Occurrence and fate of Fusarium mycotoxins during commercial processing of oats in the UK

The commercial processing of oats is different from that of other cereals, such as wheat and maize. In northwest Europe, oats also appear to be more susceptible to contamination with HT-2 and T-2 toxins than other cereals. Mycotoxins, such as deoxynivanol and zearalenone, in cereals are already controlled by EU legislation. With regard to additional, impending legislation, this study examined HT-2 and T-2 toxins together with zearalenone, deoxynivalenol and other related toxins in a commercial oat mill and how the concentrations varied from raw oats to the final prepared oat flakes. Concentrations of each Fusarium mycotoxin fell by 90-95% during the process, with the major loss being a physical distribution occurring at the de-hulling stage. Initial studies of losses occurring at other stages, such as kilning or de-branning of prepared oat groats, suggest these to be small. The use of colour sorting after kilning showed higher concentrations of each mycotoxin in the discoloured groats. The feasibility of developing a predictive tool for the oat industry is examined.     

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.     

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.     

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.     

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.     

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.     

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.     

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).     

Occurrence of Fusarium Head Blight species and Fusarium mycotoxins in winter wheat in the Netherlands in 2009

Most recent information on the occurrence of Fusarium Head Blight species and related mycotoxins in wheat grown in the Netherlands dates from 2001. This aim of this study was to investigate the incidence and levels of Fusarium Head Blight species and Fusarium mycotoxins, as well as their possible relationships, in winter wheat cultivated in the Netherlands in 2009. Samples were collected from individual fields of 88 commercial wheat growers. Samples were collected at harvest from 86 fields, and 2 weeks before the expected harvest date from 21 fields. In all, 128 samples, the levels of each of seven Fusarium Head Blight species and of 12 related mycotoxins were quantified. The results showed that F. graminearum was the most frequently observed species at harvest, followed by F. avenaceum and M. nivale. In the pre-harvest samples, only F. graminearum and M. nivale were relevant. The highest incidence and concentrations of mycotoxins were found for deoxynivalenol, followed by zearalenone and beauvericin, both pre-harvest and at harvest. Other toxins frequently found - for the first time in the Netherlands - included T-2 toxin, HT-2 toxin, and moniliformin. The levels of deoxynivalenol were positively related to F. graminearum levels, as well as to zearalenone levels. Other relationships could not be established. The current approach taken in collecting wheat samples and quantifying the presence of Fusarium Head Blight species and related mycotoxins is an efficient method to obtain insight into the occurrence of these species and toxins in wheat grown under natural environmental conditions. It is recommended that this survey be repeated for several years to establish inter-annual variability in both species composition and mycotoxin occurrence.     

Detection of trichothecenes in animal feeds and foodstuffs during the years 1997 to 2000 in Saudi Arabia

A total of 843 commercial animal feed and foodstuff samples (465 samples of agricultural commodities and 378 samples of animal feeds) from all over the Kingdom of Saudi Arabia were collected during the years 1997 to 2000 and analyzed for type A and type B trichothecenes (diacetoxyscirpenol, neosolaniol, HT-2 toxin, T-2 toxin, nivalenol, fusarenon-x, deoxynivalenol). Levels of mycotoxins detected ranged from <2 to 4,000 microg/kg deoxynivalenol, 3.25 to 500 microg/kg fusarenon-x, 3.13 to 600 microg/kg nivalenol, 3.13 to 50 microg/kg diacetoxyscirpenol, 6.25 to 200 microg/kg neosolaniol, 3.13 to 18.75 microg/kg HT-2 toxin, and 6.25 microg/kg T-2 toxin. The study reflected the need for routine surveillance of agricultural commodities to minimize potential hazards to human health.     

