Objective To develop a method for the determination of nosiheptide residue in food animal tissues （porcine muscle， porcine liver， porcine kidney， chicken muscle， chicken liver and chicken kidney） with high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）.Methods After extracted by 0.1% acidic acetonitrile， the nosiheptide extract was cleaned up by Captiva ND cartridge and determined by HPLC-MS/MS in negative electrospray ionization mode. Quantification was performed using corresponding matrix-matched calibration curves.Results Nosiheptide in corresponding matrix showed good linear relationships （r>0.99） in the calibration range of 2~200 μg/L， and the limit of detection （LOD） and limit of quantification （LOQ） of the method were 2.0 and 7.0 μg/kg， respectively. The recoveries of the method ranged from 76.27% to 92.31% at the spiking level of 3.5， 7.0 and 70 μg/kg with relative standard deviations of 2.15%~8.03%.Conclusion The proposed method is simple， rapid， accurate as well as reproducible and can be applied to monitor and determine nosiheptide residue in food animal tissues.