Chromatography is a physical technique of separation of almost all biological substances like proteins, lipids, nucleic acids, etc. by a continuous redistribution between two phases. The phases used are stationary phase and moving phase. Chromatography was first employed by the Russian scientist Mikhail Tsvet in 1900. Chromatography is based on the concept of partition coefficient. Any solute separates between two immiscible solvents. In chromatography a liquid is pumped through a bed of particles. The liquid is called the mobile phase and the particles the stationary phase. Some common terms used in chromatography, types of chromatography, and uses of chromatography are covered in this chapter.