Designation: E 260 – 96 (Reapproved 2001)

Standard Practice for

Packed Column Gas Chromatography1
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 (e) indicates an editorial change since the last revision or reapproval.

1. Scope CGA P-12 Safe Handling of Cryogenic Liquids3

1.1 This practice is intended to serve as a general guide to HB-3 Handbook of Compressed Gases3
the application of gas chromatography (GC) with packed 3. Terminology
columns for the separation and analysis of vaporizable or
gaseous organic and inorganic mixtures and as a reference for 3.1 Terms and relations are defined in Practice E 355 and
the writing and reporting of GC methods. references therein.

NOTE 1—This practice excludes any form of gas chromatography 4. Summary of Practice
associated with open tubular (capillary) columns. 4.1 A block diagram of the basic apparatus needed for a gas
1.2 This standard does not purport to address all the safety chromatographic system is as shown in Fig. 1. An inert,
concerns, if any, associated with its use. It is the responsibility pressure or flow-controlled carrier gas flowing at a measured
of the user of this standard to establish appropriate safety and rate passes to the injection port or gas sample valve. A sample
health practices and determine the applicability of regulatory is introduced into the injection port, where it is vaporized, or if
limitations prior to use. Specific hazard statements are given in gaseous, into a gas sample valve, and then swept into and
Section 8 and 9.1.3. through the column by the carrier gas. Passage through the
column separates the sample into its components. The effluent
2. Referenced Documents from the column passes to a detector where the response of
2.1 ASTM Standards: sample components is measured as they emerge from the
E 355 Practice for Gas Chromatography Terms and Rela- column. The detector electrical output is relative to the
tionships2 concentration of each resolved component and is transmitted to
E 516 Practice for Testing Thermal Conductivity Detectors a recorder, or electronic data processing system, or both, to
Used in Gas Chromatography2 produce a record of the separation, or chromatogram, from
E 594 Practice for Testing Flame Ionization Detectors Used which detailed analysis can be obtained. The detector effluent
in Gas Chromatography2 must be vented to a hood if the effluent contains toxic
E 697 Practice for Use of Electron Capture Detectors in Gas substances.
Chromatography2 4.2 Gas chromatography is essentially a physical separation
E 840 Practice for Using Flame Photometric Detectors in technique. The separation is obtained when the sample mixture
Gas Chromatography2 in the vapor phase passes through a column containing a
E 1140 Practice for Testing Nitrogen/Phosphorus Thermi- stationary phase possessing special adsorptive properties. The
onic Ionization Detectors for Use in Gas Chromatography2 degree of separation depends upon the differences in the
2.2 CGA Publications: distribution of volatile compounds, organic or inorganic, be-
CGA P-1 Safe Handling of Compressed Gases in Contain- tween a gaseous mobile phase and a selected stationary phase
ers3 that is contained in a tube or GC column. In gas-liquid
CGA G-5.4 Standard for Hydrogen Piping Systems at Con- chromatography (GLC), the stationary phase is a nonvolatile
sumer Locations3 liquid or gum coated as a thin film on a finely-divided, inert
CGA P-9 The Inert Gases: Argon, Nitrogen and Helium3 support of a relatively large surface area, and the distribution is
CGA V-7 Standard Method of Determining Cylinder Valve based on partition. The liquid phase should not react with, and
Outlet Connections for Industrial Gas Mixtures3 should have different partition coefficients for, the various
components in the sample. In gas-solid chromatography
1
This practice is under the jurisdiction of ASTM Committee E13 on Molecular (GSC), the stationary phase is a finely divided solid adsorbent
Spectroscopy and is the direct responsibility of Subcommittee E13.19 on Chroma- (see 4.4).
tography. 4.2.1 After separation in the analytical column, the compo-
Current edition approved April 10, 1996. Published June 1996. Originally
published as E 260 – 65 T. Last previous edition E 260 – 96. nents are detected, and the detector signal is related to the
2
Annual Book of ASTM Standards, Vol 3.06. concentration of the volatile components. Tentative identifica-
3
Available from Compressed Gas Association, Inc., 1725 Jefferson Davis tions can be made by comparison with the retention times of
Highway, Arlington, VA 22202-4100.

Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.

1
 E 260

FIG. 1 Block Diagram of a Basic Gas Chromatographic System

known standards under the same conditions, either on a single 4.6 Components in a mixture may be tentatively identified
column or preferably by injecting the sample onto two columns by retention time. Ideally, each substance has a unique reten-
of different selectivity. Ancillary techniques, such as mass tion time in the chromatogram for a specific set of operating
spectrometry or infrared spectrophotometry, are generally nec- conditions. However, caution is required because the GC
essary for positive identification of components in samples. separation may be incomplete and a single peak may represent
4.2.2 Prior to performing a GC analysis, the following more than one compound. This is especially true of unknown
parameters must be considered: mixtures and complex mixtures because of the very large
4.2.2.1 Sample preparation. number of possible compounds in existence and the finite
4.2.2.2 Stationary phase and loading on support. number of peaks that a chromatograph might resolve. Addi-
4.2.2.3 Column material required. tional characterization data may be provided by ancillary
4.2.2.4 Solid support and mesh size. techniques, such as spectrometry.
4.2.2.5 Column length and diameter.
4.2.2.6 Instrument and detector type that will be needed. 5. Significance and Use
4.2.2.7 Injector, column oven, and detector temperatures 5.1 This practice describes a procedure for packed-column
required for analysis. gas chromatography. It provides general comments, recom-
4.2.2.8 Injection techniques, such as flash volatilization, mended techniques, and precautions. A recommended form for
on-column technique, purge and trap, pyrolysis, etc. reporting GC methods is given in Section 14.
4.2.2.9 Carrier gas and flow rate.
4.2.2.10 Data handling and presentation. 6. Apparatus
4.3 In gas-liquid chromatography, the degree of separation
possible between any two compounds (solutes), is determined 6.1 Carrier Gas System—Common carrier gases are helium
by the ratio of their partition coefficients and the separation and nitrogen. Paragraph 7.6 provides more details on carrier
efficiency. The partition coefficient, K, is the ratio of the solute gases. Means must be provided to measure and control the flow
concentration in the liquid phase to the solute concentration in rate of the carrier gas. Any flow or pressure control and
the vapor phase at equilibrium conditions. The partition coef- measurement combination may be used that will give an
ficient is affected by temperature and the chemical nature of the accurately known and reproducible flow rate over the desired
solute (sample) and solvent (stationary phase). range.
4.4 Another mechanism for separation is gas-solid chroma- 6.1.1 The main gas supply is regulated with a two-stage
tography. With this technique there is no liquid phase, only a regulator which must have a stainless steel diaphragm. Rubber
porous polymer, molecular sieve, or solid adsorbent. Partition or plastic diaphragms permit oxygen or water to diffuse into the
is accomplished by distribution between the gas phase and the carrier gas. In addition, instruments will have a flow controller
solid phase. between the pressure regulator and column inlet to maintain a
4.5 After the sample is resolved into individual components constant flow during temperature programming. Copper or
by the chromatographic column, the concentration or mass stainless steel carrier gas lines, not plastic tubing, should be
flow of each component in the carrier gas can be measured by used to avoid diffusion of oxygen (air) into the carrier gas.
an appropriate detector which sends an electrical signal to a When using the thermal conductivity detector, variations in the
recording potentiometer or other readout device. The curve flow will change retention and response. The carrier gas line
obtained by plotting detector response against time is referred pressure must be higher than that required to maintain the
to as a chromatogram. For flame ionization and thermal column flow at the upper temperature limit for the flow
conductivity detectors, either the peak areas or the peak heights controller to operate properly. A pressure of 40 to 60 psi is
are proportional to the concentration of the components in the usually sufficient.
sample within the linear range of the detector system. How- 6.2 Column Temperature Control—Precise column tem-
ever, response fractors are not necessarily the same for all perature control is mandatory if reproducible analyses are to be
compounds, and linearity of detector response may depend on obtained. Temperature control must be within 0.1°C if reten-
operating conditions. (Testing of detector performance is tion times are to be compared with another instrument.
discussed in ASTM Standard Practices for the appropriate 6.2.1 Air Bath—The thermostated forced-air bath is gener-
detector, see 2.1). ally accepted as the best practical method of temperature

