Survey of T-2 and HT-2 toxins by LC–MS/MS in oats and oat products from European oat mills in 2005–2009

T-2 and HT-2 toxins were analysed in oats (n?=?243), oat flakes (n?=?529), oat meal (n?=?105) and oat by-products (n?=?209) from 11 European mills during 2005–2009 by high-performance liquid chromatography with a triple quadrupole mass spectrometer. Limits of quantification were 5?µg?kg^?1 for both T-2 and HT-2 toxins in oats. The incidence of T-2?+?HT-2 (>5?µg?kg^?1) in oats, oat flakes, oat meal and oat by-products was 93, 77, 34 and 99%, respectively. The mean values of T-2?+?HT-2 were 94, 17, 11 and 293?µg?kg^?1 for oats, oat flakes, oat meal and oat by-products, respectively. T-2 and HT-2 occurred together and the T-2 level was 52% of HT-2 in oats. Maximal T-2 and HT-2 concentration in oat flakes and oat meal were 197 and 118?µg?kg^?1. The toxins were reduced by 82–88% during processing, but increased 3.1 times in oat by-products.     

Fusarium mycotoxin content of UK organic and conventional oats

Every year between 2002 and 2005 approximately 100 samples of oats from fields of known agronomy were analysed by GC/MS for 10 trichothecenes: deoxynivalenol (DON), nivalenol, 3-acetylDON, 15-acetylDON, fusarenone X, T-2 toxin (T2), HT-2 toxin (HT2), diacetoxyscirpenol, neosolaniol and T-2 triol. Samples were also analysed for moniliformin and zearalenone by HPLC. Of the 10 trichothecenes analysed from 458 harvest samples of oat only three, 15-acetylDON, fusarenone X and diacetoxyscirpenol, were not detected. Moniliformin and zearalenone were absent or rarely detected, respectively. HT2 and T2 were the most frequently detected fusarium mycotoxins, present above the limit of quantification (10 µg kg^?1) in 92 and 84% of samples, respectively, and were usually present at the highest concentrations. The combined mean and median for HT2 and T2 (HT2 + T2) was 570 and 213 µg kg^?1, respectively. There were good correlations between concentrations of HT2 and all other type A trichothecenes detected (T2, T2 triol and neosolaniol). Year and region had a significant effect on HT2 + T2 concentration. There was also a highly significant difference between HT2 + T2 content in organic and conventional samples, with the predicted mean for organic samples five times lower than that of conventional samples. This is the largest difference reported for any mycotoxin level in organic and conventional cereals. No samples exceeded the legal limits for DON or zearalenone in oats intended for human consumption. Legislative limits for HT2 and T2 are currently under consideration by the European Commission. Depending on the limits set for unprocessed oats intended for human consumption, the levels detected here could have serious consequences for the UK oat-processing industry.     

Occurrence of type A trichothecenes in conventionally and organically produced oats and oat products

Among cereals, oats are known to be very frequently contaminated with type A trichothecenes and so they can play a major role in the exposition of the consumer to these mycotoxins. Seventy representative oat samples of both conventional and organic production were drawn at mills and at wholesale stage according to Commissions Regulation (EC) No 401/2006 and analyzed for nine type A trichothecenes by LC-MS/MS. High contamination rates were found for most of the toxins in conventional as well as in organic products (e. g. 100% for T-2 toxin or 99% for HT-2 toxin). The mean concentration of T-2/HT-2 (sum of the toxins) was 17 +/- 18 microg/kg (mean +/- SD) in all samples, 27 +/- 21 microg/kg in conventional, and 7.6 +/- 4.6 microg/kg in organic products, respectively. The highest T-2/HT-2 level has been determined in conventionally produced oat flakes (85 microg/kg). The mean level of T-2 tetraol (9.5 +/- 7.7 microg/kg) in all samples was found to be even higher than that of T-2 (5.1 +/- 6.0 microg/kg), whereas levels of T-2 triol, 4,15-diacetoxyscirpenol, 15-monoacetoxyscirpenol, and neosolaniol were considerably lower. For oats and oat products from organic farming contamination levels of T-2, HT-2, T-2 triol, T-2 tetraol, and neosolaniol were significantly lower. The results are discussed with respect to possible health risks for the consumer.     

