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Gas Chromatography Methods in Bacteria Identification

In addition to the use of chemical, physical and physiological characteristics of bacteria, in recent years gas chromatography has also been used to detect specific cellular components in bacteria and for rapidly identifying bacteria. Either intact bacterial cells or typical bacterial constituents from the cell wall, cell membrane or the cytoplasm are converted into volatile compounds and analysed by gas chromatography.

The patterns can be then compared for the identification of the organism. Also, the end products of metabolism in a given medium are generally characteristic of a given organism and therefore by analyzing the end products it is possible to tentatively identify the various bacteria

In principle, gas liquid chromatography (GLC) is a method of separating components in a volatile mixture by partition between two phases, a stationary phase and a moving gaseous phase. A vapourised sample is introduced into the stream of moving ,phase at the inlet end of the column which consists of metal or glass tubing containing a specific type of packing material.

The carrier gas (N2, He or Ar) flows through the column and the compo­nents are distributed between the mobile phase and the liquid stationary phase in the column. The separated components of sample emerge as more or Jess separate bands at the column outlet, which are detected either using a thermal conductivity detector (TCD) or a flame ionization detector (FID).

In the pyrolysis gas liquid chromatographic method, thermal degradation of a sample is carried out under carefully controlled conditions and the resulting breakdown products are separated by gas liquid chromatography. Various types of pyrolysers are used in combination with gas chromatography. Each bacterial strain produces a characteristic pyrochromatogram which is used as a finger print to identify the microorganisms.The basic presumption in this technique is that the chemical composition of a given cellular component in two identical bacteria grown under similar condition is identical.

Analysis of cell components such as carbohydrates, fatty acids, hydroxyacids, organic acids, amino acids and amines cam also be done by gas chromatography. A general profile of microbial cell components gives useful information for classification; however, analysis of specific cell constituents provides a mean of specific identification.

Characteristic metabolic products produced by bacteria can be analyzed by gas chromatography and used in identification. for example, gas chromatographic methods have been used routinely as aids in the identification of anaerobic bacteria through analysis of end products such as alcohols, fatty acids and other volatile compounds. Organic acids have also been analyzed from various aerobic bacterial cultures and the chromatographic patterns of organic acids produced by bacteria have been helpful in identification.

 

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