Columns, capillary

There are numerous textbooks on and guides to chromatography, it is not intended to discuss the details of this technique in this chapter. The use of capillary columns, particularly in conjunction with headspace or purge and trap analysis, and modem bench-top mass spectrometers with large desk-top computing capacity, has produced a powerful technique for the analysis of VOCs. Typically, the separation, identification and quantification of up to about one hundred compounds in a single analysis are possible and routinely carried out. 

Fused silica is the most common inert and temperature stable material used in the manufacture of capillary columns. They are flexible, robust and easy to install. Many laboratories find difficulty in separating the ever increasing list of volatile compounds requiring analysis. Many phases suffer from low thermal stability, and analysis times are unacceptably long. The most common column used in the analysis of VOCs is the 624 column, available from several suppliers worldwide. 

This method is especially suitable for thermolabile components and for components which may form artifacts at high injector temperatures. An example of this is oxazepam. Use of a split/splitless injector leads to artifact formation, but this can be largely prevented by use of a temperature-programmable injector. 

In this method, the sample is injected directly onto the column. Here, the sample is not contained in the glass insert. Columns with a small inside diameter are unsuitable for this technique of sample itroduction. As on-column injection is a splitless method, only low-concentration samples can be injected. This method is suitable for polar and thermally unstable components. 

This special injection technique has advantages over all other methods of analyzing volatile substances with respect to separating power, sensitivity, ease of handling, and automation possibilities, and the headspace technique is therefore becoming more and more widely used. 

In a static headspace, the sample is in a closed static system in which conditions are at thermodynamic equilibrium. After establishment of the equilibrium, the 

For reproducible analysis, it is important that equilibrium between the phases should first be established. This equilibrium is influenced by the conditioning time and temperature. The removal of the sample for the GC analysis can be by means of overpressure or underpressure, although care must be taken that the temperature not only of the sample vessel but also of the metering device is kept constant. The HS vessels (vials) are closed with a PTFE (polytetrafluoroethylene) or aluminum-coated silicone septum. The septum itself is protected from the overpressure of the interior space by an aluminum cap. Before each analysis, a blank determination with an empty HS vial is carried out to detect whether any constituents of the materials of the septum or vial are being emitted. 

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Used in virtually all organic chemistry analytical laboratories, gas chromatography has a powerful separation capacity. Using distillation as an analogy, the number of theoretical plates would vary from 100 for packed columns to 10 for 100-meter capillary columns as shown in Figure 2.1. 

The column is swept continuously by a carrier gas such as helium, hydrogen, nitrogen or argon. The sample is injected into the head of the column where it is vaporized and picked up by the carrier gas. In packed columns, the injected volume is on the order of a microliter, whereas in a capillary column a flow divider (split) is installed at the head of the column and only a tiny fraction of the volume injected, about one per cent, is carried into the column. The different components migrate through the length of the column by a continuous succession of equilibria between the stationary and mobile phases. The components are held up by their attraction for the stationary phase and their vaporization temperatures. 

Interest in this method has decreased since advances made in gas chromatography using high-resolution capillary columns (see article 3.3.3.) now enable complete identification by individual chemical component with equipment less expensive than mass spectrometry. 

The hydrocarbons are separated in another column and analyzed by a flame ionization detector, FID. As an example, Figure 3.13 shows the separation obtained for a propane analyzed according to the ISO 7941 standard. Note that certain separations are incomplete as in the case of ethane-ethylene. A better separation could be obtained using an alumina capillary column, for instance. 

The resolution of capillary columns enables the separation of all principal components of a straight-run gasoline. The most frequently used stationary phases are silicone-based, giving an order of hydrocarbon elution times close to the order of increasing boiling point. 

Gas chromatography is not an identification method the components must be identified after their separation by capillary column. This is done by coupling to the column a mass spectrometer by which the components can be identified with the aid of spectra libraries. However tbe analysis takes a long time (a gasoline contains aboutTwo hundred components) so it is not practical to repeat it regularly. Furthermore, analysts have developed te hpiques for identifying... 

One of the most important advances in column construction has been the development of open tubular, or capillary columns that contain no packing material (dp = 0). Instead, the interior wall of a capillary column is coated with a thin film of the stationary phase. The absence of packing material means that the mobile phase... 

Another approach to improving resolution is to use thin films of stationary phase. Capillary columns used in gas chromatography and the bonded phases commonly used in HPLC provide a significant decrease in plate height due to the reduction of the Hs term in equation 12.27. 

In gas chromatography (GC) the sample, which may be a gas or liquid, is injected into a stream of an inert gaseous mobile phase (often called the carrier gas). The sample is carried through a packed or capillary column where the sample s components separate based on their ability to distribute themselves between the mobile and stationary phases. A schematic diagram of a typical gas chromatograph is shown in Figure 12.16. 

The most common mobile phases for GC are He, Ar, and N2, which have the advantage of being chemically inert toward both the sample and the stationary phase. The choice of which carrier gas to use is often determined by the instrument s detector. With packed columns the mobile-phase velocity is usually within the range of 25-150 mF/min, whereas flow rates for capillary columns are 1-25 mF/min. Actual flow rates are determined with a flow meter placed at the column outlet. 

A chromatographic column provides a location for physically retaining the stationary phase. The column s construction also influences the amount of sample that can be handled, the efficiency of the separation, the number of analytes that can be easily separated, and the amount of time required for the separation. Both packed and capillary columns are used in gas chromatography. 

