CHAPTER 27 GAS CHROMATOGRAPHY
GAS CHROMATOGRAPHY • In gas chromatography (GC), the sample is vaporized and injected onto the head of a chromatographic column. • Elution is brought about by the flow of an inert gaseous mobile phase. In contrast to most other types of chromatography, the mobile phase does not interact with molecules of the analyte; its only function is to transport the analyte through the column.
Two types of gas chromatography are encountered: gas-solid chromatography (GSC) and gas-liquid chromatography (GLC). • Gas-liquid chromatography finds widespread use in all fields of science, where its name is usually shortened to gas chromatography (GC), despite the fact that this usage neglects gas-solid chromatography as a legitimate kind of chromatography. • Gas-liquid chromatography is based upon the partition of the analyte between a gaseous mobile phase and a liquid phase immobilized on the surface of an inert solid. • The concept of gas-liquid chromatography was first enunciated in 1941 by Martin and Synge, who were also responsible for the development of liquid-liquid partition chromatography. • Three years later, in 1955, the first commercial apparatus for gas-liquid chromatography appeared on the market. Since that time, the growth in applications of this technique has been phenomenal.
Gas-solid chromatography is based upon a solid stationary phase in which retention of analytes is the consequence of physical adsorption. Gas-solid chromatography has limited application owing to semipermanent retention of active or polar molecules and severe tailing of elution peaks (a consequence of the nonlinear character of adsorption process). Thus, this technique has not found wide application except for the separation of certain low-molecular-weight gaseous species; it is, therefore, discussed only briefly in Section 27E.
INSTRUMENTS FOR GAS-LIQUID CHROMATOGRAPHY In the last two decades, many changes and improvements in gas chromatographic instruments have appeared in the marketplace. In the 1970s, electronic integrators and computer-based data-processing equipment became common. The 1980s saw computers being used for automatic control of most instrument parameters, such as column temperature, flow rates, and sample injection; development of very high-performance instruments at moderate costs; and perhaps most important, the development of open tubular columns that are capable of separating a multitude of analytes in relatively short times.
Carrier Gas Supply Carrier gases, which must be chemically inert, include helium, nitrogen, and hydrogen. Generally, it is assumed that flow rates will be constant if the inlet pressure remains constant.
Sample InjectionSystem Column efficiency requires that the sample be of suitable size and be introduced as a “plug” of vapor; slow injection of oversized samples causes band spreadingand poor resolution. The most common method of sample injection involves the use of a microsyringe to inject a liquid or gaseous sample through a self-sealing, silicone-rubber diaphragm or septum into a flash vaporizer port located at the head of the column (the sample port is ordinarily about 50°C above the boiling point of the least volatile component of the sample). For ordinary analytical columns, sample sizes vary from a few tenths of a microliter to 20 µL. Capillary columns require much smaller samples (~10-3 µL); here, a sample splitter system is employed to deliver only a small fraction of the injected sample to the column head, with the remainder going to waste.
Column Configurationsand Column Ovens • Two general types of columns are encountered in gas chromatography, packed and open tubular, or capillary.To date, the vast majority of gas chromatography has been carried out on capillarycolumns.
Column Configurationsand Column Ovens Column temperature is an important variable that must be controlled to a few tenths of a degree for precise work. Thus, the column is ordinarily housed in a thermostated oven. The optimum column temperature depends upon the boiling point of the sample and the degree of separation required. Roughly, a temperature equal to or slightly above the average boiling point of a sample results in a reasonable elution time (2 to 30 min). For samples with a broad boiling range, it is often desirable to employ temperature programming, whereby the column temperature is increased either continuously or in steps as the separation proceeds. Figure 27-5 Effect of temperature on gas chromatograms: (a) isothermal at 45°C; (b) isothermal at 145°C; (c) programmed at 30° to 180°C.
