Fumonisin occurrence in naturally contaminated wheat grain harvested in Argentina

A survey was carried out to determine fumonisin contamination in 135 common and 40 durum wheat samples collected during 2011 harvest season (non-FHB epidemic year) in the main wheat production area of Argentina using LC–MS/MS. A 93% of total samples showed fumonisin contamination, with levels ranging from 0.16 to 680.44 ng/g in common and from 0.15 to 1304.39 ng/g in durum wheat samples, respectively. FB₁ was the fumonisin most frequently found during the evaluated year. Twenty five wheat samples (15 common and 10 durum) were selected for a deoxynivalenol (DON) analysis among all the samples analyzed for fumonisin content using different contamination levels as selection criteria. DON contamination was present in 24 out of 25 wheat samples, the levels ranging from 50.60 to 28650 ng/g. Nine out of 25 wheat samples reached values higher than 1000 ng/g. However there was no correlation between fumonisin and DON contamination. This is the first report of natural fumonisin presence in common wheat grains in Argentina, as well as of DON co-occurrence in both types of wheat.     

Simultaneous determination of Fusarium mycotoxins in wheat grain from Morocco by liquid chromatography coupled to triple quadrupole mass spectrometry

In the present study, eighteen (18) mycotoxin produced by the genus Fusarium (fumonisins (FB1, FB2 and FB3), type-B trichothecenes (NIV, DON, FUS-X, 3Ac-DON and 15Ac-DON), type-A trichothecenes (NEO, DAS, T2 and HT2), zearalenone (ZEA), beauvericin (BEA), and enniatins (ENA, ENA₁, ENB and ENB₁)) were monitored in different samples of wheat grain commercialized in Morocco. A liquid chromatography coupled to triple quadrupole mass spectrometry method previous matrix solid phase dispersion extraction was used for sample analysis. A total of eighty (80) samples of durum wheat were collected in different local markets from several areas in Morocco. Analytical results showed that 54 out of 80 total wheat samples (68%) were contaminated. The mycotoxins found were ENA, ENA₁, ENB and ENB₁, DON and BEA. The rest of investigated mycotoxins were below the limits of quantification. In positive samples, enniatins levels ranged between 2.5 and 2570 μg/kg. DON levels ranged between 121 and 1480 μg/kg and BEA levels were between 5.4 and 16 μg/kg. Among enniatins, ENB was predominant with a frequency of 61%. Co-presence of DON, enniatins and BEA from durum wheat from Morocco was found for the first time.     

Screening of deoxynivalenol contamination in durum wheat by MOS-based electronic nose and identification of the relevant pattern of volatile compounds

Fungal volatile metabolites can be used as an indicator of mycotoxins occurrence in cereals. A rapid, easy-to-perform and non-invasive method using an electronic nose based on metal oxide sensors was developed to distinguish the quality of durum wheat samples in three classes based on the content of the mycotoxin deoxynivalenol (DON): class A ([DON] < 1000 μg/kg), class B (1000 ≤ [DON] ≤ 2500 μg/kg;) and class C ([DON] > 2500 μg/kg). Two different approaches were tested by performing 525 analyses of naturally contaminated durum wheat either on whole and ground kernels. Discriminant Function Analysis (DFA) provided calibration models that permitted to classify wheat samples with mean recognition percentages (R) for whole and ground kernels of 69.3% and 86.7%, respectively. Samples belonging to class A were better recognized than those belonging to classes B and C. The e-nose method applied to the analysis of ground wheat showed highest mean recognition percentage (R = 82.1%) in validation. By considering data with recognition percentage rate (R_r) higher than 80% in validation, performances of this method were further improved in terms of either mean recognition percentage (R = 90.1%) and percentage of misclassified samples (4%). An SPME-GC–MS method was also developed that allowed to characterize the pattern of volatile compounds of durum wheat samples. A positive (trichodiene, longifolene, 3-methyl butanal, tridecane, γ-caprolactone, 6,10,14-trimethyl-2-pentadecanone) or negative (hexadecane, 2,3,7-trimethyl-decane, 4,6-dimethyl-dodecane) association with DON content was observed. These results confirm that the developed e-nose method could be used as useful tool for high throughput screening of DON contamination in durum wheat.     

