A method was evaluated for determination of twenty-one doping (including nandrolone, boldenone and methandienone) in human urine by gas chromatography-high resolution mass spectrometry.  Samples were prepared by liquid-liquid extraction, concentrated, TMS derivatization and limit of detection at ng·mL⁻¹ by MID/GC/HRMS.  According to the code of the World Anti-Doping Agency (WADA), precision and recoveries of the procedure were evaluated by replicate analysis (n = 6), the recoveries in the range of 66%−103%, with the RSD below 10.0%.  The precision within the day of the method with three different concentrations was also determined, RSD were less than 9.5%, 10.0% and 9.7%.