Stability-Indicating HPLC Method for the Determination of Metadoxine as Bulk Drug and in Pharmaceutical Dosage Form

Abstract

A sensitive and reproducible method is described for the quantitative determination of metadoxine in the presence of its degradation products. The method was based on high performance liquid chromatographic separation of the drug from its degradation products on the reversed phase, kromasil column [C₁₈ (5-micron, 25 cm × 4.6 mm, i.d.)] at ambient temperature using a mobile phase consisting of methanol and water (50: 50, v/v). Flow rate was 1.0 mL min⁻¹ with an average operating pressure of 180 kg cm⁻² and t _R was found to be 2.85 ± 0.05 min. Quantitation was achieved with UV detection at 286 nm based on peak area with linear calibration curves at concentration range 10–100 μg mL⁻¹. This method has been successively applied to pharmaceutical formulation. No chromatographic interference from the tablet excipients was found. The method was validated in terms of precision, robustness, recovery and limits of detection and quantitation. Drug was subjected to acid, alkali and neutral hydrolysis, oxidation, dry heat, wet heat treatment and photo and UV degradation. As the proposed method could effectively separate the drug from its degradation products, it can be employed as stability indicating one. Moreover, the proposed HPLC method was utilized to investigate the kinetics of the acidic, alkaline and oxidative degradation processes at different temperatures and their respective apparent pseudo first order rare constant, half-life and activation energy was calculated with the help of Arrhenius plot. In addition the pH-rate profile of degradation of metadoxine in constant ionic strength buffer solutions with in the pH range 2–11 was studied.