2
 E 260
regulation for most applications. Temperatures can be con- programmed operation.
trolled by regulators or proportionally controlled heaters using 6.3.4 Injection Port Septum:
a thermocouple or platinum-resistance thermometer as a sens- 6.3.4.1 The septum is a disc, usually made of silicone
ing element. The advantage of a forced-air bath is the speed of rubber, which seals one end of the injection port. It is important
temperature equilibration. Air bath ovens are readily adaptable to change the septum frequently after two to three dozen
to temperature programming and are capable of operating over injections, or preferably at the end of the working day. The best
a range of 35 to 450°C. This range can be extended down technique is to change the septum when the column is
to − 100°C by using cryogenic equipment. relatively cool (below 50°C) to avoid contact of stationary
6.2.2 Other Devices—Liquid baths, drying ovens, incuba- phase in a hot column with air (danger of oxidation). After the
tors, or vapor jacket enclosures are less stable, less convenient septum is changed, return the inlet temperature to that which
means of providing a source of heat to maintain or raise the was originally set. The inlet temperature should be the opti-
temperature of a chromatographic column. These devices are mum for the particular analysis, as well as within the recom-
not recommended for precision chromatographic applications. mended operating temperature of the septum. If the septum is
6.3 The Injection Port—The purpose of the injection port is punctured too many times, it will leak air into the gas
to introduce the sample into the gas chromatographic column chromatographic system, even though it is under pressure. At
by instantaneous volatilization following injection into the gas high temperatures, above 150 to 200°C, air (oxygen) in the
chromatographic system. Two sample inlet types are in com- carrier gas from a septum leak will degrade the stationary
mon use in gas chromatography: the flash vaporization and the phase. An excessive septum leak will also produce a change in
on-column injection inlets. carrier gas flow rate (a change in retention time) and loss of
6.3.1 The temperature of the flash vaporization inlet should sample (irreproducible peak heights) due to outflow from the
be above the boiling points of the sample components and is leak. When installing the septum, do not overtighten the
limited by the amount of septum bleed generated and the retaining nut. The septa will swell at high temperature and
temperature stability of sample components. It should be set at extrude out of the injection port. A snug fit at room temperature
that temperature above which no improvement in peak shape is sufficient. It is important for septum life to make sure the
occurs but should be determined by the nature of the sample injection needle is sharp with no bent tip. Fine emery cloth, or
and the volume injected, not by the temperature of the column. a fine sharpening stone, can be used to sharpen the point.
If the inlet temperature is too low, broad peak with a slowly 6.3.4.2 Ghost peaks may be observed in temperature pro-
rising front edge will result from slow vaporization of the grammed runs due to septum bleed. Septum bleed is due to the
sample. If the temperature is set far above what is necessary to thermal decomposition, 300°C or higher, of the septum that
produce fast vaporization, thermal decomposition of the produces primarily lower molecular weight cyclic dimethylsi-
sample, decreased septum life, and ghost peaks due to septum loxanes. It contributes to baseline response and is frequently
bleed may be observed. Generally, a good guideline is to observed as evenly spaced peaks in a temperature programmed
maintain the inlet temperature 25 to 30°C higher than the run in which no sample has been injected. This situation can be
highest boiling point of any sample component. demonstrated by the disappearance of ghost peaks after placing
6.3.2 A glass liner placed inside the injection port will aluminum foil (pre-cleaned with solvents such as methylene
eliminate sample contact with hot metal inner walls of the inlet, chloride or toluene) over the inner face of the septum or by
which can catalyze thermal decompositions. Any debris left in turning off the injector temperature and making several blank
the liner, especially from biological samples, can be a source of runs. Septum bleed can be decreased by using either air- or
excessive sample adsorption. If a liner is used, the debris can water-cooled septum retaining nuts, by using a septum flush
easily be removed by replacing the liner. Deactivation of the head, or by using special high-temperature septa which are
glass liner by treatment with dimethyldichlorosilane may be available from a number of gas chromatographic supply
necessary for some compounds. houses.
6.3.3 With on-column injection technique, the sample is 6.4 Detector Temperature Control—The detector tempera-
deposited in the liquid state directly on the column packing. ture should always be above that of the maximum operating
The sample must be small enough to preclude flooding of the analytical temperature, to prevent the possibility of condensa-
column, with possible detrimental effects to peak shape and tion of sample components or stationary phase bleed in the
column life. Ideally, the on-column inlet is a part of the detector and connecting line. Because there is usually some
column, so its temperature may be controlled as the column temperature gradient across a detector, the temperature should
temperature is controlled. In practice, because an on-column be set at 30 to 50°C above the maximum analysis temperature
inlet usually has a somewhat higher thermal mass than an to ensure that the entire detector is hot enough to prevent
equivalent sector of the rest of the column, the inlet must be condensation. Usually, it is neither necessary nor desirable to
heated somewhat above the maximum analysis temperature of use an excessively high temperature since this can result in
the column oven. The criteria of good peak shape and reduced sensitivity, increased noise level, frequent need to
quantitation should be used to determine the maximum re- clean the detector, and thermal decomposition of sample or
quired temperature for the inlet. One should consider the stationary phase.
temperature limit of the column packing when heating the 6.5 Measurement of Temperature—The choice of sensing
injection inlet and detector. With some samples, a nonheated elements used to measure temperature depends on the desired
injection port is adequate, especially with temperature- accuracy (control about a set point) and precision of the

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 E 260
measurements. Instrument read-outs should be verified peri- bleed. As many as forty detection systems have been reported,
odically. Some common temperature measurement devices are yet only about a dozen are commonly used. Table 1 shows
as follows: some of the more commonly used detectors. Of these, the
6.5.1 Standardized Mercury Thermometer: thermal conductivity, the flame ionization, the electron capture,
Range, °C Accuracy, °C the nitrogen-phosphorus, and the flame photometric detectors
0 to 100 60.02 are the most popular. Nondestructive detectors should be
100 to 200 60.05
200 to 400 60.50
vented to a hood to remove any toxic effluents from the
workplace. The effluent from destructive detectors may also be
6.5.2 Calibrated Platinum Resistance Thermometer: toxic. Details on detectors can be found in the applicable
Range, °C Accuracy, °C methods in Practices E 516, E 594, E 697, E 840, and E 1140.
−140 to 500 60.01
6.8 Programmed Temperature Operation— The apparatus
6.5.2 Thermocouple (iron − constantan, or other). used in programmed temperature gas chromatography differs
6.6 Analytical Column: in some respects from that normally used for isothermal work.
6.6.1 The analytical column is a length of tubing (glass, Basically, the column temperature is varied with time (program
metal, or plastic) that is filled with a packing material. It is rate) to enhance speed of separations. The advantages of using
discussed thoroughly in Section 7. programmed temperature operation include better resolution of
6.6.1.1 Column Characteristics—Specified by method. lower boiling components because of lower starting tempera-
6.6.1.2 Carrier Gas—Specified by method. ture and greater sensitivity because of sharper peaks for the
6.6.1.3 Sample Size—Variable within limits. higher boiling components.
6.6.1.4 Flow Rates of Carrier Gas and Detector Gases— 6.8.1 Column Heater and Temperature Programmer—It is
Variable within limits. of utmost importance that the column temperature program be
6.6.1.5 Column Temperature—Usually specified by method, reproducible, and that the difference between the set (desired)
and temperature and the true average column temperature be as
6.6.1.6 Physical or Chemical Characteristic of Compound small as possible. However, these requirements are difficult to
Analyzed, or both. achieve at high heating rates and with columns of large
6.6.2 Detector Characteristics—Desirable detector charac- diameter. The mass of the column and its heater should be kept
teristics should include the following: as small as possible. This will minimize thermal lag and will
6.6.2.1 Good stability (low noise level, minimum response give proportionately small variations around the set tempera-
to changes in temperature and flow rate). ture at any time. Proportional temperature controllers supply
6.6.2.2 Ruggedness and simplicity. almost full power to the heater until the set point is very closely
6.6.2.3 Sensitivity to the components for which analysis is approached.
desired. Use either a selective detector for materials of interest
6.8.2 The recirculating air bath is the recommended method
or one with a similar response for all components.
of heating in programmed temperature gas chromatography
6.6.2.4 Linearity of response versus sample concentration.
(PTGC). The obvious advantages are extremely rapid heating
Wide linear range.
(and cooling after an analysis is completed) with very little
6.6.2.5 Rapid response to changes in column effluent com-
temperature lag.
position (small internal volume or flow-through design, or
both). 6.8.3 Liquid baths may be used for very low heating rates.
6.6.2.6 Detectors, which are nondestructive and do not They are commonly contained in taped Dewar flasks.
contribute to band broadening may be used in series with other 6.8.4 No matter what type of heating device is used,
detectors. accurate control of the temperature program is necessary. This
6.7 Types of Detectors—The detector is located at the outlet is usually accomplished by appropriate electronic systems that
end of the chromatographic column and both senses and develop linear (or other) programming rates as desired.
measures the amount of components that have emerged from 6.8.5 Detectors for programmed temperature gas chroma-
the column. The optimum detector should have high sensitiv- tography should be relatively insensitive to minor temperature
ity, low noise level, a wide linearity of response, a response to and flow fluctuations and insensitive to stationary-phase bleed.
all compounds of interest, and yet be insensitive to changes in These difficulties can be overcome by operating the detector at
flow and temperature. Selective detectors are characterized as or near the upper temperature limit for the analysis and by
having selective, or greatly enhanced response to certain using adequate flow controllers. If stationary-phase bleeding is
components. Linearity is decreased for all detectors by column excessive during PTGC runs, a second conditioning procedure

TABLE 1 Applicability of Commonly Used Gas Chromatographic Detectors

Applicability Range of Minimal Detectable
DetectorA
(Type of Compound) Amounts (grams)
Thermal Conductivity All 10−6 to 10−7
Flame Ionization Organic 10−12
Electron Capture Halogenated and Oxygenated 10−12 to 10−15
Flame Photometric Sulfur, Phosphorus 10−11
Alkali Flame Nitrogen, Phosphorus 10−12 (even lower for phosphorus)
A
Further information can be found in Practices E 516, E 594 and E 697.

