Mycotoxins, toxic secondary metabolites of fungi, affect global agriculture so prolifically that they are virtually ubiquitous at some concentration in the average human diet. Studies of in vitro and in vivo toxicity are discussed, leading to investigations of co-exposed mycotoxins, as well as carcinogenic effects. Some of the most common and toxicologically significant mycotoxins, such as the aflatoxins, ochratoxins, fumonisins, deoxynivalenol, T-2 toxin, HT-2 toxin, patulin, zearalenone, and some ergot alkaloids are outlined. The wide variety of pathogenic mechanisms these compounds employ are shown capable of inducing a complex set of interactions. Of particular note are potential synergisms between mycotoxins with regard to carcinogenic attributable risk, indicating an important field for future study.
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.
An LC-MS/MS method was developed and validated for the simultaneous determination of deoxynivalenol, zearalenone, T-2-toxin, HT-2-toxin and metabolites, including 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol-3-glucoside, α-zearalenol, β-zearalenol, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate in maize, wheat, oats, cornflakes and bread. Extraction was performed with acetonitrile/water/acetic acid (79/20/1, v/v/v) followed by a hexane defatting step. After filtration, the extract was evaporated and the residue was redissolved in mobile phase for injection. The mobile phase, which consisted of a mixture of methanol and water with 10 mM ammonium acetate, was adjusted to pH 3 with glacial acetic acid. A sample clean-up procedure was not included because of the low recoveries of free and masked mycotoxins and their differences in polarity. The method allowed the simultaneous determination of 13 Fusarium mycotoxins in a one-step chromatographic run using a Waters Acquity UPLC system coupled to a Quattro Premier XE mass spectrometer. The method was validated for several parameters such as linearity, apparent recovery, limit of detection, limit of quantification, precision, expanded measurement uncertainty and specificity. The limits of detection varied from 5 to 13 ng g⁻¹; those for the limit of quantification from 10 to 26 ng g⁻¹. The results of the performance characteristics of the developed LC-MS/MS method were in good agreement with the criteria mentioned in Commission Regulation (EC) No. 401/2006. Thirty samples of a variety of food and feed matrices were sampled and analysed between July 2010 and January 2011.
A total of 174 cereal-based food products, 67 compound feeds and 19 feed raw materials were analysed for the occurrence of deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, zearalenone, α-zearalenol, β-zearalenol, and their respective masked forms, including deoxynivalenol-3-glucoside, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate. Fibre-enriched bread, bran-enriched bread, cornflakes, popcorn and oatmeal were collected in Belgian supermarkets from April 2010 to October 2011. All food samples analysed were contaminated with an average of 2 to 6 mycotoxins, including 1 to 3 masked forms. Feed raw materials that were used in the analysed compound feeds were collected by the manufacturer. Feed raw materials included were beet pulp, sunflower seed meal, soy bean, soy peel, oats, barley, maize germs, maize gluten feed, maize, wheat gluten feed, wheat bran pellets, wheat bran and wheat. Beet pulp, sunflower seed meal, soy bean and soy peel were hardly contaminated. The feed raw materials that were mostly infected with deoxynivalenol, zearalenone and derivatives were maize and its by-products. Also, the glucosylated and sulfated forms occurred in substantial amounts. As well, wheat and its by-products were contaminated with α-zearalenol (wheat gluten feed and wheat bran) and zearalenone (wheat). The contamination pattern and level of feed raw materials were reflected in the corresponding compound feeds.
The aim of this study was to determine the toxicokinetic characteristics of ZEN and its modified forms, α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalenone-14-glucoside (ZEN14G), and zearalenone-14-sulfate (ZEN14S), including presystemic and systemic hydrolysis in pigs. Crossover pig trials were performed by means of intravenous and oral administration of ZEN and its modified forms. Systemic plasma concentrations of the administered toxins and their metabolites were quantified and further processed via tailor-made compartmental toxicokinetic models. Furthermore, portal plasma was analyzed to unravel the site of hydrolysis, and urine samples were analyzed to determine urinary excretion. Results demonstrate complete presystemic hydrolysis of ZEN14G and ZEN14S to ZEN and high oral bioavailability for all administered compounds, with further extensive first-pass glucuronidation. Conclusively, the modified-ZEN forms α-ZEL, β-ZEL, ZEN14G, and ZEN14S contribute to overall ZEN systemic toxicity in pigs and should be taken into account for risk assessment.
In the present study, a quantitative dietary exposure assessment of mycotoxins and their masked forms was conducted on a national representative sample of the Belgian population using the contamination data of cereal-based foods. Cereal-based food products (n = 174) were analysed for the occurrence of deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, zearalenone, α-zearalenol, β-zearalenol, T-2-toxin, HT-2-toxin, and their respective masked forms, including, deoxynivalenol-3-glucoside, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate.Fiber-enriched bread, bran-enriched bread, breakfast cereals, popcorn and oatmeal were collected in Belgian supermarkets according to a structured sampling plan and analysed during the period from April 2010 till October 2011. The habitual intake of these food groups was estimated from a national representative food intake survey. According to a probabilistic exposure analysis, the mean (and P95) mycotoxin intake for the sum of the deoxynivalenolequivalents, zearalenone-equivalents, and the sum of HT-2-and T-2-toxin for all cereal-based foods was 0.1162 (0.4047, P95), 0.0447 (0.1568, P95) and 0.0258 (0.0924, P95) µg kg -1 body weight day -1 , respectively. These values were below the tolerable daily intake (TDI) levels for deoxynivalenol, zearalenone and the sum of T-2 and HT-2 toxin (1.0, 0.25 and 0.1 µg kg -1 body weight day -1 , respectively). The absolute level exceeding the TDI for all cereal-based foods was calculated, and recorded 0.85%, 2.75% and 4,11% of the Belgian population, respectively. KEYWORDS: mycotoxins,
Crossover animal trials were performed with intravenous and oral administration of deoxynivalenol-3-β-D-glucoside (DON3G) and deoxynivalenol (DON) to broiler chickens and pigs. Systemic plasma concentrations of DON, DON3G and de-epoxy-DON were quantified using liquid chromatography-tandem mass spectrometry. Liquid chromatography coupled to high-resolution mass spectrometry was used to unravel phase II metabolism of DON. Additionally for pigs, portal plasma was analysed to study presystemic hydrolysis and metabolism. Data were processed via tailor-made compartmental toxicokinetic models. The results in broiler chickens indicate that DON3G is not hydrolysed to DON in vivo. Furthermore, the absolute oral bioavailability of DON3G in broiler chickens was low (3.79 ± 2.68 %) and comparable to that of DON (5.56 ± 2.05 %). After PO DON3G administration to pigs, only DON was detected in plasma, indicating a complete presystemic hydrolysis of the absorbed fraction of DON3G. However, the absorbed fraction of DON3G, recovered as DON, was approximately 5 times lower than after PO DON administration, 16.1 ± 5.4 compared with 81.3 ± 17.4 %. Analysis of phase II metabolites revealed that biotransformation of DON and DON3G in pigs mainly consists of glucuronidation, whereas in chickens predominantly conjugation with sulphate occurred. The extent of phase II metabolism is notably higher for chickens than for pigs, which might explain the differences in sensitivity of these species to DON. Although in vitro studies demonstrate a decreased toxicity of DON3G compared with DON, the species-dependent toxicokinetic data and in vivo hydrolysis to DON illustrate the toxicological relevance and consequently the need for further research to establish a tolerable daily intake.
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