Ochratoxin A Contamination Research Articles

Dietary Bacillus subtilis relieved the growth retardation, hepatic failure, and antioxidative depression induced by ochratoxin A in Thinlip Mullet (Liza ramada)

Mycotoxicosis is a severe challenge in the aquafeed industry and is involved in low productivity and high economic loss. On this occasion, available dietary supplements, including probiotics, may relieve the impacts of mycotoxicosis on the performances of aquatic animals. In this study, four test diets were prepared to test the effects of Bacillus subtilis (BS), ochratoxin A (OTA), and their mixture (BS/OTA) on the performances of Mullet (Liza ramada). Fish of similar initial weight (11.38 ± 0.24 g) were divided into four groups stocked in triplicate hapas (0.5 ×0.5 ×1 m) at 15 fish per hapa. Fish fed the control diet (without BS or OTA), BS (2 × 106 CFU/g), OTA (1 mg/kg) or BS (2 × 106 CFU/g) and OTA (1 mg/kg) (BS/OTA). After eight weeks, the final weight (FBW) and specific growth rate (SGR) were markedly enhanced by dietary BS and reduced by OTA contamination, while the feed conversion ratio (FCR) was meaningfully reduced by dietary BS and increased by OTA. The hemoglobin (Hb), packed cell volume (PCV), red blood cells (RBCs), and white blood cells (WBCs) were markedly lowered by OTA toxicity. Fish fed BS, or BS/OTA diets showed no significant differences with the control in terms of Hb, PCV, RBCs, and WBCs (p ˃ 0.05). The BS-fed fish showed the same normal intestinal and liver structure with an improved appearance of intestinal villi. The OTA-exposed fish showed deterioration of intestinal mucosa and stunted growth of intestinal villi with severe liver vascular dilatation and congestion in addition to hepatocytes degeneration. The combination of BS with OTA alleviated the pathological effect of individual OTA on the intestinal villi, improved intestinal morphology, and restored the normal hepatic structures with mild periductal inflammatory reaction around the bile duct. The blood total protein and albumin levels were markedly increased by dietary BS and lowered by OTA. Fish fed BS/OTA diet had higher alanine aminotransferase (ALT), aspartate aminotransferase (AST), and urea than fish fed the control but lower than those contaminated with OTA. The catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) were markedly increased by dietary BS and lowered by OTA toxicity. However, the malondialdehyde (MDA) level was decreased by dietary BS and increased by OTA toxicity. Interestingly, fish fed the control and BS/OTA diet had similar FBW, SGR, FCR, survival rates, Hb, PCV, RBCs, WBCs, creatinine, CAT, GPx, SOD, and MDA levels. In conclusion, dietary B. subtilis relieved the negative impacts of OTA contamination via protecting fish's intestine, liver, and kidney function.

Natural Occurrence of Ochratoxin A in Spices Marketed in the Czech Republic during 2019-2020.

Spices are a popular ingredient in cuisine worldwide but can pose a health risk as they are prone to fungal infestation and mycotoxin contamination. The purpose of this study was to evaluate ochratoxin A (OTA) in 54 single-kind traditional and less traditional spices, each of which was purchased in six samples of different batches (324 samples in total) at the Czech market during 2019–2020. The HPLC-FLD method with pre-treatment by immunoaffinity columns was employed to determine OTA. The limits of detection and quantification were 0.03 ng g−1 and 0.10 ng g−1, respectively. A total of 101 (31%) samples of 19 spice kinds were positive at concentrations ranging from 0.11–38.46 ng g−1. Only turmeric was contaminated with an OTA level exceeding the European Union limits. However, most spices have no regulation, thus further extensive monitoring of various mycotoxins in various kinds of spices is necessary. Chilli and black pepper are the most studied spices for OTA contamination, however, many other kinds of spice can also be highly contaminated, but studies on them are less common, rare, or have not yet been performed. The uniqueness of this study lies in the wide range of spice types studied for the presence of OTA on the Czech market.

The Occurrence and Contamination Level of Ochratoxin A in Plant and Animal-Derived Food Commodities.

