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.     

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.     

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.     

Mycotoxin occurrence in commodities,feeds and feed ingredients sourced in the Middle East and Africa

Between February and October 2009, 324 grain, feed and feed commodity samples were sourced directly at animal farms or feed production sites in Middle East and Africa and tested for the presence of A- and B-trichothecenes, zearalenone, fumonisins, aflatoxins and ochratoxin A, or for selected groups of mycotoxins only. Samples were analyzed after clean-up by immunoaffinity or solid-phase extraction followed by HPLC with derivatization where appropriate and fluorescence, UV or mass spectrometric detection. The percentage of positive samples of B-trichothecenes ranged from 0 to 87% of tested samples. The prevalence of fumonisins in the different countries was >50% in most cases. Zearalenone was present in tested commodities from all countries except three. The presence of aflatoxin in analyzed samples varied from 0 to 94%. Ochratoxin A was present in 67% of samples in Sudan and in 100% of Nigerian samples. No A-trichothecenes were found in this survey.     

Mycotoxins in durum wheat grain: hygienic-health quality of sicilian production

ABSTRACT:  Deoxynivalenol, zearalenone, and aflatoxin concentrations in Sicilian durum wheat were determined through ELISA tests. The results highlighted the safety of the grain samples at harvest because deoxynivalenol, zearalenone, and aflatoxin levels did not exceed European legal limits. With regard to aflatoxins, reliability of the ELISA test was evaluated, comparing it with HPLC analyses. The comparison of HPLC and ELISA data showed a tendency to overestimate aflatoxin concentrations with respect to chromatographic determinations. The usefulness of ELISA was confirmed as a rapid screening method; however, when contamination levels are close to legal limits, chromatographic analyses are necessary to quantify aflatoxins with greater accuracy and specificity.     

Dietary exposure to mycotoxins of the Hong Kong adult population from a Total Diet Study

Dietary exposure of Hong Kong adults to mycotoxins and their metabolites including aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FNs), deoxynivalenol (DON), acetyldeoxynivalenols (AcDONs) and zearalenone (ZEA) was estimated using the Total Diet Study (TDS) approach to assess the associated health risk to the local people. Sixty commonly consumed food items, collected in four seasons, were sampled and prepared as consumed. These mycotoxins were primarily found at low levels. The highest mean levels (upper bound) were: AFs, 1.50 µg kg^–1 in legumes, nuts and seed; OTA, 0.22 µg kg^–1 in sugars and confectionery; FNs, 9.76 µg kg^–1 in cereals and their products; DON and AcDONs, 33.1 µg kg^–1 in cereals and their products; and ZEA, 53.8 µg kg^–1 in fats and oils. The estimated dietary exposures of Hong Kong adults to the mycotoxins analysed were well below the respective health-based guidance values, where available. For AFs, the upper-bound exposure for high consumers is 0.0049 µg kg bw^–1 day^–1, which was estimated to contribute to about 7.7 (< 1%) of liver cancer cases when compared with 1222 liver cancer cases per year in Hong Kong. The percentage contributions of the estimated 95th percentile dietary exposures (lower and upper bound) to the health-based guidance values of individual mycotoxins were: ochratoxin A, 3.6–9.2%; fumonisins, 0.04–8.5%; deoxynivalenol and acetyldeoxynivalenols, 21.7–28.2%; and zearalenone 3.3–34.5%. The findings indicate that dietary exposures to all the mycotoxins analysed in this study were unlikely to pose an unacceptable health risk to the Hong Kong population.     

油料和食用油中真菌毒素快速测定方法的研究

本研究对油料（大豆、油菜籽）以及食用植物油中黄曲霉毒素、赭曲霉毒素A和玉米赤霉烯酮的快速检测方法进行了研究,建立了快速检测的筛选方法（免疫亲和柱层析净化荧光光度法）和确证方法（免疫亲和柱层析净化高效液相色谱法）.     