Contamination of barley seeds with Fusarium species and their toxins in Spain: an integrated approach

Fusarium is a globally distributed fungal genus that includes different species pathogenic to cereals among others crops. Some of these Fusarium species can also produce toxic compounds towards animals and humans. In this work, the presence of the most important Fusarium toxins was determined in barley seeds from Spain, sampled according to European Union requirements. The results obtained were compared with the presence of mycotoxigenic species considered responsible for their synthesis by using species-specific polymerase chain reaction protocols. Fumonisins B₁ and B₂, zearalenone, trichothecenes type A (T-2 and HT-2) and trichothecenes type B (deoxynivalenol and nivalenol) were analysed by using high-performance liquid chromatography. Deoxynivalenol and zearalenone were detected in 72% and 38% of the barley samples, respectively, at levels below European Union limits in all cases. However, the co-occurrence of both toxins in 34% of the samples suggested that synergistic activity of these two mycotoxins should be evaluated. Nivalenol and HT-2/T-2 were detected at low levels in 17% and 10% of the samples, respectively. Fumonisins occurred in 34% of the samples at levels up to 300?µg/kg. This suggested that they might represent a risk in Spanish barley, and to our knowledge, this is the first report on the presence of fumonisins in barley in this country. The species-specific polymerase chain reaction assays to detect mycotoxin-producing Fusarium species showed a very consistent correlation between F. verticillioides detection and fumonisin contamination as well as F. graminearum presence and zearalenone, deoxynivalenol and nivalenol contamination in barley samples. The approach used in this study provided information of mycotoxin contamination of barley together with the identification of the fungal species responsible for their production. Detection of the species with the current polymerase chain reaction assay strategy may be considered predictive of the potential mycotoxin risk in this matrix.     

Transfer of Fusarium mycotoxins and ‘masked’ deoxynivalenol (deoxynivalenol-3-glucoside) from field barley through malt to beer

The fate of five Fusarium toxins — deoxynivalenol (DON), sum of 15- and 3-acetyl-deoxynivalenol (ADONs), HT-2 toxin (HT-2) representing the main trichothecenes and zearalenone (ZON) during the malting and brewing processes — was investigated. In addition to these ‘free’ mycotoxins, the occurrence of deoxynivalenol-3-glucoside (DON-3-Glc) was monitored for the first time in a beer production chain (currently, only DON and ZON are regulated). Two batches of barley, naturally infected and artificially inoculated with Fusarium spp. during the time of flowering, were used as a raw material for processing experiments. A highly sensitive procedure employing high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was validated for the analysis of ‘free’ Fusarium mycotoxins and DON-conjugate in all types of matrices. The method was also able to detect nivalenol (NIV), fusarenon-X (FUS-X) and T-2 toxin (T-2); nevertheless, none of these toxins was found in any of the samples. While steeping of barley grains (the first step in the malting process) apparently reduced Fusarium mycotoxin levels to below their quantification limits (5–10 µg kg^?1), their successive accumulation occurred during germination. In malt, the content of monitored mycotoxins was higher compared with the original barley. The most significant increase was found for DON-3-Glc. During the brewing process, significant further increases in levels occurred. Concentrations of this ‘masked’ DON in final beers exceeded ‘free’ DON, while in malt grists this trichothecene was the most abundant, with the DON/DON-3-Glc ratio being approximately 5:1 in both sample series. When calculating mass balance, no significant changes were observed during brewing for ADONs. The content of DON and ZON slightly decreased by a maximum of 30%. Only traces of HT-2 were detected in some processing intermediates (wort after trub removal and green beer).     

Fate of trichothecene mycotoxins during the processing: milling and baking.

Toxic secondary metabolites produced by fungi representing Fusarium genus are common contaminants in cereals worldwide. To estimate the dietary intake of these trichothecene mycotoxins, information on their fate during cereal processing is needed. Up-to-date techniques such as high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS) was used for the analysis of seven trichothecenes (deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin, 15- and 3-acetyldeoxynivalenol, and fusarenon-X) in bread production chain (wheat grains, intermediate products collected during milling and baking process, breads). Regardless of whether the grains were naturally infected or artificially inoculated by Fusarium spp. in the field, the fractions obtained from the grain-cleaning procedure contained the highest mycotoxin levels. During milling the highest concentrations of deoxynivalenol were found in the bran, the lowest in the reduction flours. Baking at 210 degrees C for 14 min had no significant effect on deoxynivalenol levels. The rheological properties of dough measured by fermentograph, maturograph, oven rise recorder, and laboratory baking test were carried out, and based on the obtained results the influence of mycotoxin content on rheological behaviour was investigated.     