To minimize the multiple path and mass transfer contributions to plate height (equations 12.23 and 12.26), the packing material should be of as small a diameter as is practical and loaded with a thin film of stationary phase (equation 12.25). Compared with capillary columns, which are discussed in the next section, packed columns can handle larger amounts of sample. Samples of 0.1-10 )J,L are routinely analyzed with a packed column. Column efficiencies are typically several hundred to 2000 plates/m, providing columns with 3000-10,000 theoretical plates. Assuming Wiax/Wiin is approximately 50, a packed column with 10,000 theoretical plates has a peak capacity (equation 12.18) of... 

A capillary column that does not contain a particulate packing material. 

Capillary Columns Capillary, or open tubular columns are constructed from fused silica coated with a protective polymer. Columns may be up to 100 m in length with an internal diameter of approximately 150-300 )J,m (Figure 12.17). Larger bore columns of 530 )J,m, called megabore columns, also are available. 

Capillary columns are of two principal types. Wall-coated open tuhular columns (WCOT) contain a thin layer of stationary phase, typically 0.25 pm thick, coated on the capillary s inner wall. In support-coated open tuhular columns (SCOT), a thin layer of a solid support, such as a diatomaceous earth, coated with a liquid stationary phase is attached to the capillary s inner wall. 

An important problem with all liquid stationary phases is their tendency to bleed from the column. The temperature limits listed in Table 12.2 are those that minimize the loss of stationary phase. When operated above these limits, a column s useful lifetime is significantly shortened. Capillary columns with bonded or... 

A technique for injecting samples onto a capillary column in which only a small portion of the sample enters the column. 

A technique for injecting a sample onto a capillary column that allows a higher percentage of the sample to enter the column. 

The direct injection of thermally unstable samples onto a capillary column. 

Environmental Analysis One of the most important environmental applications of gas chromatography is for the analysis of numerous organic pollutants in air, water, and wastewater. The analysis of volatile organics in drinking water, for example, is accomplished by a purge and trap, followed by their separation on a capillary column with a nonpolar stationary phase. A flame ionization, electron capture, or... 

Time, Cost, and Equipment Analysis time can vary from several minutes for samples containing only a few constituents to more than an hour for more complex samples. Preliminary sample preparation may substantially increase the analysis time. Instrumentation for gas chromatography ranges in price from inexpensive (a few thousand dollars) to expensive (more than 50,000). The more expensive models are equipped for capillary columns and include a variety of injection options and more sophisticated detectors, such as a mass spectrometer. Packed columns typically cost 50- 200, and the cost of a capillary column is typically 200- 1000. 

Despite their importance, gas chromatography and liquid chromatography cannot be used to separate and analyze all types of samples. Gas chromatography, particularly when using capillary columns, provides for rapid separations with excellent resolution. Its application, however, is limited to volatile analytes or those analytes that can be made volatile by a suitable derivatization. Liquid chromatography can be used to separate a wider array of solutes however, the most commonly used detectors (UV, fluorescence, and electrochemical) do not respond as universally as the flame ionization detector commonly used in gas chromatography. 

The narrow bore of the capillary column and the relative thickness of the capillary s walls are important. When an electric field is applied to a capillary containing a conductive medium, such as a buffer solution, current flows through the capillary. This current leads to Joule heating, the extent of which is proportional to the capillary s radius and the magnitude of the electric field. Joule heating is a problem because it changes the buffer solution s viscosity, with the solution at the center of the... 

Schematic diagram showing a cross section of a capillary column for capillary electrophoresis.

An injection technique in capillary electrophoresis in which pressure is used to inject sample into the capillary column. 

A means of concentrating solutes in capillary electrophoresis after their injection onto the capillary column. 

A form of capillary electrophoresis in which the capillary column contains a gel enabling separations based on size. 

Procedure. A vitamin B complex tablet Is crushed and placed In a beaker with 20.00 mL of a 50% v/v methanol solution that Is 20 mM In sodium tetraborate and contains 100.0 ppm of o-ethoxybenzamIde. After mixing for 2 min to ensure that the B vitamins are dissolved, a 5.00-mL portion Is passed through a 0.45- xm filter to remove Insoluble binders. An approximately 4-nL sample Is loaded Into a 50- xm Internal diameter capillary column. For CZE the capillary column contains a 20 mM pH 9 sodium tetraborate/sodlum dIhydrogen phosphate buffer. For MEKC the buffer Is also 150 mM In sodium dodecylsulfate. A 40-kV/m electric field Is used to effect both the CZE and MEKC separations. 

Although aimed at the introductory class, this simple experiment provides a nice demonstration of the use of GG for a qualitative analysis. Students obtain chromatograms for several possible accelerants using headspace sampling and then analyze the headspace over a sealed sample of charred wood to determine the accelerant used in burning the wood. Separations are carried out using a wide-bore capillary column with a stationary phase of methyl 50% phenyl silicone and a flame ionization detector. 

This somewhat lengthy experiment provides a thorough introduction to the use of GG for the analysis of trace-level environmental pollutants. Sediment samples are extracted by sonicating with 3 X 100-mL portions of 1 1 acetone hexane. The extracts are then filtered and concentrated before bringing to a final volume of 10 mL. Samples are analyzed with a capillary column using a stationary phase of 5% phenylmethyl silicone, a splitless injection, and an EGD detector. 

The following data were obtained for four compounds separated on a 20-m capillary column. 

Zhou and colleagues determined the %w/w H2O in methanol by GG, using a capillary column coated with a nonpolar stationary phase and a thermal conductivity detector. A series of calibration standards gave the following results. 

The following data have been reported for the gas chromatographic analysis of p-xylene and methylisobutylketone (MIBK) on a capillary column. ... 

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