Over the course of the project, extensive progress has been made in characterising human exposure to mycotoxins, their metabolism, and their potential health impacts through a combination of analytical, experimental, and epidemiological approaches. Biological samples, including urine, blood, and faeces, were successfully collected from volunteers and control subjects. These samples were analysed using UHPLC-MS/MS, followed by comprehensive data processing. This work enabled the development of detailed toxicokinetic profiles and models for each investigated mycotoxin, providing key insights into their absorption, distribution, metabolism, and excretion in humans.
In parallel, a wide range of in vitro incubation studies was conducted for the targeted mycotoxins. Both in vitro-derived samples and in vivo samples from the human studies were analysed using UHPLC-HRMS. The integration of these datasets allowed for the systematic identification and structural characterisation of a broad spectrum of mycotoxin metabolites. This resulted in a comprehensive and well-curated metabolite dataset that significantly advances current knowledge on mycotoxin biotransformation.
To support exposure assessment at the population level, an EPIC end-user database was established for dietary mycotoxin exposure calculations. This resource enabled large-scale epidemiological analyses within the full EPIC cohort, leading to the investigation of associations between mycotoxin exposure and the risk of colorectal, kidney, and hepatocellular cancers. These analyses provide important evidence linking exposure biomarkers to disease outcomes.
All necessary ethical approvals were obtained to conduct three complementary sub-studies, and biological samples were successfully transferred and processed. Multi-mycotoxin biomarker profiles were measured in case-control settings across EPIC, Malawian serum, and Groningen plasma cohorts using UHPLC-MS/MS. Subsequent data processing and statistical analyses enabled the identification of exposure patterns and their associations with disease risk across geographically and demographically diverse populations.
Mechanistic investigations further strengthened the project’s findings. In vivo analyses of ochratoxin A (OTA) were completed, including the identification of a putative mutational signature. Additionally, in vitro studies for OTA, fumonisin B1 (FB1), deoxynivalenol (DON), patulin (PAT), and citrinin (CIT) were finalised, providing mechanistic insight into their toxicological effects and supporting the interpretation of epidemiological associations.
The project has generated a unique and comprehensive body of knowledge on mycotoxin exposure, metabolism, and health effects. The analytical methodologies, toxicokinetic models, and curated metabolite datasets developed during the project represent valuable tools for the scientific community and for regulatory risk assessment.
