Abstract: Gas chromatography-mass spectrometry (GC/MS) with selective ion monitoring (SIM) mode was applied to study the detection method of metabolites of Tibolone in human urine. According to the rule of metabolism, the experimental data obtained from masculine urine and blank urine of the tested were compared. The results indicated Δ4 double-bond i-someride,3α- hydroxyl group and 3β- hydroxyl group are detected, 6- and 16-hydroxylated metabolites not detected. The excretion curve of time-concentration of 3p-hydroxylated metabolite was investigated, the study showed that Tibolone was absorbed quickly and its concentration reached to peak value after oral for 9 hours. These features can be applied to the establishment of scientific basis for the detection of this kind of doping.