Determination of mycotoxins by HPLC, LC-MS/MS and health risk assessment of the mycotoxins in bee products of Turkey.

The aim of this study is to determine the levels of deoxynivalenol (DON), HT-2 toxin (HT2), T-2 toxin (T2), and ochratoxin A (OTA) in bee products (bee pollen, propolis, honey and royal jelly) available in Turkey. In addition, exposure and health risk assessments were performed to identify the potential health risk of these mycotoxins. The mycotoxins were analyzed by high-performance liquid chromatography (HPLC) with a UV detector and positive samples were confirmed with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The most common mycotoxins in all bee products were DON and T-2 toxin, with mean concentrations of 1.601 and 0.704 µg/per kg dry sample, respectively, followed by OTA and HT-2 toxin. It was determined that the mycotoxins taken as a result of consuming bee products in specified amounts do not pose a risk to health.

Occurrence of mycotoxins in rice consumed by Iranians: a probabilistic assessment of risk to health.

Risks based on cancer and non-cancer endpoints, to Iranians from exposure to several mycotoxins (aflatoxin B1, ochratoxin, deoxynivalenol and T-2 toxin) following consumption of rice were evaluated. Point estimates of hazard were made for each mycotoxin and a hazard index (HI) and probabilistic estimates were based on results of Monte Carlo Simulations (MCS). All known 17 peer-reviewed studies, published in databases included in Science Direct, PubMed, Scopus and Web of Science, as well as grey literature published in Google Scholar from 2008 to 2017 were considered. The 95th and 50th centiles of Hazard Index (HI) in Iranians due to ingestion of rice were estimated to be 2.5 and 0.5, respectively. The 95th and 50th centiles of people with positive surface antigens for hepatitis B (HBsAg+) risk characterisation for AFB1 in Iranian consumers of rice were 81 and 79.1, respectively. The 95th and 50th centiles for risks of Iranians negative for the surface antigen of hepatitis B HBsAg (HBsAg-) were 4.4 and 2.6, respectively. Based on results of the MCS for risks to cancer effects, the 95th and 50th centiles of margins of exposure (MOE) were 233 and 231, respectively. Therefore, it is recommended to update agricultural approaches and storage methods and implement monitoring and regulations based on risks to health posed by consumption of rice by the Iranian population.

Occurrence and risk assessment of mycotoxins, acrylamide, and furan in Latvian beer.

This work reports data on the occurrence of nine mycotoxins and two food processing contaminants - acrylamide and furan - in a total of 100 beers produced in Latvia. Mycotoxins were detected by high-performance liquid chromatography (HPLC) coupled with time-of-flight mass spectrometry, acrylamide by HPLC coupled with quadrupole-Orbitrap mass spectrometry, and furan by headspace gas chromatography-mass spectrometry. The most frequently occurring mycotoxins were HT-2 and deoxynivalenol (DON), which were detected in 52% and 51% of the analysed samples. The highest content was observed for DON, reaching the maximum of 248 µg kg-1. Furan was ubiquitous, and 74% of the samples contained acrylamide. In terms of the estimated exposure, the biggest potential risk was identified for HT-2 representing more than 11% of tolerable weekly intake. The margin of exposure approach indicated the exposure to furan through beer as significant, this parameter being close to the critical limit.

Mould and mycotoxin exposure assessment of melon and bush mango seeds, two common soup thickeners consumed in Nigeria.

