substantial errors into the results.
adsorptivity, thermal sensitivity, or excessive column bleed. Methods for calibration will vary for different manufac-
If the latter is the cause, change to a more thermally stable turers' devices but may include accurate constant voltage
column or one of different polarity. Column reactivity can supplies or pulse generating equipment. The instruction
be characterized by skewed, misshaped peaks. This can be manual should be studied and thoroughly understood before
corrected by installing a fresh column of the same type that attempting to use electronic integration for peak area or peak
does not have reactive sites. Test mixtures can be used to height measurements.
demonstrate nonreactivity. Other sources of adsorptivity or 14.2 Types of Calculations:
reactivity with the sample are the injection port, connecting 14.2.1 Each method of quantitative analysis has advan-
lines to the detector, or glass wool. Each of these sources can tages and disadvantages. The four methods of quantitative
be detected by carefully troubleshooting the system. analysis are as follows:
, 14.1.4.2 The detector performance should be checked 14.2.1.1 Internal standardization,
periodically throughout the analysis. This can be done by 14.2.1.2 External standardization,
injecting one of the linearity standards and comparing it to 14.2.1.3 Normalization, and
the linearity plot. 14.2.1.4 Corrected area.
14.1.5 Peak Area or Height Measurement-Many types 14.2.2 Internal Standardization-In this technique, a
of peak area and height measurement techniques exist. The pure component (the internal standard) is added to a sample
oldest methods for calculating the peak area are manual in a known amount. The peak area, or height, of all
measurement with a ruler of the peak area using' one of the components of interest is compared to the peak area, or
following equations: height, of the internal standard. These comparisons are
peak area = Wh x h (5) referred to as response factors:
where: RF = Ae/A/s x W/s/We (7)
Wh = peak width at half height, and where:
h = peak height RF = response factor,
or Ae = peak area of component,
peak area = 1/2 Wb x h (6) A/s = peak area of internal standard,
W/s = mass of internal standard, and
where: We = mass of component.
Wb = peak width at the base of the peak, and
The amount of the component can be calculated from the
h = peak height. weights of the sample and internal, standard, the response
Another precise measurement defines the peak area as factor, and the peak areas (or heights) as follows:
retention distance (in millimetres) times the peak height (also
in millimetres). For peak height, this distance is simply %Cqnce= W/s/Ws X Ae/A/sx I/RF x 100% (8)
measured from the baseline to the apex 'of the peak. where:
However, these techniques now, for the most part, have been Conce = concentration of component in sample,
replaced by electronic integration, which is much faster. The W1S = mass internal' standard,
proper use of these devices is crucial for accurate quantitative Ws = mass sample,
analysis. The instruction manual for the particular integrator Ae = 'peak area of component,
should be studied and understood thoroughly before at- A1S = peak area of internal standard, and
tempting to use electronic integration for peak area or peak RF = response factor.
height measurement. This technique provides a correction for the relatively high
14.1.6 Data Handling: variability of syringe injection and; therefore, yields a more

