Multiple inverse isotope dilution assay for the stereospecific determination of R(+)- and S(−)-oxprenolol in biological fluids

Abstract

An isotope dilution assay has been developed for the determination of both oxprenolol enantiomers in biological samples after administration of the racemic 14C-labelled mixture. The enantiomers were reacted with optically pure S()-1-phenylethyl isocyanate and the diastereoisomeric urea derivatives formed were separated by normal-phase high-performance liquid chromatography. Quantitation was performed by on-line ultraviolet detection at 275 nm and off-line radiometry. Endogenous compounds and oxprenolol metabolites did not interfere with the assay. Analysis of water and blood, plasma and urine samples of rats and dogs spiked with [ 14C]oxprenolol hydrochloride showed mean recoveries for R(+)-oxprenolol hydrochloride of 99.2% (water), 99.3% (blood), 99.1% (plasma) and 97.9% (urine), and for S()-oxprenolol hydrochloride of 99.7% (water), 98. 1% (blood), 98.6% (plasma) and 96.9% (urine). In a pilot study, the presented method was used to investigate the metabolic fate of the enantiomers in two dogs dosed orally with racemic [ 14C]oxprenolol hydrochloride (3 mg/kg). The results show that conjugation of R(+)-oxprenolol exceeded that of S()-oxprenolol.