Ochratoxin A exposure assessment of the inhabitants of Lisbon during winter 2007/2008 through bread and urine analysis

A survey on the occurrence of ochratoxin A (OTA) in 41 bread samples was carried out in the Portuguese capital, Lisbon. Maize (5) and wheat bread (36) and 43 representative urine samples from the Lisbon region were assayed for OTA levels using immunoaffinity column cleanup (IAC) and HPLC with fluorimetric detection (LC–FD). The percentage of OTA-positive samples was slightly higher for maize bread (80%) than wheat bread (70.8%), although, due to its higher consumption, the latter contributes more to OTA exposure, featuring a higher estimated daily intake (EDI). In the urine samples analyzed, both female and male residents displayed similarly high levels of OTA frequency and average contamination. In summary, OTA is a food contaminant of concern and may constitute a hazard for public health through consumption of cereal-based products.     

Determination of Ochratoxin A in Bread: Evaluation of Microwave-Assisted Extraction Using an Orthogonal Composite Design Coupled with Response Surface Methodology

An analytical method using microwave-assisted extraction (MAE) and liquid chromatography (LC) with fluorescence detection (FD) for the determination of ochratoxin A (OTA) in bread samples is described. A 2⁴ orthogonal composite design coupled with response surface methodology was used to study the influence of MAE parameters (extraction time, temperature, solvent volume, and stirring speed) in order to maximize OTA recovery. The optimized MAE conditions were the following: 25 mL of acetonitrile, 10 min of extraction, at 80 °C, and maximum stirring speed. Validation of the overall methodology was performed by spiking assays at five levels (0.1–3.00 ng/g). The quantification limit was 0.005 ng/g. The established method was then applied to 64 bread samples (wheat, maize, and wheat/maize bread) collected in Oporto region (Northern Portugal). OTA was detected in 84 % of the samples with a maximum value of 2.87 ng/g below the European maximum limit established for OTA in cereal products of 3 ng/g.     

Levels of ochratoxin A in wheat and maize bread from the central zone of Portugal

Ochratoxigenic fungi are natural contaminants of cereal and the produced toxins are harmful to humans and animals. Ochratoxin A (OTA) is among the most important mycotoxins, and the International Agency for Research on Cancer (IARC) classifies it as possibly carcinogenic to humans (group 2B). A total of 61 samples of bread from the central zone of Portugal were analysed for OTA by liquid chromatography (LC) with fluorescence detection (FD). For confirmation two procedures were applied, methyl ester derivatization with boron trifluoride-methanol and liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI/MS/MS). As far as we know, this is the first report where on-line LC/electrospray ionization (ESI) tandem mass spectrometry (MS/MS) was used for OTA analysis in bread. Limits of detection (LOD) and quantification (LOQ) were 0.015 and 0.03 ng/g, using LC-FD, and 0.03 and 0.09 ng/g by LC-MS/MS. The incidence of OTA was 12.9% and 70.0% for wheat and maize bread, respectively. The highest OTA levels were obtained for maize bread, having one sample exceeded the European maximum limit established for OTA in cereal products. The estimate daily intake (EDI) was below the tolerable daily intake.     

Occurrence and Safety Evaluation of Ochratoxin A in Cereal‐based Baby Foods Collected from Iranian Retail Market

Contamination of agricultural commodities with ochratoxin A (OTA) is a worldwide concern in recent decades. Consumption of OTA‐contaminated baby foods exerts health implications especially in children as the most vulnerable subpopulations. In the current study, for the first time in Iran, 64 baby foods (rice, wheat, and multigrain) samples from five different brands available in the Iranian market were analyzed to determine OTA level, using a HPLC with fluorescence detector. Overall, OTA was observed in 41% of analyzed samples with a mean and maximum level of 0.42 ± 0.27 and 1.1 μg/kg, respectively. OTA levels in five of 64 samples (7.8 %) were higher than the permissible limit recommended by European Commission (permissible limit: 0.5 μg/kg) and OTA levels in two of 64 samples (3.1%) were higher than the standard set by Iranian standard organization (1 μg/kg). The highest OTA contamination was observed in rice‐based baby food cereals (1.1 μg/kg; 57% of the samples), followed by wheat‐based (23%) and multigrain (20%) samples. OTA intake in infants (≥9 months old) was more than established provisional tolerable weekly intake by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and the European Food Safety Authority (EFSA) (100 and 120 ng OTA per kg of body weight, respectively). OTA content in baby food and cereals, as well as other raw foodstuff should be investigated comprehensively to reduce the exposure rate of young children to OTA.     

