DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR DETERMINATION OF ERTUGLIFLOZIN IMPURITY PROFILE

Abstract

ABSTRACT

Several procedures were conducted to develop and validate an analytical method and impurity profile for new antidiabetic drug. In method development, reverse-phase high-performance liquid chromatography (RP-HPLC) employed a Jasco, Finepack C18 column (250 × 4.6 mm, 5 µm). The selected mobile phase consists of mobile phase A: (0.1% OPA+0.1%TFA) and mobile phase B: Mix acetonitrile and Methanol in the ratio of 40:60 (v/v), both were mixed in the proportion of 85:15, with a flow rate of 1.0 mL/min. Detection occurred at 260 nm. System appropriateness was validated with a tailing factor below 1.5 for principal peaks, theoretical plates exceeding 2000, and relative standard deviations (RSD) under 2.0% for both intra-day and inter-day precision. The validation results indicated an accuracy with recovery percentages ranging from 98.4 to 100.2. Linearity was assessed by linear regression analysis. Linearity was observed within the range of 50-150 µg/mL, with correlation coefficients of 0.999. Additionally, solution stability was confirmed, exhibiting less than a 2.0% variation in peak area over 48 hours at room temperature. The chromatographic analysis showed that Impurity I had a retention time of 5.71 minutes with a purity of 99.54%, while Impurity II exhibited a retention time of 13.47 minutes with a purity of 99.25%. whereas, the chromatographic analysis of Impurity III revealed that it had a retention time of 22.70 minutes, with a purity of 88.74%.