Fate of trichothecene mycotoxins during the processing: milling and baking

Toxic secondary metabolites produced by fungi representing Fusarium genus are common contaminants in cereals worldwide. To estimate the dietary intake of these trichothecene mycotoxins, information on their fate during cereal processing is needed. Up-to-date techniques such as high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS) was used for the analysis of seven trichothecenes (deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin, 15- and 3-acetyldeoxynivalenol, and fusarenon-X) in bread production chain (wheat grains, intermediate products collected during milling and baking process, breads). Regardless of whether the grains were naturally infected or artificially inoculated by Fusarium spp. in the field, the fractions obtained from the grain-cleaning procedure contained the highest mycotoxin levels. During milling the highest concentrations of deoxynivalenol were found in the bran, the lowest in the reduction flours. Baking at 210 degrees C for 14 min had no significant effect on deoxynivalenol levels. The rheological properties of dough measured by fermentograph, maturograph, oven rise recorder, and laboratory baking test were carried out, and based on the obtained results the influence of mycotoxin content on rheological behaviour was investigated.     

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.     

Mycotoxins in breakfast cereals from the Canadian retail market: A 3-year survey

One hundred and fifty-six samples of breakfast cereals were collected from the Canadian retail marketplace over a 3-year period. The samples were analysed for the mycotoxins deoxynivalenol, nivalenol, HT-2 toxin, zearalenone, ochratoxin A, and fumonisins B₁ and B₂ to contribute to dietary exposure estimates in support of the development of Canadian guidelines for selected mycotoxins in foods. The samples included corn-, oat-, wheat- and rice-based cereals, as well as mixed-grain cereals, and were primarily from North American processors. Overall, deoxynivalenol was the most frequently detected mycotoxin  — it was detected in over 40% of all samples analysed. Fumonisins and ochratoxin A were each detected in over 30% of all samples. Zearalenone was detected in over 20% of all samples. Nivalenol and HT-2 toxin were each detected in only one sample. The survey clearly demonstrated regular occurrence of low levels of multiple mycotoxins in breakfast cereals on the Canadian market.     

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。     

LC-MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs.

Mycotoxin analysis is usually carried out by high performance liquid chromatography after immunoaffinity column cleanup or in enzyme-linked immunosorbent assay tests. These methods normally involve determination of single compounds only. EU legislation already exists for the aflatoxins, ochratoxin A and patulin in food, and legislation will come into force for deoxynivalenol, zearalenone and the fumonisins in 2007. To enforce the various legal limits, it would be preferable to determine all mycotoxins by routine analysis in different types of matrices in one single extract. This would also be advantageous for HACCP control purposes. For this reason, a multi-method was developed with which 33 mycotoxins in various products could be analysed simultaneously. The mycotoxins were extracted with an acetonitrile/water mixture, diluted with water and then directly injected into a LC-MS/MS system. The mycotoxins were separated by reversed-phase HPLC and detected using an electrospray ionisation interface (ESI) and tandem MS, using MRM in the positive ion mode, to increase specificity for quality control. The following mycotoxins could be analysed in a single 30-min run: Aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin, alpha-zearalenol, alpha-zearalanol, beta-zearalanol, sterigmatocystin, cyclopiazonic acid, penicillic acid, fumonisins B1, B2 and B3, diacetoxyscirpenol, 3- and 15-acetyl-deoxynivalenol, zearalanone, ergotamin, ergocornin, ergocristin, alpha-ergocryptin, citrinin, roquefortin C, fusarenone X, nivalenol, mycophenolic acid, alternariol and alternariol monomethyl ether. The limit of quantification for the aflatoxins and ochratoxin A was 1.0 microg kg(-1) and for deoxynivalenol 50 microg kg(-1). The quantification limits for the other mycotoxins were in the range 10-200 microg kg(-1). The matrix effect and validation data are presented for between 13 and 24 mycotoxins in peanuts, pistachios, wheat, maize, cornflakes, raisins and figs. The method has been compared with the official EU method for the determination of aflatoxins in food and relevant FAPAS rounds. The multi-mycotoxin method has been proven by the detection of more than one mycotoxin in maize, buckwheat, figs and nuts. The LC-MS/MS technique has also been applied to baby food, which is subject to lower limits for aflatoxin B1 and ochratoxin A, ergot alkaloids in naturally contaminated rye and freeze-dried silage samples.     