An examination of the mould and fungal metabolite pattern in melon and bush mango seeds locally produced in Nigeria was undertaken in order to understand the mycotoxicological risk posed to consumers of both of these important and commonly consumed soup thickeners. The variation in mycotoxin levels in graded categories of both foodstuffs were also determined. Aspergillus, Fusarium, Penicillium, Mucorales and Trichoderma were the recovered fungi from the foodstuffs with Aspergillus species dominating (melon=97.8%; bush mango=89.9%). Among the Aspergillus species identified Aspergillus section Flavi dominated (melon: 72%; bush mango: 57%) and A. flavus, A. parasiticus, A. parvisclerotigenus and A. tamarii were the recovered species. About 56% and 73% of the A. flavus isolates from melon and bush mango seed samples, respectively were aflatoxigenic. Thirty-four and 59 metabolites including notable mycotoxins were found in the melon and bush mango seeds respectively. Mean aflatoxin levels (µg/kg) in melon (aflatoxin B1 (AFB1)=37.5 and total aflatoxins=142) and bush mango seeds (AFB1=68.1 and total aflatoxins=61.7) were higher than other mycotoxins, suggesting potential higher exposure for consumer populations. Significantly (p<0.05) higher levels of mycotoxins were found in hand-peeled melon and discoloured bush mango seeds than in machine-peeled melon and non-discoloured seeds except for HT-2 and T-2 toxins which occurred conversely. All melon and bush mango seeds exceeded the 2µg/kg AFB1 limit whereas all melon and 55% of bush mango seeds exceeded the 4µg/kg total aflatoxin EU limit adopted in Nigeria. This is the first report of (1) mycotoxin co-occurrence in bush mango seeds, (2) cyclopiazonic acid, HT-2 toxin, moniliformin, mycophenolic acid, T-2 toxin and tenuazonic acid occurrence, and (3) mycotoxin exposure assessment of both foodstuffs.

The use of immunoaffinity columns connected in tandem for selective and cost-effective mycotoxin clean-up prior to multi-mycotoxin liquid chromatographic-tandem mass spectrometric analysis in food matrices.

This paper describes the use of two immunoaffinity columns (IACs) coupled in tandem, providing selective clean-up, based on targeted mycotoxins known to co-occur in specific matrices. An IAC for aflatoxins+ochratoxin A+fumonisins (AOF) was combined with an IAC for deoxynivalenol+zearalenone+T-2/HT-2 toxins (DZT); an IAC for ochratoxin A (O) was combined with a DZT column; and an aflatoxin+ochratoxin (AO) column was combined with a DZT column. By combining pairs of columns it was demonstrated that specific clean-up can be achieved as required for different matrices. Samples of rye flour, maize, breakfast cereal and wholemeal bread were analysed for mycotoxins regulated in the EU, by spiking at levels close to EU limits for adult and infant foods. After IAC clean-up extracts were analysed by LC-MS/MS with quantification using multiple reaction monitoring. Recoveries were found to be in range from 60 to 108%, RSDs below 10% depending on the matrix and mycotoxin combination and LOQs ranged from 0.1n g/g for aflatoxin B1 to 13.0 ng/g for deoxynivalenol. Surplus cereal proficiency test materials (FAPAS(®)) were also analysed with found levels of mycotoxins falling within the satisfactory range of concentrations (Z score ≤ ± 2), demonstrating the accuracy of the proposed multi-mycotoxin IAC methods.

T-2 and HT-2 toxins in cereals and cereal-based products in South Korea.

A total of 214 samples, consisting of brown rice, barley, mixed grains, corn, wheat and wheat flour were analysed for T-2 and HT-2 toxins using high-performance liquid chromatography with fluorescence detection. Recovery and repeatability were 79.9%-107.5% and 4.9%-14.5% for T-2, and 74.0%-106.1% and 5.0%-17.9% for HT-2, respectively. T-2 toxin was detected in 11 (5.1%) of all samples. The highest incidence was found in corn (21.7%) followed by mixed grains and brown rice. Mean of all samples was 1.5-4.1 µg kg⁻¹, the maximum level being 41.5 µg kg⁻¹ in corn. HT-2 toxin was detected in 126 (58.9%) of all samples, and the mean values were 26.4-59.2 µg kg⁻¹. The estimated daily intakes for the sum of T-2 and HT-2 toxins were 2.56, 3.22, 2.53, 0.03, 0.01 and 2.45 ng (kg bw)⁻¹ day⁻¹ in brown rice, barley, mixed grains, corn, wheat and wheat flour, respectively.

Fusariotoxins in Russian Federation 2005-2010 grain harvests.

