CHROMATOGRAPHY, Vol. 25 (2004), No. 3, pp. 111-118
Use of sodium bis (2-ethyl hexyl) sulfosuccinate (AOT) anionic surfactant mobile phase systems in thin-layer chromatography of amino acids : simultaneous separation of thioamino acids
Ali Mohammad*and Sufia Hena
Analytical Research Laboratory, Department of Applied Chemistry, Z. H. College of Engineering & Technology Aligarh Muslim University Aligarh-202002, INDIA
Received for review January 23, 2004. Revised manuscript received August 23, 2004. Accepted November 15, 2004.
Silica gel has been used as stationary phase in combination with surfactant-mediated eluents for thin layer chromatography of 12 amino acids. Several combinations of mobile phase systems comprising of different constituent and ratio have been tested for rapid and reliable separation of amino acids. The results obtained with mobile phase M7 were compared with those obtained with mobile phases M2+DMSO+methanol (3:2:7; v/v), M2+DMSO+ethanol (3:2:7; v/v), M2+DMSO+1-propanol (3:2:7; v/v) Thin layer chromatography system constituting silica gel as stationary phase and 0.001 M aqueous AOT (Sodium bis(2-ethylhexyl) sulfosuccinate) and 1-butanol (3:2:7; v/v) as mobile phase was identified as most favourable system for the separation of coexisting L-methionine (L-Met), L-cystein (L-Cys) and L-cystine (L-Cys-Cys). The proposed method is rapid and suitable for identification and separation of L-Met, L-Cys and L-Cys-Cys in the presence of many common heavy metals. The separation of microgram quantities of L-Met from milligram quantities of L-Cys or vice-versa was reported. Parameter such as limits of detection was also studied. Quantitative determination of L-Cys were done.
Keywords: TLC, Separation, Amino Acids, TLC-Spectrophotometry, AOT.