Optimization and Validation of a Fluorescence Polarization Immunoassay for Rapid Detection of T-2 and HT-2 Toxins in Cereals and Cereal-Based Products

A fluorescence polarization (FP) immunoassay has been optimized and validated for rapid quantification of T-2 and HT-2 toxins in both unprocessed cereals, including oats, barley and rye, and cereal-based products for direct human consumption, such as oat flakes, oats crispbread and pasta. Samples were extracted with 90 % methanol, and the extract was filtered and diluted with water or sodium chloride solution prior to the FP immunoassay. Overall mean recoveries from spiked oats, rye, barley, oat flakes, oats crispbread and pasta ranged from 101 to 107 %, with relative standard deviations lower than 7 %. Limits of detection (LODs) of the FP immunoassay were 70 μg/kg for oats, 40 μg/kg for oat flakes and barley, 25 μg/kg for pasta and 20 μg/kg for rye and oats crispbread. The trueness of the immunoassay was assessed by using two oat and oat flake reference materials for T-2 and HT-2 toxins, showing good accuracy and precision. Good correlations (r?>?0.953) were observed between T-2 and HT-2 toxin contents in naturally and artificially contaminated samples determined by both FP immunoassay and ultra-high-performance liquid chromatography (UHPLC) with immunoaffinity column cleanup used as reference method. These results, combined with rapidity and simplicity of the assay, show that the optimized assay is suitable for high-throughput screening, as well as for reliable quantitative determination of T-2 and HT-2 toxins in cereals and cereal-based products.     

Mycotoxin contamination of cereal grain commodities in relation to climate in North West Europe

This study aimed to investigate mycotoxin contamination of cereal grain commodities for feed and food production in North Western Europe during the last two decades, including trends over time and co-occurrence between toxins, and to assess possible effects of climate on the presence of mycotoxins. For these aims, analytical results related to mycotoxin contamination of cereal grain commodities, collected in the course of national monitoring programmes in Finland, Sweden, Norway and the Netherlands during a 20-year period, were gathered. Historical observational weather data, including daily relative humidity, rainfall and temperature, were obtained from each of these four countries. In total 6382 records, referring to individual sample results for mycotoxin concentrations (one or more toxins) in cereal grains were available. Most records referred to wheat, barley, maize and oats. The most frequently analysed mycotoxins were deoxynivalenol, 3-acetyl-deoxynivalenol, nivalenol, T-2 toxin, HT-2 toxin and zearalenone. Deoxynivalenol had the highest overall incidence of 46%, and was mainly found in wheat, maize and oats. Mycotoxins that showed co-occurrence were: deoxynivalenol and 3-acetyl-deoxynivalenol in oats; deoxynivalenol and zearalenone in maize and wheat; and T-2 toxin and HT-2 toxin in oats. The presence of both deoxynivalenol and zearalenone in wheat increased with higher temperatures, relative humidity and rainfall during cultivation, but the presence of nivalenol was negatively associated with most of these climatic factors. The same holds for both nivalenol and deoxynivalenol in oats. This implies that climatic conditions that are conducive for one toxin may have a decreasing effect on the other. The presence of HT-2 toxin in oats showed a slight decreasing trends over time, but significant trends for other toxins showed an increasing presence during the last two decades. It is therefore useful to continue monitoring of mycotoxins. Obtained results can be used for development of predictive models for presence of mycotoxins in cereal grains.     

Thermal stability of T-2 and HT-2 toxins during biscuit- and crunchy muesli-making and roasting

