Question

Separation of amino acids by thin layer chromatography

Answer 1

Thin layer chromatographic (TLC) technique readily provides qualitative information and with careful attention to details, it is possible to obtain quantitative data. Thin layer chromatography is a technique used to separate and identify compounds of interest. A TLC plate is made up of a thin layer of silica adhered to glass or aluminum for support. The silica gel acts as the stationary phase and the solvent mixture acts as the mobile phase. In the ideal solvent system the compounds of interest are soluble to different degrees. Separation results from the partition equilibrium of the components in the mixture.
In the simplest form of the technique, a narrow zone or spot of the sample mixture to be separated is applied near one end of the TLC plate and allowed to dry. The strip or plate is then placed with this end dipping in to the solvent mixture, taking care that the sample spot/zone is not immersed in the solvent. As the solvent moves towards the other end of the strip, the test mixture separates into various components. This is called as the development of TLC plates. The separation depends on several factors; (a) solubility: the more soluble a compound is in a solvent, the faster it will move up the plate. (b) attractions between the compound and the silica, the more the compound interacts with silica, the lesser it moves, (c) size of the compound, the larger the compound the slower it moves up the plate.



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