DETERMINATION OF DEBRISOQUINE AND 4-HYDROXYDEBRISOQUINE IN URINE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION AFTER SOLID-PHASE EXTRACTION

A simple, selective and sensitive method has been developed to determi ne debrisoquine and 4-hydroxydebrisoquine in human urine. Separation o f the analytes was obtained using a mobile phase of 0.1 M sodium dihyd rogen phosphate-acetonitrile (87:13, v/v) and a muBondapak C18 column. The column effluent was monitored with fluorescence detection at 210 nm (ex) and 290 nm (em). Rapid sample preparation was achieved by soli d-phase extraction columns (Bond Elut CBA, 3 ml capacity) which provid ed excellent recovery values for both compounds. The cost per sample u sing this approach could be minimized by column regeneration and re-us e. The within-day and the day-to-day reproducibilities were less than 7% for both components. The method was shown to be suitable for the st udy of the debrisoquine-sparteine type genetic polymorphism in man.