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precise method of analysis. Neither the quantity of soh,ltion ties are easily determined by filling a 50-~L syringe to about
injected, nor change in detector response, will alter the area 3Q ilL, wiping the needl~j weighing ,it, expelling thesampJe,
ratio of the analyte and the internal stand~d. To achieve wiping the needle again, and rew~ighing, it.
optimum performance, the internal standard mu~t me!'t i.h.e t42.3.4 T~Ui. pe~ areas or heights of the component in
following criteria. the sample and the standard compound are measured and
14.2.2.1 The internal standard must elute in-an area of the the concentration calculated as follows:
chromatogram that is free of sample components, or possible % Conce = Ae/AEs x WES/WS x % ConcES (9)
sample components.
14.2.22 The internal standard must not react with the where:
sample or any of its components. Conce = concentration' of component,
14.2.2.3 The internal standard and the sample must be Ac = peak area of component in sample,
homogeneous. A cosolvent ma.y be .used to produce a A ES of
= peak area external standard,
homogene0us mixture.
WES = mass of external standard injected,
Ws '. = mass of sample injected, and
14.2.2.4 The internal standard must be easily and accu-
ConcES = concentration dfexternal standard in solution.
rately added.
14.2.4 Normalization-"·:'-Thls caIculationassumes ilia.t
14.2.2.5 The internal standard must be pure.
every cOnlponent elutes and thai each has' similar tespon~e
14.2.2.6 The intemalstandard should elute near the
factors. Ifis a fast procedure that reqUiI:es no weighing. The
component of interest.
sample is injecteo, and the peak areas or heights of all
14.2.2.7 The concentration oLthe· internal standard, rela- components are measured. The concentration of the' compo-
tive to that of the analyte,.should be such that these two nent of interest is calculated as follows:
peaks are within'50 to 100 % of full scale deflection with.the
same electronic attenuation and sensitivity setting in order to % Conce = Ae/AALL x 100 (10)
allow manual measurements and calculations of parameters, where: '"
if desired. Concc = concentration of component of interest,
14.2.2.8 The most common use for the internal standard Ac = area of component, and
technique in chromatography is to correct for quantitative AALL = sum of areas of all components.
variations in the injection, particularly when using syringes. Severe errors result if the components have different re-
For this. purpose, the internal standard need not be chemi- sponse factors or do not all c;:lute. , ,
cally,related to the analyte, ,but must possess the criteria cited 14.2.5 Corrected Area-This method corrects for 4ill'er-
above and may be added in the final solution. ences in response bu~ still assumes that ~ compo:nents elute
14.2.2.9 In certain applications, an internal standard with and are observed by ~he dete,c;toJ;. Response factors are used
functional groups similar to the analyte may be desirable. to correct for response differences as ;follows: ,
For instance, those with a labile proton can be' expected to % Cdiic2'= Ae/(A x RF);uL x RFc x 100 % (11)
exhibit similar adsorption isotherm behavior and to undergo
similar' physico-chemical, transformations during such pro- where:
cesses, as extraction from a complex matrix ,or deriva- Concc = concentration of component of interest,
tization, or both. ,Likewise, similar electronegative functional Ac = peak area of component, ,.' , '
groups are likely to behave' similarly towards an electron (A x RF)ALL = sum of peale areas times their respeetive
response factors relative to a standard~ and
capture detector.
14.2.3 External Standardization:
= response factor of component to the same
standard.
14.2.3.1 This method compares peak areas or heights of
components in a sample chromatogram to those in a
standard solution injected separately. It is critical that 15. Recommended Form for Writing Gas Chlomatographic
accurate amounts of sample and standard be injected for the Methods
method to be valid. Generally, the solvent flush injection 15.1 General-Not all ofthe steps outlined in this section
technique (see 10.3.1.2) or a sample valve offixed volume is may be needed to describe adequately a method. A number
preferred. of variations in procedure format are shown in the publica-
14.2.3.2, The advantages ofthis method are as follows: tion, ASTM Standards in Chromatography.7Ideally, the
, (a) Nond~teCted components ddnot bias the results. procedure should be written so that it can be followed by a
(b) It 'can be used where several known components must person with the equivalent of a high school-chemistry
be determined in a very complex sample. understanding or six to twelve months of practical laboratory
(c) It can quantitate relatively reactive components. experience. Critical steps should be identified along with any
(d) A single sample can be analyzed where maximum reasons\ that show why this step is 'necessary to achieve 'a
accuracy is not: requireq. successful analysis. Any involved ·procedures should be
(e) Nonlinearity has a minimal effect'if the external written in an Appendix So that the main points in the
standard is near the concentration of the sample. procedure can be read more easily.
14.2.3.3 The critical part of this method is .the injection. 15.2 Recommended Form:
The volume of sample in the injection syringe and standard 15.2.1 TitM-The title should be concise; but complete
must be accurately measured, allowing no bubbles in the slug
of sample or standard solution. If the sample. and standard
have different densities, a correction must be'made. Densi- 7 ASTM Standards on Chromatography, ASTM, 1981.

