Validation of a multi-mycotoxin detection method for plant-based meat alternatives using UHPLC-TOF-MS and UHPLC-QTRAP-MS/MS and its application to commercialized plant-based burger samples

The increasing global demand for sustainable and nutritious food sources has led to the rapid growth of plant-based meat alternatives (PBMAs) as eco-friendlier replacements of animal-based products. However, the widespread use of cereals, pulses, and nuts in PBMA production raises concerns about mycotoxin contamination. This study validates an analytical workflow for the extraction and determination of 20 mycotoxins in PBMAs, combining an optimized QuEChERS-based extraction method with low- and high resolution mass spectrometry techniques: UHPLC-TOF-MS and UHPLC-QTRAP-MS/MS, respectively. The method was evaluated according to the performance criteria set by the Commission Implementing Regulation (EU) 2023/2782, covering precision, recovery and limits of quantification (LOQ). The results demonstrated high sensitivity for both techniques, with LOQ values below the regulatory limits, ensuring reliable detection of mycotoxins in complex food matrices. The validated method was applied to 12 PBMA burger samples collected from supermarkets in Spain and Portugal. Several mycotoxins, including aflatoxins (B₁ and B₂), moniliformin, and tentoxin, were detected, ranging 0.3-103 ng g^-1. In conclusion, these results highlight the importance of continuous mycotoxin monitoring in PBMAs.