The limits of detection (LOD) and quantification (LOQ) were estimated as the lowest analyte concentration that produced S/N ¼ 3 and S/N ¼ 10 for the quantification transition, respectively. In addition, S/N ¼ 3 must be verified for the confirmation transition in both cases. Under these criteria LOD was 0.001 mg L1 and LOQ 0.003 mg L1. These low values clearly indicate the good capability of the method to comply with MRL specifications worldwide for AFM1 in milk (0.5 mg L1 for MERCOSUR, United States and China, and 0.05 mg kg1 for European Union). An identification and quantification criteria was established for UHPLC-MS/MS determination. Positive samples were identified and quantified when the following criteria was verified: a) the presence of both quantification (329 > 273) and confirmation (329 > 259) transitions; b) retention time tolerance of ±0.2 min for AFM1 in sample extract compared to the calibration standard and between both transitions; c) S/N 10 for quantification trace (Q) and S/N 3 for confirmation trace (q) at the same time; d) q/Q ratio tolerance of 30% for the sample extract compared to a standard in matrix