Rajesh Shrimali
Column Chromatography consists of a wide range of physical techniques used to separate and purify the complex mixture of compounds. It is used for the separation of organic compounds.
Column Chromatography is useful as the separation method for the mixture that contains a small number of components with similar physical and chemical properties. Such components cannot be separated by any other method. Only the Column chromatography technique can be used for such type of components separation.
In Industries, synthesising bio-molecules requires a special technique that does not denature the characteristics of the bio-molecules during Separation and Purification. Column Chromatography is one such technique having different modes that are used on a large scale at the Industrial level as it is bio-compatible and requires a relatively short time for separation and purification. The Column Chromatography Principle is mainly based on the sample's size, Charge, and Hydrophobicity, which in turn leads to various Chromatographic techniques such as-Size exclusion chromatography or Gel permeation chromatography, Adsorption chromatography, Ion exchange chromatography, and Affinity Chromatography. Each method is based on the biological interaction between the sample bio-molecules and the packing material, such as Alumina or Silica gel.
(Types of chromatography and their applications)
Size exclusion chromatography:
It is known as molecular sieve chromatography separates analyses according to their size as they pass on through the stationary phase, namely Silica, which is porous and acts as a hydrophilic surface.
In Adsorption chromatography:
The stationary phase is considered the Adsorbent, which is used in separating Non-volatile mixtures. In this method, the Adsorbent used can be Silica, Alumina or Cellulose.
Ion exchange chromatography:
It separates polar molecules and ions such as proteins and amino acids containing positive and negative charged chemical groups. In this method, the stationary phase is Silica or Alumina's slurry.
Affinity chromatography:
Affinity chromatography is used to separate recombinant proteins and enzymes, and here the stationary phase acts as a Support medium. Chromatography techniques are widely used-In Biochemical research applications to separate and identify the compound of biological origin. In petroleum industries, to control the production of saturated carbons, in chemical industries to purify the chemicals, and in sugar, factories refine sugar from molasses.
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