Ochratoxin A (OTA) is a highly toxic mycotoxin and poses great threat to human health. Due to its serious toxicity and widespread contamination, great efforts have been made to evaluate its human exposure. This review focuses on the OTA occurrence and contamination level in nine plant and animal derived food commodities: cereal, wine, coffee, beer, cocoa, dried fruit, spice, meat, and milk. The occurrence and contamination level varied greatly in food commodities and were affected by many factors, including spices, geography, climate, and storage conditions. Therefore, risk monitoring must be routinely implemented to ensure minimal OTA intake and food safety.

Ochratoxin A and zearalenone in poultry feed samples from South China

AbstractData regarding ochratoxin A (OTA) and zearalenone (ZEA) contamination in feed in South China are limited. Here, we evaluated OTA and ZEA contamination in 514 poultry feed samples collected from South China. We validated a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method and the mycotoxins were quantified by high‐pressure liquid chromatography–tandem mass spectrometry. OTA and ZEA were detected in 2.92% and 100% of the feed samples, respectively. Furthermore, 0%, 0.68%, and 7.0% of Hainan, Guangdong, and Guangxi feed samples were contaminated with OTA, at mean concentrations of 0.00, 1.14, and 0.88 μg/kg, respectively. All samples were contaminated with ZEA, and the mean concentrations were 2094.1, 14.3, and 8.73 μg/kg in the samples from Hainan, Guangdong, and Guangxi, respectively. The findings of this study indicate the strong need for intense monitoring of mycotoxin, especially ZEA, contamination in feed.

Determination and Comprehensive Risk Assessment of Dietary Exposure to Ochratoxin A on Fermented Teas.

A specialized method for ochratoxin A (OTA) determination on fermented teas was developed and validated using ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Methodology results showed that recovery, relative standard deviation, accuracy, and precision were qualified. The limits of detection and quantification were 0.32 and 0.96 μg/kg, respectively. Two of 158 collected samples were screened for OTA contamination. Comprehensive risk assessment based on OTA contaminations of this study and other peer-reviewed publications was performed. The highest hazard quotient (HQ) value (8.86 × 10-2) and the highest 1/MoE value (8.61 × 10-5) in probabilistic assessment were equally below the recommended non-neoplastic and neoplastic thresholds, indicating no health risks. However, the HQ and 1/MoE values of the 95th percentiles in 20-39 and ≥50 years of age were close to thresholds of 1.0 and 1.0 × 10-4, respectively. Under the extreme case, there were only a few scenarios (e.g., 40-49 years of age) of HQ values below the non-neoplastic threshold, but the 1/MoE value of each group exceeded the neoplastic threshold. This is the first extensive risk assessment on OTA from fermented teas worldwide, but the sample size is still limited, and a large number of samples is encouraged in a future study for a more accurate assessment.

Ochratoxin a levels in fermented specialty coffees from Caparaó, Brazil: Is it a cause of concern for coffee drinkers?

ABSTRACT Although postharvest coffee fruit fermentation can improve coffee flavour and quality, the mycotoxin ochratoxin A (OTA) can also be a result of microbiological activity, albeit in the later drying step of coffee processing. To evaluate the possible occurrence of OTA contamination in postharvest fruit fermentation, fourteen coffees that entailed two different postharvest fruit fermentation times were evaluated. These coffees originated in the surroundings of the village of Pedra Menina in the qualified Denomination of Origin and coffee producer region of Caparaó on the border between Minas Gerais and Espírito Santo states in Brazil. All coffees were classified according to the Specialty Coffee Association (SCA) protocol and 12 achieved specialty level. OTA was determined in all 14 coffees using immunoaffinity for sample clean-up and high-performance liquid chromatography with fluorescence detection for quantification. One sample presented an OTA concentration of 0.75 µg kg−1 and two samples showed OTA concentrations of 0.87 µg kg−1. The other samples had concentrations of OTA below the limit of quantification obtained in this work (0.64 µg kg−1). Thus, all samples showed OTA concentrations far below the most stringent maximum residue limit (MRL) of 5 µg kg−1 established for roasted coffees by European legislation. These low levels were similar to most of the previous results for Brazilian coffees listed and tabled in this work. This comparison showed that OTA contamination due to this kind of postharvest process – fruit fermentation – should not be a concern for producers and consumers of these fermented coffees.

Occurrence of Ochratoxin A in Different Types of Cheese Offered for Sale in Italy.

