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A new thin layer chromatographic system comprising of silica gel impregnated with 0.1% aqueous solution (below CMC) of nonionic surfactant Brij-35 as stationary phase and 0.1% aqueous solution (below CMC) of cationic surfactant (cetrimide) as eluent has been found most suitable for the separation of closely related amino acids (phenylalanine and tyrosine) from their mixture. Effect of substitution of Brij-35 by cetrimide and sodium cholate has been examined to assess the impact of charge of impregnant on the chromatographic behavior of analytes. The changes brought about by the impregnation in the structure and homogeneity of silica gel have been studied by scanning electron microscopy (SEM) and fourier transform infra-red spectroscopy (FTIR) techniques. Optimizations of concentration of Brij-35 and cetrimide in the stationary and mobile phases respectively and the effect of bed height on the separation have also been examined. Chromatographic parameters like ΔR F , separation factor (α) and resolution (R S ) were calculated for the separation of tyrosine from phenylalanine and separation mechanism have been proposed. The limits of detection for phenylalanine and tyrosine were found to be 0.25 and 0.23 μg respectively. The proposed method studied is simple, rapid and free from the use of volatile organic solvents.

Synergistic Effect of Cationic-Nonionic Surfactants on Simultaneous Separation of Phenylalanine and Tyrosine