Resolution of multicomponent mixture of amino acids using environmentally benign eluents: A green chromatographic...

A new green TLC has been used for identifying and monitoring the migration behavior of amino acids through silica and kieselguhr static flat bed in contact of n-butyl alcohol, ethyl acetate or ethylene glycol and their mixtures. From the point of view of chromatographic performance, a mixture of n-butyl alcohol-70% aqueous ethylene glycol-ethyl acetate ratio 5:3:2 by volume proves to be more efficient than the individual components for separation of amino acids from their binary, ternary and quaternary mixtures and the chromatographic parameters like ΔR(F) , separation factor (α) and resolution (R(S) ) for the separation were calculated. Effect of the presence of foreign substances such as metal cations, anions, vitamins and pesticides as impurities in the sample on the separation was also examined. Effect of substitution of butanol by various alcohols has been examined to assess the impact of hydrophobicity of alcohols on the separation of amino acids. The limits of detection for tyrosine, tryptophan, alanine, isoleucine, methionine and serine were found to be 0.10 μg/spot, whereas for lysine it is 0.05 μg/spot. Application of the selected TLC system for the identification and separation of amino acids present in drugs/pharmaceuticals has been performed.