Ochratoxin A in dried vine fruits from Chinese markets

A total of 56 dried vine fruits, including 31 sultanas and 25 currants, were selected from Chinese markets in 2012. All samples were analysed for Ochratoxin A (OTA) using solid-phase extraction and high-performance liquid chromatography (HPLC) with fluorescence detection. It turned out that 58.9% of the samples were positive and the OTA amount ranged from <0.07 to 12.83 μg/kg, with a mean level of 0.99 μg/kg. Only one sample exceeded the European Union (EU) maximum level of 10 μg/kg. Meanwhile, it was shown that OTA contamination increased among north-western, northern and southern China, which showed OTA means of 0.08, 0.99 and 2.01 μg/kg, respectively. Moreover, in samples of products sold in sealed plastic bags, i.e. consumer-size packages (n = 19, mean = 0.30 μg/kg) less OTA was detected when compared with sampled bulk packages (n = 37, mean = 1.67 μg/kg). In addition, sultanas (mean = 0.92 μg/kg) had less OTA contamination than currants (mean = 1.22 μg/kg).     

Occurrence of ochratoxin A and citrinin in Czech cereals and comparison of two HPLC methods for ochratoxin A detection

The aims of the study were to obtain information about the occurrence of ochratoxin A (OTA) and citrinin (CIT) in cereals harvested in the Czech Republic and to compare two analytical procedures for detecting OTA. A total of 34 cereal samples, including two matrix reference materials (R-Biopharm, Germany), were analysed. The results were compared with the limit for raw cereal grains used as a foodstuff according to Commission Regulation No. 1881/2006, which allows a maximum OTA level of 5 µg kg^?1. Compared were two methods based on the high-performance liquid chromatography principle, one using the immunoaffinity columns OchraTest? (VICAM) and the second based on solvent partition (PART), both followed by fluorescence detection. The highest OTA contents were found in two barley samples. According to the method employed, the results for the first sample (malting barley) were VICAM = 31.43 µg kg^?1 and PART = 44.74 µg kg^?1. For the second sample (feeding barley) they were VICAM = 48.63 µg kg^?1 and PART = 34.40 µg kg^?1. Two samples of bread wheat had an OTA content approaching the legal limit (VICAM = 4.71 µg kg^?1 and PART = 6.03 µg kg^?1; VICAM = 4.12 µg kg^?1 and PART = 3.95 µg kg^?1). CIT was analysed using the PART method only, and its highest content (93.64 µg kg^?1) was found for the malting barley sample with high OTA content (44.74 µg kg^?1 as analysed using PART).     

Aflatoxins and ochratoxin A in tea prepared from naturally contaminated powdered ginger

The migration of several major mycotoxins, aflatoxins B₁ (AFB₁), B₂, G₁, and G₂ (AFT, total of the aflatoxins) and ochratoxin A (OTA), from naturally contaminated powdered ginger to surrounding liquid (tea) was investigated. The toxins are commonly found in cereal grains and are toxic, carcinogenic and thermostable. Ginger root is widely used for digestive problems. Powdered ginger (2 g) found to contain AFT and OTA was placed in an empty heat sealable tea bag. The tea bag was heat-sealed and used to prepare tea under different conditions: temperature (50 and 100°C), time (5 and 10 min) and volume (100 and 200 ml). The tea bag was placed in hot water and stirred every 1 min for 5 s during the first 5 min of steeping. After steeping, the tea bag was removed and the tea and ginger residue (in the tea bag) were analysed separately for AFT and OTA. After extraction and immunoaffinity column (IAC) clean-up, the isolated AFT and OTA were separated by reversed-phase liquid chromatography and quantified using a fluorescence detector. At 100°C, approximately 30–40% of AFB₁ and AFT and 20–30% of OTA in the contaminated ginger were found in the ginger tea; the total amounts of AFT and OTA in tea varied less than 5% under the three conditions of preparation. At 50°C, about 10% of OTA and AFT were found in tea. This is the first study on the migration of AFT from botanicals to tea. It is also the first to study the distribution of AFT and OTA from powdered ginger to tea and ginger residue.     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

Validation of a high-performance liquid chromatography analytical method for ochratoxin A quantification in cocoa beans

