Chromatography according to USP can be defined as a procedure by which solutes are separated by a differential migration process in a system consisting of two or more phases, one of which moves continuously in a given direction and in which the individual substances exhibit different motilities by reason of differences in absorption, partition, solubility, vapour pressure, molecular size, or ionic charge density.
Column chromatography may be defined as the selective absorption and separation of a mixture of chemical compounds on a column of stationery phase, through which a suitable mobile phase (solvent) has been passed under the influence of gravity.
Types of Column Chromatography
1) Liquid-solid chromatography:
Here stationery phase is solid and mobile phase is liquid
2) Liquid- liquid chromatography:
Here both stationery and mobile phase is liquid
3) Thin layer chromatography:
Here stationery phase is a plane in the form of a solid supported on an inert plate.
4) High performance liquid chromatography:
This system relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material.
Principle of Column Chromatography
The principle underlying the separation of the compounds is adsorption at the solid-liquid interface. For successful separation the compounds of a mixture must show different degree of affinity for the solid support (or adsorbent) and the interaction between the compounds and the adsorbent must be irreversible. As the adsorbent is washed with fresh solvent, the various components will, therefore, move down through the column in order of their affinity for the adsorbent. Those with least affinity move down the column at a faster rate than those with the greatest affinity for the adsorbent.