Assessment of the exposure to ochratoxin A in the province of Lleida,Spain

Exposure to ochratoxin A (OTA) of 279 blood donors of nine localities of the province of Lleida (Spain) was assessed. OTA levels were detected in the blood plasma of the participants by HPLC-fluorescence detection with previous clean-up of the samples by immunoaffinity columns. Limit of detection was 0.075 ng/mL. Participants answered a questionnaire on consumption frequency of foods possibly contaminated with OTA. Foodstuffs were grouped: cereals and derived products, dried fruits and derived products, cacao and derived products, grape juice, wine, beer and coffee. The range of positive samples was 0.11–8.68 ng/mL and the median was 0.54 ng/mL. No differences were found between OTA plasma levels in men and women, neither in the different localities, but there were significant differences among three age groups. Highest consumed foods were cereals and derived products, followed by beer and wine. No correlation was found between food consumption and OTA plasma levels. OTA daily intake was estimated based on OTA plasma concentrations and on the food consumption data combined with food contamination data taken from the literature. Mean values were 1.69 and 1.96 ng/kg body weight/day, respectively. These values are below the latest proposed tolerable daily intake of 14 ng/kg body weight/day.     

Assessment of Citrinin in Spices and Infant Cereals Using Immunoaffinity Column Clean-Up with HPLC-Fluorescence Detection

Historically, the analysis of citrinin has mainly been performed on cereals such as red yeast rice; however, in recent years, more complex and abnormal commodities such as spices and infant foods are becoming more widely assessed. The aim of this study was to develop and validate clean-up methods for spices and cereal-based infant foods using a citrinin immunoaffinity column before HPLC analysis with fluorescence detection. Each method developed was validated with a representative matrix, spiked at various citrinin concentrations, based around European Union (EU) regulations set for ochratoxin A (OTA), with recoveries >80% and % RSD < 9% in all cases. The limit of detection (LOD) and the limit of quantification (LOQ) were established at 1 and 3 µg/kg for spices and 0.1 and 0.25 µg/kg for infant cereals, respectively. These methods were then tested across a variety of spices and infant food products to establish efficacy with high recoveries >75% and % RSD < 5% across all matrices assessed. Therefore, these methods proved suitable for providing effective clean-up of spices and infant cereals, enabling reliable quantification of citrinin detected. Samples such as nutmeg and infant multigrain porridge had higher levels of citrinin contamination than anticipated, indicating that citrinin could be a concern for public health. This highlighted the need for close monitoring of citrinin contamination in these commodities, which may become regulated in the future.     

Bioaccessibility of deoxynivalenol and its natural co-occurrence with ochratoxin A and aflatoxin B1 in Italian commercial pasta

Cereals products for direct human consumption are rarely contaminated by moulds, unlike raw materials, which are often infected, either in the field or during storage. In this study, 27 samples of dried pasta characterised by size, packaging and marketing intended for young children consumption were collected and analysed by liquid chromatography (LC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) for Deoxynivalenol (DON), Ochratoxin A (OTA) and Aflatoxin B(1) (AFB(1)) determination. The samples that showed the highest amounts of one of the mycotoxins were cooked for 10min, digested with an in vitro gastrointestinal protocol and bioaccessibility values were calculated. Seven of the 27 samples exceeded from 120% to 225% the legal limit of 200μg/kg for DON fixed for processed cereal-based baby foods by an European Regulation; all the collected samples were under the OTA legal limit (0.05μg/kg) fixed by the European Regulation and no sample was contaminated by AFB(1) over the instrumental limit of detection of 0.10μg/kg. The mean value of gastric bioaccessibility verified for the DON resulted of 23.1%, whereas mean duodenal bioaccessibility was 12.1%.     

Patulin and ochratoxin A co-occurrence and their bioaccessibility in processed cereal-based foods: A contribution for Portuguese children risk assessment

Patulin (PAT) and ochratoxin A (OTA) are well known enteropathogenic mycotoxins that are present in several foodstuffs. Processed cereal-based foods are among the first solid foods eaten by children, a particularly vulnerable population group. There is a lack of knowledge related to the co-occurrence of PAT and OTA in food intended for children consumption and their potential interactions during the digestion process. The present study aims to evaluate, for the first time, the co-occurrence of PAT and OTA in processed cereal-based foods for children consumption, the bioaccessibility of these two mycotoxins, and the contribution of the bioaccessibility data for human health risk assessment. PAT and OTA incidence were 75% and 50%, respectively. These mycotoxins co-occurred in 40% of analysed samples. Bioaccessibility assays revealed mean values of 52% and 56% for PAT, alone and combined with OTA; and 100% and 106% for OTA, alone and combined with PAT. Considering the human health risk assessment, and taking into account the co-occurrence and the bioaccessibility results, this study indicates a tolerable exposure to these mycotoxins representing a low risk for Portuguese children. The present work reinforces the importance of a holistic approach for risk assessment which gathers data from occurrence, exposure and bioaccessibility.     

