Stereospecific high-performance liquid chromatographic determination of an S(-)-benzopyran methyl ester derivative (CGP 50 068), its (-)-carboxylic acid metabolite (CGP 55 461) and the related (+)-enantiomer (CGP 54 228) in human and dog plasma.

The simultaneous determination of CGP 50 068, S(-)-enantiomer (I), its (-)-carboxylic acid metabolite CGP 55 461 (II) and the related (+)-enantiomer CGP 54 228 (III) by stereospecific high-performance liquid chromatography, in human plasma, is described. The three compounds and racemic acebutolol, used as internal standard, were isolated from plasma by liquid-solid extraction on disposable C18 columns. The resolution and determination of I and the two carboxylic acid enantiomers were achieved by direct chromatography using a Chiral-AGP column refrigerated at 5 degrees C. The mobile phase was tetrabutylammonium iodide in a pH 7 phosphate buffer solution used at a constant flow-rate of 0.5 ml/min. The UV detection wavelength was set at 270 nm. The reproducibility and accuracy of the method were found to be suitable over the concentration range 0.56-28.0 mumol/l for II and III and 2.0-26.7 mumol/l for I.