Trichothecenes and zearalenone production by Fusarium equiseti and Fusarium semitectum species isolated from Argentinean soybean

Fusarium equiseti and Fusarium semitectum represent the most abundant species in the Fusarium complex isolated from flowers, soybean pods and seeds in Argentina. The aim of the present study was to assess the production of major type A and type B trichothecenes (diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin, nivalenol, deoxynivalenol) and zearalenone by 40?F. equiseti and 22 F. semitectum isolates on rice culture. Mycotoxins were determined by HPLC with fluorescence detection after derivatisation with 1-anthronylnitrile for type A trichothecenes (i.e. diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin), by HPLC with UV detection for type B trichothecenes (i.e. nivalenol and deoxynivalenol), and by TLC for zearalenone. A total of 22 of 40?F. equiseti isolates produced diacetoxyscirpenol, nivalenol and ZEA alone or in combination, whereas only two of 20?F. semitectum isolates were nivalenol and ZEA producers. Both Fusarium species did not produce any deoxynivalenol, neosolaniol, T-2 toxin and HT-2 toxin. The variable retention in toxigenicity displayed by both fungal species suggests that these species have a saprophytic lifestyle in the soybean agroecosystem in Argentina.     

Trichothecenes and zearalenone production by Fusarium equiseti and Fusarium semitectum species isolated from Argentinean soybean

Fusarium equiseti and Fusarium semitectum represent the most abundant species in the Fusarium complex isolated from flowers, soybean pods and seeds in Argentina. The aim of the present study was to assess the production of major type A and type B trichothecenes (diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin, nivalenol, deoxynivalenol) and zearalenone by 40?F. equiseti and 22 F. semitectum isolates on rice culture. Mycotoxins were determined by HPLC with fluorescence detection after derivatisation with 1-anthronylnitrile for type A trichothecenes (i.e. diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin), by HPLC with UV detection for type B trichothecenes (i.e. nivalenol and deoxynivalenol), and by TLC for zearalenone. A total of 22 of 40?F. equiseti isolates produced diacetoxyscirpenol, nivalenol and ZEA alone or in combination, whereas only two of 20?F. semitectum isolates were nivalenol and ZEA producers. Both Fusarium species did not produce any deoxynivalenol, neosolaniol, T-2 toxin and HT-2 toxin. The variable retention in toxigenicity displayed by both fungal species suggests that these species have a saprophytic lifestyle in the soybean agroecosystem in Argentina.     

Studies into the occurrence of some trichothecene mycotoxins in UK home-grown wheat and in imported wheat

A total of 199 UK home-grown wheat samples collected over three harvests (1980–82 inclusive) and 33 imported wheat samples were analysed for the presence of seven trichothecene mycotoxins (nivalenol, deoxynivalenol, fusarenon-x, neosolaniol, diacetoxyscirpenol, HT-2 toxin and T-2 toxin). Analysis was performed by a gas–liquid chromatographic method and positive results greater than 0.1 mg kg^?1 were confirmed by mass spectrometry. The only mycotoxin detected in any of the samples was deoxynivalenol (vomitoxin) which occurred in 32 out of 199 UK home-grown wheats at levels ranging from 0.02 to 0.40 mg kg^?1 and 23 out of 33 imported wheats at levels ranging from 0.02 to 1.32 mg kg^?1. Microbiological evidence suggests that the lower incidence and levels of deoxynivalenol in UK, other EEC and Western Canadian wheat compared with Eastern Canadian and Midwest US wheat may be caused by a geographical variation in the distribution of Fusarium species.     

