Introduction to chromatography chromatography is a non-destructive procedure

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CHROMATOGRAPHY Introduction to chromatography Chromatography is a non-destructive procedure for resolving a multi-component mixture of trace, minor, or major constituents into its individual fractions. Different variations may be applied to solids, liquids, and gases. While chromatography may be applied both quantitatively, it is primarily a separation tools. Chromatography is relatively a new technique which was first invented by M. Tswett, a botanist in 1906 in Warsaw. In that year, he was successful in doing the separation of chlorophyll, xanthophyii and several other colored substances by percolating vegetable extracts through a column of calcium carbonate. the calcium carbonate column acted as an adsorbent and the different substances got adsorbed to different extent and this gives rise to coloured bands at different positions, ion the column. Tswett termed this system of coloured bands as the chromatogram and the method as chromatography after the Greek words chroma and graphs meaning “colour” and “writing” respectively. However, in the majority of chromatographic procedures no coloured products are formed and the term is a misnomer. Considerable advances have since been made and the method is used to separate coloured as well as colourless substances. The column of calcium carbonate, used in Tswett’s method, remains stationary and is therefore termed as the stationary phase. The solution of vegetable extracts moves or flows down the column and is therefore termed as the mobile phase. Chromatography may be regarded as a method of separation in which separation of solutes occur between a stationary phase and a mobile phase. Definition of chromatography Chromatography may be defined as a method of separating a mixture of components into individual components through equilibrium distribution between two phases. Essentially, the technique of chromatography is based on the differences in the rate at which the components of a mixture move through a porous medium (called stationary phase) under the influence of some solvent or gas (called moving phase). 1

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