Objective An isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the determination of ribavirin and adamantanes (amantadine, adamantanemethylamine, rimantadine, memantine) in poultry food. Methods After enzymatic hydrolysis and precipitated by trichloroacetic acid, the samples were centrifuged by high-speed centrifuge at low temperature, and purified by PBA/PCX composite solid-phase extraction cartridge. The separation of ribavirin was performed on UPLC system with Agilent ZORBAX SB-Aq(3.0 mm×100 mm,1.8 μm), adamantanes with Waters BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) and determinated by tandem mass spectrometry. The method was quantified by internal standard. Results The calibration curves showed a good linearity between the peak ratio and the concentrations of 1.0-100 ng/mL(ribavirin) and 0.2-20 ng/mL(adamantanes) with r=0.999. The limit of detection of ribavirin was 0.5 and 0.1 μg/kg for adamantanes. The limit of quantification of ribavirin was 1.5 and 0.3 μg/kg for adamantanes. The mean recoveries of the ribavirin ranged from 91.4%-103.7% and ranged from 94.3%-108.2% for adamantanes spiked at three concentration levels(1.5-15 and 0.3-3.0 μg/kg), with the relative standard deviations (RSD) less than 10%. Conclusion This method was simple, high sensitivity and accuracy.