ABSTRACT

The mycotoxins T-2 and HT-2 toxin are frequently occurring food contaminants which are produced by Fusarium species. Humans and animals are mainly exposed to these substances by the consumption of contaminated oats, maize and wheat. For the production of crunchy muesli, bread and bakery products, these cereals undergo multiple processing steps, including baking, roasting and extrusion cooking. However, the influence of food processing on T-2 and HT-2 toxin levels is to date poorly understood. Thus, the effects of baking and roasting on both mycotoxins were evaluated during biscuit-, crunchy muesli- and toasted oat flakes-production under precise variation of various parameters: heating time and temperature as well as recipe formulation were varied in the range they are applied in the food processing industry. Therefore, oatmeal or flaked oats were artificially contaminated individually with both toxins and processed at the laboratory scale. T-2 toxin generally showed a higher degradation rate than HT-2 toxin. During biscuit-making up to 45% of T-2 toxin and 20% of HT-2 toxin were thermally degraded, showing a dependency on water content, baking time and temperature. The preparation of crunchy muesli yielded no significant toxin degradation which is probably due to the low temperatures applied. Roasting led to a degradation of 32% of T-2 toxin and 24% of HT-2 toxin. Taken together, both mycotoxins are partially degraded during thermal food processing; the degradation rates are influenced by the food composition and processing parameters.     

Mycotoxins in infant/toddler foods and breakfast cereals in the US retail market

The objective of this study was to conduct a mycotoxin survey of commercial infant/toddler foods (cereals and teething biscuits) and breakfast cereals in the United States. A total of 215 retail samples were collected from three geographical locations and analysed for aflatoxins, fumonisins, deoxynivalenol, HT-2 toxin, ochratoxin A, T-2 toxin, and zearalenone using a stable isotope dilution liquid-chromatography tandem mass spectrometry (LC-MS/MS) method. One or more mycotoxins were found in 69% (101/147) of the infant/toddler foods and 50% (34/68) of breakfast cereals. Mycotoxin co-occurrence was observed in 12% of infant/toddler foods and 32% of breakfast cereals. However, the concentrations of detected mycotoxins were lower than the current FDA action and guidance levels. Aflatoxins and HT-2 toxin were not detected in any of the samples, while deoxynivalenol was the most frequently detected mycotoxin. Rice-based cereals appeared to be less susceptible to mycotoxin contamination than other cereal types.     

Impact of agronomic and climatic factors on the mycotoxin content of harvested oats in the United Kingdom

ABSTRACT

A survey was conducted to determine the concentration of Fusarium mycotoxins in UK oats over three seasons (2006–8). One hundred oat samples were collected each year at harvest, together with agronomic details, and analysed for 10 Fusarium mycotoxins. The incidence and concentration of most Fusarium mycotoxins, including deoxynivalenol and zearalenone, were relatively low in oats compared with values previously reported for wheat. HT-2 toxin (HT2) and T-2 toxin (T2) levels were relatively high with an overall combined (HT2+T2) mean of 450 μg kg^?1 for 2006–8. Data were combined with a previous dataset collected from 2002–5 to determine the effects of agronomic practices and climate. There was a negative relationship with late summer rainfall, indicating that drier conditions in July and August resulted in increased HT2 and T2 in UK oats. Agronomic factors that impacted upon HT2 and T2 in harvested oats were previous crop, cultivation, and variety. Analysis of the previous cropping history showed there was a stepwise increase in HT2+T2 as the cereal intensity of the rotation increased. Variety was an important factor, with higher levels and a wider range detected on winter versus spring varieties. Indicative levels for HT2 and T2 in cereals and cereal products were introduced by the EC in 2013. The indicative level for unprocessed oats for human consumption is a combined concentration (HT2+T2) of 1000 μg kg^?1. From 2002 to 2008, between 1% and 30% of samples exceeded 1000 μg kg^?1 HT2+T2 each year (overall mean, 16%). The introduction of European legislation on HT2 and T2 mycotoxins could have serious implications for UK oat production and oat-processing industries based on the levels detected within these studies.     

Effect of fenpropimorph, prochloraz and tebuconazole on growth and production of T-2 and HT-2 toxins by Fusarium langsethiae in oat-based medium

