Abstract: Objective:An analysis method for the simultaneous determination of 44 kinds of pesticide residues in wolfberry was established by high performance liquid chromatography-tandem mass spectrometry with dynamic multi-reaction monitoring. Methods:The samples were rehydrated with 10 mL uitra-pure water,extracted with acetonitrile(containing 1% formic acid),purified up with PSA,C₁₈ and MgSO₄,separated by a T3 column with 0.1% formic acid solution and acetonitrile as the mobile phases in gradient program,analyzed using HPLC-MS/MS in dynamic multiple reaction monitoring(d-MRM)mode and quantified by matrix-matched external standard method. Results:The 44 kinds of pesticides showed good linearity when they were in 0.002~0.200 mg/L(R²>0.9943). The limits of detection of the 44 kinds of pesticides were in 0.01~3.30 μg/kg,the recoveries were within 60.2%~115.3%,RSD were 0.63%~8.27%. The pesticide residues in the samples of wolfberry purchased from the market were detected by this method,a total of 14 kinds of pesticide residues were detected,and the detection rate of carbendazim,chlorphenamidine,imidacloprid were up to 95%. Conclusions:The method was rapid,simple,accurate and reliable. It was suitable for simultaneous detection of 44 kinds of pesticide residues in wolfberry.