Characteristics of the Ideal Detector • The ideal detector for gas chromatography has the following characteristics: • Adequate sensitivity. Just what constitutes adequate sensitivity cannot be described in quantitative terms. For example, the sensitivities of the detectors we areabout to describe differ by a factor of 107. Yet all are widely used and clearly adequate for certain tasks; the least sensitive are not, however, satisfactory for certain applications. In general, the sensitivities of present-day detectors lie in the range of 10-8 to 10-15 g solute/s. • Good stability and reproducibility. • A linear response to solutes that extends over several orders of magnitude. • A temperature range from room temperature to at least 400°C. • A short response time that is independent of flow rate. • High reliability and ease of use. The detector should, to the extent possible, be foolproof in the hands of inexperienced operators. • Similarity in response toward all solutes or alternatively a highly predictable and selective response toward one or more classes of solutes. • Nondestructive of sample.
Flame Ionization Detectors (FID) The flame ionization detector is the most widely used and generally applicable detector for gas chromatography. With a burner such as that shown in Figure 27-6, the effluent from the column is mixed with hydrogen and air and then ignited electrically. Most organic compounds, when pyrolyzed at the temperature of a hydrogen/air flame, produce ions and electrons that can conduct electricity through the flame. A potential of a few hundred volts is applied across the burner tip and a collector electrode located above the flame: The resulting current (~10-12 A) is then directed into a high-impedance operational amplifier for measurement. The ionization of carbon compounds in a flame is a poorly understood process, although it is observed that the number of ions produced is roughly proportional to the number of reduced carbon atoms in the flame. Because the flame ionization detector responds to the number of carbon atoms entering the detector per unit of time, it is a mass-sensitive, rather than a concentration-sensitive, device. Functional groups, such as carbonyl, alcohol, halogen, and amine, yield fewer ions or none at all in a flame. In addition, the detector is insensitive toward noncombustible gases such as H2O, CO2, SO2, and NOx. These properties make the flame ionization detector a most useful general detector for the analysis of most organic samples, including those that are contaminated with water and the oxides of nitrogen and sulfur. The insensitivity of the flame ionization detector to water makes it particularly useful for the detection of pollutants in natural water samples. The flame ionization detector exhibits a high sensitivity (~10-13 g/s), large linear response range (~107), and low noise. It is generally rugged and easy to use. A disadvantage of the flame ionization detector is that it is destructive of sample.
Thermal Conductivity Detectors (TCD) A very early detector for gas chromatography, and one that still finds wide application, is based upon changes in the thermal conductivity of the gas stream brought about by the presence of analyte molecules. This device is sometimes called a katharometer The sensing element of a katharometer is an electrically heated element whose temperature at constant electrical power depends upon the thermal conductivity of the surrounding gas. The heated element may be a fine platinum, gold, or tungsten wire or, alternatively, a semiconducting thermistor. A modulated single-filament thermal conductivity detector was introduced in 1979; this device offers higher sensitivity, freedom from baseline drift, and reduced equilibration time. Here, the analytical and reference gases are passed alternately over a tiny filament held in a ceramic detector cell, which has a volume of only 5 µL. The gas-switching device operates at a frequency of 10 Hz. The output from the filament is thus a 10Hz electrical signal whose amplitude is proportional to the difference in thermal conductivity of the analytical and reference gases. Because the amplifier circuit responds only to a 10-Hz signal, thermal noise in the system is largely eliminated.
Thermal Conductivity Detectors (TCD) The thermal conductivities of helium and hydrogen are roughly six to ten times greater than those of most organic compounds. Thus, in the presence of even small amounts of organic materials, a relatively large decrease in the thermal conductivity of the column effluent takes place; consequently, the detector undergoes a marked rise in temperature. The conductivities of other carrier gases more closely resemble those of organic constituents; therefore, a thermal conductivity detector dictates the use of hydrogen or helium in order to achieve good sensitivity. The advantage of the thermal conductivity detector is its simplicity, its large linear dynamic range (~105), its general response to both organic and inorganic species, and its nondestructive character, which permits collection of solutes after detection. A limitation of the katharometer is its relatively low sensitivity (~10-8 g solute/mL carrier gas). Other detectors exceed this sensitivity by factors as large as 104 to 107. It should be noted that the low sensitivity of thermal detectors often preclude their use with capillary columns because of the very small samples that can be accommodated by such columns.