Fusarium langsethiae has been isolated from infected cereals in central and northern Europe where it has been identified in the last decade as the main species involved in the occurrence of high levels of T-2 and HT-2 toxins, mainly in oats. The efficacy of three fungicides (prochloraz, tebuconazole, fenpropimorph) for controlling growth of two strains of F. langsethiae isolated from oats was examined at 0.96 and 0.98 a_w at 15, 20 and 25 °C on oat-based media. The concentrations necessary for 50 and 90% growth inhibition (ED₅₀ and ED₉₀ values) were determined. The effect on the trichothecene type A mycotoxins T-2 and HT-2 was also determined. Without fungicides both strains grew faster at 0.98 than at 0.96 a_w and the influence of temperature on growth rates was 25 > 20 > 15 °C. Prochloraz and tebuconazole were more effective than fenpropimorph against F. langsethiae. Strain, temperature and type of fungicide significantly influenced the ED₅₀ and ED₉₀ values for growth. The concentration ranges under different environmental conditions were: prochloraz (0.03-0.1 and 0.3-1.5), tebuconazole (0.06-0.9 and 1.3-8.2), and fenpropimorph (22-59 and 125-215 mg l⁻¹). Production of T-2 and HT-2 toxins was influenced by temperature, a_w, type of fungicide and dose. Levels of T-2 were usually higher than those of HT-2 under the same conditions. The biosynthesis of T-2 toxin increased after 10 day incubation, but was reduced with decreasing temperature and increasing fungicide dose. At 0.98 a_w T-2 levels increased in cultures containing fenpropimorph while at 0.96 a_w the toxin concentrations increased in response to the other two fungicides. Low doses of prochloraz or tebuconazole enhanced toxin production when compared with untreated cultures for strain 2004-59 at 0.96 a_w and 20-25 °C. HT-2 was hardly detectable in the treatments with prochloraz or tebuconazole at 0.98 a_w. This is the first study on the effect of these anti-fungal compounds on control of growth of F. langsethiae and on production of T-2 and HT-2 toxins in oat-based media.     

Molecular studies to identify the Fusarium species responsible for HT-2 and T-2 mycotoxins in UK oats

High levels of Fusarium mycotoxins HT-2 and T-2 have been detected in UK oats since surveys started in 2002. Fusarium langsethiae and the closely related species F. sporotrichioides have previously been associated with the contamination of cereals with type A trichothecenes HT-2 and T-2 in Nordic countries. Preliminary microbiological analysis of UK oat samples with high concentrations of HT-2 and T-2 detected and isolated F. langsethiae and F. poae but not the other type A trichothecene producing species F. sporotrichioides, F. sibiricum and F. armeniacum. Two hundred and forty oat flour samples with a known mycotoxin profile were selected from a previous four year study (2002-2005) to cover the full concentration range from below the limit of quantification (<20 μg/kg) to 9,990 μg/kg HT-2+T-2 combined. All samples were analysed for the DNA of F. langsethiae, F. poae and F. sporotrichioides based on previously published PCR assays. F. langsethiae was detectable in nearly all samples; F. poae was detected in 90% of samples whereas F. sporotrichioides was not detected in any sample. A real-time PCR assay was developed to quantify F. langsethiae DNA in plant material. The assay could quantify as low as 10(-4)ngF. langsethiae DNA/μl. Based on this assay and a previously published assay for F. poae, both species were quantified in the oat flour samples with known HT-2+T-2 content. Results showed a good regression (P<0.001, r(2)=0.60) between F. langsethiae DNA and HT-2+T-2 concentration. F. poae DNA concentration was not correlated to HT2+T2 concentration (P=0.448) but was weakly correlated to nivalenol concentration (P<0.001, r(2)=0.09). Multiple regression with F. langsethiae and F. poae DNA as explanatory variates identified that both F. langsethiae and F. poae DNA were highly significant (P<0.001) but F. poae DNA only accounted for an additional 4% of the variance and the estimate was negative, indicating that higher concentrations of F. poae DNA were correlated with slightly lower concentrations of HT2+T2 detected. A stronger regression (P<0.001, r(2)=0.77) between F. langsethiae DNA and HT-2+T-2 was obtained after extraction and quantification of DNA and mycotoxins from individual oat grains. The results from this study provide strong evidence that F. langsethiae is the primary, if not sole, fungus responsible for high HT-2 and T-2 in UK oats.     

