A micellar thin-layer chromatographic system comprising of silica layer impregnated with 0.01% sodium dodecyl sulphate (SDS) as stationary phase and 0.1% aqueous solution of Cween 80 as mobile phase was identified as the most favorable system for the mutual separation of five water-soluble vitamins (folic acid, cyanocobalamin, thiamine, pyridoxine, and riboflavin). Effects of the presence of metal cations, inorganic anions, and amines as impurities was determined. The detection and dilution limits, given in parenthesis for folic acid (1 microg and 1:0.1 x 10(4)), cyanocobalamin (0.08 microg and 1:1.25 x 10(4)), thiamine (0.05 microg and 1:2.0 x 10(4)), pyridoxine (0.5 microg and 1:0.2 x 10(4)), and riboflavin (0.5 microg and 1:0.2 x 10(4)) were determined.
Specific separation of thiamine hydrochloride from riboflavin, nicotinic acid, calcium D-pantothenate, pyridoxine hydrochloride, cyanocobalamin, and ascorbic acid has been achieved on commercial precoated silica gel 60F 254 TLC plates with dioxanewater 1:1 (v/v) as mobile phase. The spots were visualized under UV light. The effect of impurities (metal cations and inorganic anions) on the chromatography of thiamine hydrochloride was examined. The detection limit for thiamine hydrochloride was 0.09 μg per spot and the relative standard deviation of the R F value of thiamine hydrochloride in five analyses was 14.99%. The applicability of the method to the identification of thiamine in pharmaceutical formulations was also tested.
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