Bioanalytical Method Development and Validation for the Estimation of Raloxifene Hydrochloride in Human Plasma by RP-HPLC Method

Abstract

A rapid, simple, sensitive, and selective HPLC method was developed and validated for the determination of raloxifene hydrochloride (RLF) in human plasma using a liquid-liquid extraction method. Separation of RLF was achieved using a Phenomenex RP-C8 column, with acetonitrile and phosphate buffer (pH 3.5, adjusted with orthophosphoric acid) as the mobile phase in a 40:60 ratio. Detection was carried out at 240 nm. The method was validated for specificity, selectivity, sensitivity, linearity, accuracy, precision, recovery, and stability parameters. The retention times of RLF and amlodipine (Internal standard) were found to be 4.1 and 6.3 min, respectively. Linearity was observed (r > 0.999) in the concentration range of 40-200 ng/ml. The inter-day and intra-day accuracy and precision were found to be within acceptable limits. The percentage recovery ranged from 99.4% to 100.4%.