FTIR-ATR infrared spectroscopy for the detection of ochratoxin A in dried vine fruit

A method of screening sultanas for ochratoxin A (OTA) contamination, using mid-infrared spectroscopy/Golden Gate single-reflection ATR (attenuated total reflection), is described. The main spectral characteristics of sultanas from different sources were identified in a preliminary acquisition and spectral analysis study. Principal component analysis (PCA) showed that samples of various origins had different spectral characteristics, especially in water content and the fingerprint region. A lack of reproducibility was observed in the spectra acquired on different days. However, spectral repeatability was greatly improved when water activity of the sample was set at 0.62. A calibration curve of OTA was constructed in the range 10-40 µg OTA kg^-1. Samples with OTA levels higher than 20 µg kg^-1 were separated from samples contaminated with a lower concentration (10 µg OTA kg^-1) and from uncontaminated samples. The reported methodology is a reliable and simple technique for screening dried vine fruit for OTA.     

Fumonisin B1, fumonisin B2, zearalenone and ochratoxin A contamination of maize in Croatia

Mycotoxins are products of moulds that frequently contaminate maize. In this study the presence of mycotoxins fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), zearalenone (ZEA) and ochratoxin A (OTA) was determined in 49 maize grain samples collected in autumn 2002. The most frequent finding was that of FB₁(100%), followed by ZEA (84%) and OTA (39%), while FB₂ was found only in three samples. The co-occurrence of two and three mycotoxins was found in 55 and 37% of samples, respectively. The concentrations (mean ± SD) of FB₁, ZEA and OTA in positive samples were 459.8 ± 310.7, 3.84 ± 6.68 and 1.47 ± 0.38 µg kg^-1, respectively, and the concentrations of FB₂ in three positive samples were 68.4, 109.2 and 3084.0 µg kg^-1. Although such low concentrations of mycotoxins are not a significant source of exposure in countries with a European diet, a few samples with extreme values indicate that thorough control is needed.     

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.     

Incidence of toxigenic fungi and ochratoxin A in dried fruits sold in Brazil

A total of 117 dried fruit samples (black sultanas, white sultanas, dates, dried plums, dried figs and apricots) from different origins were analysed both for toxigenic fungi and for the presence of ochratoxin A. Amongst the fungi found, Aspergillus niger was predominant, with 406 isolates, of which 15% were ochratoxin A producers. They were followed by A. ochraceus, with 15 isolates and 87% ochratoxigenics, and A. carbonarius, with only five isolates of which 60% were ochratoxin A producers. The average infection rates for A. niger in black sultanas, plums, figs, dates and white sultanas were 22.0, 8.0, 4.0, 1.5 and 0.5%, respectively. The apricot samples were not contaminated by any fungi or ochratoxin A. Black sultana and dried figs contained the highest contamination with ochratoxin A, with 33 and 26.3% of the samples containing more than 5 µg kg^-1 respectively, while all the white sultanas, dates and plums had no sample that exceeded this limit.     

Consumers' ability to discriminate aflatoxin-contaminated Brazil nuts

The objectives of the study were to investigate the extent to which consumers can separate nuts with a high content of aflatoxin from sound nuts, and whether sorting results can be improved by information or whether they are affected by certain factors. A test panel consisting of 100 subjects was asked to crack 300 g Brazil nuts and to sort the nuts into those they considered edible and inedible. The test showed that consumers can, on current behaviour, discriminate aflatoxin-contaminated Brazil nuts to a significant extent. The median and the 95th percentile of the total concentrations of aflatoxins (B₁, B₂, G₁, G₂) in the samples before sorting were 1.4 and 557 µg kg^-1, respectively, and in the edible fractions after sorting 0.4 and 56 µg kg^-1, respectively. Given that levels of aflatoxins before sorting exceed either 2 µg aflatoxin B₁ kg^-1 or totally 4 µg aflatoxins kg^-1, there was no effect of aflatoxin concentrations before sorting on the probability of exceeding these thresholds in the edible fraction. This means that similar sorting results were obtained for samples with aflatoxin levels exceeding either of the two thresholds, irrespective of if the thresholds were exceeded with a few µg kg^-1 or up to more than 1000 µg kg^-1. None of the tested factors (such as sex, age, level of education, ethnic background or knowledge of mycotoxins) had any effects on the probability of exceeding either of the two aflatoxin thresholds.     

