Simultaneous determination of allopurinol and oxipurinol in human plasma and urine by high-performance liquid chromatography.

The uricostatic drug allopurinol (CAS 315-30-0) is used for treatment of hyperuricaemia and is mainly bio-transformed to the active metabolite oxipurinol (CAS 2465-59-0) in humans. A new assay was developed for the simultaneous determination of both compounds in plasma and urine using ultrafiltration and ion exchange purification steps for plasma and urine, respectively. Reversed-phase high-performance liquid chromatography with ultraviolet detection was applied for the separation and quantitation of both compounds. The limit of detection was 0.1 microgram/ml for both compounds in plasma and 0.2 and 0.5 microgram/ml for allopurinol and oxipurinol, respectively, in urine. Within-run and day-to-day precision of 3-5% and 5-7% was determined for plasma and 6-8% and 8-10% for urine analysis. The assays were further validated using liquid chromatography with photodiode array detection and by comparison with methods using protein precipitation as the purifying step. The high analytical recoveries, selectivity, sensitivity, accuracy and reproducibility were adequate for the measurement of both compounds in pharmacokinetic studies and for drug monitoring in patients on allopurinol therapy.