Limited survey of deoxynivalenol in wheat from different crop rotation fields in Yangtze-Huaihe river basin region of China

The food toxin, deoxynivalenol (DON) is frequently present in cereals such as wheat, barley and maize, which are infected by Fusarium asiaticum and Fusarium graminearium. Crop rotation and climatic conditions play a major role in the Fusarium infection in wheat. In this present study, a minor survey was conducted to find out the impact of crop rotation affecting the Fusarium infection leading to DON contamination in wheat samples from selected infected regions of Jiang su and An hui provinces, China, especially during harvest period in 2012. A total of 84 wheat samples from the highly Fusarium infected region were collected, of which 30, 8, 39 and 7 samples were from the fields where cotton, corn, rice, and soya bean was a rotation crop, respectively. DON concentration in wheat sample was analyzed by high performance liquid chromatography (HPLC) combined with ultraviolet detection. DON contamination in the wheat samples was 95% with the mean DON concentration of 3881.2 μg/kg from both the regions. Average DON contamination in wheat samples from cotton, soya beans, corn, and rice rotation fields were 2067.5, 2853.6, 3517.5, and 4899.3 μg/kg, respectively. Nearly 34% of the wheat samples from the cotton rotation fields were below the DON maximum tolerable limits of EU (1750.0 μg/kg) and only 3% samples were above 4000.0 μg/kg DON concentration. However, the average DON contamination level was the highest in wheat samples (4899.3 μg/kg) where rice was a rotation crop, with 64% of the samples were above 4000.0 μg/kg. The present study shows cotton could be a promising rotation crop in the regions where wheat is more prone to the infection of Fusarium sp, which may minimize the economic loss to the farmers in wheat.     

Distribution of deoxynivalenol and zearalenone in milled germ during wheat milling and analysis of toxin levels in wheat germ and wheat germ oil

A method consisting of solvent extraction using hexane for defatting, multifunctional cleanup column, and HPLC determination was validated for the analysis of deoxynivalenol (DON) and zearalenone (ZEA) in wheat germ and wheat germ oil. A total of 36 batches of grain wheat were subjected to industrial milling and the distribution factors in milled germ were 47% for DON and 71% for ZEA. A survey of 50 samples of germ-based dietary supplements revealed that 60% of wheat germ and 40% of wheat germ oils contained DON at mean values of 111 and 41 μg/kg, respectively, while none of germ samples and 16% oils contained ZEA (mean 6 μg/kg). Contamination levels lead to a daily intake of 1.3 μg DON and 0.03 μg ZEA, representing 1.9% and 0.23% of their respective tolerable daily intakes (TDI).     

Fusarium mycotoxins in wheat samples harvested in Serbia: A preliminary survey

The objective of this study was to determine the occurrence of trichothecenes of both the A-type and B-type, masked mycotoxin derived from DON - deoxynivalenol-3-glucoside (DON-3-Glc), 3- and 15-acetyldeoxynivalenol (ADONs), fusarenon-X (FUS-X) and nivalenol (NIV)) as well as zearalenone (ZON) in winter wheat. Total of 54 samples were collected during the harvest of 2007 representing the most important Serbian wheat-growing regions. The samples were prepared by one-step simple method and analyzed by high performance liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). The obtained recoveries proved that the used method could be successfully applied for multi-component analysis of Fusarium mycotoxins. DON, DON-3-Glc and HT-2 contents were detected approximately in 28%, 13% and 6% of the total number of samples, respectively. The amount of these toxins ranged from 17 μg/kg for DON-3-Glc to 309 μg/kg for DON. ADONs, FUS-X, NIV, T-2 toxin as well as ZON were below the limit of detection. Different susceptibility of wheat cultivars towards detected mycotoxins was observed. The results were compared to the EC Regulative and with available the literature data concerning the neighboring countries. This is first report on the simultaneous presence of 8 Fusarium mycotoxins in the wheat cultivated in the Balkan Countries region.     