坚果和干果中黄曲霉毒素的污染、检测与控制

坚果和干果在生产、加工和储藏过程中易受到黄曲霉毒素污染。黄曲霉毒素是一类毒性极强的真菌毒素, 其中黄曲霉毒素B1污染最普遍、毒性最强, 其易引发急、慢性毒性, 致癌、致畸、免疫抑制等多种危害, 严重威胁果品安全, 危害人类健康。坚果和干果中黄曲霉毒素污染状况的阐释能够更加明晰其在不同果品中的污染差异, 为有针对性地控制毒素污染奠定了基础。坚果和干果中黄曲霉毒素的准确检测是分析其污染状况的前提。另外, 如何防控黄曲霉毒素在坚果和干果中的污染, 降低黄曲霉毒素对人类健康的影响, 一直是研究的重点。本文对国内外坚果和干果中黄曲霉毒素的污染情况及限量标准, 黄曲霉毒素的检测方法和防控措施进行了综述, 以期为坚果和干果中黄曲霉毒素的污染控制提供理论基础和指导。     

Aspergillus carbonarius as the main source of ochratoxin A contamination in dried vine fruits from the Spanish market

Ochratoxin A (OTA) can occur in a wide range of foods, but unexpectedly high concentrations have been detected in dried vine fruits of various origins. The European Union has recently established a maximum OTA limit of 10 microg/kg for these foodstuffs. In order to determine the likely origin of OTA, a mycological study of 50 dried fruit samples (currants, raisins, and sultanas) representative of the Spanish market was conducted. Fungal contamination was detected in 49 of 50 (98%) samples. Black aspergilli were isolated from all of the positive samples. Aspergillus niger var. niger was isolated from 98% of the samples, and Aspergillus carbonarius was found in 58% of the samples. One hundred sixty-eight A. niger var. niger isolates and 91 A. carbonarius isolates were screened for their ability to produce OTA. Eighty-eight (96.7%) A. carbonarius isolates and one (0.6%) A. niger var. niger isolate were found to be OTA producers. Black aspergilli were the dominant fungi. Among black aspergilli, A. carbonarius has shown a consistent ability to produce OTA and is the most probable source of this mycotoxin in these substrates.     

Raisins and Currants as Conventional Nutraceuticals in Italian Market: Natural Occurrence of Ochratoxin A

The healthy consumers make a strong pressure to natural products that can prevent the chronic diseases and improve the general health status, and therefore an important aspect that have to be considered is the safe level of the nutraceuticals. This study reports the occurrence of Ochratoxin A (OTA) and associated fungal contamination in 35 samples of dried vine fruits imported in the European community potentially used for the development of new nutraceutical supplements. High pressure liquid chromatography analysis identified 18 samples as contaminated by OTA with an average level of 2.6 μg/kg. OTA was measured in 4 samples of currants (mean value of 6.6 μg/kg) and 13 samples of raisins (mean value of 1.4 μg/kg). In one sample of currants and one of raisins from Turkey OTA exceeded the limits set by European Commission of 10 μg/kg, being contaminated with 12.61 and 15.99 μg/kg, respectively. All the positive samples were confirmed by Orbitrap Q Exactive through their molecular weight and the corresponding fragmentation. The worldwide consumption of dried vine fruits contributed to OTA exposure in several group of consumers. In particular, considering the potential nutraceutical approach, this consumption may be represent a severe risk for healthy consumers that consider these products like healthy and salutistic for their contents in antioxidants, flavonoids, and polyphenols. Data reported in this study confirmed the need to regularly monitor mycotoxin levels in these food products and optimize the process of fruits drying in order to reduce the development of toxigenic molds.     

