Stability of the mycotoxin deoxynivalenol (DON) during the production of flour-based foods and wheat flake cereal

Deoxynivalenol (DON) is a mycotoxin found in cereal grains and cereal-based foods. DON concentrations in finished products are reduced under some processing conditions, but not others. DON concentrations in flour, wheat and selected foods made from them under commercially relevant conditions were compared by GC with electron capture detection. Average concentrations (n?=?9/item) in cookies, crackers and pretzels ranged from 61% (cookies) to 111% (pretzels) compared with flour (100%?=?0.46?µg?g^?1). Lesser amounts were found in donuts and bread: their respective DON concentrations were 44% and 30% that of flour. Mass balance estimates for DON (µg?g^?1 flour equivalents) ranged from 50% (bread?=?0.23?µg?g^?1 flour equivalents) to 120% (donuts), indicating that dilution by recipe ingredients contributed to DON reductions in bread and accounted for all of the apparent reduction in donuts. Mass balance estimates averaged 76% (crackers) to 107% (pretzels) for the other flour products. DON concentrations were higher in cereal flakes (0.55?µg?g^?1 in the finished product and 0.58?µg?g^?1 on a mass balance basis) than in wheat (0.40?µg?g^?1), suggesting that DON concentrations might increase during processing of wheat cereals under some conditions. In summary, DON concentrations of finished food products were reduced?≥50% only in bread and donuts. Reduction in bread resulted from a combination of DON ‘loss’ and dilution by recipe ingredients whereas the reduction in donuts was due entirely to dilution. These results are further evidence of DON stability during the preparation of popular flour or wheat-based products.     

Fate of Fusarium mycotoxins in maize flour and grits during extrusion cooking

Extrusion technology is used widely in the manufacture of a range of breakfast cereals and snacks for human consumption and animal feeds. To minimise consumer exposure to mycotoxins, the levels of deoxynivalenol (DON) and zearalenone (ZON) in cereals/cereal products and fumonisins B₁ and B₂ (FB₁ and FB₂) in maize are controlled by European Union legislation. Relatively few studies, however, have examined the loss of Fusarium mycotoxins during processing. The behaviour of FB₁, FB₂ and fumonisin B₃ (FB₃), DON and ZON during extrusion of naturally contaminated maize flour and maize grits is examined using pilot-scale equipment. DON and ZON are relatively stable during extrusion cooking but the fumonisins are lost to varying degrees. There is some loss of ZON when present in low concentrations and extruded at higher moisture contents. The presence of additives, such as reducing sugars and sodium chloride, can also affect mycotoxin levels. Moisture content of the cereal feed during extrusion is important and has a greater effect than temperature, particularly on the loss of fumonisins at the lower moistures. The effects are complex and not easy to explain, although more energy input to the extruder is required for drier materials. However, on the basis of these studies, the relationship between the concentration of Fusarium toxins in the raw and finished product is toxin- and process-dependent.     

Distribution of Fusarium mycotoxins in UK wheat mill fractions

The EU has set maximum limits for the Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZON). The maximum permitted level decreases from unprocessed wheat, through intermediary products, e.g. flour, to finished products such as bakery goods and breakfast cereals. It is, therefore, important to understand the effects of processing on the mycotoxin distribution in mill fractions. Between 2004 and 2007, samples were taken at commercial flour mills at various points in the milling process and analysed for trichothecenes and ZON. Samples with a range of mycotoxin concentrations harvested in 2004 and 2005 were processed in a pilot mill and the mycotoxins in the different mill fractions quantified. In the commercial samples, DON was the predominant mycotoxin with highest levels detected in the bran fraction. Analysis of the pilot mill fractions identified a significant difference between the two years and between mycotoxins. The proportion of DON and nivalenol in the mill fractions varied between years. DON and nivalenol were higher in flour fractions and lower in bran and offal in samples from 2004 compared to samples from 2005. This may be a consequence of high rainfall pre-harvest in 2004 resulting in movement of these mycotoxins within grains before harvest. There was no significant difference in the distribution of ZON within mill fractions between the two years. For DON, higher concentrations in the grain resulted in a greater proportion of DON within the flour fractions. Understanding the factors that impact on the fractionation of mycotoxins during milling will help cereal processors to manufacture products within legislative limits.     