Survey of Fusarium toxins in foodstuffs of plant origin marketed in Germany

A total of 219 samples of foodstuffs of plant origin, consisting of grain-based food, pseudocereals and gluten-free food as well as vegetables, fruits, oilseeds and nuts, were randomly collected during 2000 and 2001 in food and health food stores. A spectra of 13 trichothecene toxins including diacetoxyscirpenol (DAS), 15-monoacetoxyscirpenol (MAS), scirpentriol (SCIRP), T-2 and HT-2 toxins (T-2, HT-2), T-2 triol, T-2 tetraol, neosolaniol (NEO) of the A-type as well as deoxynivalenol (DON), 3- and 15-acetyl-DON (3-, 15-ADON), nivalenol (NIV), and fusarenon-X (FUS-X) of the B-type were determined by gas chromatography/mass spectrometry. Analysis of zearalenone (ZEA), alpha- and beta-zearalenol (alpha- and beta-ZOL) was made by high-performance liquid chromatography with fluorescence and UV-detection. Detection limits ranged between 1 and 19 microg/kg. Out of 84 samples of cereal-based including gluten-free foods, 60 samples were positive for at least one of the toxins DON, 15-ADON, 3-ADON, NIV, T-2, HT-2, T-2 tetraol and ZEA, with incidences at 57%, 13%, 1%, 10%, 12%, 37%, 4% and 38%, respectively, whereas SCIRP and its derivatives MAS and DAS, T-2 triol, Fus-X as well as alpha- and beta-ZOL were not detected in any sample of this subgroup. Contents of DON ranged between 8 and 389 microg/kg, for all other toxins determined concentrations were below 100 microg/kg. The pseudocereals amaranth, quinoa and buckwheat were free of the toxins investigated. Ten of 85 samples of vegetables and fruits were toxin positive. ZEA and the type A trichothecenes MAS, SCIRP, DAS, HT-2 were detected in 7, 3, 2, 1 and 1 samples, respectively. Out of 35 samples of oilseeds and nuts, 7 samples were toxin positive. HT-2, T-2 and ZEA were detected in 4, 3 and 4 samples, respectively. In vegetables and fruits as well as in oilseeds and nuts, toxin levels were below 50 microg/kg. None of the B-type trichothecenes analysed was found for both subgroups.     

Occurrence of Fusarium species and trichothecenes in Nigerian maize

A total of 180 maize samples meant for human consumption from four maize-producing states of southwestern Nigeria were screened for twelve major Fusarium mycotoxins (trichothecenes). Mycological examination of the samples showed that Fusarium verticillioides was the most commonly isolated fungi (71%), followed by F. sporotrichioides (64%), F. graminearum (32%), F. pallidoroseum (15%), F. compactum (12%), F. equiseti (9%), F. acuminatum (8%), F. subglutinans (4%) and F. oxysporum (1%). The trichothecenes include deoxynivalenol (DON), 3, mono-acetyldeoxynivalenol (3-AcDON), 15, mono-acetyldeoxynivalenol (15-AcDON), nivalenol (NIV), HT-2 toxin (HT-2), neosolaniol (NEO), T-2 toxin (T-2), T-2 tetraol and T-2 triol, diacetoxyscirpenol (DAS), MAS-monoacetoxyscirpenol (MAS) and fusarenone-X. Quantification was by high performance liquid chromatography coupled with mass spectroscopy (HPLC/MS); the detection limits for each of the mycotoxins varied between 20 and 200 microg kg(-1). Sixty six samples (36.3%) were contaminated with trichothecenes, DON (mean: 226.2 microg kg(-1); range: 9.6-745.1 microg kg(-1)), 3-AcDON (mean: 17.3 microg kg(-1); range: 0.7-72.4 microg kg(-1)) and DAS (mean: 16.0 microg kg(-1); range: 1.0-51.0 microg kg(-1)) were detected in 22%, 17% and 9% of total samples respectively. There were no 15-AcDON, NIV, HT-2, NEO, T-2, T-2 tetraol, T-2 triol, MAS and fusarenone-X detected. This is the first comprehensive report about the natural occurrence of DON, AcDON and DAS in maize for direct human consumption in Nigeria.     

