Development and Validation of Chiral HPLC Method for Identification and Quantification of (R)-Enantiomer in Ezetimibe

Abstract

A simple and sensitive normal phase isocratic high performance liq. chromatog. method was developed for the detn. of trace level enantiomer in posazonazole drug substance. The sepn. was achieved using chiral pack, IC, 250 x 4.6 mm, 5 μm using mobile phase contg. iso-Pr alc., dichloro methane and di-Et amine (50:50:0.1 vol./vol./v). The flow rate was 0.6 mL/min and column temp. was 25 °C. Enantiomer was detected using UV detector at the wave length 262 nm. The retention time was 18.5 min for enantiomer and 28 min is for posaconazole. The optimized method was validated to prove its performance characteristic by demonstrating selectivity, sensitivity (Limit of detection and Limit of quantification), linearity, precision and accuracy. The limit of detection and limit of quantification of ennatiomer was found to be 0.06 μg/mL and 0.2 μg/mL for 10 μL injection vol. The linearity of the method is found to be 0.2 μg/mL to 1.5 μg/mL. Relative std. deviation of system precision, method precision is found to be 0.3% and 0.44% resp. The accuracy of the method is found to be 96.0% to 98.1%. [on SciFinder(R)]