Mycotoxins in botanicals and dried fruits: A review

Botanicals are used in many countries for medicinal and general health-promoting purposes. Numerous natural occurrences of mycotoxins in botanicals and dried fruits have been reported. Aflatoxins or ochratoxin A (OTA) have been found in botanicals such as ginseng, ginger, liquorice, turmeric, and kava-kava in the USA, Spain, Argentina, India, and some other countries, while fumonisins have been found in medicinal wild plants in South Africa and in herbal tea and medicinal plants in Turkey. Zearalenone was identified in ginseng root. Dried fruits can be contaminated with aflatoxins, OTA, kojic acid, and, occasionally, with patulin or zearalenone. One main area of concern is aflatoxins in dried figs; bright greenish yellow fluorescence under ultraviolet light is associated with aflatoxin contamination. OTA in dried vine fruits (raisins, sultanas, and currants) is another concern. There are also reports of aflatoxins in raisins and OTA in dried figs, apricots, dried plums (prunes), dates, and quince. Maximum permitted levels in the European Union include 4 µg kg^-1 for total aflatoxins in dried fruit intended for direct consumption and 10 µg kg^-1 for OTA in dried vine fruit. This review discusses the occurrence of mycotoxins in botanicals and dried fruits and analytical issues such as sampling, sample preparation, and methods for analysis. Fungal contamination of these products, the influence of sorting, storage, and processing, and prevention are also considered.     

Effects of processing on mycotoxin stability in cereals

The mycotoxins that generally occur in cereals and other products are not completely destroyed during food‐processing operations and can contaminate finished processed foods. The mycotoxins most usually associated with cereal grains are aflatoxins, ochratoxins, deoxynivalenol, zearalenone and fumonisins. The various food processes that may have effects on mycotoxins include cleaning, milling, brewing, cooking, baking, frying, roasting, flaking, alkaline cooking, nixtamalization, and extrusion. Most of the food processes have variable effects on mycotoxins, with those that utilize high temperatures having the greatest effects. In general, the processes reduce mycotoxin concentrations significantly, but do not eliminate them completely. This review focuses on the effects of various thermal treatments on mycotoxins. © 2014 Society of Chemical Industry     

Validation of the procedure for the simultaneous determination of aflatoxins ochratoxin A and zearalenone in cereals using HPLC-FLD

Method validation for quantitative analysis of aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) in cereals using HPLC with fluorescence detector (FLD) is described. Mycotoxins were extracted with methanol?:?water (80?:?20) and purified with a multifunctional AOZ immunoaffinity column before HPLC analysis. The validation of the analytical method was performed to establish the following parameters: specificity, selectivity, linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision (within- and between-day variability), stability, robustness, measurement of performance, and measurement of uncertainty. Calibration curves were linear (r?>?0.999) over the concentration range, from the LOQ to 26, 40 and 400?ng/g for AFs, OTA and ZEA, respectively. LOD and LOQ were 0.0125 and 0.05?ng/g for aflatoxin B1 (AFB1) and G1 (AFG1), 0.0037 and 0.015?ng/g for aflatoxin B2 (AFB2) and G2 (AFG2), as well as 0.05 and 0.2?ng/g for OTA and 0.5 and 2?ng/g for ZEA, respectively. The mean recovery values were 77–104% for different concentrations of AFs, OTA and ZEA in spiked cereal samples. Both intra- and inter-day accuracy and precision were within acceptable limits. This method was successfully applied for the simultaneous determination of mycotoxins for 60 cereal samples collected from Malaysian markets. Fifty per cent of the cereal samples were contaminated with at least one of these mycotoxins, at a level greater than the LOD. Only one wheat sample and two rice samples were contaminated with levels greater than the European Union regulatory limits for AFs and OTA (4 and 5?ng/g). The means and ranges of mycotoxins obtained for the cereal samples were 0.4?ng/g and 0.01–5.9?ng/g for total AFs; 0.18?ng/g and 0.03–5.3?ng/g for OTA; and 2.8?ng/g and 2.4–73.1?ng/g for ZEA, respectively. The results indicate that the method is suitable for the simultaneous determination of AFs, OTA and ZEA in cereals and is suitable for routine analysis.     