Mycoflora and ochratoxin A producing strains of Aspergillus in Algerian wheat

Wheat is a basic staple food for very large segments of the population of Algeria. The aim of this study is to analyse ochratoxin A (OTA)-producing mould and OTA-contaminated wheat. To evaluate the mycoflora and the potential for OTA production by Aspergillus strains, a total of 85 samples of wheat destined for human consumption were collected from two regions in Algeria (Tizi Ouzou and Setif) during the following phases: preharvest, storage in silos, and after processing. The mean value counts of fungi ranged from 275 to 1277 CFU g(-1). The dominant genus was Aspergillus, predominantly A. flavus, A. niger and A. versicolor. The other isolated species were A. ochraceus, A. alliaceus, A. carbonarius, A. terreus, A. fumigatus, A. candidus and Aspergillus spp. The occurrence and the levels of the genus Penicillium, Fusarium, Alternaria and Mucor were substantially lower than those of Aspergillus. The storage in silos shows high levels of Aspergillus (66 to 84%), especially A. flavus, but A. niger and other fungi were isolated at relatively low percentages. Equal distribution of the fungal contamination into the bran, flour and semolina fractions was observed from Flour Mill and Semolina Mill. The genus Aspergillus remained present at high levels at several phases of the production process. In addition, the ability to produce OTA by 135 isolates belonging to eleven species of Aspergillus and 23 isolates of Penicillium spp. was analyzed using fluorescent detection-based HPLC. Thus, it was found that 51 isolates (32.3%) were ochratoxigenic. All isolated strains of A. ochraceus (12) and A. alliaceus (6) produced OTA at concentrations ranging from 0.23 to 11.50 microg g(-1). Most of the A. carbonarius strains (80%) were OTA producers (0.01 to 9.35 microg g(-1)), whereas A. terreus (50%), A. niger (28%), A. fumigatus (40%), A. versicolor (18%) and Penicillium spp. (21.7%) were low level producers (0.01 to 0.07 microg g(-1)). The concentration of OTA was determined in 30 samples of wheat. OTA was detected in 12 (40%) of the samples at levels ranging from 0.21 to 41.55 microg kg(-1).     

INSTRUMENTAL TEXTURE and BAKING QUALITY of HIGH-FIBER TOAST BREAD AS AFFECTED BY ADDED WHEAT MILL FRACTIONS

The effect of bran type, level of addition, particle size, addition of wheat germ, as well as other additives like improvers and dough conditioners, on the instrumental texture and baking quality of high-fiber toast bread (white pan bread) has been investigated. the specific loaf volume decreased significantly (3.45 cc/g) when the bran level was raised to 30%, but at 20% bran addition, the specific loaf volume was superior to that of the control bread. the specific loaf volume of test breads remained higher than the control bread up to a level of 7.5% wheat germ addition. Additives like ascorbic acid (50 ppm) and sodium stearoyl-2-lactylate (0.5%) further improved the baking quality of test bread samples. the objective texture values (measured as compression force, kg) indicated that the test bread with bran addition up to 20% and germ up to 7.5% possessed a softer texture (0.80 kg) than the control bread (1.02 kg). In comparison with control bread, the panelists gave higher sensory scores for all attributes of test bread samples containing up to 20% red coarse bran or up to 30% red fine bran. A similar trend in sensory quality of test samples containing up to 7.5% wheat germ was also observed. High-fiber toast bread made from white flour, equal proportions of coarse and fine bran at 20%, wheat germ at 7.5%, plus sodium stearoyl-2-lactylate at 0.5% levels, was found to possess softer texture and improved sensory quality than the whole wheat flour bread.     

Detection of deoxynivalenol contamination in wheat products in Thailand

A total of ninety samples in three kinds of wheat products (30 noodle, 30 bread, and 30 cereal samples) were collected from the supermarkets in Bangkok, Thailand, from February to April 2007. The occurrence of deoxynivalenol (DON) contamination in wheat products was investigated using high-performance liquid chromatography equipped with a UV light detector. The extraction method was performed using a multifunctional cleanup column. The limit of quantification was 0.10 microg x g(-1) from the range obtained in a linear calibration. The survey found almost 94% of the DON-contaminated samples below 1 microg x g(-1), which corresponds to the U.S. Food and Drug Administration advisory level. DON was detected in 18.9% (17 of 90) of all samples, in 6.67% (2 of 30) and 16.67% (5 of 30) of noodle and bread samples at levels from 0.17 to 0.35 and 0.14 to 1.13 microg x g(-1), respectively, while it was in 33.33% (10 of 30) of cereal samples at levels from 0.13 to 0.39 microg x g(-1). The results suggest that the exposure to DON from the consumption of wheat products, especially noodles, bread, and cereal, is at a very low risk level.     