Aflatoxins and ochratoxin A in Brazilian paprika

Seventy paprika samples collected in the city of São Paulo, Brazil, from January to April 2006 were analysed for aflatoxins and ochratoxin A (OTA) using an immunoaffinity column clean-up and HPLC-FLD. For aflatoxins, the limit of quantification (LOQ) were 0.23, 0.23, 0.45 and 0.45 μg/kg for AFB₁, AFB₂, AFG₁ and AFG₂, respectively. For OTA the LOQ was 0.80 μg/kg. Aflatoxins were found in 82.9% of samples and AFB₁ was detected in 61.4% at levels ranging from 0.5 to 7.3 μg/kg with mean concentration of 3.4 μg/kg. OTA was found in 85.7% at levels ranging from 0.24 to 97.2 μg/kg with mean concentration of 7.0 μg/kg.     

A minor survey of deoxynivalenol in Fusarium infected wheat from Yangtze–Huaihe river basin region in China

Deoxynivalenol (DON) is frequently present in infected cereals such as wheat, barley and maize. Fusarium asiaticum and Fusarium graminearium are the key fungal species which produces DON, when they infect wheat in China. Our present study is to do a minor survey on the presence of DON in the highly infected wheat regions from Jiang su and An hui province, China, harvested during the year 2010. A total of 56 wheat samples which are naturally infected by Fusarium sp., of which 35 samples from Jiang su province and 21 samples from An hui province were taken up for the study. The samples were analyzed for the presence of DON by using high performance liquid chromatography (HPLC) combined with ultraviolet detection. The recoveries of DON from the spiked samples (500–15,000 μg/kg) were obtained in the range of 85.7–90.2%, with the average relative standard deviation of 2.7%. The average percentage of contamination in all the samples was 89.3%, with the DON concentration ranging between 259 and 4975 μg/kg, and with the mean of 1962 μg/kg. According to the Chinese standard limits for DON, nearly 70% of the samples were above the maximum tolerable level of 1000 μg/kg. However, only 44% of the samples exceeded the European commission limits for unprocessed wheat of 1750 μg/kg. Regions where there was higher level of Fusarium sp. contamination due to climatic changes like heavy rainfall with high humidity, showed the presence of higher levels of DON in the samples. Cropping of Fusarium resistant wheat cultivars like Sumai 3 in the regions where the wheat is highly susceptible for the Fusarium contamination could help in the reduction of contamination in wheat.     

Fumonisin B1 and its co-occurrence with other fusariotoxins in naturally-contaminated wheat grain

The natural occurrence of fumonisin B₁ (FB₁) and its co-occurrence with zearalenone (ZEA), T-2 toxin and deoxynivalenol (DON) were surveyed in 103 winter wheat samples collected after four to six-month storage in family barns from different locations in Serbia. All 103 samples were mycotoxin positive. The mean concentrations of all mycotoxins except ZEA were greater in 2005 than in 2007. FB₁ was detected in 82.1% and 92.0% of all samples with ranges of 750-5400 μg kg⁻¹ (mean, 2079.45 μg kg⁻¹) and 750-4900 μg kg⁻¹ (mean 918.76 μg kg⁻¹) in 2005 and 2007, respectively. Moderate positive correlations were found between FB₁ and DON concentrations (r = 0.56 in 2005 and r = 0.54 in 2007) and between FB₁ and ZEA concentrations (r = 0.48 in 2005 and r = 0.60 in 2007), while a moderate negative correlation was detected between the production of FB₁ and T-2 toxin in 2007 (r = −0.33). This is the first report of FB₁ occurrence in naturally-contaminated wheat grain and its simultaneous occurrence with ZEA, DON and T-2 toxin in Serbia. Moreover, this is one of the rare reports presenting the occurrence of FB₁ on wheat in the world.     

