A hydrophilic interaction chromatography-tandem mass spectrometric（HILIC-MS/MS） method was developed for the simultaneous determination of amantadine and ribavirin in animal tissues.After added isotopically labeled standards as the internal standards,the sample was extracted with 20 g/L trichloroacetic acid and enzymed with acid phosphatase,then purified with PBA cartridge.The targeted compounds were separated on a HILIC column（100 mm×2.1 mm,3 μm） by gradient elution with 5 mmol/L ammoniumacetate（containing 0.1% formic acid） and acetonitrile as mobile phases,and detected by MS/MS in positive electrospray ionization and multiple reaction monitoring（MRM） mode.The quantification was performed by the internal standard method.The calibration curves showed good linearity in the range of 0.5-5.0 μg/L.The limits of quantitation for amantadine and ribavirin were 1.0 μg/kg and 2.0 μg/kg,respectively.The recoveries for chicken musle and liver samples at three spiked levels were in the range of 89.2%-109% with relative standard deviations of 2.0%-9.3%.This method was accurate,reliable and simple,and was suitable for the analysis of amantadine and ribavirin in animal tissues.