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The present study developed an improved analytical method for the simultaneous quantification of 17 kinds of Aspergillus, Fusarium and Penicillium mycotoxin contaminants in foods and feeds by ultra-performance liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry (UPLC–MS/MS) under the multiple reaction monitoring (MRM) mode, and especially focused on the optimization of extraction, clean-up, UPLC separation and MS/MS parameters of analytes. Homogenized samples were sequentially extracted by 84% (v/v) of acetonitrile aqueous solution with the selected internal standard (zearalanone) spiking, SPE clean-up with Mycosep 226 Aflazon+ Multifunctional cartridges, filtration, concentration and secondary filtration. Using double sample injection method, the analytes were separated by UPLC BEH C₁₈ column (100mm×2.1mm I.D., 1.7μm), and eluted …