Estimation of ochratoxin A in some Turkish populations: An analysis in urine as a simple,sensitive and reliable biomarker

In this study, ochratoxin A (OTA) occurrence and level in human urine samples, collected from four different regions of Turkey was analyzed by NaHCO₃ dilution, immunoaffinity column clean-up and high-performance liquid chromatography with fluorescent detection. For the repeatability of the method, RSD (%) values were found between 3.83 and 8.86, for the accuracy, the recovery values were found between 85.7% and 110.5% and limit of detection and limit of quantification of the method were measured as 0.006 and 0.018 ng mL⁻¹ respectively. For the analysis, first morning urine samples were collected and the results were adjusted with creatinine levels. From the total collected samples of 233 larger amounts of 83% was contaminated with OTA. Among the calculated to be OTA levels, positive sample rate, average OTA amount and the highest contamination was found in Ankara. (Positive sample rate; 90.1%, average OTA concentration; 14.34 ng g⁻¹ creatinine and highest OTA value; 75.60 ng g⁻¹ creatinine). In order to define the exposure profile of OTA in human a questionnaire was conducted among the voluntaries as well. But related to the gender, age, dietary habits, coffee consumption, smoking and alcohol habits of the volunteers, no correlation was found with the OTA exposure.     

Multi-mycotoxin determination in cereals and derived products marketed in Tunisia using ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry

The aim of the study was the use of a fast and simple method using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS) for the simultaneous determination of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), fumonisins (FB1 and FB2), deoxynivalenol (DON), T2 and HT2 toxins in 58 samples of raw wheat (n = 34), barley (n = 5), sorghum (n = 3), processed wheat (n = 13) and breakfast cereals (n = 3) from Tunisian markets. The frequency of contamination of total samples with the analyzed mycotoxins was 50%. AFG2 was the most frequently detected in 11 samples (4 wheat, 4 barley and 3 sorghum) and it was detected at 5.2–52.4 μg/kg. HT2 toxin contaminated seven samples (4 wheat and 3 barley) and it was detected at 5.0–11.1 μg/kg. FB2 was detected in one wheat, sorghum, semolina and breakfast cereal samples at 5.0–61.5 μg/kg. FB1 was detected in three samples (2 sorghum and one barley) at 6.4–120 μg/kg. AFB1 was only found in two sorghum samples at 14 and 79.9 μg/kg. OTA was detected in one sorghum sample at concentrations below limit of quantification (5 μg/kg). The analytical results also showed that all the analyzed samples were not contaminated with DON, AFG1, AFB2 and T2 toxin.     

Ochratoxin A levels in human plasma and foods in Lebanon

Ochratoxin A (OTA) is a widespread mycotoxin which contaminates food such as cereals, beer, coffee, wine and products of animal origin. OTA is known for its nephrotoxic, immunotoxic and carcinogenic properties. The prevalence of OTA in human blood and foodstuffs has been investigated in many countries. In this study, exposure of the Lebanese population to OTA was evaluated and the contamination of the most commonly consumed foods in Lebanon by OTA was assessed. Plasma samples from healthy individuals and also cereals and beer samples obtained from markets were collected from the different regions of Lebanon. OTA was detectable in 33% of tested plasma samples (n =250) with a concentration ranging from 0.1 to 0.87 ng/mL and a mean of 0.17+/-0.01ng/mL. No sex and age differences were found. The frequency of OTA-positive plasma samples obtained in the South of Lebanon and in the Bekaa valley (50 and 47%, respectively) was significantly higher compared to plasma samples obtained in the Beirut/Mount Lebanon region (19%). Food analyses showed that wheat, burghul and beer were contaminated with a mean value of 0.15+/-0.03 microg/kg, 0.21+/-0.04 microg/kg and 0.19+/-0.12 ng/mL, respectively. These data suggest that the Lebanese population is exposed to OTA through food ingestion at concentrations lower than the tolerable daily intake.     

