Benedikt Cramer

Dietary exposure to mycotoxins—including aflatoxins (AFs), ochratoxin A (OTA), and citrinin (CIT)—represents an important health risk, particularly in low-resource settings with widespread food contamination. This study investigates mycotoxin exposure among 719 participants (433 women and 286 children) of a cluster-randomized trial in rural Bangladesh, using residual blood samples collected during the 2019 endline survey. A total of 712 whole blood and 578 serum samples were analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Serum aflatoxin B1 (AFB ₁ )-lysine adducts were quantified following enzymatic digestion and solid-phase extraction. To evaluate potential health risks, we estimated probable daily intake (PDI) from blood concentrations of frequently detected mycotoxins. All whole blood samples contained at least two mycotoxins, and 84% of participants were co-exposed to at least three mycotoxins. Biomarkers for the main blood-relevant mycotoxins were frequently detected, with OTA detected in 100% of whole blood samples and its isomer, 2′R-ochratoxin A (2′R-OTA), detected in 98%, CIT in 91%, and Enniatin B (EnB) in 92%, while AFB ₁ -lysine—a biomarker of chronic exposure to the potent carcinogen AFB ₁ —was detected in nearly one-quarter of women and 4% of children with serum samples. Beauvericin was detected in 6% of samples. The PDI of OTA exceeded health-based guidance values in 80% of women and 99% of children, while the PDI of CIT was of concern for 12% of women and 27% of children. These findings demonstrate widespread co-exposure to multiple mycotoxins, with OTA, CIT, and AFB ₁ posing notable risks, underscoring the urgent need for targeted interventions to protect vulnerable populations.

Deoxynivalenol (DON) is a prevalent trichothecene mycotoxin in cereals that poses food and feed safety risks while causing important economic losses. Microbial biotransformation offers a selective, mild strategy for DON detoxification. Here, we screened aerobic soil-derived bacterial communities from diverse agricultural environments, using DON as the sole carbon source for this mycotoxin depletion. More than half of the tested enrichment samples showed a reduced DON signal, as observed by HPLC-UV. To assess the biological relevance, culture extracts were tested for cytotoxicity in HepG2 cells. Z13, a soil sample that depleted DON but produced no other detectable metabolites, showed reduced cytotoxicity, comparable to the negative control. In contrast, samples that depleted DON but produced 3-keto-DON remained toxic. High-resolution LC-MS analysis indicated the formation of metabolites putatively identified as 3-keto-DON in enrichment cultures and 3-epi-DON in a Devosia strain culture. Community composition was profiled with 16S rRNA gene amplicon sequencing, which showed that Z13 presented a remarkable drop in diversity upon microbial cultivation, and included genera such as Devosia, Nocardioides, and Pseudomonas. Together, these results provide integrated chemical, biological, and ecological evidence for aerobic DON biotransformation in soil-derived communities, identify pathway products, and highlight practical constraints related to community dependence and storage sensitivity.

Matrix effects (ME) during LC-ESI-MS analysis are a commonly acknowledged issue for a variety of matrices and analytes. Although sample preparation techniques are steadily evolving to reduce ME, the complexity and variability of the urine matrix remain a challenge, especially for multi-analyte methods. To investigate the extent of ME implications on method performance and quantification, we used stable isotope-labelled standards (SIL-IS) of 11 mycotoxins to evaluate the magnitude and variability of ME in urine samples from two cohorts: Bangladeshi adult women (n = 50) and Swedish children of both sexes (n = 340). Significant ME differences were observed between the two cohorts for eight of the 11 mycotoxins. Additionally, intra-cohort ME variability turned out to be very high with interquartile ranges (IQR) above 15% for 14 out of 22 analyte-cohort combinations. Maximum IQR values were observed for sterigmatocystin in the Bangladeshi cohort (318%), strongly impacting quantitative results obtained with matrix(-matched) calibration. Further experiments on a small German cohort of four subjects, each providing four to five urine samples, revealed high variability of ME within each individual. Factors influencing ME were investigated, showing little to no impact of sex and a moderate impact of age for some analytes in the Swedish cohort. Nonetheless, especially the more polar analytes, showing stronger signal suppression, demonstrated clear correlation of ME with density and creatinine concentration of the urine samples. As a result, urine samples with very high or low density or creatinine values require careful handling in regard to sensitivity or quantification errors when matrix(-matched) calibration without SIL-IS is applied.