Stability of the mycotoxin deoxynivalenol (DON) during the production of flour-based foods and wheat flake cereal

Deoxynivalenol (DON) is a mycotoxin found in cereal grains and cereal-based foods. DON concentrations in finished products are reduced under some processing conditions, but not others. DON concentrations in flour, wheat and selected foods made from them under commercially relevant conditions were compared by GC with electron capture detection. Average concentrations (n = 9/item) in cookies, crackers and pretzels ranged from 61% (cookies) to 111% (pretzels) compared with flour (100% = 0.46 μg g?1). Lesser amounts were found in donuts and bread: their respective DON concentrations were 44% and 30% that of flour. Mass balance estimates for DON (μg g?1 flour equivalents) ranged from 50% (bread = 0.23 μg g?1 flour equivalents) to 120% (donuts), indicating that dilution by recipe ingredients contributed to DON reductions in bread and accounted for all of the apparent reduction in donuts. Mass balance estimates averaged 76% (crackers) to 107% (pretzels) for the other flour products. DON concentrations were higher in cereal flakes (0.55 μg g?1 in the finished product and 0.58 μmg g?1 on a mass balance basis) than in wheat (0.40 μg g?1), suggesting that DON concentrations might increase during processing of wheat cereals under some conditions. In summary, DON concentrations of finished food products were reduced ≥ 50% only in bread and donuts. Reduction in bread resulted from a combination of DON ‘loss’ and dilution by recipe ingredients whereas the reduction in donuts was due entirely to dilution. These results are further evidence of DON stability during the preparation of popular flour or wheat-based products.     

Changes in ochratoxin A and type B trichothecenes contained in wheat flour during dough fermentation and bread-baking

Ochratoxin A (OTA) and type B trichothecenes are mycotoxins that occur frequently in cereals and thus can be found in cereal by-products such as bread. The aim of this work was to study the variation of the levels of OTA, deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and nivalenol (NIV) during the bread-making process. This was done by using wheat flour spiked with different levels of toxins. Mycotoxin levels were controlled after fermentation of the dough with yeasts (Saccharomyces cerevisiae) and after further baking at different temperature–time combinations. Analysis of variance (ANOVA) of the results showed a significant reduction in OTA level (p < 0.05) during fermentation of the dough. The reduction ranged between 29.8% and 33.5%, depending on the initial concentration of toxin in the flour. During this period, the level of the other mycotoxins studied was not modified. By contrast, in the baking phase there were significant changes in the levels of the four mycotoxins, although the reduction was similar under all the baking conditions. Considering all the temperature–time conditions tested, it can be concluded that during the baking period the average reduction of OTA, NIV, 3-ADON, and DON was 32.9%, 76.9%, 65.6%, and 47.9%, respectively.     

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).     

Deoxynivalenol and other Fusarium toxins in wheat and rye flours on the Danish market

Information on the contamination of Danish cereals and cereal products with Fusarium toxins is limited and the last survey is from 1984/1985. In the present study, the occurrence of deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and zearalenone (ZON) was investigated in flour of common wheat, durum wheat and rye. The samples were collected from 1998 to 2001 from both mills and the retail market in Denmark. A total of 190 flour samples were analysed for DON and NIV and about 60 samples for HT-2, T-2 toxin and ZON. DON was most frequently detected with an incidence rate of 78% over all samples for all years. The contamination level varied considerably from year to year, and for wheat and rye the highest incidence and DON concentrations were found in samples from the 1998 harvest. There were regular and heavy rainfalls in Denmark during the flowering period of the crops that year, and DON was found in all samples, with mean concentrations in wheat and rye flour of 191 μg kg^-1 (n =14) and 99 μg kg^-1 (n =16), respectively. Comparison of data from each harvest year showed higher contents of DON in samples of wheat (range 20-527 μg kg^-1) than in rye (20-257 μg kg^-1). Contents of NIV, HT-2 toxin and ZON in samples of wheat and rye were generally low, and even in positive samples the contents were close to the detection limit of the methods. The T-2 toxin was detected in only a few of the wheat samples and in low amounts. However, the toxin was found in about 50% of the rye samples collected during 1998-2000, with a mean content of 49 μg kg^-1 (n =25). Durum wheat flour showed the highest DON contamination level, and all samples (n =33) collected during 2000 and 2001 contained DON with means and medians above 1100 μg kg^-1. Over 70% of the samples contained more than 500 μg kg^-1 DON, and the highest observed concentration was 2591 μg kg^-1. The concentration of T-2 toxin in durum wheat flour was also high with five of the 10 analysed samples containing more than 100 g kg^-1.     