The detection of Ochratoxin A (OTA) in the milk of ruminants occurs infrequently and at low levels, but its occurrence may be higher in dairy products such as cheese. The aim of this study was to investigate the presence of OTA in cheeses purchased in the metropolitan city of Bologna (Italy) and the surrounding area. For the analysis, a LC-MS/MS method with a limit of quantification (LOQ) of 1 µg/kg was used. OTA was detected in seven out of 51 samples of grated hard cheese (concentration range 1.3–22.4 µg/kg), while it was not found in the 33 cheeses of other types which were also analysed. These data show a low risk of OTA contamination for almost all types of cheese analysed. To improve the safety of cheese marketed in grated form, more regulations on cheese rind, which is the part most susceptible to OTA-producing moulds, should be implemented or, alternatively, producers should consider not using the rind as row material for grated cheese. It would be interesting to continue these investigations particularly on grated hard cheeses to have more data to update the risk assessment of OTA in cheese, as also suggested by EFSA in its 2020 scientific opinion on OTA.

Inhibitory effect of allyl and benzyl isothiocyanates on ochratoxin a producing fungi in grape and maize

The purpose of this study was to evaluate the inhibitory effect of allyl-isothiocyanate (AITC) and benzyl-isothiocyanate (BITC) on fungal growth and Ochratoxin A (OTA) production by Aspergillus ochraceus, A. carbonarius and A. niger. Here, we found that spore germination and fungal growth of the three fungi were significantly inhibited when the concentration of AITC and BITC was higher than 1.25 μg/mL. The inhibitory effect of AITC or BITC on A. carbonaceus and A. ochraceus was significantly stronger than that of A. niger. Scanning electron microscopy showed that the mycelia of all three fungi were changed by AITC and BITC. Compared with A. ochraceus and A. carbonarius, the damage to A. niger was lower. For OTA production, AITC and BITC could significantly down-regulated the expression of all five OTA biosynthesis genes in A. niger and A. carbonarius. In A. ochraceus, although several OTA biosynthesis genes were up-regulated, the key PKS gene was down-regulated by AITC and BITC. Twenty-five μg/mL of AITC or BITC could reduce the infection of the three fungi on grapes with inhibition rates of 28%–36% during 14 days and prolong the shelf life of grapes. In maize, the OTA production of the three fungi was significantly reduced by 25 μg/mL of AITC and BITC with the inhibition rates 68.04%–93.49% and 65.87%–75.45%, respectively. These results suggest that AITC and BITC can be used as natural fungicides to prevent A. niger, A. carbonarius and A. ochraceus from infecting grapes and maize and control OTA contamination.

Ochratoxin contamination in pig feed from pig farming centres in Indonesia

Ochratoxin A (OTA) is the most common toxin found in nature. Ochratoxin is a metabolic product by Aspergillus spp. and Penicillium spp. OTA produced by many Aspergillus or Penicicillium species that contaminate animal feeds can cause diseases. This study aims to determine the incidence of ochratoxin contamination in pig feed such as pellets, meat bone meal (MBM), and tofu by products. The sampling method used the disease detection formula to collect 36 samples 36 taken from several regions. North Sumatra, East Nusa Tenggara, Bali, Bulan Island, Solo, Lampung, Banten, Bogor and Jakarta. Ochratoxin contamination in animal feed was detected based on the Enzyme Linked Immunosorbent Assay (ELISA) using Agraquant® ochratoxin assay following the manufacturer’s protocol. The analysis showed that 6 of the 36 samples (16.7%) were tested positive for ochratoxin contamination. Pellets and MBM were the feed and feedstuff which were found to contain ochratoxin. The concentration of ochratoxin in MBM was below the limit set by the National Agency of Drug and Food Control of the Republic of Indonesia (5 ppb). The high contamination of ochratoxin was found in pellets from Lampung (19 ppb). The fungi found to dominates the culture media and come from pellet feed were Aspergillus flavus and Aspergillus niger.

Predictive Modeling and Validation on Growth, Production of Asexual Spores and Ochratoxin A of Aspergillus Ochraceus Group under Abiotic Climatic Variables.