A validated high-performance liquid chromatography (HPLC) method with fluorescence detection for the quantitative analysis of ochratoxin A (OTA) in cocoa beans is described. OTA was extracted with methanol-3% sodium hydrogen carbonate solution and then purified with immunoaffinity columns before its analysis by HPLC. The validation of the analytical method was based on the following criteria: selectivity, linearity, limit of detection and quantification, precision (within- and between-day variability) and recovery, robustness and uncertainty. Detection and quantification limits were 0.04 and 0.1 μg kg^-1, respectively. Recovery was 88.9% (relative standard deviation = 4.0%). This method was successfully applied to the measurement of 46 cocoa bean samples of different origins. A total of 63% of cocoa bean samples was contaminated with a level greater than the limit of detection. The means and medians obtained for cocoa bean were 1.71 and 1.12 μg kg^-1, respectively. Surveillance controls should be set up in both crops and factories involved in transformation processes to avoid this mycotoxin in final products.     

Occurrence of ochratoxin A,fumonisin B2 and black aspergilli in raisins from Western Greece regions in relation to environmental and geographical factors

The presence of ochratoxin A (OTA), fumonisin B₂ (FB₂) and black aspergilli in raisins from Western Greece regions (Messinia, Corinthia, Achaia, Ilia and Zante Island) was investigated in relation to the different geographic and climatic conditions in the 2011 growing season. The biseriate species Aspergillus niger “aggregate” and A. carbonarius were mainly identified. The population of A. niger “aggregate” species occurred in all raisin samples at colony-forming units (CFU) concentrations significantly higher (mean 2.2 × 10⁵ CFU g^?1 homogenate) than those of A. carbonarius population (mean 4.9 × 10³ CFU g^?1 homogenate), which occurred in 80% of the raisin samples. OTA was found in 73% of the samples at levels ranging from 0.1 µg kg^?1 to 98.2 µg kg^?1, with the highest level occurring in a raisin sample from Ilia that also contained the highest level of A. carbonarius. The European Union legal limit for OTA was exceeded in 15% of the raisin samples. FB₂ was found in 29% of the raisin samples at levels ranging from 7.1 µg kg^?1 to 25.5 µg kg^?1, with 20% of the samples co-occurring with OTA. Principal-component analysis was applied to levels of mycotoxins, fungal contamination, geographical data and environmental conditions recorded in the harvesting (August) or drying (September) period. Principal-component analysis clearly indicated a good direct correlation of rainfall and relative humidity with OTA and A. carbonarius contamination. A lack of clustering was observed when A. niger and FB₂ contamination were considered. This is the first report on the co-occurrence of the mycotoxins OTA and FB₂ in dried vine fruits from Greece.     

Determination of nine preservatives in processed foods using a modified QuEChERS extraction and quantified by HPLC-PDA

ABSTRACT

In this study, a new method was developed for simultaneously determining nine preservatives, that is, benzoic acid (BA), sorbic acid (SOA), dehydroacetic acid (DHA) and PHBAs (methyl p-hydroxybenzoate [PHBA-me], ethyl p-hydroxybenzoate [PHBA-et], isopropyl p-hydroxybenzoate [PHBA-ipro], propyl p-hydroxybenzoate [PHBA-npro]), isobutyl p-hydroxybenzoate [PHBA-ibut] and butyl p-hydroxybenzoate [PHBA-nbut]), in processed foods, employing liquid chromatography (LC). This procedure accelerated sample preparation and improved efficiency by employing modified quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction without clean-up. Samples were prepared with 20 mL of acetonitrile/water (1:1) with the assistance of a ceramic stone. The extract solutions were diluted 10 times or according to the detection amount and then injected into an LC-PDA. This method showed good linearity, and the LOQs were 10 mg/kg for BA, SOA and DHA and 5?mg/kg for the PHBAs. When validating this method, the recoveries of the nine preservatives were in the range 77.0–99.6%, RSD_r values were in the range 0.7–5.3% and those of RSD_wr were in the range 2.3–8.4%. These results suggest that this new method is highly reproducible.     

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.     

