SIMULTANEOUS DETERMINATION OF VITAMINS B1, B2 AND B6 IN MULTIVITAMIN TABLET AND BIOLOGICAL FLUID BY RP-HPLC

Multivitamin tablets containing various substances of varying characteristics may have a problem in quantitative analysis. This research has developed HPLC method for simultaneous determination of three vitamin components that is thiamine (Vit.B1), riboflavin (Vit.B2) and pyridoxine (Vit.B3) in tablet formulation. The chromatographic separation was achieved by using a C-18 column with dimension of 4.6 mm I. D. X 250 mm and particle size of 5 mu m. A mixture of methanol: water (22:78) was used as mobile phase. The aqueous mobile phase contained O-phosphoric acid adjusted to pH 2.5, with flow rate of 1mL/min. The effluent was monitored at 290 nm at ambient temperature. Effective separation and quantification was achieved in less than 10 min. The method was simple, accurate, precise, and could be successfully applied for the analysis of thiamine, riboflavin, and pyridoxine multivitamin tablets. The HPLC results revealed that % vitamins were found to be for B1, B2 and B6 respectively in marketed multivitamin formulation. HPLC analysis of biological fluid (urine) showed that % Vit. B1 in 2, 4, 6 hrs were found to be 0.69, 0.12 and 0.015.