Anabolic androgenic steroids (AASs) are usually illegally added to animal feed because they can significantly promote animal growth and increase carcasses' leanness, which threatens the safety of animal-derived foods and indirectly hazards human health. This study aimed to establish an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous detection of twelve AAS residues in livestock and poultry meat. The homogenized samples were extracted with acetonitrile containing 1% acetic acid ( / ) and purified using the one-step extraction column. After concentration using nitrogen, the residues were redissolved in acetonitrile and then quantified with an external standard method using UHPLC-MS/MS. The results showed that the above-mentioned method had a satisfactory linear correlation ( ≥ 0.9903) with a concentration range of 1-100 μg/L, and the limits of detection (LODs) and quantification (LOQs) were 0.03-0.33 μg/kg and 0.09-0.90 μg/kg, respectively. With the intraday and interday precision less than 15%, the average recoveries of pork, beef, lamb, and chicken, at different spiked levels, ranged from 68.3 to 93.3%, 68.0 to 99.4%, 71.6 to 109.8%, and 70.5 to 97.7%, respectively. Overall, the established method is validated, precise, and capable of the high-throughput determination of the residues of twelve AASs in livestock and poultry meat.