Optimization and validation of a HPLC method for simultaneous determination of aflatoxin B1, B2, G1, G2, ochratoxin A and zearalenone using an experimental design

A reversed-phase HPLC optimization strategy is presented for investigating the separation and retention behavior of aflatoxin B₁, B₂, G₁, G_2, ochratoxin A and zearalenone, simultaneously. A fractional factorial design (FFD) was used to screen the significance effect of seven independent variables on chromatographic responses. The independent variables used were: (X1) column oven temperature (20–40°C), (X2) flow rate (0.8–1.2?ml/min), (X3) acid concentration in aqueous phase (0–2%), (X4) organic solvent percentage at the beginning (40–50%), and (X5) at the end (50–60%) of the gradient mobile phase, as well as (X6) ratio of methanol/acetonitrile at the beginning (1–4) and (X7) at the end (0–1) of gradient mobile phase. Responses of chromatographic analysis were resolution of mycotoxin peaks and HPLC run time. A central composite design (CCD) using response surface methodology (RSM) was then carried out for optimization of the most significant factors by multiple regression models for response variables. The proposed optimal method using 40°C oven temperature, 1?ml/min flow rate, 0.1% acetic acid concentration in aqueous phase, 41% organic phase (beginning), 60% organic phase (end), 1.92 ratio of methanol to acetonitrile (beginning) and 0.2 ratio (end) for X1–X7, respectively, showed good prediction ability between the experimental data and predictive values throughout the studied parameter space. Finally, the optimized method was validated by measuring the linearity, sensitivity, accuracy and precision parameters, and has been applied successfully to the analysis of spiked cereal samples.     

免疫亲和柱的制备及在真菌毒素检测中应用的研究进展

真菌毒素是真菌的次生代谢产物,是引起谷物和饲料污染的重要元凶,其检测手段日趋完善。目前免疫亲和柱在真菌毒素检测分析方面已经被广泛使用,本文综述了免疫亲和柱的原理及其制备方式,及其在几种常见真菌毒素检测方面的应用。      

Aflatoxins,ochratoxin A and zearalenone in Brazilian corn cultivars

Past surveys indicated that the occurrence of aflatoxins, zearalenone and ochratoxin A was not a problem in corn and corn products in the state of São Paulo, Brazil. However, according to recent studies, a change in pattern has been detected. To obtain a better overview, these toxins were searched for in 110 samples of freshly harvested corn, corresponding to 48 commercial cultivars planted at three different locations in the state. Aflatoxin contamination was found in 60 (54.5%) of the samples, in levels ranging from 6 to 1600 µg kg^?1 aflatoxin B₁. Insect control was exercised, so this was not the main route of corn infection. Endosperm type, germplasm type, number of days to flowering, and length of time the mature corn remained in the field had no effect on aflatoxin contamination. Ochratoxin A was found in two samples (206 and 128 µg kg^?1) and zearalenone in one sample (4640 µg kg^?1). Possible causes of the increase in aflatoxin levels may lie in the changing nature of the commercial cultivars employed, associated with the forsaking of the original landraces, and in a change in the toxigenicity pattern of the corn mycoflora Aspergillus flavus/Aspergillus parasiticus prevailing strains. © 2001 Society of Chemical Industry     

Survey of ochratoxin A and deoxynivalenol in stored grains from the 1999 harvest in the UK