Incidence of ochratoxin A in dried fruits and co-occurrence with aflatoxins in dried figs

Ninety eight dried figs, 53 sultanas and 20 dried apricots destined for export from Turkey to the European Union were tested for ochratoxin A (OTA) contamination utilizing immunoaffinity column clean-up and high-performance liquid chromatography (HPLC). While only 2 (4%) of the sultanas exceeded the 10 ng g⁻¹ maximum limit set by the EU, 28 (53%) of 53 sultana samples contained detectable levels of OTA, in the range of 0.51–58.04 ng g⁻¹. Eighteen of 98 (18%) dried figs contained detectable levels of OTA, in the range of 0.87–24.37 ng g⁻¹. Only one of the 20 dried apricots was contaminated, with 0.97 ng g⁻¹ OTA. Dried figs analyzed for OTA were also tested for aflatoxin contamination to determine the co-occurrence of both toxins. Seven samples were confirmed aflatoxin positive, in the range of 0.23–4.28 ng g⁻¹, and only 2 samples contained both toxins, with a maximum concentration of 24.37 ng g⁻¹for OTA and 1.02 ng g⁻¹ for aflatoxin B₁. The average recovery and relative standard deviation (RSD) obtained from dried fruits spiked with OTA ranged from 80.5% to 91.5% and 0.99–5%, respectively. The average recovery and relative standard deviation (RSD) obtained from dried figs spiked with aflatoxin ranged from 88.78% to 93.53% and 2.54–7.25%, respectively.     

Mycotoxin Exposure in Children through Breakfast Cereal Consumption in Chile

Mycotoxins are unavoidable contaminants produced by fungi in food, especially grains. This study aimed to measure the occurrence and levels of total aflatoxins (AFs); ochratoxin A (OTA); zearalenone (ZEN); fumonisins B1, B2, and B3 (FUM); deoxynivalenol (DON); and T-2/HT-2 toxins in the four most commonly consumed breakfast cereals in Chile and to assess mycotoxin exposure and risk in children aged 2 to 13 years due to cereal consumption. In this study, a total of 110 batches with three subsamples of the four brands were sampled in supermarkets from November 2019 to June 2021. Samples were analyzed by Veratox^® ELISA (Neogen). Exposure was assessed by estimated daily intake (EDI) considering the levels found in a modified lower bound (mLB) and upper bound (UB). Risk was estimated by margin of exposure (MOE) in the case of OTA and AFs and hazard quotient (HQ) for the rest of the mycotoxins. No T2/HT2 toxins were detected. Few samples had quantifiable levels of ZEN, FUM, and DON except for brand 1, with a mean (standard deviation, SD) of 54 (20), 1552 (351), and 706 (218) ng/g, respectively. In addition, three FUM samples and one DON sample had values over the Chilean regulation. Brands 2, 3, and 4 had quantifiable levels of AFs, with mean (SD) values of 1.3 (0.1), 2.1 (0.6), and 1.9 (0.4) ng/g, respectively. Brand 3 had quantifiable levels of OTA, with a mean (SD) of 2.3 (0.4) ng/g. Estimated exposure indicated a risk of AFs in all scenarios, and of FUM for brand 1 consumption, OTA and DON for brand 3 consumption, and OTA for brand 4 consumption in the mLB worst-case scenario. In general, mycotoxin levels were below the Chilean regulatory limits, but most of them were above the EU regulation for processed cereal-based food in young children. Because the risk was higher in the 2- to 5-year-old children, we recommend special regulations for this group in Chile.     

Comparison of Clean-Up Methods for Ochratoxin A on Wine,Beer, Roasted Coffee and Chili Commercialized in Italy