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enough to identify the component(s) analyzed, the nature of the dimensions (outer diameter and inner diameter, or wall
the method (gas chromatography), the detector, and the thickness and length). Any pretreatment of the column
materials to which it is applicable. Select words that easily material, solvent washing, or silanization should be men-
lend themselves to indexing. tioned.
15.2.2 Scope-State as clearly as possible the range of 15.2.11.2 Partitioning Phase-Solid adsorbent, if used
application of method. In a separate paragraph, note (type and mesh size). Coated support if used (liquid phase,
interferring substances or any significant limitations of the percent loading and coating procedure, support type and
method. This material could be placed in a later section pretreatment, and mesh size). Note sources for special
(15.2.S), if an involved description is necessary. materials in footnotes. Provide preparation and purification
15.2.3 Pertinent Documents or References: method for materials not commercially available. In an
15.2.3.1 ASTM Standards. Appendix, note other liquid phases that have been success-
15.2.3.2 Other Standard Methods-Include any standard fully used in this analysis.
methods. 15.2.11.3 Column Preparation-Describe the procedure
15.2.4 Summary of the Method-Describe the method in used to pack the column. Note the amount of packing in the
a general way, without going into details of the procedure. It column.
may be appropriate to touch briefly on the following points: lS.2.l1.4 Column Installation-Note if the column is set
sample introduction technique, column dimensions and type up for back-flushing, if a sample fraction is removed on a
of tubing material, nature of the packing material, mesh size pre-column, or other special column arrangement.
of support or adsorbent, liquid phase loading (if a liquid 15.2.11.5 Column Conditioning-Provide the column
phase was used), isothermal or programmed temperature conditioning procedure.
and detector type (thermal conductivity, flame ionization, lS.2.11.6 Column Evaluation-Give the procedure for
electron capture, etc.). evaluating the column. Calculation of the resolution between
IS.2.S Significance and Application-Use this section for two components in a standard mixture will often be suffi-
a more detailed discussion than can be fitted in the Scope. cient. Provide some estimate of column life and signs of
15.2.6 Definitions-Include special definitions in this sec- degrading column performance (loss of resolution, peak
tion. General chromatographic definitions are already avail- broadening, or tailing). Provide examples of good and bad
able in Practice E 355, to which reference can be made.
chromatograms.
15.2.7 Interferences-Use this section for a more detailed
15.2.12 General Apparatus..,....Volumetric flasks and pi-
discussion than can be fitted into the Scope.
pets, microsyringes for sample introduction, balance (ca-
IS.2.8 Special Comments-Use this section to include a
pacity and sensitivity), heat lamps, hot plates, etc.
description of special requirements needed to achieve a
successful analysis. 15.2.13 Reagents and Materials:
15.2.9 Safety Precautions-If the method involves haz- lS.2.13.1 Chemicals and Reagents-Include derivatizing
ards, insert a warning to this effect. Point out the nature of reagents. Note purity, or purification methods, if required.
the hazards, and describe precautionary measures which 15.2.13.2 Calibration Standards-Note purity required.
must be taken. Refer to the latest OSHA regulations re- 15.2.13.3 Gas (or Gases)-Carrier gas, fuel gases for flame
garding all materials used in this procedure. ionization detector, special gas for electron capture detector,
15.2.10 Gas Chromatographic System-List and describe etc. Note purity required.
the apparatus. Describe the essential features of the appa- lS.2.14 Calibration-Describe in detail the calibration
ratus that are necessary to achieve the desired analysis. Avoid procedure. State whether pure components or standard
the use of trade names. Include schematic drawings or mixtures are used and the basis of measurement. Include
photographs if they are needed to clarify. or supplement the equations and describe the preparation of any calibration
text. The gas chromatographic conditions can be either charts. Show the calibration curve. If a trace method is
summarized in a table, as in Table 6, or in the text as follows: described, provide a chromatogram of the lowest detectable
IS.2.1O.1 Sample Injection Port-Construction: stainless amount. Lengthy procedures, such as the development of
steel, glass liner, fitted for on-column injection with a glass or complex equations, or the preparation of standard mixtures,
metal column, etc. Temperature at which used. should be placed in a section of an Appendix.
15.2.10.2 Column Oven-Isothermal or temperature pro- 15.2.15 Procedure-Include, in proper sequence, direc-
grammed operation: give temperatures and programming tions for carrying out the method. Refer to the pertinen,t
rates required. parts of the calibration procedure in lS.2.14. Do not repeat
15.2.10.3 Detector-Type (flame ionization, thermal con- these steps here. Possible subheadings are as follows:
ductivity, etc.), temperature of operation, sensitivity re- lS.2.1S.1 Final Conditioning and Adjustment o/the Gas
quired. Detector gases used and flow rates. Chromatographic System-This section is intended to in-
15.2.10.4 Recorder-Operating range (in millivolts), chart clude adjustment and verification of the state of the chro-
speed, time for full-scale deflection of pen. matographic system before analytical use.
15.2.10.5 Integrator-Note operating characteristics of lS.2.1S.2Sampling-Careful attention must be given to
integrator and parameters used. the sampling techniques since representative samples are
15.2.11 Preparation and Installation of the Chromato- essential to achieve successful analysis. Include special direc-
graphic Column: tions that may be required for taking samples, for preserva-
lS.2.11.1 Tubing Material-Note the type of material, as tion of samples, and for special treatment of samples prior to
stainless steel, nickel, glass, or glass-lined tubing, as well as injection.

