Green analytical method for identification of amino acids, vitamins and sugars with preliminary separation on HPTLC plates

High performance thin layer chromatography (HPTLC) has been used as a green analytical method for the identification and separation of amino acids, vitamins and sugars using silica static flat bed in contact of n-butyl alcohol, ethylene glycol or ethyl acetate and their mixtures. From the point of view of chromatographic performance, a mixture of n-butyl alcohol-70% aqueous ethylene glycol-ethyl acetate in the ratio of 5:2:3 by volume proves to be more efficient than the individual components for providing useful separations of amino acids, vitamins and sugars from their multi-component mixtures. The effect of presence of inorganic ions (cations and anions) as impurities in the sample has been examined for the separation of amino acids, vitamins, and sugars. The limits of detection for different analytes are found to be 7.5 mu g spot(-1) for proline, thiamine, ascorbic acid, mannose dextrose and fructose; 1.5 mu g spot(-1) for pyridoxine and xylose; 0.75 mu g spot(-1) for leucine, methionine, histidine, maltose and galactose; and 0.15 mu g spot(-1) for glutamic acid. The worked out HPTLC procedure has been tested for its applicability in the identification and separation of amino acids and vitamins present in the commercially available drug and beverage samples respectively. The proposed method is simple, rapid and free from the use of volatile organic solvents and is therefore environmentally acceptable.