Mycotoxins in fruits,fruit juices,and dried fruits

This review gives an overview of the presence of mycotoxins in fruits. Although several mycotoxins occur in nature, very few (aflatoxins, ochratoxin A, patulin, Alternaria toxins) are regularly found in fruits. It has been shown that the presence of fungi on fruits is not necessarily associated with mycotoxin contamination. The formation of mycotoxins depends more on endogenous and environmental factors than fungal growth does. Mycotoxins may remain in fruits even when the fungal mycelium has been removed. Depending on the fruit and the mycotoxin, the diffusion of mycotoxins into the sound tissues of fruits may occur. The influence of the selection and storage of fruits and the influence of different processing steps involved in the production of fruit juices and dried fruits on possible mycotoxin contamination is described. It is shown that the careful selection, washing, and sorting of fruits is the most important factor in the reduction of mycotoxin contamination during the production of fruit juices. The processing of fruits does not result in the complete removal of mycotoxins.     

Five-year survey of ochratoxin A in processed sultanas from Turkey

The results of surveillance for ochratoxin A (OTA) in 1885 samples of sultanas taken during five crop years between 1999 and 2003 are reported. The analytical method was based on extraction with methanol + sodium bicarbonate and clean-up by immunoaffinity column chromatography followed by high-performance liquid chromatography with fluorescence detection. The limit of detection for OTA was 0.3 µg kg^-1. The results show that 9.3% of the samples contained no detectable levels of OTA, whereas 0.6% had concentrations exceeding 10 µg kg^-1; the remaining 90.3% had levels within the range 0.3-10 µg kg^-1. The overall mean OTA concentration in the total number of 1885 samples taken was 1.36 ± 2.91 µg kg^-1; the overall median was calculated as 0.90 µg kg^-1.     

Aflatoxins in ginseng roots

Ginseng roots can be infected by molds during growth, harvest and storage and result in contamination with mycotoxins. In this study, an analytical method for the determination of aflatoxins B₁, B₂, G₁ and G₂, a group of structurally similar mycotoxins, in ginseng root was developed. Test samples were extracted with methanol-water (8 + 2), diluted and passed through an immunoaffinity column packed with antibodies specific for aflatoxins. The purified extract was then derivatized with a mixture of water, trifluoroacetic acid and acetic acid. Aflatoxins were then separated and quantified by reverse phase liquid chromatography (LC) with fluorescence detection. Recoveries of total aflatoxins at 2, 4, 8 and 16 ng/g added to toxin-free 4 to 5-year old dried sliced Wisconsin ginseng were 92, 77, 91 and 83% respectively; and relative standard deviations were 3.6, 8.0, 6.9 and 2.0% respectively. A total of 11 wild simulated and 12 cultivated ginseng root samples were analysed for aflatoxins. All cultivated roots were found to be free of aflatoxin contamination. Two of the wild simulated roots contained total aflatoxins B₁, B₂, G₁ and G₂ at 15.1 and 15.2 ng/g. One moldy ginseng root purchased from a grocery store was found to be contaminated with aflatoxins at 16 ng/g.     

European research on ochratoxin A in grapes and wine

European wine production represents about 70% of world production and thus is an important export commodity. Ochratoxin A (OTA) was first detected as a wine contaminant in 1996 and the role of Aspergillus section Nigri and A. carbonarius in OTA production discovered in Europe in 1999. Subsequently Europe-wide surveys have shown that A. carbonarius is predominantly responsible for OTA contamination of grapes, wine and vine fruits. Analyses of wine samples throughout Europe have shown that there is a gradient in OTA concentration with a decrease from red, to rose and to white wines. The latitude of production is an important factor in determining risk from OTA contamination. Some geographic regions in Southern Europe are more prone to contamination with the toxigenic species and OTA. Ochratoxin A has also been found in much higher concentrations (max. 53 mug/kg) in dried vine fruit than in wine suggesting that A. carbonarius can dominate the drying vine fruit ecosystem. There is a significant lack of knowledge in Europe on conducive climatic conditions preharvest and their relationship with levels of risk from OTA contamination in grapes and their fate in wine production. This needs to be integrated with cultivation system to maximise the prevention of OTA entering this food chain.     

