RP-HPLC Method Validation for Estimation of related Substances of Empagliflozin

In this study, researchers aimed to develop and validate a reproducible and precise Reverse Phase High-Performance Liquid Chromatography technique for accurately estimating a chemical related to empagliflozin. The primary objective was to establish a method suitable for quality control of empagliflozin batches and its impurities. To achieve this, effective chromatographic separation has done by using two mobile phases: Mobile phase A, which comprised water adjusted to pH 3.9±0.5 using glacial acetic acid, and Mobile phase B, consisting of pure acetonitrile. The separation was conducted on a Phenomenex (Kinetex) C18 stationary phase (250mm × 4.6mm, particle size 5μm) at a flow rate of 1.5mL/min. Detection occurred at 240nm at 50°C. An injection volume of 50μL was used for analysis. Empagliflozin was quantified at a concentration of 0.03g/mL, while impurities were measured between 0.052-0.090g/mL. The results indicated successful chromatographic separation and accurate quantification. The proposed method demonstrated its efficacy for quality monitoring of bulk samples containing Empagliflozin, ensuring the reliability and consistency necessary for routine quality control purposes in the pharmaceutical industry.