Multi-mycotoxin biomonitoring in Italian adults: Revealing the connection between diet habits and mycotoxin exposure

This study investigated the relationship between food consumption patterns and mycotoxin exposure. We analyzed 300 spot urine samples from Italian adults for 24 mycotoxins and their metabolites using enzymatic hydrolysis followed by ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-ESI-MS/MS). Target mycotoxins were aflatoxins (AFs), fumonisins B1 and B2 (FB1, FB2), alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), ochratoxin A (OTA), ochratoxin-alpha (OT-alpha), zearalenone (ZEN), zearalanone (ZAN), α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), T-2/HT-2 toxin, deoxynivalenol (DON), deepoxy-deoxynivalenol (DOM-1), enniatins B, B1, A, A1 (ENNs), and beauvericin (BEA). Participants were scored according to three dietary patterns: the Mediterranean Diet Score, the Plant-based Diet Index, and the general Pro-vegetarian Food Pattern. For each index, low- and high-adherence groups were defined based on score distribution. Emerging mycotoxins, such as Alternaria toxins, ENNs, and BEA showed the highest prevalence (>90%), while DON (34%), OTA (33%), and ZEN (25%) had lower prevalence, with DON showing the highest mean concentration (7.10 μg/g creatinine). Cereals, nuts, legumes, fruits, and vegetables were significantly associated with increased mycotoxin exposure, especially AFs, OTA, ENNs, and DON. Estimated daily intakes for DON were negligible, whereas some individuals classified as high adherers to one or more of these dietary patterns approached or exceeded the safe threshold for ZEN. Exposure to OTA represented a potential health risk across all individuals. These findings support periodic updating of EU maximum-level frameworks for contaminants in food and related exposure and risk assessments so that increasingly consumed plant-based raw materials and composite foods are better represented in monitoring and risk characterization. Such updates would support a safe and effective transition toward sustainable diets in line with public health goals.