Green and Sustainable Analytical Chemistry-Driven Chromatographic Method Development for Stability Study of Apixaban Using Box–Behnken Design and Principal Component Analysis.

This article focuses on the development of a green and sustainable stability-indicating high-performance thin-layer chromatography (HPTLC) method for the estimation of apixaban (APX), a USFDA-approved anticoagulant drug. The method was designed using the Analytical Quality by Design (AQbD) approach, incorporating chemometric tools such as principal component analysis (PCA) and design of experiments (DoE) to optimize critical method variables while minimizing the use of toxic organic solvents. Unlike previously published chromatographic methods that rely on hazardous solvents like acetonitrile and methanol, this method employs safer class 3 solvents—n-butanol, ethanol, and glacial acetic acid—aligned with International Council for Harmonization (ICH) guidelines, thereby enhancing analyst safety and environmental protection. Validation studies confirmed the method’s specificity, accuracy, precision, robustness, and sensitivity, and forced degradation tests demonstrated its stability-indicating capability. The developed method showed a superior greenness profile compared to existing methods, making it a more eco-friendly and economical option for quality control and stability studies of apixaban pharmaceutical formulations.