Characteristics of commercially heat-treated oat groats

Oat groats, sampled at various steps in the commercial production of oat flour, were examined to determine the effects of heat treatment on oat quality. The samples were: original cleaned milling oats, conditioned groats, dried groats and cutmeal. The amount of oat protein solubilized by different concentrations of sodium dodecanoate was indicative of the degree of heat treatment received during processing. Pasting characteristics of conditioned groats were lower than those of other oat samples. Dry groats and cutmeal samples had similar protein solubility and pasting characteristics. Lipase activity of grain samples, except cutmeal, were lower than those of flours obtained from them; that of dried groats was negligible. Differences were observed in the three endothermic transitions (starch, protein and amylose-lipid complex) in the differential scanning calorimetry (DSC) thermograms of oat samples.     

Less Fusarium infestation and mycotoxin contamination in organic than in conventional cereals

A total of 602 samples of cereals, consisting of organically and conventionally produced barley, oats and wheat, were collected at harvest during 2002–2004 in Norway. Organic and conventional cereals were sampled in comparable numbers regarding cereal species, localisation and harvest time, and analysed for Fusarium mould and mycotoxins. Fusarium infestation and mycotoxin content were dependent on cereal species and varied year-by-year. However, in all cereal species, Fusarium infestation and levels of important mycotoxins were significantly lower when grown organically than conventionally. Concerning the most toxic trichothecenes, HT-2 and T-2 toxin, lower concentrations were found in organic oats and barley. Wheat was not contaminated by HT-2 and T-2, but lower concentrations of deoxynivalenol (DON) and moniliformin (MON) were found when organically produced. For mycotoxins considered to constitute the main risk to humans and animals in Norwegian cereals, i.e. HT-2 in oats and DON in oats and wheat, the median figures (mean levels in brackets) were as follows: HT-2 in organic and conventional oats were <20 (80) and 62 (117) µg/kg, DON in organic and conventional oats were 24 (114) and 36 (426) µg/kg, and DON in organic and conventional wheat were 29 (86) and 51 (170) µg/kg, respectively. Concentrations of HT-2 and T-2 in the samples were strongly correlated (r = 0.94). Other mycotoxins did not show a significant correlation to each other. Both HT-2 and T-2 concentrations were significantly correlated with infestation of F. langsethiae (r = 0.65 and r = 0.60, respectively). Concentrations of DON were significantly correlated with F. graminearum infestation (r = 0.61). Furthermore, nivalenol (NIV) was significantly correlated with infestation of F. poae (r = 0.55) and MON with F. avenaceum (r = 0.37). As lower Fusarium infestation and mycotoxin levels were found in organic cereals, factors related to agricultural practice may reduce the risk of contamination with Fusarium mycotoxins. Studies of these issues will be presented separately.     

Study of the natural occurrence of T-2 and HT-2 toxins and their glucosyl derivatives from field barley to malt by high-resolution Orbitrap mass spectrometry

This paper reports a new method for the determination of T-2 and HT-2 toxins and their glucosylated derivatives in cereals, and some survey data aimed at obtaining more comprehensive information on the co-occurrence of T-2 and HT-2 toxins and their glucosylated derivatives in naturally contaminated cereal samples. For these purposes, barley samples originating from a Northern Italian area were analysed by LC-HRMS for the presence of T-2, HT-2 and relevant glucosyl derivatives. Quantitative analysis of T-2 and HT-2 glucosides was performed for the first time using a recently made available standard of T-2 glucoside. The glucosyl derivative of HT-2 was detected at levels up to 163 µg kg^–1 in 17 of the 18 analysed unprocessed barley grains, whereas the monoglucosyl derivative of T-2 toxin was detected in only a few samples and at low µg kg^–1 levels. The ratio between glucosylated toxins (sum of T-2 and HT-2 glucosides) and native toxins (sum of T-2 and HT-2) ranged from 2% to 283%. Moreover, taking advantage of the possibility of retrospective analysis of full-scan HRMS chromatograms, samples were also screened for the presence of other type-A trichothecenes, namely neosolaniol, diacetoxyscirpenol and their monoglucosyl derivatives, which were detected at trace levels. A subset of nine different samples was subjected to micro-maltation in order to carry out a preliminary investigation on the fate of T-2, HT-2 and relevant glucosides along the malting process. Mycotoxin reduction from cleaned barley to malt was observed at rates ranging from 4% to 87%.     

