Identification and Validation of Genotoxic Impurity in Ezetimibe by Reverse Phase High-Performance Liquid Chromatography

Abstract

The method emphasizes identification and validation of potential Genotoxic impurity in pharmaceutical drug substances of Ezetimibe by Reverse Phase High-Performance Liquid Chromatography (HPLC). The impurity was separated by using the Zorbax Rx Octylsilane (C8) HPLC column with 250 cm length and internal diameter of 4.6 mm with pore size 5 μm. The partition of impurity was operated at a significant pH 3.0 was maintained by buffer of 10% potassium dihydrogen phosphate and Acetonitrile with 80:20 ratio and the mobile phase is Acetonitrile with a gradient inflow of 1.5 mL/min. The UV absorption maximum were observed at 258 nm. The proposed approach shows the results of linear boundaries in between 0.16 μg/g to 7.5 μg/g with correlation coefficient is lower than 0.999. The method was further evident by accuracy results are in the region of 98.82% to101.04% for Genotoxic impurity of (5R, 6S)-1-(4-fluorophenyl)-5-((S)-3-(4-fluorophenyl)-3-hydroxypropyl)-3-(2-hydroxy-1-phenylethyl)-6-(4hydroxyphenyl)di-hydropyrimidine-2,4(1H,3H)-dione. The approach was shown acceptable results as per International Council of Harmonisation (ICH) guidelines and the method was operated even at lower concentrations.