Indian Journal of Chemical Technology, Vol.15, No.6, 598-603, 2008
Stability-indicating HPLC method for determination of vitamins B-1, B-2, B-3 and B-6 in pharmaceutical liquid dosage form
A simple, selective, precise and stability-indicating high-performance liquid chromatographic (HPLC) method for determination of vitamins B-1, B-2, B-3 and B-6 in pharmaceutical liquid dosage form was developed and validated. The chromatographic conditions comprised a reversed-phase C-18 column (250 x 4.6 mm), 5 mu with a mobile phase consisting of a mixture of solution (0.015 M-1-hexane sulphonic acid sodium salt, pH 3.0 +/- 0.05) and methanol in gradient elution. The flow rate was kept at 1.5 mL/min. and the detection was carried out at 280 nm. The retention times of vitamins B-3, B-2, B-6 and B-1 were 6.3, 15.1, 19.9 and 42.7 min. respectively. The linear regression analysis data for the calibration plots showed good linear relationship with coefficient of correlation values, r = 0.999 for vitamins B-1, B-2, B-3 and B-6 in the concentration ranges of 23.08-42.85, 9.61-17.84, 116.36-216.09 and 7.01-13.02 mu g/mL respectively. The method was validated for precision, recovery and robustness. The vitamins undergo degradation under acidic, basic, peroxide, photochemical and thermal conditions. Statistical analysis proves that the method is reproducible and selective for the estimation of vitamins under study. As the method could effectively estimate the vitamins in presence of their degradation products, it can be employed as a stability-indicating method.