4
 E 260
(Section 9) might improve the situation. Alternately, a dupli- the observed results will include a drifting baseline or exces-
cate analysis column can be used on the reference side of the sive spiking on the baseline. Under these conditions, the liquid
detector. By equalizing substrate bleed on both sides of the phase will decompose or volatilize, and thus be removed from
detector, the baseline drift can be substantially compensated. the column. This situation will eventually lead to decreased
However, this technique does not improve column life and is retention times with broader peaks resulting in poorer resolu-
detrimental to detector linearity. If at all possible, operate the tion of very close peaks. Peak tailing will also be observed as
column within its recommended temperature range. the uncoated surface becomes exposed by removal of liquid
6.8.6 When using the temperature programming technique, phase, thus shortening column life. Bleeding also can expose
the resistance to carrier gas flow in the gas chromatographic bare support surface that can adsorb molecules being analyzed
column increases with increasing temperature. The flow con- and reduce column efficiency. In extreme cases, phase bleeding
trollers need a positive pressure of 10 psi to operate properly. will result in fouling the detector and connecting lines. The
By setting the second stage of the regulator to 40 to 60 psi, observed maximum temperature will depend upon many ex-
there will usually be sufficient excess pressure to maintain a perimental variables, such as type of liquid phase column,
constant gas flow through the column. Higher pressures might conditioning, phase-loading level, column temperature, sensi-
be required to maintain flow when using relatively long tivity setting of the detector, and purity of the carrier gas. In
columns of 10 ft, or longer, or packings finer than 120 mesh. programmed temperature runs, the column can sometimes be
operated for short periods about 25°C above maximum tem-
7. Materials perature. However, column bleed should be minimized for
7.1 Stationary Phases—The stationary phases (partitioning quantitative results since it decreases the linear range of all
agents) that have been successfully used for specific separa- detectors.
tions are found most quickly by a literature search. Many 7.2 Active Solids:
phases are listed in ASTM publications AMD-25A and AMD- 7.2.1 Molecular Sieves—The synthetic zeolite molecular
25A-51.4 The most desirable stationary phases do not volatilize sieve sorbents separate molecules by size and structural shape.
(bleed) significantly from the solid supports at temperatures Isomers with a more round shape, as branched versus straight
required to elute the sample. chain molecules, diffuse in and out of the zeolite structure more
7.1.1 The polarity of the stationary phases is currently best easily than isomers with the long chain structures. Separations
characterized by McReynolds Constants.5 The higher the are affected by the differences in times required for molecules
McReynolds Constant, the more polar the phase. Rohr- of different sizes to find their way into and out of the sieve-like
schneider constants can also be used to measure the polarity of structure of the adsorbent. Molecular sieves are most useful for
stationary phases.6 separating H2, O2, N2 , CO, and CH4. Carbon molecular sieves
7.1.2 The effects of using polar and nonpolar stationary are also available, and can be used to separate O2, N2, CO, CO2
phases are summarized as follows: , H 2 O, and C1 to C4 hydrocarbons.
7.1.2.1 Nonpolar stationary phases separate compounds pri- 7.2.2 Porous Polymers:
marily by order of relative volatility or boiling point. 7.2.2.1 One type of porous polymer used in gas chromatog-
7.1.2.2 Polar stationary phases separate compounds by or- raphy is available in the form of microporous cross-linked,
der of both relative volatility and relative polarity. With polar polymer beads produced by copolymerizing styrene and divi-
phases, nonpolar compounds will elute before polar com- nylbenzene or more polar copolymers, or both. These materials
pounds of the same boiling point. are generally used as received without coating with any liquid
7.1.2.3 Polarity alone is insufficient to describe the separa- phase. They provide symmetrical peaks for polar, hydrogen-
tion power of a column. One must consider the overall bonding compounds such as water, alcohols, free acids,
selectivity of a column towards a set of analytes. This amines, ammonia, hydrogen sulfide, etc., and organic com-
selectivity is a summation of the effects of dispersive interac- pounds up to molecular weights corresponding to about 170.
tions, acid-base interactions and the dipole interactions offered 7.2.2.2 Another porous polymer is poly(2,6-diphenyl-p-
by the various pendent groups in the stationary phase. phenylene oxide). This material is useful for the analysis of
7.1.3 The stationary phases used in gas chromatographic amines, alcohols, and hydrogen-bonding compounds. It is also
columns have both minimum and maximum temperature used as an adsorbent for trapping trace organic compounds in
limits. The chromatographer must be aware of the limits for the water and air.
phase being used. Below the minimum temperature, the phase
7.2.3 Silica Gel, Alumina, and Carbon— Among the active
will behave as either a viscous liquid or a solid. Less efficient
solid adsorbents are silica gel, alumina, and activated carbon.
separation will be observed, and the chromatographic results
They are useful for low-boiling hydrocarbons.
will be exhibited as broader peaks in the gas chromatogram due
to poor mass transfer of components in the stationary phase. 7.2.4 Solid adsorbents modified by low concentrations of
7.1.3.1 Above the maximum temperature limit, the phase liquid phases may retain the advantageous properties of both.
will begin to bleed off the column at an accelerated rate, and Some solid adsorbents can be modified by the addition of
surface activating compounds such as wetting agents, silver
nitrate, and the metal salts of fatty acids.
4
Gas Chromatographic Data Compilation, ASTM, 1981.
7.3 Diatomaceous Earth Supports—The most popular gas
5
McReynolds, W. O., Journal of Chromatography Science, Vol 8, 1970, p. 685. chromatographic supports are those prepared from diatoma-
6
Supina, W. R., and Rose, L. P., Journal of Chromatography, Vol 8, 1970, p. 214. ceous earth, also called diatomaceous silica or kieselguhr. The

5
 E 260
two main types are white and pink in color. The white supports a silanized diatomaceous earth, even coating of the most polar
are recommended over the pink supports because of their more liquid phases is difficult to achieve. Acid-washed, silanized
inert surface. The former are, however, very friable and must grades of white diatomaceous earths are recommended as
be handled very carefully when preparing packings and loading supports for the polar liquid phases, such as polyesters and
into gas chromatographic columns. Before using these sup- silicones of high cyano-group content.
ports, check the manufacturer’s literature for comments on 7.3.5 If the column is 6 ft (2 m), or less, use particle size of
their use. 100 to 120 mesh (125 to 149 µm) for highest efficiency under
7.3.1 The white-colored supports are produced by calcina- isothermal conditions. If the column is longer than 6 ft, use 80
tion of diatomaceous earth with sodium carbonate as a flux. In to 100 mesh (149 to 177 µm) particles. If temperature pro-
this process, the diatomaceous earth fuses, due to formation of gramming is used, 80 to 100 mesh particles should be used to
a sodium silicate glass. The product is white in color due to lessen resistance to carrier gas flow.
conversion of iron oxide into a colorless complex of sodium 7.3.6 Further information concerning the liquid phase load-
iron silicate. These white materials are used to prepare the ing is given in 9.3.
more inert gas chromatographic supports. However, they are 7.4 Halocarbon Supports—The two types of halocarbon
fragile and subject to abrasion from excessive handling in the supports are those prepared from poly(tetrafluoroethylene) and
course of sieving, packing, or shipping. Abrasion will produce poly(chlorotrifluoroethylene). These supports are relatively
finer particles, or fines, which will decrease column efficiency. inert and are nonpolar. They eliminate peak tailing observed in
7.3.2 The pink-colored supports are prepared by crushing the analysis of organic compounds capable of hydrogen bond-
diatomaceous earth firebrick that has been calcined with a clay ing, such as water, alcohols, amines, etc. They are the preferred
binder. The metal impurities remaining form complex oxides supports in the analysis of corrosive halogen compounds such
that contribute to the pink color of the support. These pink as HF, BCl3, UF6, COCl2, F2, and HCl.
supports are denser than the white supports because of the 7.4.1 Poly(tetrafluoroethylene) supports require special han-
greater destruction of the diatomite structure during calcina- dling procedures. When used as received, they are soft and tend
tion. They are harder and less friable than the white supports to form gums upon handling. They can also build up a static
and are capable of holding larger amounts of liquid phase (up charge and spray out of the column during the packing
to 30 %) without becoming too sticky to flow freely. Their operation. These problems can be virtually eliminated by
surface is generally more adsorptive than white supports. For cooling the support to 0°C before coating with liquid phase and
this reason, they are not recommended for use in the gas by avoiding the use of glass vessels. Rinsing poly(tetrafluoro-
chromatographic analysis of polar compounds. However, pink ethylene) with methanol and drying before use is another way
supports provide excellent efficiencies for the analysis of to eliminate the static-charge problem.
hydrocarbons and organic compounds of low polarity.
7.4.2 Supports prepared from poly(chlorotrifluoroethylene)
7.3.3 Chemical Treatment of Diatomaceous Earth are structurally harder and are much easier to handle and to
Supports—Neither the pink nor the white materials give pack into a column.
generally acceptable analysis of more polar compounds with-
out further treatment. With these compounds, severe peak 7.5 Tubing Materials—Tubing materials should be chosen
tailing is often observed, especially with the dense pink on the basis of the following criteria:
supports. This tailing is due to the presence of adsorptive and 7.5.1 They should be nonreactive with the stationary phase,
catalytic centers on all diatomaceous earth supports. The sample solvent, and carrier gas.
adsorptive sites are attributed to metal oxides (Fe, Al) and 7.5.2 They should possess physical properties to withstand
surface silanol groups, -SiOH, on the support surface. The temperature and pressure of operating conditions, and
latter are capable of forming hydrogen bonds with polar 7.5.3 They can be shaped to fit in the column oven of the
compounds. chromatograph.
7.3.3.1 Metal impurities are removed by washing with 7.5.4 Satisfactory materials include glass, nickel, stainless
hydrochloric acid, which leaches out iron and aluminum and steel, and glass-lined stainless steel. Glass is the material of
renders the surface both less adsorptive and less catalytically choice, unless conditions prohibit its use. Nickel tubing is more
active. However, even with acid washing, the pink supports are inert than stainless steel in most applications. Less frequently
still more adsorptive toward polar compounds than the white- used column materials are poly(tetrafluoroethylene), alumi-
type supports. Acid washing is sometimes followed by base num, and copper.
washing, which seems to remove only minor amounts of metal 7.6 Carrier Gas—The use of an impure carrier gas will
impurities, but is a good pretreatment for supports that are to be produce problems in gas chromatography. Trace water and
used for the analysis of basic compounds. oxygen can cause decomposition of the liquid phase coated on
7.3.3.2 Neither acid or base washing is effective in reducing the support. The common carrier gases, helium and nitrogen,
peak tailing due to hydrogen bonding with the surface silanol should contain less than 5 ppm water and less than 1 to 2 ppm
groups, -SiOH. These groups are most effectively masked by oxygen by volume. An oxygen adsorption trap can be used to
treatment with dimethyldichlorosilane. remove trace oxygen, while trace amounts of water and
7.3.4 Acid-washed silanized grades of white diatomaceous hydrocarbons with molecular weights higher than methane, can
earths are recommended as supports for nonpolar and medium be trapped on a molecular sieve trap. Place the molecular sieve
polarity liquid phases. Because of the hydrophobic character of drier nearest the gas supply. Calcium sulfate has been used in

