*2.3. Mycotoxin LC-MS*/*MS Analyses*

Determination of mycotoxins with the LC-MS/MS method in wheat grains and wheat malt was performed as previously described [31]: In brief, 5.00 g of ground wheat was extracted with 20 mL of extraction solvent composed of acetonitrile (AcN):water (W):acetic acid (HAC) = 79:20:1 (*v*:*v*:*v*) on a rotary shaker (GFL 3017, GFL; Burgwedel, Germany) for 90 min at room temperature in a horizontal position. After extraction, 500 μL of the extract was diluted with 500 μL of dilution solvent composed of AcN:W:HAC = 20:79:1 in vials. Finally, 5 μL was injected into an LC-MS/MS system composed of a QTrap 5500 MS/MS (Sciex, Foster City, CA, USA) coupled with an Agilent 1290 series UHPLC system (Agilent Technologies, Waldbronn, Germany). The separation of analytes was performed on a Gemini C18 column (150 × 4.6 mm i.d., 5 μm particle size) with a 4 × 3 mm precolumn with the same characteristics (Phenomenex, Torrance, CA, USA). The eluents used were composed of methanol (MeOH):W:HAC = 10:89:1 (*v*:*v*:*v*) as eluent A, and MeOH:W:HAC = 97:2:1 (*v*:*v*:*v*) as eluent B. The analysis was performed on the fully validated method described in detail by Sulyok et al. (2020) for measurement of 500+ mycotoxins and other secondary metabolites.