LC-MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs

Mycotoxin analysis is usually carried out by high performance liquid chromatography after immunoaffinity column cleanup or in enzyme-linked immunosorbent assay tests. These methods normally involve determination of single compounds only. EU legislation already exists for the aflatoxins, ochratoxin A and patulin in food, and legislation will come into force for deoxynivalenol, zearalenone and the fumonisins in 2007. To enforce the various legal limits, it would be preferable to determine all mycotoxins by routine analysis in different types of matrices in one single extract. This would also be advantageous for HACCP control purposes. For this reason, a multi-method was developed with which 33 mycotoxins in various products could be analysed simultaneously. The mycotoxins were extracted with an acetonitrile/water mixture, diluted with water and then directly injected into a LC-MS/MS system. The mycotoxins were separated by reversed-phase HPLC and detected using an electrospray ionisation interface (ESI) and tandem MS, using MRM in the positive ion mode, to increase specificity for quality control. The following mycotoxins could be analysed in a single 30-min run: Aflatoxins B1, B2, G1 and G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 toxin, HT-2 toxin, alpha-zearalenol, alpha-zearalanol, beta-zearalanol, sterigmatocystin, cyclopiazonic acid, penicillic acid, fumonisins B1, B2 and B3, diacetoxyscirpenol, 3- and 15-acetyl-deoxynivalenol, zearalanone, ergotamin, ergocornin, ergocristin, alpha-ergocryptin, citrinin, roquefortin C, fusarenone X, nivalenol, mycophenolic acid, alternariol and alternariol monomethyl ether. The limit of quantification for the aflatoxins and ochratoxin A was 1.0 microg kg(-1) and for deoxynivalenol 50 microg kg(-1). The quantification limits for the other mycotoxins were in the range 10-200 microg kg(-1). The matrix effect and validation data are presented for between 13 and 24 mycotoxins in peanuts, pistachios, wheat, maize, cornflakes, raisins and figs. The method has been compared with the official EU method for the determination of aflatoxins in food and relevant FAPAS rounds. The multi-mycotoxin method has been proven by the detection of more than one mycotoxin in maize, buckwheat, figs and nuts. The LC-MS/MS technique has also been applied to baby food, which is subject to lower limits for aflatoxin B1 and ochratoxin A, ergot alkaloids in naturally contaminated rye and freeze-dried silage samples.     

Fate of trichothecene mycotoxins during the processing: Milling and baking

Toxic secondary metabolites produced by fungi representing Fusarium genus are common contaminants in cereals worldwide. To estimate the dietary intake of these trichothecene mycotoxins, information on their fate during cereal processing is needed. Up-to-date techniques such as high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS) was used for the analysis of seven trichothecenes (deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin, 15- and 3-acetyldeoxynivalenol, and fusarenon-X) in bread production chain (wheat grains, intermediate products collected during milling and baking process, breads). Regardless of whether the grains were naturally infected or artificially inoculated by Fusarium spp. in the field, the fractions obtained from the grain-cleaning procedure contained the highest mycotoxin levels. During milling the highest concentrations of deoxynivalenol were found in the bran, the lowest in the reduction flours. Baking at 210°C for 14 min had no significant effect on deoxynivalenol levels. The rheological properties of dough measured by fermentograph, maturograph, oven rise recorder, and laboratory baking test were carried out, and based on the obtained results the influence of mycotoxin content on rheological behaviour was investigated.