进口玉米酒糟粕中15种真菌毒素的检测和风险分析

建立了液相串联质谱检测玉米酒糟粕中15种真菌毒素的检测方法,并对结果分布进行了讨论。检测的真菌毒素包括黄曲霉毒素B1等15种真菌毒素。这些毒素采用溶剂提取,多功能净化柱净化,液相串联质谱检测。方法均经过优化和验证,满足进口玉米酒糟粕的检测要求。对67个进口玉米酒糟粕样品的检测结果表明,玉米赤霉烯酮、脱氧雪腐镰刀菌烯醇、T-2毒素、HT-2毒素等真菌毒素均有检出。本试验对这些数据进行了统计分析,提出了对进口玉米酒糟粕的真菌毒素风险分析结果。     

A new solid phase extraction clean-up method for the determination of 12 type A and B trichothecenes in cereals and cereal-based food by LC-MS/MS

A new reliable and cost-efficient solid phase extraction-based clean-up method for the determination of 12 type A and B trichothecenes [deoxynivalenol (DON), nivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, fusarenon-X, T-2 toxin, HT-2 toxin, neosolaniol, monoacetoxy-scirpenol, diacetoxyscirpenol, T-2 triol and T-2 tetraol] in cereals and cereal-based food is presented. Furthermore, the suitability for the simultaneous determination of zearalenone is examined. Toxins were extracted from cereal samples using ACN/water (80/20, v/v), purified by means of a new Bond Elut Mycotoxin column and analyzed via liquid chromatography-electrospray ionization tandem mass spectrometry. Limits of detection were calculated for the matrix wheat and ranged from 0.3 to 5 ng/g, depending on the toxin. Average recovery rates for the tested compounds in seven cereal-based matrices have been determined ranging from 65 to 104%. The relative standard deviations of the complete method ranged from 2.67 (DON, wheat) to 20.0% (T-2 toxin, oats).     

Mycotoxin contamination of sorghum and its contribution to human dietary exposure in four sub-Saharan countries

This research aimed at evaluating the safety, and the type, level and prevalence of mycotoxins in grain sorghum of four sub-Saharan African (SSA) countries (Burkina Faso, Ethiopia, Mali and Sudan). A multi-analyte LC-MS/MS method for quantification of 23 mycotoxins (nivalenol, deoxynivalenol, fusarenon X, neosolaniol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, roquefortine C, HT-2 toxin, alternariol, T-2 toxin, FB1, FB2, FB3, zearalenone, aflatoxin G₁, aflatoxin G₂, aflatoxin B₁, aflatoxin B₂, sterigmatocystin, OTA, altenuene, alternariol monomethylether) was applied to different sorghum matrices. Of the 1533 analysed samples, 33% were contaminated with at least one of the following mycotoxins: aflatoxins, fumonisins, sterigmatocystin, Alternaria toxins, OTA and zearalenone. Country of origin, colour, source and collection period of sorghum samples significantly influenced the type, level and prevalence of mycotoxins. Sterigmatocystin (15%), fumonisins (17%) and aflatoxins (13%) were the most prevalent. FB1 (274 ± 585 µg/kg) had the highest mean concentration followed by FB2 (214 ± 308 µg/kg) while diacetoxyscirpenol (8.12 ± 19.2 µg/kg) and HT-2 (11.9 ± 0.00 µg/kg) had the lowest concentrations. Neosolaniol, fusarenon-X, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, T-2 toxin, nivalenol and roquefortine C were not detected in any of the samples. Sudan had the lowest prevalence and mean concentration of all mycotoxins. Pink sorghum had the highest concentrations of fumonisins and aflatoxins. Mycotoxins from Aspergillus spp. and Alternaria spp. are the mycotoxins of concern in SSA grain sorghum with regard to prevalence, concentration and possible health risk from exposure. Based on the performed risk characterisation, daily consumption of sorghum containing aflatoxins, alternariol, alternariol monomethyl ether, sterigmatocystin and OTA could result in exceeding the established health-based guidance values for these toxins.     

Production of trichothecene mycotoxins by Australian Fusarium species.

Australian isolates of Fusarium species were grown on potato dextrose agar. Trichothecenes produced by these species were extracted by ethyl acetate followed by methanol and a silica gel column was used to clean-up the extract. The extracted samples were derivatized by acetylation with trifluoroacetic anhydride and the derivatives analysed by gas chromatography/mass spectrometry (GC/MS). Multiple ion detection was used to trace ions characteristic of the trichothecenes expected to be present. Quantitation of those found was based on a known mass of pentabromophenol that was added as an internal standard. Eight species of Fusarium (nineteen strains) were surveyed, of which three species, F. acuminatum, F. equiseti and F. sporotrichioides, produced the trichothecenes scirpentriol, diacetoxyscirpenol, neosolaniol, HT-2 toxin, T-2 toxin, T-2 tetraol and deoxynivalenol. Wheat samples were inoculated with four different species of Fusarium, F. acuminatum, F. equiseti, F. graminearum and F. sporotrichioides, and in these samples diacetoxyscirpenol, neosolaniol, HT-2 toxin and T-2 toxin were found.     