This study aimed to generate predictive models for growth, sporulation, and ochratoxin A (OTA) production under abiotic climatic variables, including temperatures (15–35 °C) and water activity levels (0.99–0.90 aw) by Aspergillus ochraceus group. The data were divided into three sets: one for training, one for testing, and the third one for model validation. Optimum growth occurred at 0.95 aw and 25 °C and 0.95 aw and 30 °C for A. westerdijkiae and A. steynii, respectively. Significantly improved A. westerdijkiae and A. steynii spore production occurred at 0.95 aw and 20 °C and 0.90 aw and 35 °C, respectively. A. steynii and A. westerdijkiae produced the majority of OTA at 35 °C and 0.95 aw and 25–30 °C at 0.95–0.99 aw, respectively. The accuracy of the third-order polynomial regression model reached 96% in growth cases, 94.7% in sporulation cases, and 90.9% in OTA production cases; the regression coefficients (R2) ranged from 0.8819 to 0.9978 for the Aspergillus ochraceus group. A reliable agreement was reached between the predicted and observed growth, sporulation, and OTA production. The effects of abiotic climatic variables on growth, sporulation, and OTA production of A. ochraceus group have been effectively defined, and the models generated were responsible for adequately predicted and validated models against data from other strains within A. ochraceus group that had been published in the literature under the current treatments. These models could be successfully implemented to predict fungal growth and OTA contamination on food matrices for these strains under these conditions.

Environmental Conditions Affecting Ochratoxin A during Solar Drying of Grapes: The Case of Tunnel and Open Air-Drying.

Drying optimization, to mitigate fungal growth and Ochratoxin A (OTA) contamination is a key topic for raisin and currant production. Specific indicators of environmental conditions and drying properties were analyzed using two seedless grape varieties (Crimson—red and Thompson—white), artificially inoculated with Aspergillus carbonarius under open air and tunnel drying. The air temperature (T), relative humidity, grape surface temperature (Ts) and water activity throughout the drying experiment, the grapes’ moisture content and the fungal colonization and OTA contamination during the drying process and their interactions were recorded and critically analyzed. Drying properties such as the water diffusivity (Deff) and peel resistance to water transfer were estimated. The grapes Ts was 5–7 °C higher in tunnel vs. open air–drying; the infected grapes had higher maximum Ts vs. the control (around 4–6 °C). OTA contamination was higher in tunnel vs. open air–dried grapes, but fungal colonies showed the opposite trend. The Deff was higher in tunnel than in the open air–drying by 54%; the infected grapes had more than 70% higher Deff than the control, differences explained by factors affecting the water transport. This study highlighted CFU and OTA indicators that affect the water availability between red and white grapes during open air and tunnel drying, estimated by the Deff and peel resistance. This raises new issues for future research.

Mycotoxins in red wine: Occurrence and risk assessment

Mycological contamination of food products is a common problem in the food industry, associated with implications for consumer health. Red wine is regarded as an alcoholic beverage with health benefits, notably for the circulatory system. However, in spite of its health-promoting properties, it can also be a source of toxic substances. Wine is a documented source of ochratoxin A, which is one of the ubiquitous, highly toxic secondary metabolites produced by filamentous fungi. It is the only mycotoxin for which a regulatory maximum level in wine has been established. There are no legal regulations on the content of other mycotoxins in wine, as a product with an increased risk of mycological contamination, and so their levels are not monitored. The aim of this study was to determine the concentrations of ochratoxin A (OTA), deoxynivalenol (DON) as well as T-2 and HT-2 toxins in dry red wines. The study included 19 wines originating from Spain, France and Poland, from both conventional and organic viticulture. Mycotoxin levels were determined using immunoenzymatic assays (ELISA). The pH values were determined by potentiometric method. In all of the wines included in the study, mycotoxin levels exceeded the applicable limits. The levels of OTA and DON contamination were associated with the country of origin, and ochratoxin A was additionally linked to the farming method and certification status. The content of T-2 and HT-2 toxins was the only parameter to be negatively correlated with pH. In conclusion, wine may provide a source of many mycotoxins, not only OTA but also DON T-2 and HT-2 toxins, which was shown for the first time, and it appears further research is required to establish reference values for various mycotoxins in a wider range of foodstuffs, including wine.

