Abstract: A multi-residue method based on ultra performance liquid chromatography tandem mass spectrometry was developed for the determination of ten anticholinergic drugs residues in animal tissues. In this method,analytes were extracted by acidified acetonitrile,and purified by Oasis PRiME HLB SPE column. Ten anticholinergic drugs were analyzed by Acquity UPLC BEH C₁₈ column(50 mm×2.1 mm,1.7 μm)using methanol and 0.1% formic acid as the mobile phases. The signals were acquired through ESI+and MRM mode. All analytes could be well-separated by a gradient program during 10.5 minutes. The calibration curves of ten anticholinergic drugs were linear in a concentration range of 1.0~50.0 μg/L(r>0.9951),and the limits of quantitation were all less than 1.0μg/kg in animal tissues. The recoveries of 1.0,2.0,10 μg/kg fortified pork samples ranged from 64.7%~108.2% with the intra-assay coefficient of variation was 1.09%~10.5%,and the inter-assay coefficient of variation was 2.31%~11.4%. The recoveries fortified liversamples ranged from 62.6%~108.0% with the intra-assay coefficient of variation was 0.38%~10.7%,and the inter-assay coefficient of variation was 2.24%~11.2%.The method could meet the requirements of the drug residue detection in animal tissues,which played a certain technical support role in effectively preventing drug abuse,illegally adding behavior and combating water injection.