果品中真菌毒素的污染、毒性、生物合成及影响因素研究进展

真菌毒素是危害果品质量的重要因子。果品中重点关注的真菌毒素有展青霉素、链格孢霉毒素、赭曲霉毒 素A和黄曲霉毒素,其形成、转化、消解规律和控制技术是果品质量安全研究的重要内容。本文从果品中真菌毒素 的污染种类、产毒菌株、毒性毒理、产毒基因、合成途径、影响因素等方面综述了相关研究进展。     

Determination of mycotoxins in biscuits,dried fruits and fruit jams: an assessment of human exposure

ABSTRACT

A reliable, fast and simple method using UHPLC-MS/MS was developed for the determination of aflatoxins B₁ (AFB1), G₁ (AFG1), B₂ (AFB2) and G₂ (AFG2), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEA), HT-2 toxin and T-2 toxin in crude extracts of biscuits with fruit filling, cookies, dried fruits and fruit jams. The method was successfully demonstrated on 39 samples of biscuits with fruit filling, 34 cookies, 14 dried fruits and 10 fruit jams. The mycotoxins detected in biscuits samples were ZEA, OTA, T-2 and AFB1 with an average concentrations of positive samples of 2.64, 4.10, 8.13 and 1.32 µg kg^?1, respectively; while the mycotoxins detected in jam samples were AFB1, OTA, T-2 and AFB2 with an average concentrations of positive samples of 2.00, 17.7, 4.37 and 1.15 µg kg^?1, respectively. The results showed that the majority of samples were in compliance with relevant regulations. However in eight samples of biscuits and three samples of fig jam the contents of OTA were higher than the existing OTA limits. The combined dietary exposure of selected mycotoxins was estimated for the first time for children, adolescents and adults. The estimated combined dietary exposures were all lower than the proposed value assumed to predict a possible risk scenario.     

Determination of selected mycotoxins in mould cheeses with liquid chromatography coupled to tandem with mass spectrometry

A simple and feasible method is described for analysing nine mycotoxins in cheese matrix. The method involves liquid extraction followed by high performance liquid chromatographic separation and mass spectrometric detection of the analytes, and allows the determination of aflatoxins B1, B2, G1, G2 and M1, ochratoxin A, mycophenolic acid, penicillic acid and roquefortine C simultaneously. Average recoveries of the mycotoxins from spiked samples at concentration levels of 5-200 µg kg^-1 ranged from 96-143%. Within-day relative standard deviations at these concentration levels varied from 2.3-12.1%. The limit of quantification for aflatoxin M1 was 0.6 µg kg^-1 and for the other compounds 5 µg kg^-1. The method developed was applied for analysing these mycotoxins in blue and white mould cheeses purchased from Finnish supermarkets. Roquefortine C was detected in all of the blue mould cheese samples in concentrations of 0.8-12 mg kg^-1. One blue cheese contained also 0.3 mg kg^-1 mycophenolic acid. The other investigated mycotoxins were absent in the samples.     

Heat of sorption of water in dried fruits

The heats of moisture adsorption and desorption in four dried fruits (sultana raisins, figs, prunes and apricots) were estimated from equilibrium sorption data, using the Clausius-Clapeyron equation in the temperature range 15–60°C. The net isosteric heat of sorption ( q st) decreased sharply from about 20 kJ mol⁻¹ water to near zero when the moisture content was increased from 0.05 to 0.50kg water kg⁻¹ dry matter. An exponential function was fitted to the experimental q st values at various moisture contents (X) , yielding two characteristic constants ( q 0 and X 0) for each fruit. Mean and total heats of sorption were calculated from the proposed empirical equation, which are useful for enthalpy prediction in food dehydration. The heats of desorption were higher than the heats of adsorption, indicating significant hysteresis in the sorption of water, especially in dried apricots.     