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drying tubes, but cannot dry carrier gas to the same level as 9.1.1 Glass columns should be cleaned and deactivated, first
molecular sieve. by rinsing with 30 mL acetone and then 30 mL toluene. Next,
7.6.1 For some applications, hydrogen may be the preferred fill the column with 10 volume % solution of dimethyldichlo-
carrier gas. However, additional safety precautions are required rosilane in toluene. Allow the solution to stand in the column
due to hydrogen’s explosive nature. for 30 min. Finally, rinse the column with anhydrous toluene
7.6.2 Air (oxygen) can leak into the gas chromatographic and then anhydrous methanol to cap unreacted DMDCS CL
system through loose fittings or a septum, that has been groups. Dry the column by passing a stream of dry nitrogen
punctured too many times, even though the carrier gas is under through it. Cap both ends of the column until such time that it
a pressure of 40 to 60 psi. Keep all fittings on the gas delivery can be packed.
lines tight, and check them at periodic intervals. Change the 9.1.2 Metal columns should be cleaned thoroughly before
septum in the injection port frequently. Plastic tubing should packing by rinsing with methanol, acetone, and chloroform.
never be used for carrier gas, hydrogen fuel (for FID), or The column should be dried by passing nitrogen or dry air
make-up gas lines due to the possibility of oxygen or moisture through it. Do not blow house air through the column since this
diffusing through the tubing wall. compressed air usually contains an oil aerosol from the pump.
7.6.3 Each cylinder of carrier gas has its own impurity level.
NOTE 3—Most chromatographic supply houses provide metal tubing
Occasional tanks contain large amounts of impurities which that has been washed with solvents and is ready for use.
might overcome a low-capacity oxygen adsorption trap and
destroy a gas chromatographic column at high temperature. A 9.1.3 An alternative procedure is recommended for nickel
new tank or a fresh oxygen adsorption unit, or both will tubing and can be used to clean stainless steel tubing. Rinse the
improve this situation. nickel tubing with ethyl acetate, methanol, and distilled water.
7.6.4 Always change the tank when the pressure is less than Then fill the tube with 20 volume % nitric acid and let it stand
200 psi. As the total pressure in the cylinder decreases, there is for 10 min. (Warning: Work in a hood and wear safety
an increase in the partial pressure of the water and other equipment when using nitric acid.) Next, rinse the tube with
impurities adsorbed on the inner walls of the gas cylinder. As distilled water to neutrality and then rinse with methanol and
a result, the last amounts of gas delivered from the gas cylinder acetone. Finally, dry the column by blowing nitrogen or helium
contain high levels of impurities. through it.
7.6.5 Carrier Gas for Instruments with Thermal Conductiv- 9.1.4 The column length is generally 3 to 6 ft (1 to 2 m).
ity Detectors—A major factor in sensitivity is the difference in Shorter columns can be used to decrease the time of analysis or
thermal conductivity of the compound being analyzed and the to separate high boiling compounds. Longer columns are used
thermal conductivity of the carrier gas. Helium (thermal to improve resolution, but have longer analysis times. (Col-
conductivity = 33.60 cal/cm-s-°C) is usually the carrier gas of umns longer than 20 ft (6.1 m) require excessive pressures to
choice. maintain the proper carrier gas flow.) A compromise is usually
7.6.6 Carrier Gas for Instruments with Flame Ionization made between analysis time and resolution. As a general rule,
Detectors—The most commonly used carrier gases are nitro- an increase or decrease of column length by a factor of 3 to 4
gen or helium. A maximum impurity level of 0.05 volume % is necessary to see a significant change in peak separation.
does not generally interfere with most applications. Hydrogen 9.1.5 The diameter of the column can be 1⁄8 in. (3.2 mm) or
1⁄4 in. (6.4 mm) outside diameter. The 1⁄8-in. column has less
is less commonly used in the US but is more popular in Europe
because of availability and relatively low cost. sample capacity, but greater efficiency, and is the most com-
mon type. Glass columns are generally 2 mm or 4 mm inside
NOTE 2—If hydrogen is used, special precautions must be taken due to
diameter. Some analysts have found that 3⁄16 in. (4.8 mm
its explosive nature, to ensure that the system is free from leaks and that
the effluent is properly vented. outside diameter) metal columns are the ideal combination
between the capacity of 1⁄4 in. (6.4 mm outside diameter)
7.6.7 Carrier Gas for Instruments with Electron-Capture columns and the efficiency of 1⁄8 in. (3.2 mm) outside diameter
Detectors—Users should follow the manufacturers’ recom- columns.
mendations for the choice of carrier gas. Some common ones 9.2 Choice of Diatomaceous Earth Support for Packed
are nitrogen or 95 % argon/5 % methane. When using a tritium Columns—See 7.3.
source in the detector, do not use hydrogen as the carrier gas. 9.3 Phase Loading on Diatomaceous Supports—For pre-
Hydrogen will replace tritium in the source. parative work and analysis of substances boiling below room
8. Hazards temperature, use 15 wt % loadings for white-type supports and
8.1 Gas Handling Safety—The safe handling of compressed 30 wt % for pink-type supports. For general work, use loadings
gases and cryogenic liquids for use in chromatography is the of the range of 3 to 15 wt %. For highest efficiency, shortest
responsibility of every laboratory. The Compressed Gas Asso- retention times, and the least amount of bleed during high-
ciation, a member group of specialty and bulk gas suppliers, temperature operation, use 3 wt. % loadings. The lower phase
publishes the following guidlines to assist the laboratory loadings have lower sample capacity and elute components
chemist to establish a safe work environment: CGA P-1, CGA more rapidly and at lower temperatures. Always check the
G-5.4, CGA P-9, CGA V-7, CGA P-12, and HB-3. manufacturers’ literature for suggested phase loadings for a
particular support. For some applications (especially headspace
9. Preparation of Packed Gas Chromatographic Columns analysis) loadings as low as 0.2 wt. % are used which result in
9.1 Preparation of the Tubing Material: very narrow peaks and short analysis times. High phase

7
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loadings tend to produce less reactive packings. TABLE 2 Solvent for Liquid Phases
9.4 Preparation of the Gas Chromatographic Packing— Liquid Phase
Solvent
The following procedures describe the coating of a solid TypeA

support with stationary phase. The following four methods are Dimethyl Silicone Toluene
Phenylmethyl Silicone Ethyl Acetate
commonly used to prepare gas chromatographic packings: (a) Cyanopropylphenyl Silicone Methylene Chloride
Filtration or Solution Coating Method, (b) Rotating Evaporator Trifluoropropyl Silicone Ethyl Acetate
Method (c) Evaporative Method, and (d) Vacuum Evaporative Polyethylene Glycol Methylene Chloride
Cyanopropyl Silicone Methylene Chloride
Method. When preparing packings with loadings in the range Use solvent as recommended
of less than 5 wt %, the Filtration or Solution Coating Method Other Liquid Phases
by supply house.
is recommended. This method is preferred because it provides
minimum handling of the friable white-type supports. For
loadings of more than 5 wt. %, other methods can be used. The (Most catalogs of gas chromatography equipment suppliers
Rotating Evaporator Method is recommended, but should only contain lists of suitable solvents.)
be used if a rotating evaporator is available, which turns very 9.4.2.2 Gradually add the support to the solution with gentle
slowly at 20 to 30 rev per min. swirling or agitation but with no mechanical stirring. (Sug-
gested solvents are given in Table 2.) The amount of solution
NOTE 4—A 5 wt. % loading of stationary phase consists of 5 g
should be just enough to wet the solid support and form a slurry
stationary phase added to 95 g of support.
with little excess solvent.
9.4.1 Filtration or Solution Coating Method—Prepare 100 9.4.2.3 Evacuate the flask briefly several times to remove air
mL of a solution of the desired phase in a vacuum filter flask. bubbles from the pores of the support. Be prepared to release
Use a suitable high boiling solvent (boiling point more than the vacuum if the slurry foams excessively.
60°C). The actual loading of the liquid phase on the support 9.4.2.4 Transfer the slurry to a large flat borosilicate glass
will depend upon both the viscosity of the phase solution and dish, and slowly evaporate the solvent in a hood with no further
the density and mesh size of the support. handling. The dish must be of a size that the packing is spread
9.4.1.1 Add 20 g of support to the filter flask. Reduce the on the bottom in a thin layer, no more than about 1⁄4-in. thick.
pressure in the filter flask for a few minutes with a water A borosilicate glass baking dish makes a suitable container.
aspirator, then release the vacuum. Repeat this procedure for 9.4.2.5 The critical stage occurs when excess solvent has
several cycles in order to remove air bubbles from the pores of evaporated, but the bed is still quite damp with a slight excess
the support particles. Be prepared to release the vacuum if the of solvent. Break up the damp bed by gently raking it with a
slurry foams excessively. spatula. As the solvent evaporates from the surface of a static
9.4.1.2 Allow the slurry to stand for several minutes. Pour bed of support, it leaves a higher concentration of phase at the
the slurry into a coarse-frit sintered-glass filter funnel, and bed surface. Therefore, the bed must be broken up frequently
allow the solvent to drain freely until the support settles. during the final stages of solvent evaporation to prevent
9.4.1.3 Apply vacuum cautiously and stop instantly when formation of an unevenly coated support.
the solvent stops dripping. Dump the support into a flat 9.4.2.6 Continue to air-dry the material in the hood until the
borosilicate glass dish, and allow it to dry. Do not scrape the last traces of solvent are gone. Avoid excessive handling of the
particles out of the funnel, since this might crush the particles. particles to prevent formation of fines due to abrasion, espe-
Do not resieve before use. cially in the case of the white-type supports.
9.4.1.4 The actual phase loading will depend upon the 9.4.3 Rotating Evaporator Method—Prepare the slurry of
viscosity of the phase solution and both the density and particle support and phase as described in 9.4.2.1 to 9.4.2.3, except in
size of the support. For example, a 2 % solution of dimethyl an indented, round-bottom flask. Connect the flask to a rotating
silicone gum liquid phase will give a 3.8 wt % loading on evaporator. Rotate the flask very slowly (less than 20 to 30
white-type supports. A10 wt % solution of a less viscous liquid revolutions per minute) and evaporate the solvent under
phase will give a 5.5 wt % loading on white-type supports and reduced pressure (water aspirator). Very slow rotation is
7.5 wt % on pink-type supports. Loadings obtained with other necessary to prevent the particles from abrading against each
phases on other supports are best determined by experimenta- other. Use of a heat lamp increases the evaporation rate. This
tion. method is not recommended for fluorocarbon supports.
9.4.1.5 The best way to determine the percent loading is to 9.4.4 Vacuum Evaporative Method—Prepare a slurry of
extract it from the support by extraction in a Soxhlet apparatus support and phase in a filtration flask of sufficient capacity.
and determine the weight loss. Alternatively, measure the (Suitable solvents are given in Table 2.) Attach the flask to a
volume of solution recovered and calculate the volume of vacuum source (water aspirator) and apply vacuum briefly. (Be
solution held up by the support. Calculate the approximate prepared to release the vacuum if the slurry foams excessively.)
percent loading on the support by assuming that the concen- Repeat this procedure several times in order to remove the air
tration of the solution does not change. bubbles from the pores in the support.
9.4.2 Evaporative Method: 9.4.4.1 Apply the vacuum for a longer period, and swirl the
9.4.2.1 Weigh out the desired amounts of support and phase. contents of the flask occasionally until all the solvent is almost
Use a larger amount than that required to account for attrition, evaporated. This is the critical stage.
spills, etc. Dissolve the liquid phase in a chemically inert, 9.4.4.2 Now shake the contents of the flask by gently
low-boiling solvent contained in a filtration flask (see Table 2). bumping the flask on a wood or plastic board. This will break