Analysis of trichothecenes in barley tea and beer by liquid chromatography/tandem mass spectrometry

A simple method for analysis of trichothecenes [Type A: diacetoxyscirpenol, neosolaniol, HT-2 toxin, and T-2 toxin, Type B: deoxynivalenol, nivalenol, fusarenon-X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivarenol] in barley tea and beer using liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed. Trichothecenes were extracted with ethyl acetate-methanol (19:1). The solvent was evaporated to dryness and the residue was dissolved in water-methanol (3:1) for injection into the LC/MS/MS. The LC separation was performed with an octadecylated silica column at a flow-rate of 0.2 mL/min, using a mobile phase consisting of water, methanol and acetonitrile. MS/MS was used in multiple reaction monitoring, employing electrospray ionization (ESI-MRM). The recoveries of trichothecenes from drinks at 1 microg/L (Type A) and 10 microg/L (Type B) were 52.5-115.2% (barley tea) and 68.1-127.5% (beer). Five barley tea and ten beer samples were analyzed by this method. Trichothecenes were not detected in them. This method may have applications in quality assurance.     

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.     

Incidence of trichothecenes and zearalenone in poultry feed mixtures from Slovakia

A total of 50 samples of poultry feed mixtures of Slovakian origin were analyzed for eight toxicologically significant Fusarium mycotoxins, namely zearalenone (ZON), A-trichothecenes: diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2) and B-trichothecenes: deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON) and nivalenol (NIV). The A-trichothecenes and the B-trichothecenes were detected by means of high pressure liquid chromatography with tandem mass spectrometry detection (HPLC-MS/MS) and gas chromatography electron capture detection (GC-ECD), respectively. Reversed phase-high performance liquid chromatography with a fluorescence detector (RP-HPLC-FLD) was used for ZON detection. The most frequent mycotoxin detected was T-2, which was found in 45 samples (90%) in relatively low concentrations ranging from 1 to 130 microg kg(-1) (average 13 microg kg(-1)), followed by ZON that was found in 44 samples (88%) in concentrations ranging from 3 to 86 microg kg(-1) (average 21 microg kg(-1)). HT-2 and DON were detected in 38 (76%) and 28 (56%) samples, respectively, in concentrations of 2 to 173 (average 18 microg kg(-1)) for HT-2 and 64 to 1230 microg kg(-1) sample (average 303 microg kg(-1)) for DON. The acetyl-derivatives of DON were in just four samples, while NIV was not detected in any of the samples investigated. In as many as 22 samples (44%), a combination of four simultaneously co-occurring mycotoxins, i.e. T-2, HT-2, ZON and DON, was revealed. Despite the limited number of samples investigated during this study poultry feed mixtures may represent a risk from a toxicological point of view and should be regarded as a potential source of the Fusarium mycotoxins in Central Europe. This is the first reported study dealing with zearalenone and trichothecene contamination of poultry mixed feeds from Slovakia.     

Fusarium mycotoxin content of UK organic and conventional barley

Each year (2002–2005), approximately 100 samples of barley 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, T-2 toxin (T2), HT-2 toxin (HT2), diacetoxyscirpenol, neosolaniol, and T-2 triol. Samples were also analysed for moniliformin and zearalenone by high-performance liquid chromatography (HPLC). Of the ten trichothecenes analysed from 446 harvest samples of barley, only two, diacetoxyscirpenol and neosolaniol, were not detected. The concentrations of type A trichothecenes were similar to those that occurred in wheat over the same period, whilst those of type B trichothecenes were markedly lower. Deoxynivalenol was the most frequently detected Fusarium mycotoxin, present above the limit of quantification (10 µg kg^?1) in 57% of samples, and was usually present at the highest concentration. A single sample (0.2%) exceeded the legal limit for DON in unprocessed barley over the 4-year period. Moniliformin and zearalenone were both rarely detected (2% of samples greater than 10 µg kg^?1 for both toxins) with maximum concentrations of 45 and 44 µg kg^?1, respectively. Year and region had a significant effect on DON and HT2 + T2, but there was no significant difference in the concentration of these mycotoxins between organic and conventional samples. Overall, the risk of UK barley exceeding the newly introduced legal limits for Fusarium mycotoxins in cereals intended for human consumption is very low, but the percentage of samples above these limits will fluctuate between years.     