Fusarium mycotoxins in cereals harvested from Hungarian fields

The Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEN) and T-2 frequently contaminate grain crops in Middle and Eastern Europe. In this survey, 116 cereal samples (maize, wheat, barley and oat) were examined for DON, ZEN and T-2 mycotoxins. Samples were collected from different areas in two Hungarian regions (North and South Transdanubia). The method of analysis was indirect competitive ELISA. Maize was the most contaminated grain regarding DON (86%), ZEN (41%) and T-2 (55%) toxins. The average results of the deoxynivalenol and zearalenone tests of maize proved to be significantly higher than those of barley or oat. DON was the most represented Fusarium mycotoxin followed by T-2 and ZEN. The examination of these mycotoxins would be necessary at a larger scale as to re-evaluate permissible levels, so increase of the monitoring programme would be advisable for the future.     

Occurrence of mycotoxins in spelt and common wheat grain and their products

Organic farming does not allow the use of conventional mineral fertilizers and crop protection products. As a result, in our experiments we chose to grow different species of cereals and to see how cereal species affect mycotoxin accumulation. This study describes the occurrence of deoxynivalenol (DON), zearalenone (ZEA) and T-2/HT-2 toxin in a survey of spelt and common wheat and their bran as well as flour. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method. The concentrations of DON, ZEA and T-2/HT-2 in Triticum spelta and T. aestivum were influenced by species, cereal type and year interaction. The highest concentrations of these mycotoxins were found in spelt grain with glumes, in spelt glumes and in spring wheat. These results show significantly higher concentrations of Fusarium toxins in glumes than in dehulled grain, which indicates the possible protective effect of spelt wheat glumes. The lowest DON, ZEA and T-2/HT-2 concentrations were determined in spelt grain without glumes. The research shows that it is potentially risky to produce bran from grain in which mycotoxin concentrations are below limits by European Union Regulation No. 1881/2006, since the concentration of mycotoxins in bran can be several times higher than that in grain. As a result, although bran is a dietary product characterised by good digestive properties, it can become a harmful product that can cause unpredictable health damage.     

Fate of deoxynivalenol,T-2 and HT-2 toxins and their glucoside conjugates from flour to bread: an investigation by high-performance liquid chromatography high-resolution mass spectrometry

Deoxynivalenol, T-2 and HT-2 toxins are mycotoxins frequently occurring in cereals and cereal-based products along with their conjugated forms. In this paper, we provide insights into the fate of deoxynivalenol, T-2 and HT-2 toxins and their glucoside derivatives during bread making, using naturally contaminated wheat flour. High-resolution mass spectrometry was used to assess the extent of degradation of the three mycotoxins during bread baking and to identify some glucoside conjugates, namely deoxynivalenol, T-2 and HT-2 mono-glucosides, detected both in the flour and in the respective breads. Our findings show deoxynivalenol's levels markedly increased upon baking, whereas those of HT-2 and T-2 toxins were decreased in the final bread with special regard to the T-2 toxin.     

Fusarium langsethiae: Storage environment influences dry matter losses and T2 and HT-2 toxin contamination of oats

The aim of this study was to determine the environmental conditions over which Fusarium langsethiae species can (a) grow, (b) produce T-2 and HT-2 toxins in oats, and (c) to determine the relationship between dry matter losses (DMLs) and mycotoxin accumulation in stored oats for the first time. Oats were stored under different combinations of water activity (0.89–0.97 a_w) and temperature (15–30 °C) and the amount of CO₂ produced was measured on a daily basis by gas chromatography (GC). These data were used to quantify the effect of storage on dry matter losses. Results showed that the optimum conditions for colonisation of oats and T-2 and HT-2 toxin production by F. langsethiae were at 0.97 a_w and 25 °C. T-2 and HT-2 contamination exceeded 4 mg/kg and 0.75 mg/kg respectively after 10 days storage at these conditions and the calculated DMLs were also higher, exceeding 2%. At intermediate (0.945) and suboptimal (0.89) a_w levels for growth of F. langsethiae, DMLs were higher at 30 °C than 25 °C. Models were developed to relate quality losses and toxin production to the different storage conditions. This allows the identification of high and low risk conditions for storage of oats. A good positive correlation was obtained between combined T-2 and HT-2 toxin production and DMLs. This suggests that in situ measurements of CO₂ production during storage may be useful in predicting toxin accumulation in stored oats.     