Impact of non-selective fungicides on the growth and production of ochratoxin A by Aspergillus ochraceus and A. carbonarius in barley-based medium

The aim of this study was to assess the influence of the non-selective fungicides mancozeb, copper oxychloride, and sulfur on the growth and capability for producing ochratoxin A (OTA) of ochratoxigenic isolates of Aspergillus carbonarius and A. ochraceus in barley-based medium. Lag phases and growth rates were determined for each fungicide at different doses, at 15°C and 25°C and at 0.97?a_w . Mancozeb at 40?mg?l^?1 inhibited fungal growth and provided lag phases >24 days at 10–20?mg?l^?1 and 15°C. OTA was observed only at 25°C and doses <10?mg?l^?1. At 15°C, copper oxychloride proved inhibitory at 800?mg?l^?1, while at 25°C growth was not delayed and only high doses decreased OTA levels. Sulfur was inhibitory or provided large lag phases at 5–8?g?l^?1 (at 15°C) while at 25°C growth took place even at 8?g?l^?1, although OTA levels were low or undetectable. The antifungal activity decreased in the order mancozeb?>?copper oxychloride?>?sulfur, and was lower at 25°C than at 15°C. OTA accumulation was affected by the type of fungicide, dose, temperature and time. The efficacy of these fungicides on the growth of A. carbonarius and A. ochraceus and OTA production in barley-based medium is assessed for the first time.     

Monitoring of free glutamic acid in Malaysian processed foods,dishes and condiments

A study to quantify the free glutamic acid content of six processed foods, 44 dishes and 26 condiments available in Malaysia was performed using high-performance liquid chromatography with a fluorescence detector (HPLC-FRD). Recovery tests were carried out with spiked samples at levels from 6 to 31 mg g^?1. High recovery in different matrices was achieved ranging from 88% ± 13% to 102% ± 5.12%, with an average of 97% ± 8.92%. Results from the study revealed that the average free glutamic acid content ranged from 0.34 ± 0.20 to 4.63 ± 0.41 mg g^?1 in processed foods, while in prepared dishes it was as low as 0.24 ± 0.15 mg g^?1 in roti canai (puffed bread served with curry or dhal) to 8.16 ± 1.99 mg g^?1 in dim sum (a small casing of dough, usually filled with minced meat, seafood, and vegetables, either steamed or fried). Relatively, the content of free glutamic acid was found to be higher in condiments at 0.28 ± 0 mg g^?1 in mayonnaise to 170.90 ± 6.40 mg g^?1 in chicken stock powder.     

Effects of different drying treatments on fungal population and ochratoxin A occurrence in sultana type grapes

This study aimed to determine the changes in mould and ochratoxin A (OTA) occurrence in sultanas under three different conventional drying conditions. Five different vineyards were chosen, and the three different treatments were applied to these grapes while drying. At the end of the drying process, total mould and black aspergilli (BA) populations in the samples varied from 2.45 to 5.61 log colony-forming units (CFU) g^–1 and from 0 to 4.92 log CFU g^–1, respectively. Significant increases (p < 0.05) occurred in mould loads depending on the extending drying period. However, independent of vineyard location, all the samples treated with cold dipping solution showed the lowest fungal loads. These results indicate that dipping solution treatment was the most effective drying method to minimise fungal infection of grapes. The expected results could not be achieved by drying grapes artificially contaminated with ochratoxigenic Aspergillus carbonarius spores. Seventy-one of 96 isolates (73.95%) obtained during drying were Aspergillus spp., and the remaining (n = 25, 26.05%) belonged to other genera, such as Penicillium, Trichoderma and Cladosporium. Grape juice-based agar medium was used to determine the realistic OTA production capacities of the isolated mould strains. The highest OTA production capacities were 809.70 ± 9.19, 87.58 ± 16.89 and 45.44 ± 18.78 ng g^–1 in 50% grape juice agar (GJ50), all five of which were from A. niger isolates. OTA was not present in any sample during the drying period; however, OTA was detected in two samples at 0.32 ± 0.15 and 0.52 ± 0.36 µg kg^–1 after the end of the drying process. The limit of detection (LOD) and limit of quantitation (LOQ) of the method used for detecting OTA in samples were 0.1 and 0.3 µg kg^–1, respectively.     

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.     

