Thin layer chromatographic studies of amino acids were performed on three differentially charged surfaces of silica gel, alumina and cellulose with 40% aqueous solution of five carbohydrates namely dextrose, fructose, maltose, lactose and sucrose. 40% dextrose-alumina and 40% dextrose-cellulose TLC systems were identified as most favorable for selective separation of glutamic acid and tryptophan from the mixture of other amino acids. In addition to this, several combinations of amino acids have been resolved on silica gel and alumina layers with 40% dextrose as eluent. The lowest detectable limit of glutamic acid and tryptophan, stability of mixtures of amino acids and reproducibility of RF values were determined. The proposed method is environmentally acceptable because of the use of non-toxic nature of eluents used.