8
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up the bed of packing. Do not allow the solvent to evaporate acidic compounds, as organic acids and phenols, adsorption
from the surface of the support bed. Otherwise, the solvent will can be decreased by using phosphoric acid-treated glass wool
evaporate and leave a higher concentration of phase at the bed to plug the column ends. Wear safety glasses when pressure-
surface. packing columns.
9.4.4.3 Continue to apply vacuum until the packing is a 9.5.3.1 Attach the end of the empty column to an apparatus
freely flowing powder, then transfer it to a tray for air-drying in similar to that shown in Fig. 2. Add to the reservoir sufficient
a hood. packing material to fill the column, plus about 30 %. Attach the
9.4.5 Fluidized Drying Technique—This technique has been upper end of the reservoir to a nitrogen supply line controlled
used to produce efficient, uniformly coated packings. During to provide approximately 40 psi. Check that all connections are
the drying stages of methods 9.4.1 to 9.4.4, when the packing tightened, place a safety shield in front of the setup, and apply
has reached the consistency of a wet sand, add it to a fluidizer. 40 psi to the system.
Then dry the packing by passing a flow of inert, warmed gas 9.5.3.2 As the stationary phase starts to fill the column,
(nitrogen or helium) through the bed of packing. gently tap the column with a wood rod (handle of spatula or
9.5 Packing the Gas Chromatographic Column—The pur- screwdriver) or an electrical vibrator set at a very low vibration
pose in packing a gas chromatographic column is to fill the level. Continue tapping until the packing shows no voids and
column with packing as completely as possible, leaving no the level of packing in the reservoir remains constant.
empty spaces. Two variations are noted in 9.5.3 and 9.5.4 (a 9.5.3.3 Shut off the nitrogen supply and wait for the
pressure-fill procedure and a vacuum fill procedure). pressure to dissipate. Disconnect the column from the reser-
9.5.1 It is preferable to coil the column before packing to voir. Do not disconnect the column while it is under pressure.
prevent crushing of the support particles. Metal columns can be Have a clean beaker available to collect excess packing
coiled after loading to meet equipment requirements. Bends in material that will fall from the opened reservoir. Tap out about
the packed region must never be made with radii less than 1⁄8 in. (3 mm) of column packing, and replace it with a silanized

those specified in 9.5.2, to avoid crushing the packing in the glass wool plug. Affix a metal column tag engraved with a
column. description of the stationary phase, loading, support, and the
9.5.2 Right-angle bends are often necessary to make con- assigned column number.
nections to injection and detection systems, and must be made 9.5.4 Vacuum-Fill Procedure:
before packing the column since some tubing deformation will 9.5.4.1 Clamp the column so that the detector and injector
occur, which will crush some of the solid support. Bends for ends point upward. Plug the detector end of the column with a
such purposes should be within 4 in. (10 cm) of the column 1⁄4-in. plug of silanized glass wool. Use phosphoric-acid treated

ends. For coiled columns, minimum diameter mandrels should glass wool when analyzing for trace organic acids and phenols.
be as follows: for 1⁄8 in. (3.2 mm) OD column use a 11⁄2-in. 9.5.4.2 Attach a small funnel to the injection port end of the
(38-mm) mandrel; for 1⁄4 in. (6.4 mm) OD column use a 2-in. column. Attach the detector end of the column to a vacuum
(51-mm) mandrel. These configurations do not preclude the source, either a vacuum pump (preferably) or a water aspirator.
use of U- or W-shaped columns. If a U- or W-shaped column (If a water aspirator is used, a 500-mL filter flask, or the device
is to be used, the minimum 180° bend diameter must be at least shown in Fig. 3, should be placed in the line between the pump
that given for the above mandrel sizes. and the column.) Do not turn on the vacuum yet.
9.5.3 Pressure Fill Procedure—To each end of the column 9.5.4.3 Add 1 to 2 mL of packing to the funnel, and tap the
to be filled, fit a nut, a back ferrule, and a suitable front ferrule. column gently to settle the packing. A pencil or a wooden
Place a small plug of silanized glass wool into the detector end spatula handle can be used. Alternatively, the column can be
of the column, and cap the column by screwing in a metal cap stroked with a plastic saw. The use of an electric vibrator is not
with a 1⁄16-in. vent hole drilled into it. When analyzing trace recommended. Excessive vibration will cause the particles to

FIG. 2 Vacuum Fill Apparatus

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9.6.3 Heat the column at a rate of 2°C/min to the condition-
ing temperature. The latter temperature should be at least 25°C
higher than the analytical temperature but 25°C lower than the
maximum operating temperature recommended for the liquid
phase. Maintain this temperature overnight with carrier gas
flow.
9.6.4 The next day cool the column and connect it to the
detector. Detectors operated in very sensitive modes, particu-
larly the electron capture detector, might require two or more
days of conditioning at the higher temperatures before a
satisfactory baseline is obtained. (Other sources for baseline
drift and noise are impurities in the carrier gas, a dirty detector,
air leaks in the gas-line fittings, insufficient carrier gas pres-
sure, a much-punctured septum, chemical decomposition of the
phase (due to presence of traces of acid or base on the support,
in the phase, or on the inner column walls, and incorrect fuel
gas ratios to the flame ionization detector.)
9.6.5 There is a special “no-flow” conditioning procedure
which can be used with certain silicone phases, as methyl and
methylphenyl silicones with or without low vinyl content. It
has been reported to improve analysis for drug compounds.
The procedure starts by conditioning the column for 1⁄2 h as
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 310°C for 1.5
FIG. 3 Pressure Fill Apparatus
h with no carrier gas flow. Cool the column to 100°C. Uncap
the oven, turn on the carrier gas flow, and continue the regular
abrade against each other, producing fines and newly fractured conditioning procedures listed in paragraph 7.6.3.
surfaces that are not coated with stationary phase.
NOTE 5—This no flow conditioning procedure may damage or destroy
9.5.4.4 Turn on the vacuum source. Continue to add the
non-silicone containing stationary phase or silicone containing functional
packing in small increments with tapping until the column is groups other than phenyl or methyl.
full. Finally, apply pressure to the head of the column to pack
it a little tighter. However, take care to make sure the pressure 9.6.6 Many of the liquid phases are commercial-grade
is equalized slowly, because packing will be blown out of the material, and conditioning might require several days before
column if the pressure is released too suddenly. the noise level is low enough to provide usable baselines at
9.5.4.5 Next, tap out enough packing to create a 1⁄8 in. (3 high sensitivity. The use of gas chromatographic-grade phases
mm) void space at the injector port end of the column. Plug this is recommended since they have been carefully purified and
end with a silanized glass wool plug. Do not pack the plug too long periods of conditioning are usually not necessary.
tightly. This will either impede the carrier gas flow or crush the
packing particles. 10. Sample Injection Procedures
9.5.4.6 Higher efficiencies are always observed if the col- 10.1 Injection Technique—Useful analyses are obtained
umn is packed for on-column injection. In this technique, the only by injecting representative samples into the gas chromato-
column is packed so that there is space at the injection port end graph. Since chromatographic samples are small, the best
of the column, which is then placed inside the injection port. practices and procedures must be followed.
This void space should be of such a length that the injection 10.1.1 If elution is to be otherwise, the sample injection
needle just reaches, or slightly penetrates, only the glass wool must be almost instantaneous in order to introduce the sample
plug, not the packing, when the column is installed. Thus the as a plug. Avoid unnecessary sample dilution and inadvertent
sample is injected almost directly onto the column. trapping.
9.6 Conditioning of Packed GC Columns: 10.2 Sample Size—Sample sizes used for analysis with 2- to
9.6.1 The purpose of the conditioning process is to remove 5-mm ID packed columns are in the range of 0.1 to 10 µL
extraneous material (solvent and adsorbed material) from the multi-component liquids. Gas samples usually range from 10
column before analytical usage. Since the column is heated, the µL to 2 mL.
liquid phase also redistributes itself over the support surface to 10.2.1 There is frequently a reduction in retention times
provide a more even coating. with sample size when the column is overloaded. Therefore,
9.6.2 Install the column into the gas chromatograph at the when identifying by comparison with a known sample, the
injection side only. Do not connect the column to the detector same amount of each component must be used in an amount
during the conditioning stage. Any column bleed might foul the that does not overload the column. The sample size that
detector and the connection lines between the column and overloads the column is that size which decreases the efficiency
detector. Turn on the normal analytical carrier gas flow and (Section 11) by 10 % compared to a smaller sample size.
flush air out of the column at ambient temperature for 30 min. Sample overload is sometimes shown by peaks with sloping

