Its underlying principle rests on the fact that single compound from mixture can be separated on the basis of differential adsorption on the adsorbate while moving through the column at different rates. The compound with the strongest affinity for the adsorbent is adsorbed at the top while the compound with least affinity is adsorbed at the bottom of column. Different grade series of adsorbents have been identified like starch, inulin, tale, magnesium carbonate, etc. Complete description of the preparation of the adsorption columns and different solvents with experimental layout have been discussed in this chapter. Column chromatography finds various applications like analytical chemistry, separation of geometrical isomers, separation of diasteromers, separation of tautomeric mixtures, separation of racemates, etc.