Abstract: An analytical method for rapid screening of 184 pesticide residues in tobacco was established based on filter solid phase extraction method by gas chromatography-quadrupole time of flight mass spectrometry (GC-QTOF/MS). Single factor examination combined with the Box-Behnken response surface analysis experiments was used to investigate the effects of solvent volume, water volume and purification volume on the recovery of target pesticides. The optimal sample preparation procedure was as follows:the tobacco sample was extracted with 15 mL acetonitrile. After shaking and centrifugation, 1 mL supernatant was cleaned up by Cleanert NANO purification columns. After dried by parallel concentrator, the extract residues were reconstituted with n-hexane-acetone(1/1,V/V) and then detected by GC-QTOF/MS. The linearity of the method was good. At the addition levels of 0.05, 0.25 and 0.5 μg/g, the average recovery rates were 62.05% to 122.19%, 65.18% to 120.21% and 68.55% to 119.41%, except for some pesticides. The relative standard deviation (RSD) were 0.16% to 14.04%, 0.15% to 12.06% and 0.03% to 14.92% (n=3). At the addition levels of 0.05, 0.25 and 0.5 μg/g, the proportion of pesticides detected by the workflow software was 71.74%, 92.39% and 98.91%, respectively. The convenient, rapid and accurate method can be applied for qualitative and quantitative detection of multiple pesticide residues in tobacco.