Fate of deoxynivalenol and nivalenol during storage of organic whole-grain wheat flour

This study was carried out to determine the level of retention of the mycotoxins deoxynivalenol (DON) and nivalenol (NIV) during 120 days of storage (aging) of flours produced from organic wheat grain naturally infected with Fusarium fungi. Three types of flour (standard white flour prepared by a roller-grinder mill - IRG, whole-grain flour produced by a hammer-crusher mill - IHC and whole-grain flour prepared by a millstone - OMS) were packaged in food-grade paper or polypropylene plastic bags and stored at two different storage temperatures (constant 10 °C or 25 °C). The concentrations of DON and NIV were measured prior to and after storage by means of HPLC-UV detection methods. After 120 days of storage, the concentrations of DON and NIV decreased between 0% and 29% compared to the initial measurements, depending on the combination of experimental factors. The greatest decrease in mycotoxin concentration was observed in the IHC and OMS flours packaged in paper bags and stored at 25 °C. The smallest decrease in mycotoxin concentration was observed in the IRG flours packaged in sealed plastic bags and stored at 10 °C. Statistical analysis showed that the level of retention of DON and NIV depended significantly on the type of packaging material, but did not depend on the type of flour or the storage temperature.     

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.     

Fate of the fusarium mycotoxins, deoxynivalenol, nivalenol and zearalenone, during extrusion of wholemeal wheat grain

In the European Union, deoxynivalenol in cereals and cereal products is controlled by recent legislation with the objective of minimizing consumer exposure to this mycotoxin. Relatively few studies have examined the loss of Fusarium mycotoxins during processing and whether this is accurately reflected by the processing factors. The behaviour of deoxynivalenol, nivalenol and zearalenone during extrusion of naturally contaminated wholemeal wheat flour has been examined using pilot-scale equipment. Factors examined were temperature and moisture content. Concentrations of the three mycotoxins were little changed by extrusion although the amount of deoxynivalenol decreased at the lowest moisture content. However, this effect did not appear to be temperature-dependent, suggesting that the apparent loss is either due to binding or inability to extract the residue. Under some conditions, concentrations of the mycotoxins, particularly nivalenol, were higher after extrusion.     

Exposure of consumers to deoxynivalenol from consumption of white bread in Hungary

In view of the frequent occurrence of mycotoxins in cereals, a study was initiated to assess the exposure of the Hungarian adult population. Consumption data for 1360 individuals, based on a 3-day questionnaire, indicated that white bread accounted for the major intake of cereal-based products. Various cereal products were analysed for 16 mycotoxins by a LC/MS/MS multi-toxin method with LOD of 16?µg?kg^?1 and LOQ of 50?µg?kg^?1. Deoxynivalenol (DON) was most frequently detected, but no acetyl-deoxynivalenol was present in detectable concentrations. Consumer exposure was calculated with standard Monte Carlo probabilistic modelling and point estimates, taking into account bread consumption and DON contamination in independently taken wheat flour and wheat grain samples. Over 55% of cases the DON intake were below 15% of the provisional maximum tolerable daily intake (PMTDI) of 1?µg/(kg?bw)/day. However, in 5–15% of cases, the intake from bread consumption alone exceeded the PMTDI. Wheat grain data led to the higher percentage. Intakes estimated from both data sets were at or below the acute reference dose (ARfD) of 8?µg/(kg?bw)/day in 99.94–99.97% of cases.     

Development and application of analytical methods for the determination of mycotoxins in organic and conventional wheat.

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 microg kg-1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75 microg kg-1 in conventional samples against 0.063, 285 and 19 microg kg-1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).     

Fusarium DNA traceability along the bread production chain

Wheat grain contamination with toxigenic Fusarium spp. is of great economic concern to cereal producers and to the grain processing industry and is of great relevance for the quality and safety of the final products. In particular, the bread production chain can potentially be a vehicle for mycotoxin ingestion above the tolerable total daily intake. A quantitative polymerase chain reaction‐based analytical approach has been developed as a possible tool to estimate and control the risk of mycotoxins, particularly deoxynivalenol (DON). This DNA‐based analytical method has been applied to detect and quantify contamination levels of Fusarium graminearum and Fusarium culmorum in naturally infected wheat grain samples. The persistence of Fusarium contamination was also detected along the bread production chain in wholemeal, flour and bread. A significant correlation was found between Fusarium DNA and DON concentration in all samples.     