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fronts and backs that are almost perpendicular to the baseline. chamber. The ampule is physically broken to release the
(Another reason for this peak shape is insufficient vaporization sample, which is then swept into the gas chromatographic
of the sample. Either the injection port or the column tempera- column by the carrier gas.
tures, or both, are too low.) 10.4 Sample Container—Care must be taken in the design
10.3 Sample Injection Devices—Samples may be intro- and construction of sample containers so that none of the
duced by syringes, automatic sample injectors using syringes, components of interest are in any way changed or removed
or sample valves. (There are also devices that introduce from the sample by reaction, diffusion, or adsorption. Stopcock
capsules containing sample into the injection port.) For rigor- grease and material desorbed from plastic cap liners are
ous quantitative work, any sample introduction device should frequent sample contaminants.
be flushed and filled at least three times with the sample
11. Evaluation of Column Performance
immediately before use.
10.3.1 The most common method for liquids is the use of a 11.1 Make a test mixture which contains a normal aliphatic
syringe injection technique through a self-sealing septum. In hydrocarbon and the compound being analyzed, or one similar
the usual 10-µL syringe, there is approximately 0.8 µL of dead in structure. The aliphatic hydrocarbon peak should appear in
volume in the syringe needle. This volume is additional to the the chromatogram near the second component in the test
volume in the syringe barrel and can be measured by with- mixture. (Suggested aliphatic hydrocarbon mixtures are shown
drawing the entire sample volume into the syringe barrel. in Table 3.) The peaks of both components should be about the
same size. The analytical conditions should be chosen so that
10.3.1.1 First, pump the sample back and forth vigorously
both chromatographic peaks will appear about 4 to 6 times the
in the syringe to wet the plunger and to remove air bubbles.
retention time (distance) of the solvent peak. (If a selective
Then withdraw the sample back into the syringe so that the
detector is being used, choose a compound that will give a
entire volume can be read on the volume marks of the syringe
response to that detector.)
barrel. When preparing the syringe for injection, never leave
11.2 The different methods of determining column effi-
the sample solution in the needle. This technique will minimize
ciency are shown in Eq 1-3. Figs. 4 and 5 show the measure-
sample boiling out of the needle when it is inserted into the hot
ments made on a chromatographic peak which are used to
injection port.
determine column efficiency.
10.3.1.2 In an alternative procedure, called the solvent-flush
technique, load the syringe in the following order: solvent, air, N 5 16 ~tR/wb!2 (1)
sample solution, and air, with only air remaining inside the
needle. When the sample is injected, the solvent is the last to N 5 5.54 ~t R/wh!2 (2)
leave the syringe, and it rinses out sample residue in the needle.
10.3.1.3 Wipe the syringe needle off before injection. Insert H 5 L/N (3)
the needle carefully through the GC septum, inject the sample
at once, and withdraw the needle in one continuous motion.
where:
10.3.1.4 Often the tip of the needle becomes bent, forming
N = number of theoretical plates,
a fish hook that will tear the septum on subsequent injections. tR = retention time, or distance, measured in mm,
This can be detected by brushing the end of the needle gently wb = width of the peak at base, measured in mm, (Deter-
over the end of a finger. A few strokes on a sharpening stone mined by extrapolating, as shown in Fig. 4 and Fig.
will remove the fish hook. 5.)
10.3.1.5 Syringes should be cleaned with a solvent that will wh = width of the peak at one-half the peak height, h, all
remove all traces of contamination. Consult the manufacturer’s measured in mm, (See Note 6.)
literature for further information. Many chromatographic sup- H = height equivalent to a theoretical plate, HETP, and
ply vendors sell suitable cleaning solutions and kits. Liquid L = length of the column in millimetres, or in centime-
sample valves, in both automated and manual versions, are also tres.
available.
NOTE 6—The peak width may be measured to the nearest 0.1 mm by a
10.3.1.6 Gas samples are most conveniently injected using magnifying loupe fitted with a scale graduated in 0.1 mm increments. The
gas-tight syringes. These devices are quite satisfactory for peak to be so measured should be at least 10 mm wide; this is arranged by
survey work because the sample size can easily be verified. choosing an adequately large chart speed.
However, the syringe needle is more likely to clog with pieces 11.3 Eq 1 is often used. However, it involves an extrapola-
of septum material when gas samples are injected than when tion of the baseline, shown in Fig. 4 and Fig. 5, which can be
liquids are injected. If no chromatographic peaks are observed,
test the syringe by injecting air into a liquid. If no bubbles are TABLE 3 n-Alkanes Used for Column Evaluation at 200°C
seen, unclog the needle with a wire or by filling the syringe 1 m 10 % Dimethyl silicone C18, C20, C22
without the needle and forcing solvent through the needle. 3 m 10 % Dimethyl silicone C14, C16, C18
1 m 10 % Phenylmethyl silicone C20, C22, C24
However, repeatable results in gas analysis will only be 3 m 10 % Phenylmethyl silicone C16, C18, C20
obtained using gas sample valves that have a fixed sample 1 m 10 % Cyanopropyl silicone C24, C26, C28
loop. The latter valves can easily be automated. 3 m 10 % Cyanopropyl silicone C20, C22, C24
1 m 10 % Polyethylene glycol C24, C26, C28
10.3.1.7 A sealed, friable, or puncturable ampule containing 3 m 10 % Polyethylene glycol C18, C20, C22
a weighed amount of sample may be placed in the injection

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FIG. 4 Use of the Chromatogram to Calculate Column Efficiency

efficiency of the second component is about 25 % lower than

that of the hydrocarbon, the column might not be the best
choice for analyzing the compound(s) of interest. A strong
indication of nonsuitability is shown by comparatively greater
tailing of the nonhydrocarbon component. Tailing can indicate
strong interaction with the column packing (phase or support,
or both), column tubing, or adsorption in the injection port or
in lines leading from the column to the detector.
FIG. 5 Shows the Procedure to Measure the Peak Width, at wh, to 11.6 Typical efficiencies are shown in Table 5. Not all gas
Account for the Thickness of the Pen. Use the Average of the chromatographic packings are capable of excellent efficiencies.
Two Determinations, w*h and w**h, shown here
For example, porous polymers, Teflon supports, and some
liquid phases, such as trifluoropropyl silicone can give effi-
in error. The use of Eq 2 is preferred because the term, wh can
ciencies less than 500 plates per foot.
be determined directly from the chromatogram without ex-
trapolation. However, the width of the recorder pen line can be 12. Use of the Gas Chromatographic Packed Column
variable which can lead to difficulties in determining the true
12.1 Certain precautions and preventive maintenance are
value of the peak width. Use a pen that writes with a sharp, thin
necessary to obtain the best column performance. Some of
line. The peak width should be measured from the leading edge
these points have been made before and will be referred to in
of one line to the leading edge of the other line, as shown in
this section. Further precautions will also be discussed.
Fig. 4. For accuracy, determine the peak width twice (see Fig.
12.2 Carrier Gas Purity—Trace, or adventitious, oxygen
4), and use the average value to calculate “N”. To further
and water in the carrier gas can produce degradation of liquid
minimize errors in determining the peak width, use a recorder
phases on the support. Purification of the carrier gas and
chart speed that will give a minimum peak width of 4 to 5 cm.
necessary precautions are discussed in 7.6.
The peak height should be about 40 to 80% full-scale chart
12.3 The Injection Port—The injection port is often a
deflection. This can be achieved by either changing the
source of trouble. The temperature can be either too low or too
detector sensitivity, or changing the sample size. However, be
high. See 6.3. Another problem is a leaking septum. See 6.3.4.
sure that the sample size does not exceed the capacity of the
12.4 Column Care:
column.
12.4.1 Never heat a gas chromatographic column with air in
11.4 The optimum efficiency occurs at the optimum carrier
it. When any column is first placed in a gas chromatograph,
gas flow which can be determined by plotting the efficiency of
flush any air (oxygen) out of it by flushing with carrier gas at
the hydrocarbon peak versus carrier gas flow. General optimum
normal flow rates for 15 to 30 min. at ambient temperatures.
values of the carrier gas flow rates are shown in Table 4, but
Then heat the column to the desired operating temperature.
should be determined for the particular column.
11.5 Calculate the efficiency of the second component. The
results should be about the same for both components. If the TABLE 5 Typical Efficiencies and Ratings of Gas
Chromatographic Columns
TABLE 4 General Optimum Values of Carrier Gas Flow Rates Plates/ft Plates/metre HETP Rating
General Optimum Carrier Gas Flow 500 1640 0.6 mm Excellent
Column Dimensions
Helium Nitrogen 400–500 1300–1640 0.60–0.76 mm Good
⁄ in. OD or 2 mm ID
18 30 mL/min 20 mL/min 300–400 980–1300 0.76–1.0 mm Fair
⁄ in. OD or 4 mm ID
14 60 mL/min 40 mL/min 300 980 1.0 Poor