Design and implementation of an integrated management system for ochratoxin A in the coffee production chain

Coffee is an important export product of Ecuador. Producers are challenged by the implementation of regulatory limits for ochratoxin A. Ecuador has four coffee production areas and the potential for mycotoxin contamination varies due to different environmental conditions and cultural differences in harvesting, storage, processing and commercialization. The major contributors to contamination are the lack of selection during harvesting, delays in drying or rewetting, the lack of proper drying and storage conditions, the mixing of products with different levels of moisture, and the potential for cross-contamination. The long commercialization chain involves different intermediaries that use foreign materials to increase the weight of the product without consideration of quality. An integrated mycotoxin management system using the Hazard Analysis Critical Control Point Systems (HACCP) principles was developed to prevent mycotoxin contamination at each stage of production. Critical control points were developed based on the resources available at the different stages of the production chain. Training programmes helped increase awareness about the impact of contamination, but failed to transform knowledge into improved practices. Thus, different demonstrative models specific for each productive region at all production levels were developed to show the application of prevention mechanisms using limited resources and to demonstrate the increased commercial value of coffee produced using good practices throughout the chain so producers have a better disposition to adopt improved practices. Preliminary results show that coffee managed using the models had a better quality, a lower contamination, a higher yield and better commercial value. The use of local resources and low-cost technology was important in demonstrating the practical approach.     

Detection of ochratoxin A in tropical wine and grape juice from Brazil

BACKGROUND: Ochratoxin A (OTA) is the main mycotoxin found in grapes, wines and grape juices and is considered one of the most harmful contaminants to human health. In this study, samples of tropical wines and grape juices from different grape varieties grown in Brazil were analysed for their OTA content by high‐performance liquid chromatography. RESULTS: The detection and quantification limits for OTA were 0.01 and 0.03 µg L^?1 respectively. OTA was detected in 13 (38.24%) of the samples analysed, with concentrations ranging from < 0.03 to 0.62 µg L^?1. OTA was not detected in any of the grape juice samples. Most of the red wine samples proved to be contaminated with OTA (75%), while only one white wine sample was contaminated. However, the OTA levels detected in all samples were well below the maximum tolerable limit (2 µg L^?1) in wine and grape juice established by the European Community and Brazilian legislature. CONCLUSION: The results of this study indicate a low risk of exposure to OTA by consumption of tropical wines and grape juices from Brazil. © 2012 Society of Chemical Industry     

Agaritine content of 53 Agaricus species collected from nature

Fifty-three different species of the genus Agaricus were collected in the Czech Republic during the period 1998–2001 and identified by an experienced mycologist. The samples were analysed for agaritine (N ²-(γ-L-glutamyl)-4-hydroxymethylphenylhydrazine) content, a precursor to a suspected rodent carcinogen. There was a huge variation in agaritine content between species, but less variation between samples of a species. Whereas the cultivated mushroom Agaricus bisporus commonly contain 200–500 mg agaritine kg^?1 fresh weight, no less than 24 of the 53 species contained agaritine levels above 1000 mg kg^?1 fresh weight. The highest level was found in A. elvensis containing up to 10, 000 mg kg^?1 fresh weight. Twenty species contained intermediate levels (100–1000 mg kg^?1), and nine species were below 100 mg kg^?1. Some of the species producing low levels of agaritine might be candidates for future strain development of Agaricus mushrooms for cultivation. No correlation could be observed between agaritine content and size of the mushroom, week of the year when collected, year of collection, or site of collection. Besides occurring in the genus Agaricus, some species of the genera Leucoagaricus and Macrolepiota were also shown to contain agaritine.     

Simple liquid chromatography assay for analyzing ochratoxin A in bovine milk

Ochratoxin A (OTA) is a mycotoxin with teratogenic and carcinogenic properties. Animal intake of feedstuffs contaminated with OTA may cause that some residues may be found in bovine milk, therefore, its analysis requires a highly sensitive, simple and precise technique.     

