tBoldenone (BOLD), an anabolic steroid, is likely to be abused in livestock breeding and in sports. Althoughsome of BOLD metabolites in human urine, such as 5-adrost-1-en-17-ol-3-one (BM1), have beendetected, investigations on their excretion patterns for both genders are insufficient. Moreover, littleresearch on 17-BOLD glucuronide as a metabolite in human urine has been reported. The aim of thisstudy is to make a contribution to the knowledge of 17-BOLD metabolism in humans. Three male andthree female volunteers were orally administrated with 30 mg 17-BOLD. Urine samples were collectedand analyzed with gas chromatography–tandem mass spectrometry. The data proved that 17-BOLD,BM1, and 17-BOLD were excreted in urine in both free and glucuronic conjugated forms after admin-istration of 17-BOLD. For most subjects, the urinary concentrations of BM1 were higher than that of17-BOLD. 17-BOLD was excreted in small amounts. 17-BOLD, 17-BOLD, and BM1 were presentnaturally in urine with low concentrations. Administration of 30 mg 17-BOLD could not influence theexcretion profiles of urinary androsterone, etiocholanolone, and testosterone/epitestosterone ratio. Therewere no differences in BOLD metabolic patterns between man and woman.