Molecularly imprinted polymer film based plasmonic sensors for detection of ochratoxin A in dried fig

Ochratoxin A (OTA), a group of highly toxic fungal secondary metabolites, is one of the most common mycotoxins that contaminate a wide range of agricultural products. Herein, we designed a label-free and selective surface plasmon resonance (SPR) based sensor for detection of OTA contamination in dried fig. The molecularly imprinted polymer film (MIP) was fabricated onto the SPR sensor chip by using light-initiated polymerization of N-methacryloyl-(L)-phenylalanine and 2-Hydroxyethyl methacrylate in the presence of OTA as a template. SPR chips were characterized by using contact angle, atomic force microscopy, and ellipsometry. OTA imprinted sensor was displayed a broad linear range, between 0.1 ng/mL and 20 ng/mL. The limit of detection was calculated as 0.028 ng/mL. The selectivity studies of the OTA imprinted SPR sensor was performed in the presence of aflatoxin M1, aflatoxin B1, and citrinin. The imprinting factor was found to be 2.85. The adsorption behavior of the OTA imprinted sensor was found as suitable for the Langmuir model. Results displayed that the OTA imprinted SPR sensor was successfully applied to determine OTA in dried fig samples.

Smartphone biosensor for point-of-need chemiluminescence detection of ochratoxin A in wine and coffee

Exposure to mycotoxins, which may contaminate food and feed commodities, represents a serious health risk for consumers. Ochratoxin A (OTA) is one of the most abundant and toxic mycotoxins, thus specific regulations for fixing its maximum admissible levels in foodstuff have been established. Lateral Flow ImmunoAssay (LFIA)-based devices have been proposed as screening tools to avoid OTA contamination along the whole food chain. We report a portable, user-friendly smartphone-based biosensor for the detection and quantification of OTA in wine and instant coffee, which combines the LFIA approach with chemiluminescence (CL) detection. The device employs the smartphone camera as a light detector and uses low-cost, disposable analytical cartridges containing the LFIA strip and all the necessary reagents. The analysis can be carried out at the point of need by non-specialized operators through simple manual operations. The biosensor allows OTA quantitative detection in wine and coffee samples up to 25 μg L−1 and with limits of detection of 0.3 and 0.1 μg L−1, respectively, which are below the European law-fixed limits. These results demonstrate that the developed device can be used for routine monitoring of OTA contamination, enabling rapid and reliable identification of positive samples requiring confirmatory analysis.

Exposure Biomarkers and Histopathological Analysis in Pig Liver After Exposure to Mycotoxins Under Field Conditions: Special Report on Fumonisin B1

Liver samples from finisher pigs were collected at the slaughterhouses for the analysis of zearalenone (ZEA), alfa-/beta-zearalenone (α-ZE, β-ZE), zearalanone (ZA), alfa-/beta-ZA (α-ZA, β-ZA), aflatoxin B1 (AFB1) and aflatoxin M1, fumonisin B1 (FB1), ochratoxin A (OTA) and ochratoxin B, deoxynivalenol and deepoxi-deoxynivalenol (DOM-1). For the analysis liquid chromatography-triple quadrupole coupled with mass spectrometry was applied. Liver samples with detected FB1 were further histopathologically evaluated after hematoxylin and eosin staining. Various levels of liver mycotoxins were detected in all farms. Pig livers with 2.91-8.30 μg/kg of FB1 were detected in three farms, estimate of 850-2400 μg/kg of FB1 intake, whereas 0.54 μg/kg of OTA was detected in one farm, estimate of 75 μg/kg of OTA intake. Moreover, pig livers with 0.30 μg/kg of ZEA, 1.87 μg/kg of α-ZE, and 0.63 μg/kg of β-ZE were detected in one farm, estimate with of 300 μg/kg of ZEA intake. The histopathological analysis revealed that the lesions' grading and necrosis grading were analogously increased when FB1 concentration increased from 2.91 to 4.36-8.30 μg/kg. The severity of megalocytosis was analogously increased with FB1 detection levels and particularly in levels of 4.36-8.3 μg/kg. However, the increased FB1 detection levels did not show analogous behavior with the severity of hepatic cell vacuolization. Results showed that FB1 remained the most critical risk factor in the Greek pig industry, whereas ZEA and AFB1 were also prevalent. The OTA contamination in pig farms raised a high risk for animal and human health.