Carotenoid composition and vitamin A value in ají (Capsicum baccatum L.) and rocoto (C. pubescens R. & P.), 2 pepper species from the Andean region

The carotenoid patterns of fully ripe fruits from 12 Bolivian accessions of the Andean peppers Capsicum baccatum (ají) and C. pubescens (rocoto) were determined by high-performance liquid chromatography (HPLC)-photodiode array detector (PDA)-mass spectrometry (MS). We include 2 California Wonder cultivars as C. annuum controls. A total of 16 carotenoids were identified and differences among species were mostly found at the quantitative level. Among red-fruited genotypes, capsanthin was the main carotenoid in the 3 species (25% to 50% contribution to carotenoid fraction), although ajíes contained the lowest contribution of this carotenoid. In addition, the contribution of capsanthin 5,6-epoxide to total carotenoids in this species was high (11% to 27%) in comparison to rocotos and red C. annuum. Antheraxanthin and violaxanthin were, in general, the next most relevant carotenoids in the red Andean peppers (6.1% to 10.6%). Violaxanthin was the major carotenoid in yellow-/orange-fruited genotypes of the 3 species (37% to 68% total carotenoids), although yellow rocotos were characterized by lower levels (<45%). Cis-violaxanthin, antheraxanthin, and lutein were the next most relevant carotenoids in the yellow/orange Andean peppers (5% to 14%). As a whole, rocotos showed the highest contributions of provitamin A carotenoids to the carotenoid fraction. In terms of nutritional contribution, both ajíes and rocotos provide a remarkable provitamin A activity, with several accessions showing a content in retinol equivalents higher than California Wonder controls. Furthermore, levels of lutein in yellow/orange ajíes and rocotos were clearly higher than California Wonder pepper (≥1000 μg·100/g). Finally, the Andean peppers, particularly red ajíes, can be also considered as a noticeable source of capsanthin, the most powerful antioxidant compound among pepper carotenoids. Practical Application: Capsicum peppers are known for their content in carotenoids, although there is no information about 2 species with Andean origin: ajíes and rocotos. Due to their relevance for the Andean cuisine and increasing importance in ethnic restaurants in Europe, we studied their carotenoid pattern and vitamin A contribution.     

Ochratoxin A in dried vine fruits on the Canadian retail market

Between 1998 and 2000, 151 samples of raisins and sultanas and two samples of currants were collected from retail outlets across Canada and analysed for ochratoxin A. Samples were extracted with methanol-sodium bicarbonate, and the extracts were cleaned-up by immunoaffinity column chromatography. Ochratoxin A was quantified by liquid chromatography with fluorescence detection. The minimum quantifiable level was 0.1 ng g^-1. Ochratoxin A was present, above the minimum quantifiable level, in 67 (79%) of 85 samples of raisins, in 39 (59%) of 66 samples of sultanas, and in both samples of currants. The overall mean level of ochratoxin A was 1.8 ng g^-1 in both the raisins and sultanas, and 2.8 ng g^-1 in the currants.     

Occurrence of patulin and its dietary intake through pear,peach, and apricot juices in Italy

Pb, Cd and Hg in muscles of flathead mullet (Mugil cephalus), collected from Bocca di Magra, La Spezia (Ligurian Sea, Mediterranean Sea, Italy), were determined using graphite furnace atomic absorption spectrometry after microwave digestion for Pb and Cd and direct mercury analyser for Hg. Average Pb concentrations varied in the range 0.20–0.24 mg/kg, whereas Cd and Hg levels were negligible. None of the tested 200 samples exceeded the European regulatory limits as set by EC 1881/2006 and 420/2011. Metal concentrations in fish muscles were assessed for human consumption according to provisional tolerable weekly intake. The estimated values of Pb, Cd and Hg in M. cephalus’s edible parts in this study were below the values established by the Joint WHO/FAO Expert Committee on Food Additives. Therefore, it can be concluded that there is no health problem in human consumption.     

Determination of ochratoxin A in eggs and target tissues of experimentally drugged hens using HPLC–FLD

Ochratoxins are fungal secondary metabolites that may contaminate various foods and beverages. The intake of ochratoxins by humans may result in typical syndromes (nefrotoxicity, carcinogenity, teratogenicity and immunotoxicity) and has been associated with Balkan Endemic Nephropathy (BEN). In this study we evaluated the effects of accumulation of ochratoxin A throughout the chain production of eggs, by investigating the dynamics of OA accumulation in eggs placed by laying hens experimentally exposed to OA.