Atropine is an anticholinergic drug from natural plants, and has been widely used in the clinical applications of animals and humans. However, in livestock production, excessive or improper use of atropine will lead to atropine residues in meat. When people eat animal meat from these sources, it will pose a potential threat to human health. Thus, in production practice, atropine residues in meat are usually determined quantitatively. In this study, high performance liquid chromatography (HPLC), a simple but potent method was used to quantify atropine residues in beef. The results indicated that the method could measure atropine residues accurately, with the average recoveries between 82.37 and 88.31 % when the additional concentration ranged from 0.5 to 5.0 mg/kg, and a detection limit of 0.25 mg/kg, providing an effective and reliable method for the detection of atropine residues in beef.