The Aflatoxin M1 Content of Cow Milk in Cyprus as Affected by Season and Year of Production: A Five-Year Survey

Aflatoxin M1 (AFM1) is of primary importance to the food industry, state inspection authorities and consumers alike due to the carcinogenic nature of this toxin and the respective health risk associated with its presence in dairy products. In the present work, 1197 raw cow milk samples were collected and analyzed for AFM1 in the Republic of Cyprus during a five-year period (2021–2025) using the enzyme-linked immunosorbent assay (ELISA), confirmed by liquid chromatography-mass spectrometry (UPLC-MS/MS). Health exposure assessment and risk characterization were also performed for the Cypriot population (men, women and children) regarding milk consumption through the calculation of Estimated Daily Intake (EDI), Hazard Index (HI) and Margin of Exposure (MoE). Results showed that AFM1 in cow milk varied with season: 7.34 ± 8.59 ng/kg for winter, 6.86 ± 10.00 ng/kg for spring, 6.43 ± 7.44 ng/kg for summer and 5.36 ± 7.11 ng/kg for autumn. Among the analyzed years, 2022 and 2025 showed wider concentration ranges, with several samples presenting elevated AFM1 levels compared to the other years. Of the 1197 milk samples analyzed for AFM1 content, 633 were <LOD, 284 < LOQ, 278 at concentrations of 10–50 ng/kg, and only two samples exceeded the limit set by the EU (50 ng/kg). The average amount of AFM1 exposure EDI ranged between 0.026 ng/kg b.w./day for men and 0.061 ng/kg b.w./day for children. HI was <1, recording values of 0.130 for men, 0.155 for women and 0.305 for children. Finally, MoE recorded values > 10,000 (15,385 for men and 12,903 for women) and 6557 for children, indicating that the amount of AFM1 consumed through milk by children may comprise a considerable risk for this population group and consequently AFM1 contamination of milk demands its regular monitoring and evaluation of the respective risk involved.