Development, Validation and Pharmacokinetic Application of a Simple and Robust RP-HPLC Method for Quantitation of Raloxifene in Rat Plasma

SUMMARY. A simple and sensitive HPLC method has been developed and validated for the quantification
of raloxifene in rat plasma. Liquid- liquid extraction procedure was employed for extracting raloxifene
from rat plasma sample. Separation of raloxifene was achieved through a RP- C18 column with a mobile
phase consisted of phosphate buffer and acetonitrile (66:34 %, v/v). The method was validated for specificity,
selectivity, sensitivity, linearity, accuracy, precision, recovery and stability parameters. A linear response
(R2 value 0.9991) was found over the calibration range of 50 to 500 ng/mL. The accuracy for intra
and inter day run varied between 86.73 to 102.30 % and 91.11 to 95.56%, respectively. The corresponding
precision (% CV) were within 0.82 to 9.43% and 6.23 to 8.33%. The method was also found to be specific
and stable. The applicability of the method was established through a single dose oral pharmacokinetic
study of raloxifene in rat.