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E 260
TABLE 6 Summary of Gas Chromatographic Conditions
Column Length, outside diameter, and internal oiameter (or outside diilnleter and wall
thickness); weight percent of specified stationary phase; specified solid suppOrt
(mesh size and treatment; for example, acid-washed, silanized); apprqximate
amount of total col!lmn packing in a given column'length.
Temperatures:

Sample inlet system

Detector
Column

If Isothermal
If programmed

Initial °C (note any hold time at initial temperature)

Final °C
Heating rate °C/min
(State specific temperatures and times if isothermal operation and
temperature programming are combined)
Carrier gas (specify)
Flow rate cm3/min A
Detector TCD, FlO, ECD . ,
Detector Sensitivity Flame ionization detector-amps full scale deflection, AFSs
Detector voltage or bridge current, if appliCable.
Recorder characteristics mV range; speed of full-scale deflection
Chart speed cm (or In.)/min
Sample size J.LL (or cm3 , if gas)
A Flow rate Is best measured 'at the Column or detector outlet, at the anaiytical temperature and flow rate. .
S A flame ionization detector is assumed to' be operated under optimum hydrogen and air flow rates, unless otherwise specified.

15.2.15.3 The remainder of the steps leading to the 15.2.17 Report-Show limit~ tobe reported.,
chromatogram. 15.2.18 Ptedsion~Lfmiting v::tlues" for precision shOUld
15.2.15.4 Typical Chromatogram-Show, in a figure, a be based on cooperative test, results., Judgment as to the
plot of the retention time (in minutes) versus the detector acceptability"ofresUlts (9~ % probability) shoUld be based on
response. Label the known peaks (including the dead volume the following criteria: ",; "
or unadsorbed gas peak) and indicate in' parentheses the , 15:2.18.1 Repeatability-ThefollCiWing wording shoUld
attenuation for each peak. be .used:' Duplicate resUlts'by the sallie operator shoUld not
be consi~e~ed sllspect unles,s they ~~r.~y :t;n6re than the
NOTE 8-When determining the retention time of the unadsorbed
peak, the retention time of air is used for thermal conductivity detectors,
following amounts:, (insert' cte'tenirined limits in tabular
fortri):' , ", . ' '
methane for flame ionization detectors, and methylene chloride lead
space vapors Jor ECDs.' ' 15.2.18.2 ReproduCibility.:......The followirtlfwoi'ding shoUld
be used: The resUlt submitted by 'each of rlv() hiboratories
15.2.15.5' Retention Time Data"':"Include a table listing shoUld not be considered susp'ect unlessth~two resUlts differ
retention times and, relative retention times for all com- by mote than the following amoun'ts:"(in,seit determined
pounds, of interest, for all recommended columns. Identify amounts in tabUlar form).' . .
the umiCisorbed peak and the reference material used for 15.2.19 Appendixes':"""Supplementary information may
relative retention time calculations. be induded in one. 'br more Appendixes to the report.
15.2.16 (;alculation.,-State the reference point on Examples of such' 'information are: technique' tb improve
which"tlie calculations are based (for example, sample as column life, directions to c1dm' the apparatiis, leak check
received), the terms in 'Whi<;:h the results' ate finally obtained procedures, procedures to, optimiZe' ,cblllmri,performance,
(weigh!, vQIUfJ1e, or mole perceDt)~ and whether or no~ these development of' equations "used in? \he ';calcUlations, and
values are normaliZed.. Present the calculations in equation precautions to avoid coni~on causes or e~or~,r'etc. '
form, using letter symbols to designate vari:ibie values and .ti.,

nlnuerical values of constantS. Define the letter symbols in a 16. Keywords

legend immediately following'the calculation equation. '16.1 gas chromatography; GC; packed col1.lmns

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