The most common technique used to detect ochratoxin A (OTA) in food matrices is based on extraction, clean-up, and chromatography detection. Different clean-up cartridges, such as immunoaffinity columns (IAC), molecular imprinting polymers (MIP), Mycosep™ 229, Mycospin™, and Oasis^® HLB (Hydrophilic Lipophilic balance) as solid phase extraction were tested to optimize the purification for red wine, beer, roasted coffee and chili. Recovery, reproducibility, reproducibility, limit of detection (LOD) and limit of quantification (LOQ) were calculated for each clean-up method. IAC demonstrated to be suitable for OTA analysis in wine and beer with recovery rate >90%, as well as Mycosep™ for wine and chili. On the contrary, MIP columns were the most appropriate to clean up coffee. A total of 120 samples (30 wines, 30 beers, 30 roasted coffee, 30 chili) marketed in Italy were analyzed, by applying the developed clean-up methods. Twenty-seven out of 120 samples analyzed (22.7%: two wines, five beers, eight coffees, and 12 chili) resulted positive to OTA. A higher incidence of OTA was found in chili (40.0%) more than wine (6.6%), beers (16.6%) and coffee (26.6%). Moreover, OTA concentration in chili was the highest detected, reaching 47.8 µg/kg. Furthermore, three samples (2.5%), two wines and one chili, exceeded the European threshold.     

Ochratoxin a producing species in the genus penicillium

Ochratoxin A (OTA) producing fungi are members of the genera Aspergillus and Penicillium. Nowadays, there are about 20 species accepted as OTA producers, which are distributed in three phylogenetically related but distinct groups of aspergilli of the subgenus Circumdati and only in two species of the subgenus Penicillium. At the moment, P. verrucosum and P. nordicum are the only OTA producing species accepted in the genus Penicillium. However, during the last century, OTA producers in this genus were classified as P. viridicatum for many years. At present, only some OTA producing species are known to be a potential source of OTA contamination of cereals and certain common foods and beverages such as bread, beer, coffee, dried fruits, grape juice and wine among others. Penicillium verrucosum is the major producer of OTA in cereals such as wheat and barley in temperate and cold climates. Penicillium verrucosum and P. nordicum can be recovered from some dry-cured meat products and some cheeses.     

Determination of aflatoxins,deoxynivalenol, ochratoxin A and zearalenone in wheat and oat based bran supplements sold in the Spanish market

The aflatoxins (AFs), deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEA) are mycotoxins produced by fungal species which can contaminate, alone or simultaneously, cereal-based raw materials. Usually, the higher mycotoxins concentrations in cereals are found in the external layers of the grain (bran). Nowadays bran is increasingly consumed for its high fibre concentration. The objectives of this study were determining the concentration of these mycotoxins in bran samples intended for direct human consumption and to study the influence of some characteristics of the samples that may affect the mycotoxins content, there are not studies about fibre for direct human consumption. 67 bran samples from shops and supermarkets from two different Spanish cities were analyzed, being 37 samples of wheat bran and the remaining of oat bran. The results showed a major presence of DON in the analyzed samples, with levels above the EU legislation in some samples. Presence of DON was more frequent in wheat samples, compared to oats ones (p < 0.05). Extruded or toasted samples, subjected to a heat treatment during processing, presented a significantly lower concentration of OTA, and differences between the organically and conventionally produced samples were also detected in OTA, which showed higher levels in the organic samples. Co-occurrence was frequently found between the Fusarium mycotoxins (ZEA and DON).Due to the high levels of DON in the analyzed samples, a calculation of DON intake has been made and it has been demonstrated that bran can account for an important percentage of DON exposure in the total diet.     

Capillary electrophoresis determination of glucosamine in nutraceutical formulations after labeling with anthranilic acid and UV detection

A new robust CE method for the determination of the glucosamine (GlcN) content in nutraceutical formulations is described after its derivatization with anthranilic acid (2-aminobenzoic acid, AA). The CE separation of derivatized GlcN with AA was performed on an uncoated fused-silica capillary tube (50 μm I.D.) using an operating pH 7.0 buffer of 150 mM boric acid/50 mM NaH₂PO₄ and UV detection at 214 nm. The method was validated for specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ). The detector response for GlcN was linear over the selected concentration range from 240 to 2400 pg (40–400 μg/mL) with a correlation coefficient greater than 0.980. The intra- and inter-day variations (CV%) were between 0.5 and 0.9 for migration time, and between 2.8 and 4.3 for peak area, respectively. The LOD and the LOQ of the method were approximately 200 and 500 pg, respectively. The intra- and inter-day accuracy was estimated to range from 2.8% to 5.1%, while the percent recoveries of GlcN in formulations were calculated to be about 100% after simple centrifugation for 10 min, lyophilization and derivatization with AA. The CE method was applied to the determination of GlcN content, in the form of GlcN–hydrochloride or GlcN–sulfate, of several nutraceutical preparations in the presence of other ingredients, i.e. chondroitin sulfate, vitamin C and/or methylsulfonylmethane (MSM) as well as salts and other agents. The quantitative results obtained were in total conformity with the label claims.     