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Always cool the column to room temperature before removing (tailing) during elution will produce a decrease in retention as
it from the gas chromatograph. If the column is to be used the concentration increases; while negative, or anti-Langmuir-
again, cap the ends with metal caps to prevent diffusion of air type fronting will produce an increase in retention time with
(oxygen) into the column during storage. Contact of the increased concentration.
stationary phase with oxygen when hot or for prolonged 13.3.1 The logarithm of the retention time of members of a
periods of time at room temperature cause degradation of the homologous series run isothermally is usually a linear function
stationary phase. This is particularly the case with polyglycols of the number of carbon atoms of a molecule. Using this
and polyester type phases and to a lesser extent cyanosilicones. characteristic, two or three reference compounds can provide
Other phases are affected to a varying degree. sufficient information to prepare a plot of the logarithm of the
12.4.2 After long periods of use, column performance may retention time versus carbon number, and they can identify
degrade as shown by peak broadening, tailing, or gradual other members of the series. Retention time, adjusted or not, is
merging of adjacent peaks. Often the problem lies in the front of little value in comparing the results from various instru-
end of the column. The injection port temperature might have ments. The use of Kovats retention index, based on the relative
been too high and destroyed the initial section of the liquid retention of a compound to the retention of normal paraffins,
phase on the column packing. Residues or decomposition provides a more reliable means of comparing the results
products might have built up on the glass wool plug. These obtained from different instruments (see Practice E 355).
problems can be remedied by repacking the first few inches of 13.4 Absolute compound identification or characterization,
a glass column, or cutting off the first few inches of a metal must be made with ancillary techniques such as mass or
column. Use fresh silanized glass wool to close the end of the infrared spectrometry, nuclear magnetic resonance, chemical
column in both cases. analysis of the effluent, or spot tests for functional groups.
13.4.1 The samples for the analyses in 13.1-13.4 may be
13. Methods of Qualitative Analysis obtained by trapping components as they emerge from the
13.1 Identification of compounds by gas chromatography chromatograph. A trap, glass capillary, or U-tube, is cooled
alone cannot be absolute, and the results must be considered with ice or dry ice, and placed in the effluent stream of the
with care. Elution of a compound is dependent upon carrier gas column. Several collections may be required to obtain a
flow rate, column temperature, support size, amount and type sufficiently large sample.
of liquid phase, column dimensions, instrument dead volume, 13.4.2 The collection of effluent is easiest with nondestruc-
and column pressure drop. These parameters must be stable to tive detectors, see 6.7. In the case of destructive detectors, a
obtain reproducible results. The recommended format for a gas split is made for the collection just before the detector.
chromatographic method is given in Section 15. 13.4.3 Apparatus is also available so that the effluent from
13.2 Tentative identification of a compound can be made by the gas chromatographic column can be analyzed directly by
comparing its adjusted retention time against those of known mass spectrometers or infrared spectrophotometers.
standards using exactly the same chromatographic parameters.
14. Methods of Quantitative Analysis
13.2.1 The retention time is the time interval measured from
the point of injection to maximum peak height of the sample. 14.1 Gas chromatography can be used to determine quanti-
Adjusted retention time, t8R, is derived by subtracting the time tatively the composition of complex samples. There are several
required for an unabsorbed gas, like air, or methane, tM, to factors that must be considered before the sample is analyzed.
traverse the column (also called the gas holdup time) from the The recommended format for gas chromatographic methods is
retention time, tR. given in Section 15.
14.1.1 The Chemistry of the Sample—The chemistry of the
NOTE 7—On some solid adsorbent columns, such as molecular sieves, sample, if known, allows a chromatographer to select more
there is no nonadsorbed component.
accurately a column compatible with the sample and to
anticipate potential interferences from reaction by-products.
t8R 5 t R 2 tM (4) 14.1.2 The Choice of a Detector—A detector must be
13.2.2 Retention times are affected by all chromatographic chosen with the needed selectivity and sensitivity. If compo-
parameters. As a result, direct comparison of retention times of nents will be analyzed at low levels, an electrolytic conductiv-
the same components on different instruments or between ity electron capture, nitrogen phosphorus, microcoulometric,
laboratories should be done with caution. Use of relative ionization, or flame photometric detector should be selected.
retention time is an easy practical technique for providing The detector may be limited to these lower concentrations and
elution data. The retention of a component is expressed relative not applicable to high concentrations.
to the retention of a known reference standard. The reference 14.1.3 Initial Separation of Components— Next, a column
standard should possess structural or chemical similarity to the must be chosen that will resolve the components of interest in
compounds being analyzed. the sample within a reasonable amount of time. First, a rough
13.3 The retention of a given weight of compound is usually separation should be achieved with known standards. Next,
independent of its concentration if the compound does not actual samples should be analyzed to determine if there are any
overload the column producing skewed peaks. The retention of interferences. A second column, or an ancillary technique
the compound is also independent of other substances present (GC/mass spectrometry, GC/infrared spectrometry, etc., should
if there is no appreciable overlap with another compound. be used to verify that additional components are not eluting
Substances that exhibit positive, or Langmuir-type, skewing with the component of interest. Each new sample adds the

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possibility of an interference eluting with the component of where:
interest; therefore this should be checked often. If an interfer- wb = peak width at the base of the peak, and
ence is detected, the chromatographer must change the method h = peak height.
to remove it. The several options for doing this are as follows: Another precise measurement defines the peak area as
14.1.3.1 Select a column stationary phase with a greater retention distance (in millimetres) times the peak height (also
selectivity for either the interference or the component of in millimetres). For peak height, this distance is simply
interest. measured from the baseline to the apex of the peak. However,
14.1.3.2 Choose a different type of detector that would these techniques now, for the most part, have been replaced by
detect the component of interest but not the interference. electronic integration, which is much faster. The proper use of
Examples would be water not being detected by a flame- these devices is crucial for accurate quantitative analysis. The
ionization detector, or hydrocarbons not detected by an elec- instruction manual for the particular integrator should be
trolytic conductivity or electron capture detector. studied and understood thoroughly before attempting to use
14.1.3.3 Consider other types of chromatographic separa- electronic integration for peak area or peak height measure-
tion such as capillary gas chromatography for more efficient ment.
separation of volatile compounds, liquid chromatography for 14.1.6 Data Handling:
separation of non-volatile compounds, or another appropriate 14.1.6.1 All manufacturers supply an integral electrometer
separation technique. to allow the small electrical current changes to be coupled to
14.1.4 Detector Sensitivity and Linearity— Once the chro- recorders/integrators/computers. The preferred system will in-
matographic separation has been optimized, the detector can be corporate one of the newer integrators or computers that
optimized and calibrated. Gas flows should be adjusted to the converts an electrical signal into clearly defined peak area
optimum levels to get peak sensitivity at the concentration counts in units such as microvolt-seconds. These data can then
range of the components of interest. The detector must also be be readily used to calculate the linear range.
clean and leak-tight. (See the manufacturer’s manual for 14.1.6.2 Another method uses peak height measurements.
suggested procedures.) This method yields data that are very dependent on column
14.1.4.1 The linearity of the detector over the desired performance and, therefore, not recommended.
concentration range of the component(s) of interest is deter- 14.1.6.3 Regardless of which method is used to calculate
mined using prepared standards. This step will determine what linear range, peak height is the only acceptable method for
the response is to increasing amounts of component. The peak determining minimum detectability.
area or height should be plotted versus the concentration for 14.1.7 Calibration— It is essential to calibrate the measur-
about five concentrations near the expected sample concentra- ing system to ensure that the nominal specifications are
tion. There should be a linear correlation. Nonlinearity may be acceptable and particularly to verify the range over which the
caused by reactivity, adsorptivity, thermal sensitivity, or exces- output of the device, whether peak area or peak height, is linear
sive column bleed. If the latter is the cause, change to a more with respect to input signal. Failure to perform this calibration
thermally stable column or one of different polarity. Column may introduce substantial errors into the results. Methods for
reactivity can be characterized by skewed, misshaped peaks. calibration will vary for different manufacturers’ devices but
This can be corrected by installing a fresh column of the same may include accurate constant voltage supplies or pulse gen-
type that does not have reactive sites. Test mixtures can be used erating equipment. The instruction manual should be studied
to demonstrate nonreactivity. Other sources of adsorptivity or and thoroughly understood before attempting to use electronic
reactivity with the sample are the injection port, connecting integration for peak area or peak height measurements.
lines to the detector, or glass wool. Each of these sources can 14.2 Types of Calculations:
be detected by carefully troubleshooting the system. 14.2.1 Each method of quantitative analysis has advantages
14.1.4.2 The detector performance should be checked peri- and disadvantages. The four methods of quantitative analysis
odically throughout the analysis. This can be done by injecting are as follows:
one of the linearity standards and comparing it to the linearity
14.2.1.1 Internal standardization,
plot.
14.2.1.2 External standardization,
14.1.5 Peak Area or Height Measurement— Many types of
14.2.1.3 Normalization, and
peak area and height measurement techniques exist. The oldest
methods for calculating the peak area are manual measurement 14.2.1.4 Corrected area.
with a ruler of the peak area using one of the following 14.2.2 Internal Standardization—In this technique, a pure
equations: component (the internal standard) is added to a sample in a
known amount. The peak area, or height, of all components of
peak area 5 wh 3 h (5)
interest is compared to the peak area, or height, of the internal
standard. These comparisons are referred to as response
where: factors:
wh = peak width at half height, and R F 5 AC/AIS 3 W IS/WC (7)
h = peak height
or
where:
peak area 5 ½ wb 3 h (6)
RF = response factor,

14
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AC = peak area of component, components in a sample chromatogram to those in a standard

AIS = peak area of internal standard, solution injected separately. It is critical that accurate amounts
WIS = mass of internal standard, and of sample and standard be injected for the method to be valid.
WC = mass of component. Generally, the solvent flush injection technique (see 10.3.1.2)
The amount of the component can be calculated from the or a sample valve of fixed volume is preferred.
weights of the sample and internal standard, the response 14.2.3.2 The advantages of this method are as follows:
factor, and the peak areas (or heights) as follows: (a) (a) Nondetected components do not bias the results.
% ConcC 5 WIS/WS 3 A C/AIS 3 1/RF 3 100 % (8) (b) (b) It can be used where several known components
must be determined in a very complex sample.
(c) (c) It can quantitate relatively reactive components.
where: (d) (d) A single sample can be analyzed where maximum
ConcC = concentration of component in sample,
accuracy is not required.
WIS = mass internal standard,
WS = mass sample, (e) (e) Nonlinearity has a minimal effect if the external
AC = peak area of component, standard is near the concentration of the sample.
AIS = peak area of internal standard, and 14.2.3.3 The critical part of this method is the injection. The
RF = response factor. volume of sample in the injection syringe and standard must be
This technique provides a correction for the relatively high accurately measured, allowing no bubbles in the slug of sample
variability of syringe injection and, therefore, yields a more or standard solution. If the sample and standard have different
precise method of analysis. Neither the quantity of solution densities, a correction must be made. Densities are easily
injected, nor change in detector response, will alter the area determined by filling a 50-µL syringe to about 30 µL, wiping
ratio of the analyte and the internal standard. To achieve the needle, weighing it, expelling the sample, wiping the
optimum performance, the internal standard must meet the needle again, and reweighing it.
following criteria. 14.2.3.4 The peak areas or heights of the component in the
14.2.2.1 The internal standard must elute in an area of the sample and the standard compound are measured and the
chromatogram that is free of sample components, or possible concentration calculated as follows:
sample components. % Conc C 5 AC/AES 3 W ES/WS 3 % Conc ES (9)
14.2.2.2 The internal standard must not react with the
sample or any of its components.
where:
14.2.2.3 The internal standard and the sample must be
ConcC = concentration of component,
homogeneous. A cosolvent may be used to produce a homo- AC = peak area of component in sample,
geneous mixture. AES = peak area of external standard,
14.2.2.4 The internal standard must be easily and accurately WES = mass of external standard injected,
added. WS = mass of sample injected, and
14.2.2.5 The internal standard must be pure. ConcES = concentration of external standard in solution.
14.2.2.6 The internal standard should elute near the compo- 14.2.4 Normalization—This calculation assumes that every
nent of interest. component elutes and that each has similar response factors. It
14.2.2.7 The concentration of the internal standard, relative is a fast procedure that requires no weighing. The sample is
to that of the analyte, should be such that these two peaks are injected, and the peak areas or heights of all components are
within 50 to 100 % of full scale deflection with the same measured. The concentration of the component of interest is
electronic attenuation and sensitivity setting in order to allow calculated as follows:
manual measurements and calculations of parameters, if de- % ConcC 5 A C/AALL 3 100 (10)
sired.
14.2.2.8 The most common use for the internal standard
technique in chromatography is to correct for quantitative where:
variations in the injection, particularly when using syringes. ConcC = concentration of component of interest,
For this purpose, the internal standard need not be chemically AC = area of component, and
related to the analyte, but must possess the criteria cited above AALL = sum of areas of all components.
and may be added in the final solution. Severe errors result if the components have different re-
14.2.2.9 In certain applications, an internal standard with sponse factors or do not all elute.
functional groups similar to the analyte may be desirable. For 14.2.5 Corrected Area—This method corrects for differ-
instance, those with a labile proton can be expected to exhibit ences in response but still assumes that all components elute
similar adsorption isotherm behavior and to undergo similar and are observed by the detector. Response factors are used to
physico-chemical transformations during such processes, as correct for response differences as follows:
extraction from a complex matrix or derivatization, or both. % ConcC 5 AC/~A 3 R F!ALL 3 RFC 3 100 % (11)
Likewise, similar electronegative functional groups are likely
to behave similarly towards an electron capture detector. where:
14.2.3 External Standardization: ConcC = concentration of component of interest,
14.2.3.1 This method compares peak areas or heights of AC = peak area of component,