Three hundred and twenty samples from the 1999 UK harvest comprising wheat (201 samples), barley (106) and oats (13) were analysed for ochratoxin A and deoxynivalenol. A small number of organic samples was also obtained. Samples were collected from farms, central stores, mills, maltings and ports from across the UK from February to April 2000. Ochratoxin A and deoxynivalenol analysis was by affinity column clean up and high-performance liquid chromatography with fluorescence and ultraviolet light detection, respectively, with limits of detection of 0.2 and 20 μg kg^-1. The survey found ochratoxin A at below 5 μg kg^-1 in 97% of the samples indicating satisfactory storage conditions. The remaining 3% of the samples contained ochratoxin A at levels between 5.2 and 231 μg kg^-1, but none of these samples was intended for human consumption. Deoxynivalenol was detected in 88% of all samples, with 83% below 100 μg kg^-1; the maximum level was 600 μg kg^-1. Twenty samples containing deoxynivalenol at or above 150 μg kg^-1 by high-performance liquid chromatography were all confirmed by gas chromatography/mass spectrometry. Nivalenol was also detected by gas chromatography/mass spectrometry at levels of 50 μg kg^-1 or higher in 18 of 20 samples where deoxynivalenol was confirmed.     

高效液相色谱法同时检测粮食中常见8 种真菌毒素的含量

建立免疫亲和柱净化-高效液相色谱法同时测定粮食中黄曲霉毒素B1（aflatoxins,AFB1）、黄曲霉毒素B2（AFB2）、黄曲霉毒素G1（AFG1）、黄曲霉毒素G2（AFG2）、赭曲霉毒素A（ochratoxin A,OTA）、玉米赤霉烯酮（zearalenone,ZEN）、呕吐毒素（deoxynivalenol,DON）和T-2毒素的检测方法。样品经乙腈-水提取后,用免疫亲和柱净化,Agilent Elipse Plus C18（100 mm×4 mm,3.5 μm）色谱柱分离,以甲醇-乙腈-水-乙酸为流动相,流速1 mL/min,柱温35 ℃,进样量20 μL,检测系统为可变波长检测器串联光化学衍生器串联荧光检测器。根据信噪比为3的峰响应值,确定各真菌毒素的检出限为：AFB1 0.446 ng/mL、AFB2 0.152 ng/mL、AFG1 0.523 ng/mL、AFG2 0.334 ng/mL、ZEN 7 ng/mL、OTA 0.7 ng/mL、DON 200 ng/mL、T-2毒素100 ng/mL。样品中各真菌毒素的平均加标回收率,玉米为80.0%～104.5%,小麦为83.2%～102.8%,方法精密度为2.6%～10.2%。从样品前处理到分析整个过程耗时约2 h。本方法简便、快速、灵敏度高,适用于粮食中多种真菌毒素的快速测定。     

Simultaneous determination of 10 mycotoxins in grain by ultra-high-performance liquid chromatography–tandem mass spectrometry using 13C15-deoxynivalenol as internal standard

An ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method for simultaneous determination of 10 mycotoxins in grain was developed. The selected mycotoxins were: deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, fusarenon X, moniliformin, zearalenone, zearalanone, ochratoxin A and ochratoxin B. The samples were extracted with aqueous acetonitrile (84?:?16,?v/v) and purified by reliable laboratory-made mixed cartridges. The analytes were separated on an Acquity UPLC HSS T3 column (100?×?2.1?mm,?1.8?µm) and eluted with a mobile phase of water containing 0.2% aqueous ammonia and acetonitrile/methanol (90?:?10,?v/v). All mycotoxins were detected with a Waters Micromass Quattro Ultima Pt tandem quadrupole mass spectrometer operating in negative electrospray ionization using multiple reaction monitoring mode. Accurate determination was achieved by employing commercial ¹³C₁₅-deoxynivalenol as internal standard, which compensated for target loss and eliminated matrix effects. The established method was further validated by determining the linearity (R ^2?>?0.9990), average recovery (75.8–106.5%), sensitivity (limit of quantitation 0.09–8.48?µg?kg^?1) and precision (relative standard deviation?≤?6.9%). It was shown to be a suitable method for simultaneous determination of 10 mycotoxins in grain. Finally, a total of 69 corn samples randomly collected from eastern and northern China were analyzed. The results showed that deoxynivalenol was the most frequently detected contaminant, whilst 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol, zearalenone, zearalanone, fusarenon X and moniliformin also occurred frequently. Ochratoxin A and ochratoxin B were present only in trace amounts in a small number of samples.     

