A method had been developed for determination of residues of 10 anabolic steroids (ASs) in animal muscle tissues by liquid chromatography tandem mass spectrometry (LC/MS/MS). After enzymolysis, the sample was extracted with tert-butyl methyl ether, cleaned up through reverse solid-phase extraction and further determined by LC/MS/MS under multiple reaction monitoring (MRM) mode. The limits of detection (LOD) of LC/MS/MS method used for testing epitestosterone (ETS), nandrolone (17 beta-NT), 17 alpha-methyl-testosterone (MTS), testosterone 17-propionate (PTS), medroxyprogesterone (MED), progesterone (PG), estrone (ESN), 17 beta-estradiol (17 beta-ES), 17alpha-ethynylestradiol (EES) and estriol (EST) in animal muscle ranged from 0.06 to 0.22 microg/kg, and the limits of quantification (LOQ) were from 0.12 to 0.54 microg/kg. Experiments on spiked samples of pork, beef, chicken and fish showed that at addition level of 1.0 microg/kg, the average recoveries of the ASs ranged from 64% to 77%, and coefficients of variation from 7.1% to 20.3%, while at addition level of 2.0 microg/kg, the average recoveries ranged from 70% to 89%, and coefficient of variation from 7.1% to 19.1%.