ANALYTICAL QUALITY BY DESIGN-ASSISTED DEVELOPMENT AND VALIDATION OF A RAPID STABILITY-INDICATING GRADIENT ULTRA-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS ESTIMATION OF EMPAGLIFLOZIN, SACUBITRIL, AND VALSARTAN IN PHARMACEUTICAL DOSAGE FORMS

Abstract

Objectives: Simultaneous estimation of empagliflozin, sacubitril, and valsartan in combined pharmaceutical formulations is analytically challenging due to their differing polarity, physicochemical properties, and chromatographic retention behavior. Conventional high-performance liquid chromatography methods often require longer analysis times and greater solvent consumption. To develop, optimize, and validate a rapid, robust, and stability-indicating gradient ultra-performance liquid chromatography (UPLC) method for the simultaneous determination of empagliflozin, sacubitril, and valsartan using the analytical quality by design (AQbD) approach. Methods: Separation was performed on an ACQUITY UPLC BEH C18 column using a gradient of 0.1% formic acid in water and acetonitrile. A Box– Behnken design was employed to optimize critical method parameters. The method was validated according to ICH Q2 guidelines for specificity, linearity, precision, accuracy, robustness, limit of detection, limit of quantification, and forced degradation. Results: Complete separation was achieved within 6 min, with retention times of 1.42, 2.67, and 4.26 min for empagliflozin, sacubitril, and valsartan, respectively. The method exhibited excellent linearity (R2 > 0.999), recoveries of 99.10–100.96%, and precision with percentage relative standard deviation <2%. Forced degradation studies confirmed its stability-indicating capability through effective separation of analytes from degradation products. Conclusion: The developed AQbD-based gradient UPLC method is rapid, precise, accurate, robust, and suitable for routine quality control and stability studies of pharmaceutical formulations containing empagliflozin, sacubitril, and valsartan.