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CHROMATOGRAPHY

Smt Shilpa Bandrad

Dept of Microbiology

Smt V. G. Degree College for Women

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INTRODUCTION

The term “chromatography” was coined by a Russian botanist Tswett in 1906.
It includes all those techniques in which various components in a mixture get seperated during their passage through a porous media due to the differences in their migration rates.

Components:

mobile phase: a solvent that flows through the supporting medium
stationary phase: a layer or coating on the supporting medium that interacts with the analytes
supporting medium: a solid surface on which the stationary phase is bound or coated

In chromatograhy, the sample is applied to one end of a porous support which holds the stationary phase & mobile phase is then made to flow over it.
The various constituents in a sample get seperated essentially due to differences in their partition or distribution behaviour between the stationary phase & the mobile phase.
The partition or the distribution co-efficient (Kd), is defined as the ratio of concentration of a compound in two phases (say A&B) at equilibrium.
Kd = concn of the compound in solvent A

concn of the compound in solvent B

APPLICATIONS

The chromatographic technique is used for the separation of amino acids, proteins & carbohydrates.
Used for analysis of drugs, hormones & vitamins.
Helpful for qualitative and quantitative analyses of complex mixtures.
Useful for the determination of the molecular weight of proteins.

TYPES OF CHROMATOGRAPHY