Electron-Capture Detectors (ECD) The electron-capture detector has become one of the most widely used detectors for environmental samples because this detector selectivity detects halogen containing compounds, such as pesticides and polychlorinated biphenyls. As shown in Figure 27-B, the effluent from the column is passed over a β emitter, usually nickel-63. An electron from the emitter causes ionization of the carrier gas (often nitrogen) and the production of a burst of electrons. In the absence of organic species, a constant standing current between a pair of electrodes results from this ionization process. The current decreases markedly, however, in the presence of those organic molecules that tend to capture electrons. The response is nonlinear unless the potential across the detector is pulsed. The electron-capture detector is selective in its response, being highly sensitive to molecules containingelectronegative functional groups such as halogens, peroxides, quinones, and nitro groups. It is insensitive to functional groups such as amines, alcohols, and hydrocarbons. An important application of the electrón-capture detector has been for the detection and determination of chlorinated insecticides. Electron-capture detectors are highly sensitive and possess the advantage of not altering the samplesignificantly (in contrast to the flame detector). On the other hand, their linear response range is usually limited to about two orders of magnitude.
Atomic Emission Detectors (AED) The atomic emission detector is available commercially. In this device (see Figure 27-9), the eluent is introduced into a microwave-energized helium plasma that is coupled to a diode array optical emission spectrometer. The plasma is sufficiently energetic to atomize all of the elements in a sample and to excite their characteristic atomic emission spectra. These spectra are then observed with a spectrometer that employs a movable, flat diode array capable of detecting emitted radiation from about 170 to 780 nm. As shown on the right of the figure, the positionable diode array is capable of monitoring simultaneously two to four elements at any given setting. At the present time, the software supplied with the detector allows measurement of the concentration of 15 elements. Presumably, future software will permit detection of other elements.
Atomic Emission Detectors (AED) Figure 27-10 illustrates the power of this type of detector. The sample in this case consisted of a gasoline containing a small concentration of methyl tertiary butyl ether (MTBE), an antiknock agent, as well as several aliphatic alcohols in low concentrations. The upper spectrum, obtained by monitoring the carbon emission line at 198 nm, consists of a myriad of peaks that would be impossible to sort out and identify. In contrast, whenthe oxygen line at 777 nm is used to obtain the chromatogram (Figure 27-10b), peaks for the various alcohols and for MTBE are clearly evident and readily identifiable.
GAS CHROMATOGRAPHICCOLUMNS AND STATIONARYPHASES Historically, all of the pioneering gas-liquid chromatographic studies in the early 1950s were carried out on packed columns in which the stationary phase was a thin film of liquid adsorbed on the surface of a finely divided, inert solid support. From theoretical studies made during this early period, it became apparent, however, that unpacked columns having inside diameters of a few tenths of a millimeter should provide separations that were much superior to packed columns in both speed and column efficiency. In such capillary columns, the stationary phase was a uniform film of liquid a few tenths of a micrometer thick that coated the interior of a capillary tubing uniformly.
Open Tubular Columns Table 27-1 compares the performance characteristics of fused-silica capillary columns with other types of wallcoated columns as well as with support-coated and packed columns.
The Stationary Phase Desirable properties for the immobilized liquid phase in a gas-liquid chromatographic column include: low volatility (ideally, the boiling point of the liquid should be at 100°C higher than the maximum operating temperature for the column); (2) thermal stability; (3) chemical inertness; (4) solvent characteristics such that k' and αvalues for the solutes to be resolved fall within a suitable range.
The Stationary Phase • The retention time for a solute on a column depends upon its distribution constant, which in turn is related to the chemical nature of the stationary phase. Clearly, to be useful in gas-liquid chromatography, the immobilized liquid must generate different distribution constants for different solutes. In addition, however, these constants must not be extremely large or extremely small because the former lead to prohibitively long retention times and the latter result in such short retention times that separations are incomplete.
