ABSTRACT. A multiclass methodology to determine deoxynivalenol, zearalenone, and 13 widely used fungicides to control the main diseases in barley and wheat crops was optimized and validated. Three QuEChERS methods were compared in terms of recovery and repeatability. The recoveries of the validated method were in the 67-128% range for the selected compounds and relative standard deviations were below 20% according to Document SANTE No. 12682/2019. The limits of quantification (LOQs) were between 10 and 100 μg kg-1. The method was applied for the analysis of barley and wheat grains obtained under production conditions. These samples contained mycotoxins and fungicide residues at concentrations below the LOQs, except for carbendazim in wheat, which exceeded the Codex maximum residue levels. The results of this study provide a fast and simple analytical tool to evaluate the primary production of these grains and to identify possible risks to human health. © 2022 American Chemical Society.