ELISA法检测赭曲霉毒素A的改进研究

针对ELISA法检测赭曲霉毒素A(OTA),使用微振荡酶标板可以缩短免疫反应检测时间约30min。采用ELISA方法检测小麦样品中的OTA时,使用空白样品提取液来配制OTA标准溶液可以消除样品基质造成的干扰。同时对提取溶剂加以改进,采用NaCl-水-甲醇(10%:70:30)作为提取溶剂的提取效率最高。0.1、1、10μg/kg三个浓度水平的加标回收率为95%～110%,方法的重现性很好,变异系数小于10%。     

Dietary intake of ochratoxin A from conventional and organic bread

Ochratoxin A (OTA) was extracted from 100 bread samples by using accelerated solvent extraction (ASE) and analyzed with liquid chromatography coupled with fluorescence detection. The presence of OTA was confirmed by methyl-ester derivatization. Bread samples were bought from different bakeries and supermarkets, 74 of non-organic and 26 of organic bread. The incidence of OTA varied between 20.3% and 23.0% for non-organic and organic bread, respectively. The highest values were obtained with non-organic versus organic products, five samples exceeded the European maximum permitted limit of OTA (3 ng/g) for this product. Estimated daily intake of OTA in this study was 1.6 ng/kg b.w./day. This value represents 32% and 10% of the tolerable daily intake (TDI) according to the Scientific Committee on Food of the European Commission and the FAO/WHO Committee of Experts on Food Additives, respectively. The daily intake estimated from this study reflects the necessity to take a vigilant attitude to guarantee food safety.     

The effect of enzymic modification on the foaming, water absorption and baking quality of defatted soya flour

Defatted soya flour slurries were treated for 0,5,10,15 and 30 min with papain at 50°C and tested for whipability, water absorption and baking quality at replacement levels in wheat flour of 10,20 and 30% (w/w). When whipped, all modified suspensions showed volume increases exceeding 250% with lower stability ratings than the unmodified samples. The 30 min enzyme treated product exhibited excellent water absorption. Use in bread resulted in a depression of loaf volume, development of a granular texture, off-colour and flavour. As was expected, all favourable characteristics decreased upon increasing the percentage of soya product in the dough formulation. Loaf volumes of breads containing 10% modified soya flour (MSF) ranged from 444 to 527 cc; with 20% MSF between 374 and 428 cc, and with 30% MSF between 383 and 409 cc. Loaf volume of all wheat flour bread was 861cc. Hunter colour difference meter readings indicated samples at all replacement levels, regardless of the enzyme modification time, exhibited higher levels of visual lightness than all wheat flour bread. All experimental breads exhibited a higher level of yellow tones and lower level of green tones compared to the all wheat flour bread.     

Occurrence of ochratoxin A in Canadian wheat shipments, 2010–12

Randomly selected domestic and export shipments (n = 1907) of Canadian durum and other wheat that occurred between 1 January 2010 and 31 December 2012 were analysed for ochratoxin A (OTA). The majority of samples did not contain OTA above the LOQ of 1 μg kg^–1. Only 37% of samples analysed contained quantifiable OTA; the median OTA of the positive results was 2.10 μg kg^–1. Canada Western Amber Durum shipments contained OTA more frequently, and at slightly higher concentrations, than Canada Western Red Spring wheat. For both wheat classes the frequency of OTA occurrence and mean concentrations appeared to increase in the lower grades, but these increases were not statistically significant. A periodic trend of a late summer increase of mean monthly OTA concentrations in shipments appears tied to the cycle of producer deliveries of wheat to primary grain elevators.     