Monitoring results of food grain contamination with fusariotoxins-deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FB1&FB2), T-2 and HT-2 toxins-are presented. Harvests of 2005-2010 in different regions of Russia were investigated. The occurrence of DON in wheat was 8%, barley 9%, oats 4%, rye 2% and maize 2%. The highest frequency of ZEN contamination was found in oats, the lowest in wheat. Calculated average daily intake of DON varied from 0.066 to 0.096 µg/kg body weight, the highest being found in the Southern region, but substantially lower than the provisional maximum tolerable daily intake. The results of enzyme-linked immunosorbent assay and high-performance liquid chromatography-mass spectrometry analysis demonstrated the presence of T-2 toxin in 14% and HT-2 toxin in 17% of all samples. The maximum level of T-2 toxin was exceeded only in one sample of barley. Relatively high frequency and levels of FB1&FB2 contamination were found in maize.

Dietary exposure to mycotoxins and health risk assessment in the second French total diet study.

Mycotoxins are produced in plants by micro-fungi species, and naturally contaminated the food chain. In the second French total diet study (TDS), mycotoxins were analyzed in 577 food samples collected in mainland France to be representative of the population diet and prepared ((as consumed)). Highest mean concentrations were found in wheat and cereal-based products (bread, breakfast cereals, pasta, pastries, pizzas and savoury pastries…). Exposure of adult and child populations was assessed by combining national consumption data with analytical results, using lowerbound (LB) and upperbound (UB) assumptions for left-censorship management. Individual exposures were compared with available health-based guidance values (HBGV). Only the exposure to deoxynivalenol (DON) and its acetylated derivatives was found to significantly exceed the HBGV in LB in adults (0.5% [0.1; 0.8]) and children (5% [4; 6]). HBGV was exceeded in UB only for T-2 and HT-2 toxins by, respectively, 0.2% [0.02; 0.05] and 4% [3; 5] of adults, and 11% [9; 12] and 35% [32; 37] of children. Although the exposures assessed were generally lower than the previous French TDS, the results indicated a health concern for trichothecenes and a need to reduce dietary exposure as well as analytical limits.

A rapid fluorescence polarization immunoassay for the determination of T-2 and HT-2 toxins in wheat.

A rapid fluorescence polarization (FP) immunoassay has been developed for the simultaneous determination of T-2 and HT-2 toxins in naturally contaminated wheat samples. Syntheses of four fluorescein-labelled T-2 or HT-2 toxin tracers were carried out and their binding response with seven monoclonal antibodies was evaluated. The most sensitive antibody-tracer combination was obtained by using an HT-2-specific antibody and a fluorescein-HT-2 tracer. The developed competitive FP immunoassay in solution showed high cross-reactivity for T-2 toxin (CR% = 100%) while a very low CR% for neosolaniol (0.12%) and no cross-reactivity with other mycotoxins frequently occurring in wheat. A rapid extraction procedure using 90% methanol was applied to wheat samples prior to FP immunoassay. The average recovery from spiked wheat samples (50 to 200 µg kg(-1)) was 96% with relative standard deviation generally lower than 8%. A limit of detection of 8 µg kg(-1) for the combined toxins was determined. Comparative analyses of 45 naturally contaminated and spiked wheat samples by both the FP immunoassay and high-performance liquid chromatography/immunoaffinity clean-up showed a good correlation (r = 0.964). These results, combined with the rapidity (10 min) and simplicity of the assay, show that this method is suitable for high throughput screening as well as for quantitative determination of T-2 and HT-2 toxins in wheat.

[Dietary exposure assessment of some important Fusarium toxins in cereal-based products in China].

