Fundamentals Of Chromatography

Chromatography is a science of study that involves the separation of molecules in a mixture based on differences in their structure and/or composition

. It is one of the most common physical methods performed in laboratories to separate the most complex mixtures containing complex molecules. The procedure consists of two parts viz. the mobile phase and the stationery phase. The mobile phase is the mixture to be separated dissolved in a solvent and the stationery phase is the layer of a certain material in its solid form through which the mobile phase passes through and disintegrates into separate components at various stages. Adsorptive materials are used in the stationery phase.

If you notice the functions of chromatography procedure you would know what exactly it is useful for. Its application implies on to chemical or bioprocessing industries where they are faced with the challenge to separate and purify from a complex mixture a particular compound that forms an important factor in the production line. The method is so precise that it can even successfully separate proteins that vary in a single amino acid from a complex protein mixture. Also the conditions under which chromatography is performed are not severe thus it can be used to separate products with higher sensitivity as well.

Some of the conventional equipments involved in chromatography were paper and thin layer (glass and silica gel). Modern chromatographic techniques involve the use of columns. These columns are made up of glass or a metal tube that are strong enough to withstand the pressure of the materials that may be applied to it in the process. These tubes contain the stationery phase, while the mobile phase is made to pass through it. The stationery phase absorbs the solvent in the mobile phase and efficiently separates the mixture into its different components. The stationary phase columns can be of two types, viz Packed Bed Column and Open Tubular Column. While packed bed column would consist of a granular form of the stationery phase packed closely into the column and open tubular column consists of a thin film or layer on the column wall.

A broad category of chromatographic techniques performed in laboratories today are Gas chromatography and Liquid chromatography. These are classified in terms of the materials used in mobile and the stationery phases. The mobile phase is the carrier fluid for the target samples and the stationery phase normally consists of materials that have a varying affinity for the components present in the sample mixture. When the mobile phase is in the form of a gaseous substance, especially inert gas, and the stationery phase is an adsorbent or liquid that is supported on a porous support that is also inert, the process is known as gas chromatography. When both the mobile and the stationery phase are liquids and the liquid in the stationery phase is bonded to the surface of an adsorbent, the chromatography type is liquid chromatography.

by: Cliton Hitchner

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