Principle of column chromatography

The column chromatography is a broadly used technique for the separation or purification of the sample components. Solute adsorption of solution through a stationary phase and separates the mixture into each analyte, it depends on the affinity between the mobile phase and the stationary phase. The molecules which are less affinity towards the stationary phase early separated and molecules have more affinity which separated later. The mobile phase and stationary phase are the two phases of column chromatography in which the mobile phase is liquid and solid stationary phase is used. The sample mixture travels through the stationary phase with the mobile phase and separates the sample based on the affinity of each component toward the silica. The wet and dry method used for the preparation of the column.

Dry method:
In the dry method, the column is filled with the dry silica, and the mobile phase passes through it till silica is properly wet and settled. In this method of column chromatography, the column should always wet with the solvent until the end of the work.
Wet method:
In the wet method slurry of silica and solvent is first made and then using a funnel is inserted on the column it also needs to settle and wet until the end.
In some applications, different types of column chromatography such as adsorption chromatography, ion-exchange chromatography, partition chromatography, and gel chromatography are used, and they operate under the same principle of column chromatography (adsorption).


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