Risk assessment of PCDD/PCDFs and indicator PCBs contamination in Spanish commercial baby food

Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) as well as polychlorinated biphenyls (PCBs) are ubiquitous highly toxic environmental pollutants which exhibit a potential risk for human health. PCDD/Fs and PCBs contamination has been measured in samples of commercial baby food products: processed cereal and meat-and-fish-based baby food, which were made of individual samples collected from Spanish markets and pharmacies. They all presented a low dioxin content with a mean concentration ranging between 0.014 pg WHO-TEQ g⁻¹ product for fish-based baby food and 0.089 pg WHO PCDD/Fs-TEQ g⁻¹ product for processed cereal containing gluten. The mean concentration of the sum of the seven indicator PCBs was between 0.03 ng g⁻¹ product for fish-based baby food and 0.29 ng g⁻¹ product for gluten-free cereals. The estimated PCDD/Fs and indicator PCBs mean daily intake through the consumption of this kind of food has been calculated taking into account body weight and food consumption data for children aged 6–12 months. In order to assess the health risk derived from the exposure to these pollutants in children during the first year of life, data concerning infant formulae contamination has been also considered.     

Ochratoxin A in human blood in Abidjan, Côte d''Ivoire

Ochratoxin A (OTA) produced by Aspergillus and Penicillium genera contaminates a diversity of foods including cereals; cereals-derived foods; dry fruits; beans; cocoa; coffee; beer; wine; and foodstuffs of animal origin mainly poultry, eggs, pork and milk, including human breast milk. OTA is nephrotoxic to all animal species studied so far and most likely to humans, who show the longest half-life for elimination of this toxin among all species examined. Among other toxic effects, OTA is teratogenic, immunotoxic, genotoxic, mutagenic and carcinogenic, all of which lead to life-threatening pathologies through several molecular pathways.

In Côte d'Ivoire, preliminary surveys conducted by us have proven from 1998 to 2004 the reality of ochratoxin A-contamination of foodstuffs.

To assess OTA in human blood, the immunoaffinity columns were used along with HPLC for separation and fluorimetric quantification of blood samples collected in Abidjan from two categories of people: apparently healthy donors (n=63) and nephropathy patients undergoing dialysis (n=39).

Among healthy donors, 34.9% show OTA concentrations ranging from 0.01 – 5.81 μg/l with a mean value of 0.83 μg/l, whereas, among nephropathy patients undergoing dialysis 20.5% are OTA positive in a range of 0.167–2.42 μg/l and a mean value of 1.05. Although the sex ratio is 0.82 (46 females for 56 males) ochratoxin A contamination is equally distributed in both sexes. Nephropathy patients undergoing dialysis appear, however, less frequently contaminated than healthy donors (20.5 versus 34.9%) and show higher OTA concentrations (higher mean value, p=0.01). Ochratoxin A concentrations found in human blood reflect concentrations previously detected in cereals and peanuts according to the eating habits and diets of people in Côte d'Ivoire. But, the prevalence of ochratoxin A in blood of nephropathy people undergoing dialysis appears lower than expected from the frequency of OTA contamination in cereals and peanuts. Pearson χ²-test indicates that among OTA-positive individuals renal dialysis and age are important modalities for consideration.     

Aflatoxin M1 cytotoxicity against human intestinal Caco-2 cells is enhanced in the presence of other mycotoxins

Aflatoxin M1 (AFM1), a class 2B human carcinogen, is the only mycotoxin with established maximum residue limits (MRLs) in milk. Toxicological data for other mycotoxins in baby food, containing cereals and milk, either in isolation or in combination with AFM1, are sparse. The aim of this study was to investigate the cytotoxicity of AFM1, ochratoxin A (OTA), zearalenone (ZEA), and α-zearalenol (α-ZOL), individually and in combinations, in human Caco-2 cells. The tetrazolium salt (MTT) assay demonstrated that (i) OTA and AFM1 had similar cytotoxicity, which was higher than that of ZEA and α-ZOL, after a 72 h exposure; and (ii) the quaternary combination had the highest cytotoxicity, followed by tertiary and binary combinations and individual mycotoxins. Isobologram analysis indicated that the presence of OTA, ZEA, and/or α-ZOL with AFM1 led to additive and synergistic cytotoxicity in most combinations. The cytotoxicity of OTA was similar to that of AFM1, suggesting that OTA in food poses a health risk to consumers. Furthermore, AFM1 cytotoxicity increased dramatically in the presence of OTA, ZEA, and/or α-ZOL (p < 0.01), indicating that the established MRLs for AFM1 should be re-evaluated considering its frequent co-occurrence with other mycotoxins in baby food which contains milk and cereals.     

