SIMULTANEOUS DETERMINATION OF THE SULFONYLUREA GLIMEPIRIDE AND ITS METABOLITES IN HUMAN SERUM AND URINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AFTER PRECOLUMN DERIVATIZATION

A sensitive and selective high-performance liquid chromatographic method has been developed for a new sulphonylurea, glimepiride, and its metabolites. The assay involves extraction with diethyl ether, thermolysis of the sulphonylureas at 100°C and trapping of the resulting amines with 2,4-dinitrofluorobenzene. The derivatives were quantitated on a reversed-phase column by absorbance at 350 nm using a step gradient for the three compounds in serum and an isocratic run for the metabolites in urine. Analogous compounds were used as internal standards. The detection limit was 5 ng/ml for glimepiride and metabolite II and 10 ng/ml for metabolite I using 1 ml of serum. The method has been applied to the analysis of serum and urine samples from pharmacokinetic studies in humans. © 1990.