Abstract: An analytical method for simultaneous determination of 131 pesticide residues in tobacco was established by QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry ( UPLC-MS/MS). Tobacco samples were extracted with 1%(V/V) acetic acid-acetonitrile, salted out with sodium chloride and anhydrous magnesium sulfate, cleaned up with primary secondary amine PSA solide-phase dispersion, the extracts were then determined by UPLC-MS/MS in positive and negative ion modes and multiple reaction monitoring (MRM) mode, quantified by the external standard method. The results showed that all the 131 pescitides had good linearity in the ranges of 1-200 μg/L, with the coefficients of determination (r²) higher than 0.99. The limits of detection (LOD) were from 0.05 to 24.50 μg/kg. The average recoveries of all the pesticides were in the range of 65.3%-112.4% with the relative standard deviations between 3.2% to 7.3%. The method is simple, rapid, sensitive and reliable for determination of the 131 pesticide residues in tobacco.