Development and Validation of a Stability-Indicating Liquid Chromatographic Method with Mass-Spectrometric Detection for the Analysis of Apixaban and Determination of Production Related Impurities

摘要

Apixaban is a novel anticoagulant drug acting as a direct, selective and reversible inhibitor of coagulation factor Xa. The purpose of this work was to develop and validate a sensitive, stability-indicating LC-MS method for the analysis of apixaban and estimation of production-related substances in the active pharmaceutical ingredient (API) and pharmaceutical dosage forms. The analyses were carried out on a Macherey-Nagel Nucleodur C18 column (250 x 4.6 mm, 5 m particle size) with a mobile phase containing acetonitrile and water in gradient program at a flow rate of 0.8 mL/min, the column oven temperature 25 degrees C, and the UV detection tuned to 270 nm. Apixaban was subjected to stress conditions of oxidative, acid, base, hydrolytic, thermal and photolytic degradation, and found susceptible to acid and base hydrolysis conditions, while it was stable under thermal, photolytic, and oxidative degradation conditions. The degradation products were well resolved from the main peak and its impurities, proving the stability-indicating power of the proposed method. The proposed method was validated as per International Conference on Harmonization (ICH) guidelines. The method is suitable for practical routine analysis and it has been successfully applied to analyze apixaban in eliquis brand preparations. All analytical results were acceptable according to the pharmaceutical requirements.