This review summarises developments published in the period from mid-2024 to mid-2025 on the quantitative and qualitative determination of mycotoxins in various matrices. Important developments in all aspects of mycotoxin analysis, from sampling and quality assurance/quality control of analytical results, to the various detection and quantitation technologies ranging from biosensors to comprehensive instrumental methods are presented and discussed. This non-exhaustive summary and associated discussion covers such technology as chromatography coupled to (high resolution) mass spectrometry, detection other than mass spectrometry such as ultraviolet or fluorescence detection, biosensors, assays using alternatives to antibodies, as well as methods using predictive modelling and incorporating machine learning. This collaborative critical review intends to guide readers to relevant research by briefly presenting the most important developments in mycotoxin determination published in the past year. This review also relays limitations of the presented methodologies, in order to provide a fulsome assessment of the analytical developments.

Citrinin (CIT) is a nephrotoxic mycotoxin commonly found in a broad range of foods, including cereals, spices, nuts, or Monascus fermentation products. Analyses have shown that CIT is present in processed foods in significantly lower concentrations than in unprocessed materials. Modified forms of CIT arising during food processing may provide an explanation for the discrepancy. This study deals with the thermal stability of CIT and the formation of reaction products of CIT with carbohydrates, followed by toxicological evaluations using cell culture models. HPLC-HRMS degradation curves of CIT heated in different matrix model systems were recorded, and the formation of decarboxycitrinin (DCIT), the main degradation product, was quantified. Additionally, chemical structures of reaction products of CIT with carbohydrates were tentatively identified using MS/MS spectra and stable isotope labelling. Subsequently, the degradation of CIT during biscuit baking was studied, and carbohydrate-bound forms of CIT were detected after enzymatic starch digestion. The formation of DCIT could explain the majority of CIT degradation, but, depending on the process, covalent binding to carbohydrates can also be highly relevant. Cytotoxicity of DCIT in IHKE-cells was found to be lower compared to CIT, while the toxicity as well as the intestinal metabolism of carbohydrate-bound CIT was not evaluated.

This review summarises developments published in the period from mid-2023 to mid-2024 on the quantitative and qualitative determination of mycotoxins in a various matrices. Important developments in all aspects of mycotoxin analysis, from sampling and quality assurance/quality control of analytical results, to the various detection and quantitation technologies ranging from biosensors to comprehensive instrumental methods are presented and discussed. This non-exhaustive summary and associated discussion covers such technology as chromatography with targeted or non-targeted high resolution mass spectrometry, detection other than mass spectrometry such as fluorescence or diode array detection, biosensors, assays using alternatives to antibodies, as well as methods using predictive modelling and incorporating machine learning. This collaborative critical review intends to guide readers to relevant research by briefly presenting the most important developments in mycotoxin determination published in the past year. This review also relays limitations of the presented methodologies, in order to provide a thorough assessment of the analytical developments.

Cow’s milk contains A1- and A2-β-caseins. The breakdown of A1-β-casein produces β-casomorphin-7 (BCM-7), a peptide with opioid-like properties that is associated with health aspects. In addition, A1- and A2-β-casein have different technological properties. The aim of the present study was to investigate whether cheese produced from the milk of homozygous A1A1 and A2A2 cows varies in terms of its physicochemical parameters and BCM-7 concentration. These parameters were analyzed during initial cheese processing, six weeks of ripening and 84 days of storage, including additional microbiological analyses during the storage period. The pH values of the A1A1 cheeses were higher than those of the A2A2 cheeses from the beginning of production until the starter culture bacteria were added. The yellowness values of the A1A1 cheeses were lower until the salt bath treatment. Water activity, lightness, hardness, fat, protein, NaCl and dry matter content, as well as color and microbiological parameters, were not affected by the β-casein genotype. BCM-7 concentrations were higher in the A1A1 cheeses after pressing and during ripening. We found mainly comparable quality characteristics and slightly different BCM-7 levels in the A1A1 and A2A2 cheeses. From this point of view, both varieties are equally suitable for cheese production.