ECO-ENGINEERED STABILITY-INDICATING RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF EMPAGLIFLOZIN AND VILDAGLIPTIN WITH AGREE–GAPI VALIDATION

Abstract

Background: A novel and reliable reverse-phase high-performance liquid chromatography (RP-HPLC) method was established and validated to allow for the simultaneous determination of empagliflozin (EMPA) and vildagliptin (VILDA) in combination. Methods: The chromatographic separation was carried out on a C18 column (250 mm × 4.6 mm, 5 µm) using a mobile phase consisting of water (pH 4.5, adjusted with 0.01% orthophosphoric acid) and acetonitrile in a 45:55 (v/v) ratio. Detection was performed at 212 nm, and the retention times for EMPA and VILDA were observed to be approximately 4.6 and 5.4 minutes, respectively. Results: The method demonstrated good linearity for EMPA and VILDA in the ranges of 1–150 ppm and 2–300 ppm, correspondingly, with correlation coefficients (r²) of 0.999 for both analytes. The regression equations for EMPA and VILDA were y = 7688.6x − 215 and y = 5749.7x − 30204, respectively. Precision, expressed as %RSD, was found to be below 2%. The method also exhibited suitable accuracy, with recovery rates between 98.5% and 101.3% for EMPA and 98.7% and 101.2% for VILDA. The detection and quantification limits (LOD and LOQ) were 2.9 ppmand 1.9 ppmfor EMPA, and 8.82 ppmand 5.76 ppmfor VILDA, respectively. The method was robust and resilient to minor variations in conditions. Conclusion: Forced degradation studies revealed that EMPA and VILDA were stable under different stress environments, with no interference from degradation products at their respective retention times. This developed RP HPLC method is suitable for the routine quality control of EMPA and VILDA in combined pharmaceutical preparations, confirming its specificity and reliability. The greenness of the developed method was assessed using AGREE and GAPI tools, yielding a high sustainability score of 0.73.