To date, the use of biomarkers has become generally accepted. Biomarker-driven research has been proposed as a successful method to assess the exposure to xenobiotics by using concentrations of the parent compounds and/or metabolites in biological matrices such as urine or blood. However, the identification and validation of biomarkers of exposure remain a challenge. Recent advances in high-resolution mass spectrometry along with new analytical (postacquisition data-mining) techniques will improve the quality and output of the biomarker identification process. Chronic or even acute exposure to mycotoxins remains a daily fact, and therefore it is crucial that the mycotoxins' metabolism is unravelled so more knowledge on biomarkers in humans and animals is acquired. This review aims to provide the scientific community with a comprehensive overview of reported in vitro and in vivo mycotoxin metabolism studies in relation to biomarkers of exposure for deoxynivalenol, nivalenol, fusarenon-X, T-2 toxin, diacetoxyscirpenol, ochratoxin A, citrinin, fumonisins, zearalenone, aflatoxins, and sterigmatocystin.

In the present study, metabolites of T-2 toxin in in vivo and in vitro systems of Wistar rats were identified and elucidated by ultraperformance liquid chromatography-quadrupole/time-of-flight tandem mass spectrometry (UPLC-Q/TOF-MS). Expected and unexpected metabolites were detected by Metabolynx(XS) software, which could automatically compare MS(E) data from the sample and control. A total of 19 metabolites of T-2 toxin were identified in this research, 9 of them being novel, which were 15-deacetyl-T-2, 3'-OH-15-deacetyl-T-2, 3',7-dihydroxy-T-2, isomer of 3',7-dihydroxy-T-2, 7-OH-HT-2, isomer of 7-OH-HT-2, de-epoxy-3',7-dihydroxy-HT-2, 9-OH-T-2, and 3',9-dihydroxy-T-2. The results showed that the main metabolic pathways of T-2 toxin were hydrolysis, hydroxylation, and de-epoxidation. In addition, the results also revealed one novel metabolic pathway of T-2 toxin, hydroxylation at C-9 position, which was demonstrated by the metabolites 9-OH-T-2 and 3',9-dihydroxy-T-2. In addition, hydroxylation at C-9 of T-2 toxin was also generated in in vitro of liver systems. Interestingly, several metabolites of hydroxylation at C-7 of T-2 toxin were also detected in in vivo male Wistar rats, but they were not found in in vivo female rats and in in vitro systems of Wistar rats.

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Ochratoxin A (OTA) is a mycotoxin that frequently contaminates a wide variety of food and feedstuffs. The metabolism of OTA greatly affects fate and toxicity in humans and animals, because of its possible carcinogenic character (International Agency for Research on Cancer (IARC), group 2B). To completely characterize the metabolites of OTA, the metabolism of OTA in liver microsomes of rats, chickens, swine, goats, cows, and humans was investigated using ultra-performance liquid chromatography-quadrupole/time-of-flight hybrid mass spectrometry (UPLC-Q/TOF-MS). In addition, an in vivo comparative metabolism study of OTA was performed among rats and chickens after oral administration of OTA. As a result, a clear metabolic profile of OTA in different species was proposed, and a total of eight metabolites were identified, of which three hydroxylated metabolites at the phenylalanine moiety were discovered for the first time (preliminarily identified as 9'-OH-OTA, 7'-OH-OTA, and 5'-OH-OTA). Considerable amounts of 7'-OH-OTA were detected in different species' liver microsomes, especially in chickens and humans. Moreover, the metabolism of OTA in chickens was elucidated for the first time in the present study. The 7'-OH-OTA proved to be the main metabolite in vitro and in vivo in chickens. Furthermore, the 4(S)-OH-OTA isomer was the major one, and 4(R)-OH-OTA the minor metabolite in chickens, which was different from others where 4R was the major. OTA undergoes metabolism via three different pathways, namely hydroxylation, dechlorination, and conjugation. The proposed metabolic pathways of OTA in various species provide the scientific community useful data for the toxicological safety evaluation of OTA among different species, and will further facilitate the food safety evaluation of OTA.

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This research produced a highly-specific and sensitive anti-T-2 toxin monoclonal antibody (mAb), and developed a rapid and sensitive competitive indirect enzyme-linked immunosorbent assay (ELISA) method for monitoring T-2 toxin in rice. The mAb showed a negligible cross-reactivity value (CR) to most of the mycotoxins, and it could specifically bind to T-2 toxin without other mycotoxins, including HT-2 toxin (CR value at 3.08%), which exhibited a similar structure to T-2 toxin. The limit of detection (LOD) value, measured by IC10, was 5.80 μg/kg. In spiked samples, mean recoveries ranged from 72.0% to 108.5% with intraday and interday variation less than 16.8 and 13.7%. This proposed protocol was significantly confirmed by a reliable ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method and significant correlation was obtained.