Monitoring of Aflatoxin M1 in Various Origins Greek Milk Samples Using Liquid Chromatography Tandem Mass Spectrometry

Abstract

Aflatoxin M1(AFM1), a major metabolite of Aflatoxin B1(AFB1), has been identified as a potential contaminant in dairy products. Because of its possible carcinogenicity, the legislation limits as set by Commission Regulation (EC) No. 1881/2006 are very strict, namely 0.050 μg kg−1 in milk and 0.025 μg kg−1 in infant formulas. To meet these requirements, a sensitive and accurate method was developed, employing liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Ιmmunoaffinity columns (R-Biopharm) were used for sample purification and preconcentration of the analyte of interest. The quantification of AFM1 was conducted using fortified milk samples, while Aflatoxin B2 (AFB2) was used as an internal standard (IS). The method was validated in terms of linearity, precision, trueness, limits of detection and quantification and uncertainty. The performance criteria for the method were evaluated based on European Commission Regulation (EC) No. 401/2006 and its most recent amendment, as well as the suggested criteria for revision by the EU Reference Laboratory for Mycotoxins and Plant Toxins. The recovery was in the range of 77.9–81.0% for all fortification levels (0.025–0.050–0.075 μg kg−1), with RSDR values (Relative Standard Deviation of intermediate precision) ranging from 6.1% to 12%. The method’s detection and quantification limits were 0.0027 μg kg−1 and 0.0089 μg kg−1, respectively. The occurrence of AFM1 was investigated in 40 samples of different animal origin (cow, goat and sheep milk) provided by Greek producers.

Occurrence of Aflatoxin M1 and Risk Assessment in Bovine Milk from the Amazon Region

“…Other authors have identified the occurrence of AFM1 in bovine milk in countries such as Nigeria, United Arab Emirates, Mexico and in derived products in Greece, China and West Asian countries (Mukhtar et al, 2022;Mohamadin et al, 2022;Álvarez-Dí as, 2022;Panara et al, 2022;Xiong et al, 2022). However, the lack of investigations into the occurrence of AFM1 in bovine milk in northern Brazil makes it difficult to monitor contamination in the state of Amazonas, despite reports of the presence of the mycotoxin in other Brazilian regions.…”

Incidence and Levels of Aflatoxin M1 in Artisanal and Manufactured Cheese in Pernambuco State, Brazil

“…Given the potential harm of AFM 1 , Brazil has adopted regulatory limits for this mycotoxin, with maximum permissible limits (MPL) of 0.5 µg/L for fluid milk and 2.5 µg/kg for cheese [ 15 ]. The occurrence and levels of AFM 1 in milk products have been monitored [ 14 ] using several analytical methods, comprising the popular enzyme-linked immunosorbent assay (ELISA) [ 16 ], high-performance liquid chromatography (HPLC) alone [ 17 ], and HPLC combined with tandem mass spectrometry (MS/MS) [ 18 ]. In this context, monitoring AFM 1 concentrations in milk and its derived products is an essential aspect of food safety efforts, aimed at protecting consumers’ health protection [ 19 ].…”

Simultaneous Detection and Quantification of Aflatoxin M1, Eight Microcystin Congeners and Nodularin in Dairy Milk by LC-MS/MS

“…Reliable, accurate, and sufficiently sensitive methods are required to detect and quantify cyano-and myco-toxins. A wide range of extraction and analytical methods have been developed [19,[27][28][29][30][31]. The first part of the sample treatment is the extraction.…”