Simultaneous analysis of twenty-six mycotoxins in durum wheat grain from Italy

A multi-mycotoxin analysis method based on liquid chromatography coupled to triple quadrupole mass spectrometry was validated and applied for the determination of twenty-six mycotoxins, including eight trichothecenes, three fumonisins, sterigmatocystin, ochratoxin A, four aflatoxins, zearalenone, five “emerging” mycotoxins and three Alternaria mycotoxins in 74 durum wheat samples from central Italy. Eighty-eight percent of the analysed samples contained one or more mycotoxins, with enniatin B showing the highest contamination levels (78% of samples with levels ranging from 23 to 1826 ng/g), followed by enniatin B1 (10–1384 ng/g) and deoxynivalenol (48–2267 ng/g). This work gives an important contribution in terms of mycotoxin analysis and occurrence in durum wheat and in assessing the mycotoxicological risk posed by both traditional and emerging mycotoxins for the final consumers of durum wheat derived products.     

Presence of trichothecenes and co-occurrence in cereal-based food from Catalonia (Spain)

The most important trichothecenes are HT-2 toxin (HT2) and T-2 toxin (T2) from type A and deoxynivalenol (DON) from type B. Thus, the aim of the current study was to assess the occurrence of these trichothecenes in the Catalonian market. 479 food samples were taken from the most susceptible to trichothecenes contamination and most commonly consumed in Catalonia commodities. DON, T2 and HT2 toxin were determined in breakfast cereals, snacks and pasta samples following extraction, clean-up, derivatization and finally analysis by GC–ECD. Moreover, these mycotoxins were determined in sliced bread, sweet corn and beer by LC–DAD. Our results showed that DON was the main trichothecene present in the cereal-based food from Catalonian market with percentages of positive samples ranging from 1.4 to 100.0%. Despite the high incidence of DON, only five samples were above EU limits. Concerning HT2 toxin, it was present in a low percentage of samples of sliced bread (15.3%), wheat flakes (14.8%), pasta (10.0%), corn snacks (8.5%), sweet corn (6.9%) and corn flakes (6.2%), while the T2 toxin only was quantified in 5 samples out of the total 479.     

Effect of processing stages of apple juice concentrate on patulin levels

The effects of different stages of apple juice concentrate production on patulin levels were investigated. Patulin was detected in all samples analyzed in concentrations ranging from 56 to 653 μg/L. Apple paste resulted from milling process had high levels of patulin. The results of this study indicate that it is possible to reduce patulin level in apple juices. After pasteurization, enzymatic treatment, microfiltration and evaporation processes, the mean loss of patulin was 39.6, 28.3, 20.1 and 28.4%, respectively. When apple juices concentrate were diluted from 69 to 12°Brix to consume, patulin content ranged from 15 to 46 μg/L. Patulin content in all juice samples was lower than the limit of 50 μg/L considered acceptable by the Codex Alimentarius Commission. But if consider the maximum permitted concentration established for apple products intended for infants and young children by The Commission of the European Communities all samples were found to exceed patulin concentration of 10 μg/L.     

The occurrence of the selected fusarium mycotoxins in Czech malting barley

In 2008–2011, the occurrence of deoxynivalenol (DON), zearalenone (ZON), T-2 toxin (T-2), and HT-2 toxin (HT-2) was studied in 325 malting barley samples collected from various regions of the Czech Republic. The highest occurrence of fusarium mycotoxins was recorded in crop 2009 (2213.5, 59.4 and 145.0 μg/kg for DON, ZON and ∑T-2, HT-2, respectively). Only one measured value exceeded the maximum allowable limit for DON set by the European Union.The validated method of high-performance liquid chromatography coupled with mass spectrometry was used for the analysis of the above mentioned toxins. Limits of detection were 1.5 μg/kg for DON, ZON and HT-2 and 0.3 μg/kg for T-2, the limits of quantification were 5.0 μg/kg for DON, ZON and HT-2 and 1.0 μg/kg for T-2.     

