Stereospecific determination of an HIV aspartyl protease inhibitor, PNU-103017, in rat, dog and human plasma using a Pirkle-concept high-performance liquid chromatographic column

A sensitive stereospecific high-performance liquid chromatographic assay for the quantitation of the enantiomers of 4-cyano-N-(3-(cyclopropyl-(5,6,7,8,9,10-hexahydro-4-hydroxy-2-oxo-2H- cycloocta(b)pyran-3-yl)methyl)phenyl)benzenesulfonamide (PNU-103017) (I), an HIV Protease Inhibitor, in plasma of rat, dog and human was developed. The procedure involved an acetonitrile-aided protein precipitation followed by solid-phase extraction (SPE) of I from plasma into ethanol. Stereospecific separation was accomplished on a Pirkle-concept chiral column (Regis S,S-Whelk-01, 250x4.6 mm I.D.) with a mobile phase of absolute ethanol-0.1% acetic acid in hexane (30:70, v/v). The eluate was monitored by UV absorbance (295 nm). Linear calibration curves were obtained in the range of 0.2 to 500 microM, with a lower limit of quantitation of 0.1-0.2 microM for both enantiomers in either rat, dog or human plasma. Intra- and inter-assay precision and assay accuracy were demonstrated to be acceptable for the stereoselective pharmacokinetic analysis of I in plasma.