Emerging risk of infestation and contamination of dried fruits by mites in the Czech Republic

The introduction of live insects into human food is rare in developed countries. However, we report, for the first time, an emerging risk that exists from dried fruit in Central Europe. Recently, massive and frequent infestation of dried fruit imported from the Mediterranean region by the mite, Carpoglpyhus lactis L. (Acarina: Carpoglyphidae), has been found. In 180 samples taken from supermarkets, 13% were contaminated; the contamination levels ranged from 0 to 660 mites per g of dried fruit. The contamination was found in dried apricots, figs, plums and raisins. To estimate the risks and food preferences of C. lactis, its growth rate was examined under laboratory conditions. Starting with a hypothetical population of 10 mites per g of dried fruit, the risk level of 1000 mites per g of dried fruit is reached at 42 days for dried figs, 49 days for dried pineapple and 63 days for dried apricots, dates and plums at 25 °C and 85% relative humidity. We found that mites are able to enter every dried fruit packing material tested, including polypropylene and aluminum foils. This indicates that mites can move from package to package in supermarkets. Mites are known as allergen producers and vectors of mycotoxin-producing fungi. These findings indicate that an increased risk of C. lactis contamination exists in dried fruit.     

Emerging risk of infestation and contamination of dried fruits by mites in the Czech Republic

The introduction of live insects into human food is rare in developed countries. However, we report, for the first time, an emerging risk that exists from dried fruit in Central Europe. Recently, massive and frequent infestation of dried fruit imported from the Mediterranean region by the mite, Carpoglpyhus lactis L. (Acarina: Carpoglyphidae), has been found. In 180 samples taken from supermarkets, 13% were contaminated; the contamination levels ranged from 0 to 660 mites per g of dried fruit. The contamination was found in dried apricots, figs, plums and raisins. To estimate the risks and food preferences of C. lactis, its growth rate was examined under laboratory conditions. Starting with a hypothetical population of 10 mites per g of dried fruit, the risk level of 1000 mites per g of dried fruit is reached at 42 days for dried figs, 49 days for dried pineapple and 63 days for dried apricots, dates and plums at 25°C and 85% relative humidity. We found that mites are able to enter every dried fruit packing material tested, including polypropylene and aluminum foils. This indicates that mites can move from package to package in supermarkets. Mites are known as allergen producers and vectors of mycotoxin-producing fungi. These findings indicate that an increased risk of C. lactis contamination exists in dried fruit.     

Fumonisin contamination and fumonisin producing black Aspergilli in dried vine fruits of different origin

Aspergillus niger isolates are able to produce fumonisins in high quantities on agar media with a low water activity. Several agricultural products fit this criterion, including dried vine fruits, dates and figs. Data on the occurrence and role of this species in fumonisin contamination of agricultural products with high sugar content are needed to clarify the importance of A. niger in human health. The mycobiota and fumonisin contamination of various dried vine fruit samples collected from different countries were examined to clarify the role of black Aspergilli in fumonisin contamination of such products. All except two of the examined samples were contaminated with black Aspergilli. Species assignment of the isolates was carried out using sequence analysis of part of the calmodulin gene. The range of fumonisin isomers present in the raisin samples, and produced by A. niger isolates collected from dried vine fruits was also examined using reversed-phase high-performance liquid chromatography/electrospray ionization-ion trap mass spectrometry (RP-HPLC/ESI-ITMS). Among the 30 A. niger/A. awamori isolates identified, 20 were found to be able to produce fumonisins (average contamination: 5.16 mg/kg; range: 0.017-19.6 mg/kg). The average fumonisin content of the 7 dried vine fruit samples which were found to be contaminated by potential fumonisin producing black Aspergilli was 7.22 mg/kg (range: 4.55-35.49 mg/kg). The isolates produced several fumonisin isomers also present in the dried vine fruit samples, including fumonisins B_1-4, 3-epi-FB₃, 3-epi-FB₄, iso-FB₁, and two iso-FB_2,3 forms. Fumonisin B₁ was detected for the first time in A. niger cultures. Most of these isomers have previously only been identified in Fusarium species. Our data indicate that A. niger and A. awamori are responsible for fumonisin contamination of dried vine fruits worldwide. The observed levels of contamination are alarming and pose a new threat for food safety.     

Mycoflora and natural occurrence of aflatoxin, cyclopiazonic acid, fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B(1) (FB(1)) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range = 0.1-15.3 ng g(-1)), 74.7% of the samples had FB(1) (range = 0.05-3.65 mg kg(-1)), 10.0% of the samples had aflatoxin (range = 0.1-763.2 ng g(-1)) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range = 25-187 ng g(-1)). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.