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A method based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the determination of 15 water-holding functional drugs in animal tissues.The analytes were extracted with acetonitrile containing 1.0%(v/v) methanol, purified by Oasis PRiME HLB SPE column, and analyzed by Acquity UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) using methanol and 0.1%(v/v) formic acid aqueous solution as the mobile phases.The analytes were detected using an electrospray ionization (ESI) source under the MRM mode.The calibration curves of the analytes were linear in the range of 1.0-50.0 μg/kg ( r ≥ 0.9949), and the limits of quantification were all less than 1.0 μg/kg in animal tissues.The recoveries of the 15 water-holding functional drugs ranged from 60.0%-111.0% in animal tissue samples, with the intra and inter RSDs of 0.56%-11.5% and 2.31%-14.8%, respectively.The method can meet the requirements for the determination of the drug residues in animal tissues.It provides a new idea to identify potential hazards in animal-derived foods and to monitor illegal addition.