HPLC on Chiralcel OJ-R for Enantiomer Separation and Analysis of Ketoprofen, from Horse Plasma, as the 9-Aminophenanthrene Derivative

Abstract

Abstract Racemic ketoprofen is a non-steroidal anti-inflammatory drug used to treat musculoskeletal and colic conditions in horses. The enantioselective chiral inversion of ketoprofen administered to horses has been studied by use of cellulose tris(4-methylbenzoate), also known as Chiralcel OJ-R, as chiral stationary phase; acetonitrile − 0·02 m perchlorate buffer (pH 2·0) - methanol, 60:15:25 (v/v/v) was used as mobile phase. Before chromatography, to effect adequate chiral interaction with the chiral stationary phase ketoprofen was derivatized with 9-aminophenanthrene, under acid conditions, after solid-phase (C18) extraction and then liquid–liquid extraction, to ensure effective removal of endogenous plasma materials. The 9-aminophenanthrene derivative of S-ibuprofen was used as internal standard. The enantiomers of ketoprofen were separated to baseline (Rs = 6·44, α = 1·76) within a short analysis time. The results indicate that the bio-inversion of R-ketoprofen to the S isomer is significant in equine species. However, considerable differences in pharmacokinetic parameters were observed, indicating large inter-animal variation.