The Fate of Mycotoxins During the Processing of Wheat for Human Consumption

Mycotoxins are a potential health threat in cereals including wheat. In the European Union (EU), mycotoxin maximum levels are laid down for cereal raw materials and final food products. For wheat and wheat‐based products, the EU maximum levels apply to deoxynivalenol (DON), zearalenone, aflatoxins, and ochratoxin A. This review provides a comprehensive overview on the different mycotoxins and their legal limits and on how processing of wheat can affect such contaminants, from raw material to highly processed final products, based on relevant scientific studies published in the literature. The potential compliance with EU maximum levels is discussed. Of the four mycotoxins regulated in wheat‐based foods in the EU, most data are available for DON, whereas aflatoxins were rarely studied in the processing of wheat. Furthermore, available data on the effect of processing are outlined for mycotoxins not regulated by EU law—including modified and emerging mycotoxins—and which cover DON derivatives (DON‐3‐glucoside, mono‐acetyl‐DONs, norDONs, deepoxy‐DON), nivalenol, T‐2 and HT‐2 toxins, enniatins, beauvericin, moniliformin, and fumonisins. The processing steps addressed in this review cover primary processing (premilling and milling operations) and secondary processing procedures (such as fermentation and thermal treatments). A special focus is on the production of baked goods, and processing factors for DON in wheat bread production were estimated. For wheat milling products derived from the endosperm and for white bread, compliance with legal requirements seems to be mostly achievable when applying good practices. In the case of wholemeal products, bran‐enriched products, or high‐cereal low‐moisture bakery products, this appears to be challenging and improved technology and/or selection of high‐quality raw materials would be required.     

Fate of ochratoxin A in the processing of whole wheat grains during milling and bread production.

Batches of whole wheat contaminated with ochratoxin A were produced by inoculation with Penicillium verrucosum under controlled conditions in the laboratory. The fate of ochratoxin was followed through initial cleaning, abrasive scouring of the outer grain coat, milling into wholemeal wheat or into 10 milled fractions. Bread was baked from both wholemeal flour and straight-run white flour. Concentrations of ochratoxin A in the cleanings, scourings, and the bran and offal fractions were increased, but reduced in the white flour. Scouring removed up to 44% of the ochratoxin A present, but only a small further loss occurred in the bread-making process. An overall reduction of about 75% could be achieved in white bread using a combination of cleaning scouring and removal of the bran and offal fractions. Maximum overall reduction in producing wholemeal bread was about 40%. The reduction in ochratoxin A that can be achieved must be considered in relation to economic constraints concerning the disposal of wasted grain. Appropriate strategies for the use or disposal of potentially highly contaminated cleanings, scourings, bran or offal must be established.     

Fate of the fusarium mycotoxins,deoxynivalenol, nivalenol and zearalenone,during extrusion of wholemeal wheat grain

In the European Union, deoxynivalenol in cereals and cereal products is controlled by recent legislation with the objective of minimizing consumer exposure to this mycotoxin. Relatively few studies have examined the loss of Fusarium mycotoxins during processing and whether this is accurately reflected by the processing factors. The behaviour of deoxynivalenol, nivalenol and zearalenone during extrusion of naturally contaminated wholemeal wheat flour has been examined using pilot-scale equipment. Factors examined were temperature and moisture content. Concentrations of the three mycotoxins were little changed by extrusion although the amount of deoxynivalenol decreased at the lowest moisture content. However, this effect did not appear to be temperature-dependent, suggesting that the apparent loss is either due to binding or inability to extract the residue. Under some conditions, concentrations of the mycotoxins, particularly nivalenol, were higher after extrusion.     