OBJECTIVE: To elucidate the dietary exposure of Chinese populations to deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN). METHODS: Ten Fusarium toxins including DON, NIV, ZEN in domestic wheat flour and corn-based products harvested and collected in 2009 were analyzed by UPLC-MS/MS. Dietary intake assessments of human exposure to DON, NIV and ZEN were carried out in combination of national food consumption data with toxin concentration data by deterministic estimate method. RESULTS: (1) There are 2.5% adults and 10% children with the dietary exposure to DON exceeding the tolerable daily intake (TDI) on the basis of the average food consumption. At the 75th percentile food consumption level, the dietary exposure of populations to DON was higher than its TDI, 1.72 and 2.02 times (adults) as well as 1.19 and 1.09 times higher than TDI (children), respectively, based on the higher DON exposure (adults : P90 for wheat flour and P97. 5 for corn-based products, children: P50 for wheat flour and P75 for corn-based products). At the average toxin concentration, children with the high consumption level (90th, 97. 5th and 99th percentile) of either wheat flour or corn-based products, the dietary exposure to DON exceed the TDI, 1.81 to 3.17 times (wheat flour) and 1.47 - 3.97 times (corn-based products) higher than TDI, respectively. The dietary exposure of adults to ZEN exceed the TDI, based on the average food consumption data and higher ZEN level (P99), the 75th consumption data combined with P97. 5th toxin concentration, as well as 90th percentile of both food consumption data and ZEN concentration, respectively. There are 1%, 2.5%, 25%, 25% and 50% populations of children exposed ZEN higher than TDI, respectively at the 50th, 75th, 90th, 97. 5th and 99th percentile of food consumption data. (2) The level of concern (LOC) of DON in wheat flour calculated with high consumption data (90th, 97. 5th and 99th percentile) for all populations as well as DON and ZEN in corn-based products calculated with high consumption data (adults: 99th percentile, children: 97. 5th and 99th percentile) were lower than their average concentrations in above cereals. (3) No matter adults or children, the maximum daily safe intake of both wheat flour and corn-based products with high concentration of DON (97. 5th and 99th percentile, and 90th else for children) were lower than their own average food consumption data. For adults, the maximum daily safe intake of corn-based products with high ZEN contamination level (99th percentile) was lower than the average amount of corn-based products consumption. While, the maximum daily safe intake of both wheat flour and corn-based products derived from high ZEN concentration (97.5th and 99th percentile) for children were lower than their own average food consumption. CONCLUSIONS: Children dietary exposure to DON, NIV and ZEN was higher than adults. Children are the populations at the high risk of dietary exposure to these three mycotoxins. The risk to health caused by long-term consumption of wheat flour and corn-based products heavily contaminated with such high concentration of DON and ZEN was relatively high. The present tolerance limit for DON and ZEN in foods implemented in China should be revised on the basis of assessment results.

Assessment of immunochemical methods for the analysis of trichothecene mycotoxins in naturally occurring moldy corn.

The efficacy of various immunochemical methods for simultaneous analysis of different trichothecene mycotoxins in corn samples naturally contaminated with various Fusarium molds was evaluated. Antibodies against either T-2 tetraol tetraacetate (T-2-4ol-4Ac) or deoxynivalenol triacetate (Tri-Ac-DON) were used in the enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) for total type A and total DON-related trichothecenes (TCTCs), respectively. Specific antibodies against T-2 toxin were used in an RIA for analysis of T-2 toxin alone. Analytical recoveries of T-2 toxin analyzed as T-2-4ol-4Ac by ELISA and T-2 toxin by RIA at 10-200 ppb were 83.5-123% and 81.8-110.6%, respectively. T-2 toxin accounts for about 30% of total type A TCTCs by both liquid chromatography (LC)-ELISA and RIA methods. The correlation coefficients (r) of total type A TCTC levels versus T-2 toxin level by RIA was 0.725 (p < 0.0001). A good correlation (r = 0.84 and p = 0.04) also was found for T-2 toxin levels obtained by LC-ELISA and RIA methods. In addition to T-2 toxin, HT-2 toxin, diacetoxyscirpenol, and T-2 tetrol were found in samples containing high levels of type A TCTCs by LC-ELISA. For analysis of DON alone, samples were treated with a Sep-Pak C18 cartridge to remove other acetylated DONs before RIA. Recovery of DON after the C18 cartridge treatment was 90%. Overall analytical recoveries of DON as Tri-Ac-DON (sample with no C18 treatment) and DON alone (with C18 treatment) at 10-500 ppb were 81.3-94% and 72.7-104%, respectively. RIA revealed that DON accounts for about 50% of total DON-related TCTCs. The coefficient of correlation between total DON-related TCTCs and DON levels was 0.84 (p < 0.0001).