Occurrence of Fusarium T-2 and HT-2 toxins in oats from cultivar studies in Germany and degradation of the toxins during grain cleaning treatment and food processing

For the Fusarium trichothecene mycotoxins T-2 and HT-2, a combined (T-2 + HT-2) temporary tolerable daily intake (tTDI) of 0.06 µg kg^?1 body weight day^?1 was proposed at the European level in 2001 (Opinion of the Scientific Committee on Food). In the near future, maximum levels for these trichothecenes will be regulated by the European Commission as announced in EU (VO) 1881/2006. For the implementation of these maximum levels, more data on occurrence and behaviour of T-2 and HT-2 toxins in primary agricultural products as well as during cleaning treatment and food processing are needed. In the current work, we determined the T-2/HT-2 concentrations in four oat cultivars (Aragon, Dominik, Ivory, Pergamon) from ten different agricultural sites in Germany, grown in cultivar studies in 2007. The grains were de-hulled, oat meal was prepared, and bread with 20% oat meal and 80% wheat flour was baked. In the cereal-processing chain, samples were taken at various steps and subsequently analysed for their T-2/HT-2 content. We employed liquid chromatography-mass spectrometry (LC-MS) and an immunological screening method (enzyme-linked immunoabsorbant assay (ELISA)) for T-2/HT-2 determination. Detection limits were between 1 and 10 µg kg^?1 in different matrices. T-2/HT-2 concentrations determined by ELISA in oat samples from ten different agricultural sites in Germany were between 9 and 623 µg kg^?1. The median and 90th percentile were 48 and 191 µg kg^?1 T-2/HT-2, respectively. One site showed six times higher T-2/HT-2 levels than the other sites, where concentrations ranged from 322 to 623 µg kg^?1. In 80% of the samples the cultivars Pergamon and Ivory had the lowest concentration of T-2 and HT-2 toxins. Using LC-MS for T-2/HT-2 determination, cleaning of the raw material did not lead to significant reductions of T-2 and HT-2 levels, whereas de-hulling led to a reduction of over 90%. Boiling of oat meal produced from cleaned raw material to yield ‘porridge’ resulted in varying T-2/HT-2 levels in experimental replicates. No major reduction of T-2/HT-2 levels in cooked porridge was obtained. Standardized baking experiments using 20% oat meal showed that T-2 and HT-2 toxins are relatively stable during the baking process, probably due to their temperature stability.     

Fusarium mycotoxin content of UK organic and conventional wheat

Each year (2001–2005), 300 samples of wheat from fields of known agronomy were analysed for ten trichothecenes by gas chromatography-mass spectrometry (GC/MS) including deoxynivalenol (DON), nivalenol, 3-acetyl-DON, 15-acetyl-DON, fusarenone X, T2 toxin, HT2 toxin, diacetoxyscirpenol, neosolaniol and T-2 triol and zearalenone by high-performance liquid chromatography (HPLC). Of the eleven mycotoxins analysed from 1624 harvest samples of wheat, only eight were detected, and of these only five–deoxynivalenol, 15-acetyl-DON, nivalenol, HT-2 and zearalenone–were detected above 100 µg kg^?1. DON was the most frequently detected Fusarium mycotoxin, present above the limit of quantification (10 µg kg^?1) in 86% of samples, and was usually present at the highest concentration. The percentage of samples that would have exceeded the recently introduced legal limits varied between 0.4% and 11.3% over the five-year period. There was a good correlation between DON and zearalenone concentrations, although the relative concentration of DON and zearalenone fluctuated between years. Year and region had a significant effect on all mycotoxins analysed. There was no significant difference in the DON concentration of organic and conventional samples. There was also no significant difference in the concentration of zearalenone between organic and conventional samples, however organic samples did have a significantly lower concentration of HT2 and T2. Overall, the risk of UK wheat exceeding the newly introduced legal limits for Fusarium mycotoxins in cereals intended for human consumption is low, but the percentage of samples above these limits will fluctuate between years.     

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.