Maternal dietary habits and mycotoxin occurrence in human mature milk

During 2006, 82 samples of human mature milk were collected at Italian hospitals and checked for aflatoxin M1 (AFM1) and ochratoxin A (OTA) by immunoaffinity column extraction and HPLC. AFM1 was detected in four (5%) of milk samples (ranging from < 7 ng/L to 140 ng/L; mean level: 55.35 ng/L); OTA was detected in 61 (74%) of milk samples (ranging from < 5 ng/L to 405 ng/L; mean level: 30.43 ng/L. OTA levels were significantly higher (p less, not double equals 0.05) in milk of habitual consumers of bread, bakery products and cured pork meat. No other statistically significant differences were observed although habitual consumers of pasta (p = 0.059), cookies (p = 0.061) and juices (p = 0.063) had mean contamination values of OTA higher than the moderate consumer. The very few AFB1 positive samples did not allow statistical comparisons. The present study confirms that the occurrence of OTA in human milk is related to maternal dietary habits. The findings support the possibility of dietary recommendations to woman, during pregnancy and lactation, aimed to tentatively reduce the OTA contamination of human milk.     

谷物中赭曲霉毒素A化学发光酶免疫分析法的建立

建立了间接竞争化学发光酶免疫法检测赭曲霉毒素A(ochratoxin A,OTA),该方法IC50为112 pg/mL,检出限是2.47 pg/mL,平均批内和批间变异系数分别为7.03%和14.7%。在大米和小麦样本中添加浓度1.5～6μg/kg的OTA标品,平均回收率在66.97%～97.96%之间,与其他常见真菌毒素未见交叉反应。将该方法应用于30份谷物样本(包括20份大米样本和10份小麦样本)中OTA的检测,检测结果与商品化ELISA试剂盒的相关系数R2=0.942 4。该方法简单、灵敏、快速、准确适用于谷物中OTA的检测。     

Occurrence of Penicillium verrucosum in retail wheat flours from the Spanish market

In Spain, low ochratoxin A (OTA) levels have been detected in wheat and different wheat products but no information has been published about the fungi involved in this OTA contamination. Some species of the genera Penicillium and Aspergillus are known to form OTA but few of them are known to contaminate foods with this mycotoxin. Penicillium verrucosum, an important OTA producer typical of temperate and cold climates, is much more frequently found on cereals in countries where they occasionally have OTA problems as in North European countries compared with South Europe, where levels of OTA generally seem to be lower or is not detected. The aim of this study was to determine, identify and characterize the occurrence of potential OTA-producing Aspergillus spp. and Penicillium spp. from retail wheat flours purchased in the Spanish market and used for human consumption. A total of 105 Aspergillus isolates were analyzed in order to know whether they are able to produce OTA and/or citrinin (CIT). None of these isolates were able to produce these mycotoxins. However, 17 suspected P. verrucosum isolates were recovered and confirmed by RAPD analyses. Eleven isolates were OTA producers and 14 isolates produced CIT. Our results confirm the potential risk of OTA and CIT production in wheat flours if stored improperly and the occurrence of P. verrucosum in South European countries. This was the only species able to produce these mycotoxins.     

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).     

Exposure assessment to ochratoxin A in Catalonia (Spain) based on the consumption of cereals, nuts, coffee, wine, and beer

Ochratoxin A (OTA) was analysed in composite samples of cereal-based baby foods, beer, breakfast cereals (corn- and rice and wheat-based), loaf bread, peanuts and pistachios. Foodstuffs were collected in hypermarkets and supermarkets from 12 cities in the Spanish region of Catalonia, and composite samples were prepared for analysis involving liquid-liquid extraction, followed by immunoaffinity column clean-up and HPLC with fluorescence detection. Consumption data for the selected foodstuffs were collected by means of a food-frequency questionnaire. The studied population was grouped by age in infants, children, adolescents and adults; and exposure to OTA through the specified foodstuffs, and through wine and coffee, was assessed. Exposure assessment was done through deterministic and probabilistic modelling of the contamination and consumption data. OTA occurrence and mean of positive samples (ng g(-1) or ng ml(-1), for beer) were the following: 8.7% and 0.233 in baby foods; 88.7% and 0.022 in beer; 2.8% and 0.728 in corn-based breakfast cereals; 25% and 0.293 in wheat-based breakfast cereals; 12.9% and 0.283 in loaf bread; 41.7% and 0.241 in peanuts; and 2.9% and 0.228 in pistachios. The median estimated daily intake of OTA through the foodstuffs by each age group were below the latest provisional tolerable daily intakes (PTDIs) of 17 and 14 ng kg(-1)?bw day(-1) recommended by the European Food Safety Authority (EFSA) in 2006 and the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 2007, respectively, ranging from 1% and 2% of those values in adolescents and children, to 3% and 11% in adults and infants.     