粮食中主要真菌毒素危害及联合毒性研究进展

真菌毒素由丝状真菌代谢产生,具有很强的毒性。基于人类肝癌细胞系（HepG2细胞）能形成稳定的细胞系,分泌多种血浆蛋白,可以有效评估真菌毒素的体外毒性。文章综述了主要真菌毒素对粮食的危害,以及真菌毒素间联合毒性对人体细胞产生的副作用,对未来预测和预防粮食中真菌毒素对人体和动物健康影响意义重大。     

Geostatistical analysis of the spatial distribution of mycotoxin concentration in bulk cereals

Deoxynivalenol (DON) and ochratoxin A (OTA) in agricultural commodities present hazards to human and animal health. Bulk lots are routinely sampled for their presence, but it is widely acknowledged that designing sampling plans is particularly problematical because of the heterogeneous distribution of the mycotoxins. Previous studies have not explicitly looked at the interactions between the spatial distribution of the mycotoxin and the strategy used to take samples from bulk. Sampling plans are therefore designed on the assumption of random distributions. The objective of this study was to analyse the spatial distribution of DON and OTA in bulk commodities with geostatistics. This study was the first application of geostatistical analysis to data on mycotoxins contamination of bulk commodities. Data sets for DON and OTA in bulk storage were collected from the literature and personal communications, of which only one contained data suitable for geostatistical analysis. This data set represented a 26-tonne truck of wheat with a total of 100 sampled points. The mean concentrations of DON and OTA were 1342 and 0.59 µg kg^–1, respectively. The results showed that DON presented spatial structure, whilst OTA was randomly distributed in space. This difference between DON and OTA probably reflected the fact that DON is produced in the field, whereas OTA is produced in storage. The presence of spatial structure for DON implies that sampling plans need to consider the location of sample points in addition to the number of points sampled in order to obtain reliable estimates of quantities such as the mean contamination.     

Aflatoxin and ochratoxin A content of spices in Hungary

In October and November 2004, 91 spice samples (70 ground red pepper, six black pepper, five white pepper, five spice mix and five chilli samples), the majority of which originated from commercial outlets, were analysed for aflatoxins B₁, B₂, G₁ and G₂ (AFB1, AFB2, AFG1, AFG2) and ochratoxin A (OTA) content by high-performance liquid chromatography (HPLC) after immunoaffinity column clean-up. Eighteen of the 70 ground red pepper samples contained AFB1, seven of them in a concentration exceeding the 'maximum level' of 5 µg kg^-1 (range 6.1-15.7 µg kg^-1). Of the other spices assayed, the AFB1 contamination of one chilli sample exceeded 5 µg kg^-1 (8.1 µg kg^-1). Thirty-two of the 70 ground red pepper samples contained OTA, eight of them in a concentration exceeding the 10 µg kg^-1 'maximum level' (range 10.6-66.2 µg kg^-1). One chilli sample was contaminated with OTA at 2.1 µg kg^-1. The AFB1 and OTA contamination of ground red pepper exceeding the 'maximum level' (5 and 10 µg kg^-1, respectively) was obviously the consequence of mixing imported ground red pepper batches heavily contaminated with AFB1 and OTA with red pepper produced in Hungary. This case calls attention to the importance of consistently screening imported batches of ground red pepper for aflatoxin and ochratoxin A content and strictly prohibiting the use of batches containing mycotoxin concentrations exceeding the maximum permitted level.