Abstract: Objective:A method for simultaneous determination of 388 kinds of pesticide residues in grapes was established. Method:The tested sample was extracted with 1% glacial acetic acid acetonitrile. A mixture of anhydrous magnesium sulfate and anhydrous sodium acetate was added,and then purified with 300 mg silica gel,300 mg C₁₈,600 mg N-propyl ethylenediamine(PSA),and 900 mg anhydrous magnesium sulfate. The supernatant was concentrated under nitrogen. The analytes were dissolved in acetonitrile-0.1 mol/L ammonium acetate solution(1:1)and analyzed by UPLC-MS/MS with positive ion mode of electrospray ionization(ESI)under multiple reaction mode(MRM). Matrix matching internal standard method was used for the quantitative analysis. Results:The linearity was satisfied with the correlation coefficients more than 0.99. The recoveries of three spiked levels were 60%~120% with the standard deviations(RSD)less than 20%(n=7). The limits of detection(LOD)were 0.005 mg/kg. Conclusion:This sensitive,reliable and accurate method can be applied for the determination of 388 kinds of pesticide residues in grape and is suitable for the multi-pesticide residues in the routine analysis.