Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING REVERSE PHASE HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ALPRAZOLAM AND IMIPRAMINE IN PHARMACEUTICAL DOSAGE FORM

V. Mounika, P. Nagaraju* and G. Indira Priyadarshini

ABSTRACT

A simple, precise and stability indicating reversed phase liquid chromatographic method was developed and validated for simultaneous estimation of Alprazolam and Imipramine in bulk and tablet formulation. The separation was achieved on BDS C18 (150mm x 4.6mm, 5μm) analytical column with mobile phase comprising of Buffer: Acetonitrile (pH 4.8) (55:45v/v) at isocratic flow of 1.0ml/min with UV detection at 222 nm. The retention times of Alprazolam and Imipramine was found to be 5.3 and 2.3 minutes respectively. The method was successfully validated in accordance to ICH guidelines for accuracy, precision, specificity, linearity, ruggedness and robustness. The linear regression analysis data for calibration plots showed good linear relationship in the concentration range 50-300 μg/mL for Imipramine and 0.5-3 μg/mL for Alprazolam. The drugs were exposed to acidic, basic, oxidation, thermal and photolytic stress degradation conditions. The resultant stressed samples were analyzed by the proposed method and was established to provide high resolution among the degradation products and the analytes. All the peaks of degraded product were resolved from the active pharmaceutical ingredient with significantly different retention time and the peak purity of analyte peaks in the stressed samples was confirmed by photodiode array detector. The method could effectively separate the drug from its degradation product; it can be employed as a stability- indicating one.

Keywords: Alprazolam, Imipramine, RP-HPLC, validation, Stability.