Abstract:Tripamide is a drug widely used in clinical practice for the treatment of hypertension and edema. This work evaluated a screening method for Tripamide and its urinary metabolites in human urine, using high‐performance liquid chromatography diode‐array detection (HPLC/DAD). Identification of these metabolites was investigated by high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI‐MS/MS) after dosing with 15 mg Tripamide. Acid hydrolysis showed that Tripamide is conjugated in the body. Two suspected metabolites were detected by HPLC/DAD. HPLC/ESI‐MS/MS analysis suggested that these metabolites were probably hydroxylated together with loss of the ‐NH2 group and dehydrogenation. These results will be useful in confirmation methods for Tripamide in doping control. Copyright © 2003 John Wiley & Sons, Ltd.