Abstract—A high performance thin layer chromatography system (HPTLC) for the separation of vitamin B2 and B12 has been developed. The separation was successfully using a solvent system of methanol, water, ammonia 7.3.1 (V/V) as mobile phase on HPTLC plates impregnated with boric acid. The effect of other mobile phases on the separation of vitamins was also examined. The method is based on different behavior of investigated compounds in impregnated TLC plates with different amount of boric acid. The Rf values of vitamin B2 and B12 are considered on non impregnated and impregnated silica gel HPTLC plate with boric acid. The effect of boric acid in the mobile phase and on HPTLC plates on the RF values of the vitamins has also been studied. Keywords—High performance thin layer chromatography
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