Aflatoxin B1 and sterigmatocystin in wheat and wheat products from supermarkets in China

Wheat is an important cereal but it is often contaminated with mycotoxins. The natural occurrence of aflatoxin B₁ (AFB₁) and sterigmatocystin (STC) was determined in 178 food samples (32 wheat samples and 146 wheat products) purchased from Chinese supermarkets. The methodology was validated, the wheat and wheat products samples were treated with a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). From these samples 18.8% of wheat and 8.2% of cracker samples were contaminated with AFB₁. Mean levels were 0.06 µg/kg and 0.05µg/kg, respectively. There was no AFB₁ contamination in white bread or whole meal bread. Meanwhile 53.1% of wheat, 59.2% of crackers, 20.8% of white bread and 16% of whole meal bread samples were contaminated with STC. The mean levels were 0.07, 0.79, 0.12 and 0.12 µg/kg respectively. Although the levels were low, this demonstrates the need for more comprehensive surveys for these two mycotoxins in wheat and wheat products from China.     

Aflatoxins and ochratoxin A in flour: a survey of the Serbian retail market

This report presents data on the occurrence of aflatoxins (AF) and ochratoxin A in different types of flour marketed in Serbia. A total of 114 samples of wheat, buckwheat, rye, oat, barley, rice, millet and corn flour were collected in the period 2012–2016 and analysed using high performance liquid chromatography with fluorescence detection. Among flours other than corn, AFB1 was quantified only in rice, while ochratoxin A (OTA) was found in 29% of the samples. In corn flours the percentage of positive samples varied greatly over the years: AFB1 7.1–80.0%, OTA 30.0–40.6%, with a co-occurrence of 7.1–34.4%. Overall 5.2% of flours other than corn and 10.7% of corn flours exceeded the maximum levels (MLs) for AFB1 and/or OTA. The highest recorded levels were 8.80 μg kg^?1 of AFB1 (corn) and 23.04 μg kg^?1 of OTA (rye). Overall mean contamination levels of corn flours were 0.53 μg kg^?1 of AFB1 and 0.46 μg kg^?1 of OTA.     

小麦麸皮的品质改良及含麸皮面包焙烤品质的研究

本研究采用微波辐照联合复合酶法对小麦麸皮进行改良,将改良后的小麦麸皮添加至高筋粉中得到不同小麦麸皮含量的含麸皮面粉并制作含麸皮面包。结果表明:改良后小麦麸皮中脂肪酶残余酶活和脂肪氧化酶残余酶活降低至0和10.34%、粗纤维含量降低至3.21%、还原糖含量上升至3.79 g/100 g。与改良前的全麦面包相比,改良后的全麦面包组织结构得以改善、全麦面包储藏7d后吸热焓值降低了14.70%、全麦面包中酮基、羰基、醛基含量增多。改良后的全麦面包比容增加了26.40%、硬度降低了32.90%、弹性增大了6.90%。随着改良麸皮添加量的增加,含麸皮面包组织结构变得粗糙多孔,含麸皮面包吸热焓值逐渐降低,酮基、羰基、醛基含量增多,含麸皮面包芯亮度逐渐变暗、比容逐渐降低、硬度逐渐增加、弹性逐渐减小、感官品质降低。与未稳定化麸皮含量16%的全麦面包相比,稳定化全麦面包的稳定性、贮藏性和质构得到明显提高。     

Application of isotope dilution mass spectrometry: determination of ochratoxin A in the Canadian Total Diet Study

Analytical methods are generally developed and optimized for specific commodities. Total Diet Studies, representing typical food products 'as consumed', pose an analytical challenge since every food product is different. In order to address this technical challenge, a selective and sensitive analytical method was developed suitable for the quantitation of ochratoxin A (OTA) in Canadian Total Diet Study composites. The method uses an acidified solvent extraction, an immunoaffinity column (IAC) for clean-up, liquid chromatography-tandem mass spectrometry (LC-MS/MS) for identification and quantification, and a uniformly stable isotope-labelled OTA (U-[(13)C(20)]-OTA) as an internal recovery standard. Results are corrected for this standard. The method is accurate (101% average recovery) and precise (5.5% relative standard deviation (RSD)) based on 17 duplicate analysis of various food products over 2 years. A total of 140 diet composites were analysed for OTA as part of the Canadian Total Diet Study. Samples were collected at retail level from two Canadian cities, Quebec City and Calgary, in 2008 and 2009, respectively. The results indicate that 73% (102/140) of the samples had detectable levels of OTA, with some of the highest levels of OTA contamination found in the Canadian bread supply.