The present work developed an analytical method for simultaneous determination of fumonisins B1, B2 and B3 residues in maize by ultra-high-performance liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry (UHPLC–MS/MS) under the multiple reaction monitoring (MRM) mode, and especially focused on the optimization of extraction, clean-up, UHPLC separation and MS/MS parameters.