15
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(A 3 RF) ALL = sum of peak areas times their respective 15.2.10 Gas Chromatographic System—List and describe
response factors relative to a standard, the apparatus. Describe the essential features of the apparatus
and that are necessary to achieve the desired analysis. Avoid the use
RF C = response factor of component to the of trade names. Include schematic drawings or photographs if
same standard. they are needed to clarify or supplement the text. The gas
chromatographic conditions can be either summarized in a
15. Recommended Form for Writing Gas
table, as in Table 6, or in the text as follows:
Chromatographic Methods
15.2.10.1 Sample Injection Port—Construction: stainless
15.1 General—Not all of the steps outlined in this section steel, glass liner, fitted for on-column injection with a glass or
may be needed to describe adequately a method. A number of metal column, etc. Temperature at which used.
variations in procedure format are shown in the publication, 15.2.10.2 Column Oven—Isothermal or temperature pro-
ASTM Standards in Chromatography.7 Ideally, the procedure grammed operation: give temperatures and programming rates
should be written so that it can be followed by a person with required.
the equivalent of a high school-chemistry understanding or six 15.2.10.3 Detector—Type (flame ionization, thermal con-
to twelve months of practical laboratory experience. Critical ductivity, etc.), temperature of operation, sensitivity required.
steps should be identified along with any reasons that show Detector gases used and flow rates.
why this step is necessary to achieve a successful analysis. Any 15.2.10.4 Recorder—Operating range (in millivolts), chart
involved procedures should be written in an Appendix so that speed, time for full-scale deflection of pen.
the main points in the procedure can be read more easily.
15.2.10.5 Integrator—Note operating characteristics of in-
15.2 Recommended Form:
tegrator and parameters used.
15.2.1 Title—The title should be concise, but complete
15.2.11 Preparation and Installation of the Chromato-
enough to identify the component(s) analyzed, the nature of the
graphic Column:
method (gas chromatography), the detector, and the materials
15.2.11.1 Tubing Material—Note the type of material, as
to which it is applicable. Select words that easily lend
stainless steel, nickel, glass, or glass-lined tubing, as well as the
themselves to indexing.
dimensions (outer diameter and inner diameter, or wall thick-
15.2.2 Scope—State as clearly as possible the range of
ness and length). Any pretreatment of the column material,
application of method. In a separate paragraph, note interfer-
solvent washing, or silanization should be mentioned.
ring substances or any significant limitations of the method.
This material could be placed in a later section (15.2.5), if an 15.2.11.2 Partitioning Phase—Solid adsorbent, if used
involved description is necessary. (type and mesh size). Coated support if used (liquid phase,
15.2.3 Pertinent Documents or References: percent loading and coating procedure, support type and
15.2.3.1 ASTM Standards. pretreatment, and mesh size). Note sources for special materi-
15.2.3.2 Other Standard Methods—Include any standard als in footnotes. Provide preparation and purification method
methods. for materials not commercially available. In an Appendix, note
15.2.4 Summary of the Method—Describe the method in a other liquid phases that have been successfully used in this
general way, without going into details of the procedure. It may analysis.
be appropriate to touch briefly on the following points: sample 15.2.11.3 Column Preparation—Describe the procedure
introduction technique, column dimensions and type of tubing used to pack the column. Note the amount of packing in the
material, nature of the packing material, mesh size of support column.
or adsorbent, liquid phase loading (if a liquid phase was used), 15.2.11.4 Column Installation—Note if the column is set up
isothermal or programmed temperature and detector type for back-flushing, if a sample fraction is removed on a
(thermal conductivity, flame ionization, electron capture, etc.). pre-column, or other special column arrangement.
15.2.5 Significance and Application—Use this section for a 15.2.11.5 Column Conditioning—Provide the column con-
more detailed discussion than can be fitted in the Scope. ditioning procedure.
15.2.6 Definitions—Include special definitions in this sec- 15.2.11.6 Column Evaluation—Give the procedure for
tion. General chromatographic definitions are already available evaluating the column. Calculation of the resolution between
in Practice E 355, to which reference can be made. two components in a standard mixture will often be sufficient.
15.2.7 Interferences—Use this section for a more detailed Provide some estimate of column life and signs of degrading
discussion than can be fitted into the Scope. column performance (loss of resolution, peak broadening, or
15.2.8 Special Comments—Use this section to include a tailing). Provide examples of good and bad chromatograms.
description of special requirements needed to achieve a suc- 15.2.12 General Apparatus—Volumetric flasks and pipets,
cessful analysis. microsyringes for sample introduction, balance (capacity and
15.2.9 Safety Precautions—If the method involves hazards, sensitivity), heat lamps, hot plates, etc.
insert a warning to this effect. Point out the nature of the 15.2.13 Reagents and Materials:
hazards, and describe precautionary measures which must be 15.2.13.1 Chemicals and Reagents—Include derivatizing
taken. Refer to the latest OSHA regulations regarding all reagents. Note purity, or purification methods, if required.
materials used in this procedure. 15.2.13.2 Calibration Standards—Note purity required.
15.2.13.3 Gas ( or Gases)—Carrier gas, fuel gases for
7
ASTM Standards on Chromatography, ASTM, 1981. flame ionization detector, special gas for electron capture

16
 E 260
TABLE 6 Summary of Gas Chromatographic Conditions
Column Length, outside diameter, and internal diameter (or outside diameter
and wall thickness); weight percent of specified stationary phase;
specified solid support (mesh size and treatment; for example, acid-
washed, silanized); approximate amount of total column packing in a
given column length.
Temperatures:

Sample inlet system °C

Detector °C
Column

If isothermal °C
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/minA
Detector TCD, FID, ECD
Detector Sensitivity Flame ionization detector—amps full scale deflection, AFSB
Detector voltage or bridge current, if applicable.
Recorder characteristics mV range, speed of full-scale deflection
Chart speed cm (or in.)/min
Sample size µL (or cm3, if gas)
A
Flow rate is best measured at the column or detector outlet, at the analytical temperature and flow rate.
B
A flame ionization detector is assumed to be operated under optimum hydrogen and air flow rates, unless otherwise specified.

detector, etc. Note purity required. 15.2.15.5 Retention Time Data—Include a table listing re-
15.2.14 Calibration—Describe in detail the calibration pro- tention times and relative retention times for all compounds of
cedure. State whether pure components or standard mixtures interest, for all recommended columns. Identify the unadsorbed
are used and the basis of measurement. Include equations and peak and the reference material used for relative retention time
describe the preparation of any calibration charts. Show the calculations.
calibration curve. If a trace method is described, provide a 15.2.16 Calculation—State the reference point on which the
chromatogram of the lowest detectable amount. Lengthy pro- calculations are based (for example, sample as received), the
cedures, such as the development of complex equations, or the terms in which the results are finally obtained (weight, volume,
preparation of standard mixtures, should be placed in a section or mole percent), and whether or not these values are normal-
of an Appendix. ized. Present the calculations in equation form, using letter
15.2.15 Procedure—Include, in proper sequence, directions symbols to designate variable values and numerical values of
for carrying out the method. Refer to the pertinent parts of the constants. Define the letter symbols in a legend immediately
calibration procedure in 15.2.14. Do not repeat these steps following the calculation equation.
here. Possible subheadings are as follows: 15.2.16 Report—Show limits to be reported.
15.2.15.1 Final Conditioning and Adjustment of the Gas 15.2.17 Precision—Limiting values for precision should be
Chromatographic System—This section is intended to include based on cooperative test results. Judgment as to the accept-
adjustment and verification of the state of the chromatographic ability of results (95 % probability) should be based on the
system before analytical use. following criteria.
15.2.15.2 Sampling—Careful attention must be given to the 15.2.17.1 Repeatability—The following wording should be
sampling techniques since representative samples are essential used: Duplicate results by the same operator should not be
to achieve successful analysis. Include special directions that considered suspect unless they differ by more than the follow-
may be required for taking samples, for preservation of ing amounts: (insert determined limits in tabular form).
samples, and for special treatment of samples prior to injection. 15.2.17.2 Reproducibility—The following wording should
15.2.15.3 The remainder of the steps leading to the chro- be used: The result submitted by each of two laboratories
matogram. should not be considered suspect unless the two results differ
15.2.15.4 Typical Chromatogram—Show, in a figure, a plot by more than the following amounts: (insert determined
of the retention time (in minutes) versus the detector response. amounts in tabular form).
Label the known peaks (including the dead volume or unad- 15.2.18 Appendixes—Supplementary information may be
sorbed gas peak) and indicate in parentheses the attenuation for included in one or more Appendixes to the report. Examples of
each peak. such information are: technique to improve column life,
NOTE 8—When determining the retention time of the unadsorbed peak,
directions to clean the apparatus, leak check procedures,
the retention time of air is used for thermal conductivity detectors, procedures to optimize column performance, development of
methane for flame ionization detectors, and methylene chloride lead space equations used in the calculations, and precautions to avoid
vapors for ECDs. common causes of errors, etc.

17
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16. Keywords
16.1 gas chromatography; GC; packed columns

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