A method was developed for the rapid and simultaneous determination of 11 penicillins in fishery products by pass-through SPE purification with liquid chromatography tandem mass spectrometry（LC-MS/MS）．The samples were extracted with acetonitrile-water（80∶20，by volume），and then cleaned up with an Oasis PRiME HLB pass through SPE column.The separation of 11 penicillins were performed on a Kinetex XB-C18（2.1 mm×100 mm，2.6 μm） column by using a mixture of 0.05% formic acid and acetonitrile containing 0.05% formic acid as mobile phase.Qualitative and quantitative analyses of the analyte were carried out under the multiple reaction monitoring（MRM） mode with positive electrospray ionization．And the isotope internal standards were employed for the quantification.The calibration curves were linear well in corresponding concentration ranges,with correlation coefficient over 0.99.The detection limits ranged from 0.30 μg/kg to 1.5 μg/kg.The average spiked recoveries for 11 penicillins were between 85.5%and 110% with relative standard deviations（RSDs） of 5.9%-14.3%．The proposed method is accurate，sensitive and high-efficient，and is practical for the determination of penicillins in fishery products．