真菌毒素在赤霉病小麦研磨及其湿法加工中的分布研究

小麦赤霉病不仅会导致粮食减产,更会引起多种真菌毒素的高污染风险。将染病小麦进行实验室制粉并湿法分离统粉中的粗淀粉、谷朊粉,采用酶联免疫吸附测定法(Enzyme-Linked Immuno Sorbent Assay,ELISA)测定各系统的呕吐毒素(Deoxynivalenol,DON)、黄曲霉毒素B1(Aflatoxin B1,AFB1)、玉米赤霉烯酮(Zearalenone,ZEN)以及赭曲霉毒素A(Ochratoxin A,OTA)的含量,以探究赤霉病小麦在制粉及其湿法加工中四种常见真菌毒素的分布变化规律。结果显示:研磨制粉及湿法加工对真菌毒素的分布影响显著。制粉加工后,皮磨和心磨系统粉的DON、AFB1、ZEN和OTA含量比原粮显著降低,降低率分别为1.38~16.24%、20.47~71.77%及26.71~69.51%;湿法加工产物中,DON消减为相应统粉的4.88~12.11%,AFB1与OTA浓缩富集,富集率分别可达统粉的2.55、3.65倍,粗淀粉中ZEN消减为统粉的12.70~15.83%,谷朊粉则富集为统粉的4.11倍。研究表明,在工业生产中,可根据赤霉病小麦的感染类型及程度,适当选用研磨或湿法加工等合适的方法加工处理。     

Occurrence of Fusarium mycotoxins in maize imported into the UK, 2004–2007

This study examined a total of 82 consignments of French and Argentinean raw maize as received at maize mills in the UK between 2004 and 2007. Samples were analysed for deoxynivalenol (DON), nivalenol (NIV), other trichothecenes, zearalenone (ZON), and fumonisins B₁, B₂, and B₃ (FB₁, FB₂, and FB₃) using fully validated analytical methods with limits of quantification of 10 µg kg^?1 for DON, NIV, and each fumonisin mycotoxin and 3 µg kg^?1 for ZON. All samples except two containing fumonisins met the European Commission statutory maximum permissible levels for DON, ZON, and FB₁ + FB₂ as operating in 2007. The maximum concentrations found for DON, NIV, ZON, and FB₁ + FB₂ were 444, 496, 165 and 5002 µg kg^?1, respectively. Fumonisins were detected in almost every sample with 65% of Argentinean maize containing more than 1000 µg kg^?1 of FB₁ + FB₂. In contrast, ZON was not detectable in almost 50% of consignments. During this period there was a distinct difference in mycotoxin concentrations between harvests and geographic origin. Flint maize from Argentina usually contained lower concentrations of DON and related trichothecenes and higher levels of fumonisins than maize from France, although concentrations of fumonisins up to 2000 µg kg^?1 or greater occurred in samples from both regions. The incidence and concentrations of fumonisins were similar to those in a similar previous survey, while zearalenone concentrations were lower. The distribution of mycotoxins in multi-hold ships was also investigated showing that fumonisins were much more evenly distributed than DON, thus indicating their general level in the ship as a whole. The effect of cleaning regimes was found to be very variable, especially for DON, ranging from no removal of mycotoxins to greater than 50% in some instances, but was not related to concentration. Evidence here suggests that while cleaning is essential for removing foreign bodies before milling, it cannot be used as a reliable tool for reducing mycotoxins.     

Determination of trichothecenes and zearalenones in grain cereal,flour and bread by liquid chromatography tandem mass spectrometry

Although analytical methods have been already reported for legislated mycotoxins as trichothecenes and zearalenone (ZON) separately, we describe the optimization of a simple and rapid multimycotoxin method for the determination of a total of 12 mycotoxins simultaneously, nine trichothecenes (NIV, DON, FUS-X, DAS, 15-AcDON, 3-AcDON, NEO, HT-2, T-2 T2), and zearalenone and its metabolites (ZON, α-ZOL, β-ZOL), of different origin (wheat, oat, barley and spelt) and in three different products where these substance can be present (grain, flour and bread) reach the food chain and cause toxic effect either in humans or animals. The extraction procedure was based on a mixture of acetonitrile/water (84/16, v/v), which provided the highest recoveries and the lowest matrix effect. DON-d1 was used as internal standard (I.S.) which helped to compensate the significant matrix effect observed for some matrices, and to obtain high success in the method validation and to reach the parameters compiled in Commission Decision, 2002/657/EC. Analytes were determinate by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). Relative recoveries obtained were higher than 70% for the studied mycotoxins the four cereal. Good linearity (r² > 0.992) was obtained and quantification limits (2.5–25 ng/g) were below European regulatory levels. Repeatability, expressed as relative standard deviation, was always lower than 11%, whereas interday precision was lower than 11% for the developed method.