Investigations on the occurrence of mycotoxins in beer

The occurrence of several mycotoxins, including ergot alkaloids, alternariol (AOH), deoxynivalenol (DON), and zearalenone (ZEA) in beer (n = 44) from the German market was studied by using enzyme immunoassay methods. Detection limits in beer were 2.1 μg/L (DON), 0.14 μg/L (ZEA), 0.06 μg/L (ergometrine equivalents), and 0.18 μg/L (AOH). DON was detected in 75% of the samples (2.2–20 μg/L, median 3.7 μg/L). All samples were positive for ZEA (0.35–2.0 μg/L, median 0.88 μg/L) and AOH (0.23–1.6 μg/L, median 0.45 μg/L). Most samples (93%) were positive for ergot alkaloids (0.07–0.47 μg/L, median 0.15 μg/L). Correlating toxin levels in beer with European Union tolerable daily intake (TDI) levels for DON (1 μg/kg b.w.), ZEA (0.25 μg/kg b.w.), and ergot alkaloids (0.6 μg/kg b.w.), beer does not represent a major source of intake of these toxins. No TDI data are available for AOH, but considering toxin levels in other foods, beer does not seem to be a major source of intake of this toxin either. Nevertheless, the frequency of their occurrence warrants further study of ergot alkaloids and AOH in raw materials used for beer brewing.     

Natural occurrence of ochratoxin A in Tunisian cereals

Ochratoxin A (OTA) is a mycotoxin produced by several fungal species from Aspergillus and Penicillium genera. It is widespread in food and feed and its occurrence has been reported in cereals, cereal-derived products, dried fruits and spices. This mycotoxin was implicated in several human and animal pathologies such as the Balkan Endemic Nephropathy (BEN) and the Tunisian Chronic Interstitial Nephropathy (CIN) of unknown cause. In Tunisia, a clear correlation has been established between the consumption of OTA contaminated food and the induction of specific pathologies. Thereby, OTA was detected in human blood and tissues. The aim of our study was to investigate the presence of OTA in widely consumed cereals commercialized in Tunisia. The analytical methods used in our study involved the extraction of OTA by acidified toluene, immunoaffinity (IAC) clean-up and HPLC quantification with fluorescence detection. Levels and percentages of OTA contamination in different types of cereals, 110 wheat, 103 barley, 113 sorghum and 96 rice samples, were evaluated with incidences of 38%, 40%, 38% and 28%, respectively. The average of contamination by OTA found were 55, 96, 44 and 117 μg/kg, respectively, for wheat, barley, rice and sorghum. Our results showed that contamination percentages and levels in the period from 2004 to 2005 were higher then usual norms (5.0 μg OTA/kg) established by the European commission in 2002. The present report is the first one ever carried out on the natural occurrence of OTA in cereals, largely consumed by the Tunisian population.     

Determination of fumonisin B1 in bovine milk by LC–MS/MS

The aim of the present work was to develop a sensitive and selective method for identification and quantification of fumonisin B1 (FB1) in bovine milk. FB1 was isolated by immunoaffinity column and was detected using liquid chromatography coupled with tandem mass spectrometry in positive electrospray ionization (ESI+).The LOQ of the method was 0.1 μg/kg that was lower than the others reported in the literature. The high coefficient of determination (R² > 0.99) obtained in the range of 0.1–10.0 μg/kg, the good recovery (84%) and relative standard deviation (7%) of the proposed method ensure correct fumonisin detection in milk even at relatively low concentrations.The developed method was applied on different commercial samples in order to test its efficacy. FB1 was found above the LOQ in eight out of 10 samples analysed and the average level of contamination found was 0.26 μg/kg.     

Retention of deoxynivalenol and its derivatives during storage of wheat grain and flour

Fate of deoxynivalenol (DON) and its derivatives—deoxynivalenol-3-glucoside (D3G), 15-acetyldeoxynivalenol (15-AcDON) and 3-acetyldeoxynivalenol (3-AcDON) during six-month storage of wheat grain and flour was investigated. Wheat grain and flour polluted by DON, D3G, 15-AcDON and 3-AcDON were packaged in polyethylene bags, kraft paper bags and cloth bags, and stored at room temperature and 4–6 °C, and mycotoxin levels were analyzed by UPLC-MS/MS after storing for 30, 90 and 180 days. Levels of DON, 15-AcDON and 3-AcDON in wheat grain showed a generally decrease during the storage duration, and DON concentrations averagely decreased by 40–50%. An obviously increase of DON (more than 70%) and decrease of 15-AcDON and 3-AcDON were observed after storage in wheat flour. There were no significant differences of D3G levels in most of wheat grain and wheat flour samples after storage, but some increased dramatically with retention level up to 240%. The results suggested conversion of DON and its derivatives may occur during wheat grain and flour storage.     