An LC-IT-MS/MS-Based Method to Determine Trichothecenes in Grain Products

The aim of this work was to evaluate the usefulness of the ion trap mass spectrometry coupled to high-performance liquid chromatography for simultaneous determination of selected trichothecenes (nivalenol, deoxynivalenol, fusarenon-X, neosolaniol, 3-acetyl-deoxynivalenol, diacetoxyscirpenol, HT-2 and T-2 toxins) in grain products. These compounds were extracted from the grain products and then cleaned up with the developed, simple and robust procedure using some mixture of neutral alumina, charcoal and diatomaceous earth. Method recovery was 88–125 % depending on combination of the analysed mycotoxins, sample matrix and the fortification level. Method precision expressed by relative standard deviation ranged from 2.6 to 27.4 %. The concentrations of the selected trichothecenes have been determined in 94 samples of cereal-based products. Maize-based next to wheat-based products were the most contaminated with deoxynivalenol, neosolaniol, 3-acetyl-deoxynivalenol, diacetoxyscirpenol and HT-2 toxin. In 83 % of wheat-based products, deoxynivalenol was determined at the average level of 249 μg kg^?1. The highest concentration of deoxynivalenol—2,026 μg kg^?1 (476?±?471 μg kg^?1 on the average)—was found in the maize-based product. Other mycotoxins were found much less frequently: 3-acetyl-deoxynivalenol in only one sample at the concentration of 59 μg kg^?1, neosolaniol, HT-2 toxin and diacetoxyscirpenol in a few samples on average concentrations close to respective limits of quantification.     

Determination of trichothecenes in duplicate diets of young children by capillary gas chromatography with mass spectrometric detection.

Trichothecenes are mycotoxins produced by several fungal genera, mainly Fusarium species, that can contaminate a wide range of cereals used for human and animal consumption. They are associated with various adverse health effects in animals and humans such as feed refusal, vomiting and immunotoxic effects. A method based on capillary gas chromatography with mass spectrometric detection was developed and validated in-house for the determination of nine trichothecenes in duplicate diets of young children. The trichothecenes were extracted from the sample matrix by water/ethanol (90/10). The extracts were cleaned by means of ChemElut and Mycosep columns. The cleaned extracts were evaporated to dryness and derivatized to trimethylsilyl ethers at room temperature. The residues were dissolved in iso-octane and washed with water. The final extracts were analysed for trichothecenes by GC-MS. The response was linear in the range tested (1-10 microg kg(-1)). Recoveries for the trichothecenes were between 70 and 111%, with the exception of nivalenol, which had a low recovery (34%). The limit of quantification for all trichothecenes was below 0.4 microg kg(-1). Seventy-four food samples from young children collected by 74 respondents in a duplicate diet study were analysed for trichothecenes with the developed method. The mean levels of deoxynivalenol, nivalenol, HT-2 toxin and T-2 toxin were 5.8, 0.3, 0.3 and 0.1 microg kg(-1), respectively. Based on the individual results, dietary intake calculations were made. For deoxynivalenol, the tolerable daily intake of 1 microg kg(-1) body weight was exceeded by nine respondents. For the combined intake of T-2 and HT-2 toxin, the temporary tolerable daily intake of 0.06 microg kg(-1) body weight was exceeded by nine respondents.     