Biomarkers of Deoxynivalenol,Citrinin, Ochratoxin A and Zearalenone in Pigs after Exposure to Naturally Contaminated Feed Close to Guidance Values

This study applied multi-mycotoxin liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) methods to determine the biomarkers of exposure in urine and serum samples from a dose-response study with pigs. The 24 studied pigs were divided into three groups: a control and two experimental ones (with different levels of feed contamination). They were exposed to feed prepared from cereals contaminated with deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA) and citrinin (CIT) for 14 days. After that, both experimental groups received the same feed as the control group for the next 14 days to determine the kinetics of the disappearance of mycotoxin biomarkers. Urine samples were collected daily in the morning and blood samples—eight-times during the experiment. The study reported herein was the first prolonged exposure experiment for multiple mycotoxins like OTA and CIT in pigs. The urinary and serum levels of all biomarkers correlated well with the respective toxin intake; thereby demonstrating that they are suitable biomarkers of exposure in pigs. Urine is a good candidate to monitor DON, ZEN, OTA, CIT exposure while serum may be used to monitor DON, OTA and CIT. Additionally, OTA has even been quantified in both matrices in the experimental groups two weeks after changing the contaminated feed back to the control, this result differed from those produced by the other mycotoxins which were only quantified during the first two weeks. Therefore both matrices are suitable candidates to monitor prolonged OTA exposure in pigs.     

Urinary ochratoxin A and ochratoxin alpha in pregnant women

This study determined exposure of pregnant women to ochratoxin A (OTA). Forty samples of first-void urine samples from Croatian women in the third trimester of pregnancy were analyzed for OTA and its major metabolite ochratoxin alpha (OTα). The subjects filled a short food frequency questionnaire (FFQ). Analysis was performed by HPLC-FLD following liquid–liquid extraction. All samples were subjected in parallel to enzymatic treatment (β-glucuronidase/aryl sulfatase) to release OTA and OTα from the conjugates. The median urinary levels of OTA and OTα before treatment were 0.02 (range: nd–1.07) ng/mL and 0.16 (nd–1.86) ng/mL; the concentrations after enzyme hydrolysis were 0.02 (nd–1.11) ng/mL and 1.18 (0.11–7.57) ng/mL. While OTα levels increased significantly following enzymatic treatment, evidence for OTA conjugation was weak. The ratio of urinary OTα medians after and before hydrolysis was 1.5 times higher than previously reported for nonpregnant female subjects, possibly indicating upregulated metabolism and/or elimination of the mycotoxin and metabolites in pregnancy. The mean daily dietary OTA intake calculated from FFQs (1.08 ± 0.57 ng/kg body weight) was well below the provisional tolerable daily intake and the greatest contributors to intake were cereal products, fruit juices, chocolate and coffee.     

Ochratoxin A and its metabolite ochratoxin alpha in urine and assessment of the exposure of inhabitants of Lleida, Spain

Ochratoxin A (OTA) as well as its metabolite ochratoxin α (OTα) were detected in human urine in order to assess the exposure to OTA of a group of 72 adult inhabitants of the city of Lleida (Spain). Urine samples were enzymatically treated; OTA and OTα were separated by liquid-liquid extraction, and detected by HPLC-fluorescence. Exposure to OTA was also evaluated by the estimation of its daily intake from food contamination data from the literature and from food consumption data provided by the participants, who filled in a food frequency questionnaire (FFQ) and a three-day food consumption record (3DR). OTA occurrence (12.5%, limit of detection = 0.034 ng/mL) was lower than OTα occurrence (61.1%, limit of detection = 0.023 ng/mL). The range of concentrations was 0.057-0.562 ng/mL and 0.056-2.894 ng/mL for OTA and for OTα, respectively. It could be observed for positive samples that the FFQ data were related to the OTA concentration in urine, whereas the 3DR data were related to the OTα levels in urine. The OTA estimated daily intake of the participants was lower than 30% of the latest provisional tolerable daily intake of 14 ng/kg body weight/day in the worst cases of exposure.