Natural occurrence of zearalenone in Tunisian wheat grains

Fusarium mycotoxins are worldwide occurring in cereals and they are frequently reported in fresh or stored grains. Cereals represent a staple food for the Tunisian population; it therefore has a high social, economic and nutritional relevance. Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by a variety of Fusarium fungi in temperate and warm countries. Fungi-producing ZEN contaminates corn, barley, wheat, sorghum and rice. A total of 205 samples of wheat were collected during the harvest year of 2010 from the major cropping areas in Tunisia and they were analyzed for zearalenone contamination. The aim of this study was to investigate for the first time the presence of ZEN in widely-consumed cereals in Tunisia, especially durum and tender wheat, to compare the levels of contamination by ZEN with the European norms and to suggest some factors that can promote the production of ZEN in Tunisia. To perform this study, we developed and validated in our laboratory conditions an HPLC method for quantitative analysis of ZEN in solid cereal samples. Our results showed that the incidence of ZEN contamination was 75%. The levels of contamination determined in the positive samples ranged between 3 and 560 μg/kg with a mean value of 60 μg/kg. These important amounts of ZEN in wheat can be attributed to the Tunisian climate, warm temperature and prolonged wetness witch are favor to Fusarium growth and mycotoxin production during the cultivation and the final ripening period of wheat grains.     

Principal mycotoxins in wheat flour from the Serbian market: Levels and assessment of the exposure by wheat-based products

The presence of eleven principal mycotoxins from the wheat flour bought in supermarkets in Novi Sad, the capitol of the northern Serbian province of Vojvodina, was determined. The samples were prepared by simple one-step method and analyzed by ultra-high performance liquid chromatography with heated-electrospray ionization triple quadrupole mass spectrometry (UHPLC/HESI-MS/MS). Deoxynivalenol (DON) was the predominant mycotoxin for all analyzed samples followed by zearalenone (ZON) and T-2 toxin, with frequency of occurrence: 88.7%, 33.3% and 26.7%, respectively. Aflatoxins (AFs), ochratoxin A (OTA), HT-2 toxin, fumonisins B1 (FB1) as well as B2 (FB2) were below the limit of detection. All the samples complied with current European/Serbian legislation, except one sample that exceeded the DON maximum level of 750 μg/kg. In addition, mycotoxin intakes through consumption of wheat-based products were estimated for average adult consumers based on Serbian market basket and then compared with the tolerable daily intake (TDI) proposed by Scientific Committee on Food of the European Union. The calculated intakes of ZON and T-2 were lower than the respective TDIs. However, intakes of DON were assessed to be close to the level of TDI for adults. This is the first study on the intake assessment for mycotoxins present in the wheat flour through the consumption of wheat-based products on the Serbian market.     

Determination of Fusarium mycotoxins enniatins,beauvericin and fusaproliferin in cereals and derived products from Tunisia

In this study, 51 samples of cereals (wheat, Barley, maize and Sorghum) and by-products (mainly pasta and couscous) purchased from Tunisian supermarkets were examined for contamination with the emerging Fusarium mycotoxins: Enniatins ENs (EN A, EN A₁, EN B and EN B₁), beauvericin (BEA) and fusaproliferin (FUS).The extraction of the samples was performed with methanol using an Ultra-turrax homogenizer. Mycotoxins were analyzed with a liquid chromatography (LC) coupled to a diode array detector (DAD).The frequencies of contamination of total samples with ENs were 96%. EN A₁ was the most common EN found with the highest prevalence of 92.1%, levels ranged between 11.1 and 480 mg/kg. EN B was evidenced in 35 samples and levels ranged from 1.5 to 295 mg/kg. EN B₁ was detected in 20 samples (39.2%) and levels varied from 4.8 to 120.1 mg/kg and EN A was detected in 14 samples with contamination levels ranging between 19.6 and 121.3 mg/kg. The maximum concentration of total ENs in a single sample was 683.9 mg/kg (sorghum). The analytical results also showed that all the analyzed samples were free of BEA and FUS.The present work is the first one ever drafted on the presence of the emerging Fusarium mycotoxins in Tunisian cereals and derived products.