Natural occurrence of zearalenone and trichothecene toxins in maize-based animal feeds in zambia

A year-long collection of maize-based animal feed samples from the National Milling Company and mouldy maize collected from farmers fields near Lusaka were analysed for Fusarium mycotoxins. In the survey, 148 samples were tested for zearalenone, deoxynivalenol and nivalenol, and 60 samples for T-2 toxin and diacetoxyscirpenol. Zearalenone was present up to 0.6 mg kg^?1 in 17% of the feed samples, and deoxynivalenol was found at I-0 mg kg^?1 in 1.4 % of these samples. This is the first report of these toxins in animal feeds in Zambia. Zearalenone was also found in 57.6 % of the 33 mouldy maize samples collected at levels ranging from 0.08 to 6.0 mg kg^?1 (mean concentration 1.11 mg kg^?1), and 49.5% of these samples contained deoxynivalenol at levels ranging from 0.5 to 16.0 mg kg^?1 (mean concentration 5.56 mg kg ^?1). T-2 toxin and diacetoxyscirpenol were not detected.     

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.     

A sensitive enzyme-linked immunosorbent assay for detection of T-2 toxin with monoclonal antibodies

Six monoclonal antibodies (mAbs, T-2.1, 2, 3, 4, 5, 6) which react with a trichothecene mycotoxin, T-2 toxin (T-2), were prepared. All antibodies specifically reacted with T-2 but less (0.5% of T-2) with the metabolites such as HT-2 toxin and 3'-hydroxy-T-2 toxin. Significant but less than 0.02% cross-reactivity was observed with T-2 triol, 3'-hydroxy-HT-2 toxin and neosolaniol. No significant reaction with other trichothecenes such as deoxynivalenol, nivalenol, fusarenon-X, crotocin, or roridin A was observed. The least detectable amount of T-2 with the best mAb T-2.1 was 2.5 pg T-2 per assay. This specific and highly sensitive assay for T-2 was applied for the quantitation of T-2 in wheat flour spiked with mycotoxin, with combination of a simple extraction procedure.     

A rapid multiresidual determination of type A and type B trichothecenes in wheat flour by HPLC-ESI-MS

A new, rapid and sensitive method is reported for the multiresidual determination of type A (diacetoxyscirpenol, HT-2 toxin, T-2 toxin) and type B (nivalenol, deoxynivalenol, fusarenon X, 15-O-acetyl-4-deoxynivalenol) trichothecenes in wheat flour samples. Sample extraction was performed with acetonitrile/water mixtures. Mycosep^© columns were used for a fast and effective clean-up procedure. The analytes were separated by HPLC with a RP C18 column by means of a gradient elution and detected in an ESI-interfaced single quadrupole mass spectrometer. Type B and type A trichothecenes were monitored in the negative and in the positive ion mode, respectively. The method performance is reported in terms of linearity (r² = 0.999), specificity, accuracy (recoveries from 70-120%) and precision (CV% = 5), the LOQs are in the range 10-20 µg/Kg.     

Development of a new method for the simultaneous determination of 21 mycotoxins in coffee beverages by liquid chromatography tandem mass spectrometry

A new method for the simultaneous detection of 21 mycotoxins (ochratoxin A, aflatoxin B₁, aflatoxin B_2, aflatoxin G₁, aflatoxin G₂, sterigmatocystin, nivalenol, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, diacetoxyscirpenol, neosolaniol, HT-2 toxin, T-2 toxin, fumonisin B₁, fumonisin B₂, enniatin A, enniatin A₁, enniatin B, enniatin B₁, and beauvericin) in coffee beverages was internally validated. The method is based on liquid/liquid extraction with a mixture of ethyl acetate/formic acid (95:5 v/v) and detection using triple quadrupole (QqQ) and ion trap (IT) liquid chromatography tandem mass spectrometry. The limits of detection and quantification were 0.02 to 39.64 μg/kg, respectively, and the correlation coefficients were optimal for all mycotoxins (R² ≥ 0.992). The recovery values ranged from 72% to 97%. The developed method was demonstrated in six real samples of roasted and instant coffee, caffeinated and decaffeinated coffee, and coffee with sugar added. The analyses indicate the presence of the studied mycotoxins in coffee beverages at μg/kg concentrations. Ochratoxin A, a mycotoxin that is regulated in coffee, was detected in two samples under the maximum limit established by a European legislation (CE1881/2006).