Bazedoxifene acetate quantification in rat serum with the aid of RP-HPLC: Method development and validation

Abstract

Development of simple, sensitive, accurate and reproducible RP-HPLC method for Bazedoxifene acetate estimation in rat serum using ratio of peak area of analyte to internal standard (Raloxifene) for a pharmacokinetic study is the objective of the study. The procedure involves simple  liquid-liquid extraction of BAZ and IS from rat serum directly into acetonitrile which is injected onto a Hypersil BDS C₈ reverse phase stainless steel analytical column (4.6 × 150mm, 5µm) at 30⁰C and detected using a PDA detector set at 290nm. Mobile phase consisting of buffer (Potassium dihydrogen orthophosphate) and acetonitrile (60:40, v/v) was used at a flow rate of 1.0mL/min. Retention times of BAZ and IS were 5.852 and 4.052 min respectively. The standard curve for BAZ was linear (r² >0.99) in the concentration range of 0.1 - 32μg/mL.  Absolute recoveries of BAZ and IS were 90 - 110% and >95% respectively from rat serum. The lower limit of detection and quantitation of BAZ was 0.25μg/mL and 0.5μg/mL. The intra, inter–day precisions and accuracy in terms of % error were all found within the acceptable range. Stability studies results indicated that both analyte and IS were stable on bench top, autosampler and freeze-thaw cycles.