Stability indicating RP-HPLC method for methylcobalamin determination in different dosage forms: Application to photodegradation kinetics and pH rate profiling.

Abstract

A stability-indicating RP-HPLC method for methylcobalamin determination was developed. Stress degradation under variable conditions was carried out. Methylcobalamin had pronounced susceptibility to hydrolysis under acidic, alkaline, and photolytic conditions; further study of photolytic degradation kinetics and pH rate profiling over pH range 2-11 was carried out. Photodegradation of methylcobalamin followed zero-order kinetics with half-life 0.99h equivalent to 1971.53 lux. Methylcobalamin followed pseudo-first-order kinetics upon exposure to acidic and alkaline hydrolysis with highest stability at pH 5 and least stability at pH 2. Optimization of chromatographic conditions was performed using two level full factorial design, and chromatographic analysis was executed using Inertsil column (250×4.6mm, 5μm) maintained at 25◦ C. Elution was carried out using 25mM potassium dihydrogen phosphate (pH adjusted with phosphoric acid to 3.8): methanol:acetonitrile (55:35:10, v/v) as mobile phase. The flow rate was 1.0ml/min. Detection was carried out at 220nm using diode array detector. The method was validated as per ICH guidelines; the linearity was over concentration range 2-160μg/ml with coefficient of determination 0.9995. The method was effectively applied for determination of methylcobalamin in Cobalvex ampoule, Cobal tablet, Cobal-F tablet, and Methyltechon oral dissolvable film without interfering from excipients within run time 6 min.