Development and validation of a liquid chromatographic method for the simultaneous determination of aniracetam and its related substances in the bulk drug and a tablet formulation

Abstract

Simultaneous determination of aniracetam and its related impurities (2-pyrrolidinone, p-anisic acid, 4-p-anisamidobutyric acid and (p-anisoyl)-4-methyl-2-pyrrolidinone) was accomplished in the bulk drug and in a tablet formulation using a high performance liquid chromatographic method with UV detection. Separation was achieved on a Hypersil BDS-CN column (150mm×4.0mm, 5μm) using a gradient elution program with solvent A composed of phosphate buffer (pH 4.0; 0.010M) and solvent B of acetonitrile–phosphate buffer (pH 4.0; 0.010M) (90:10, v/v). The flow rate of the mobile phase was 1.0mLmin−1 and the total elution time, including the column re-equilibration, was approximately 20min. The UV detection wavelength was varied appropriately among 210, 250 and 280nm. Injection volume was 20μL and experiments were conducted at ambient temperature. The developed method was validated in terms of system suitability, selectivity, linearity, range, precision, accuracy, limits of detection and quantification for the impurities, short term and long term stability of the analytes in the prepared solutions and robustness, following the ICH guidelines. Therefore, the proposed method was suitable for the simultaneous determination of aniracetam and its studied related impurities.