The Stationary Phase • To have a reasonable residence time in the column, a species must show some degree of compatibility (solubility) with the stationary phase. Here, the principle of“like dissolves like” applies, where “like” refers to the polarities of the solute and the immobilized liquid.
The Stationary Phase • Polarity is the electrical field effect in the immediate vicinity of a molecule and is measured by the dipole moment of the species. Polar stationary phases contain functional groups such as—CN,—CO, and—OH. Hydrocarbon-type stationary phases and dialkylsiloxanes are nonpolar, whereas polyester phases are highly polar. • Polar solutes include alcohols, acids, and amines; solutes of medium polarity include ethers, ketones, and aldehydes. Saturated hydrocarbons are nonpolar. • Generally, the polarity of the stationary phase should match that of the sample components. When the match is good, the order of elution is determined by the boiling point of the eluents.
Some Widely Used Stationary Phases Five of the liquids listed in Table 27-2 are poly dimethyl siloxanes that have the general structure In the first of these, polydimethylsiloxane, the —R groups are all —CH3 giving a liquid that is relatively nonpolar, in the other polysiloxanes shown in the table, a fraction of the methyl groups are replaced by functional groups such as phenyl (—C6H5), cyanopropyl (-C3H6CN), and trifluoropropyl (—C3H6CF3 ). The percentage descriptions in each case gives the amount of substitution of the named group for methyl groups on the polysiloxane backbone. Thus, for example, 5% phenyl polydimethylsiloxane has a phenyl ring bonded to 5% by number of the silicon atoms in the polymer. These substitutions increase the polarity of the liquids to various degrees.
Film Thickness Commercial columns are available having stationary phases that vary in thickness from 0.1 to 5 µm. Film thickness primarily affects the retentive character and the capacity of a column. Thick films are used with highly volatile analytes because such films retain solutes for a longer time, thus providing a greater time for separation to take place. Thin films are useful for separating species of low volatility in a reasonable length of time. For most applications with 0.26- or 0.32- mm columns, a film thickness of 0.26 µm is recommended.
Qualitative Analysis Gas chromatograms are widely used as criteria of purity for organic compounds. Contaminants, if present, are revealed by the appearance of additional peaks; the areas under these peaks provide rough estimates of the extent of contamination. The technique is also useful for evaluating the effectiveness of purification procedures. In theory, retention times should be useful for the identification of components in mixtures. In fact, however, the applicability of such data is limited by the number of variables that must be controlled in order to obtain reproducible results. Nevertheless, gas chromatography provides an excellent means of confirming the presence or absence of a suspected compoundin a mixture, provided an authentic sample of the substance is available. No new peaks in the chromatogram of the mixture should appear upon addition of the known compound, and enhancement of an existing peak should be observed. The evidence is particularly convincing if the effect can be duplicated on different columns and at different temperatures.
Selectivity Factors We have seen that the selectivity factor a for compounds A and B is given by the relationship If a standard substance is chosen as compound B, then α can provide an index for identification of compound A, which is largely independent of column variables other than temperature; that is, numerical tabulations of selectivity factors for pure compounds relative to acommon standard can be prepared and then used for the characterization of solutes. Unfortunately, finding a universal standard that yields selectivity factors of reasonable magnitude for all types of analytes is not possible. Thus, the amount of selectivity factor data available in the literature is presently limited.
Quantitative Analysis • The detector signal from a gas-liquid chromatographic column has had wide use for quantitative and semi-quantitative analyses. • An accuracy of 1% relative is attainable under carefully controlled conditions. • As with most analytical tools, reliability is directly related to the control of variables; the nature of the sample also plays a part in determining the potential accuracy.
Gas chromatography/mass spectrometry instruments have been used for the identification of hundreds of components that are present in natural and biological systems. For example, these procedures have permitted characterization of the odor and flavor components of foods, identification of water pollutants, medical diagnosis based on breath components, and studies of drug metabolites.