A PCR method to identify ochratoxin A-producing Aspergillus westerdijkiae strains on dried and aged foods

Ochratoxins are a group of mycotoxins that frequently occur as contaminants in agricultural commodities and foods, including dry-cured meats and cheeses. The fungus Aspergillus westerdijkiae is frequently isolated from aged foods and can produce ochratoxin A (OTA). However, individual strains of the fungus can have one of two OTA production phenotypes (chemotypes): OTA production and OTA nonproduction. Monitoring and early detection of OTA-producing fungi in food are the most effective strategies to manage OTA contamination. Therefore, we examined genome sequence data from five A. westerdijkiae strains isolated from the surface of cheese from southern Italy to identify genetic markers indicative of the twoOTA chemotypes. This analysis revealed a naturally occurring deletion of the OTA regulatory gene, otaR, in an OTA-nonproducing isolate.We used this information to design a polymerase chain reaction (PCR) method that could identify A. westerdijkiae and distinguish between the two OTA chemotypes. In this method, the PCR primers were complementary to conserved sequences flanking otaR and yielded different-sized amplicons from strains with the different chemotypes. The primers did not yield ota-region-specific amplicons from other OTA-producing species. Because the method is specific to A. westerdijkiae and can distinguish between the two OTA chemotypes, it has potential to significantly improve OTA monitoring programs.

Quantification of aflatoxin and ochratoxin contamination in animal milk using UHPLC-MS/SRM method: a small-scale study.

The online version contains supplementary material available at (10.1007/s13197-021-04986-w).

Influence of an industrial dry-fermented sausage processing on ochratoxin A production by Penicillium nordicum

Dry-fermented sausages are prone to be colonised by Penicillium nordicum, which is one of the main ochratoxin A (OTA)-producing species. Its ability to produce this mycotoxin on dry-fermented sausages has been reported. However, the influence of the conditions of a traditional processing of a Spanish dry-fermented sausage and the intrinsic physicochemical parameters of this product such as water activity (aw) and pH on OTA production has not been studied yet. Thus, the aim of this study was to evaluate the influence of traditional processing (interaction of relative humidity (RH) x temperature x ripening days) on the evolution of pH and aw during maturation of dry-fermented sausage “salchichón” and its relationship with OTA synthesis by P. nordicum. The expression of otapks and otanps genes, both involved in the biosynthesis of the mycotoxin, was also assessed. For this, 27 raw sausages were inoculated with P. nordicum and ripened for 26 days in a drying chamber (3 days at 5 °C and 84% RH, 17 days at 12 °C and 84% RH, and 6 days at 12 °C and 80% RH). From results, although it seems that the pH slightly influenced on OTA biosynthesis, the aw had a great impact on this mycotoxin production. In fact, the two highest OTA concentrations found coincided with a dramatic rise of the aw value (0.92 aw) by day 18 of incubation when the RH of the drying chamber was still 84% and at the end of the incubation time when the aw decreased noticeably (0.87 aw). The expression of the otapks and otanps genes correlated with the OTA produced by P. nordicum. Results from this work confirm that the traditional processing of Spanish dry-fermented sausages favours itself OTA synthesis by P. nordicum. Our findings may help in informed decision-making in relation to RH/temperature of drying chambers and shortening of the ripening process. This may be then effectively incorporated into the hygienic production system in the framework of HACCP together with other measures including the use of Penicillium nalgiovense as protective culture or the monitoring of otapks gene expression, and aw during the processing of dry-fermented sausages. All these strategies together may put ochratoxigenic Penicillia at a disadvantage and minimise OTA contamination risks in dry-fermented sausages.

Immunoanalytical methods for ochratoxin A monitoring in wine and must based on innovative immunoreagents

Immunochemical methods are highly deployed in analytical laboratories worldwide for monitoring the incidence of mycotoxins in the food chain. Nevertheless, most conventional immunoassays for ochratoxin A (OTA), including commercial kits, show limitations to robustly determine this mycotoxin in grape-derived products below regulated levels (2 ng/mL). Herein, two rapid tests for sensitive OTA determination in wine and must were developed capitalizing on a collection of bioconjugates from innovative synthetic haptens and monoclonal antibodies with subnanomolar affinity. The ELISA (LOD = 8 pg/mL) showed excellent performance in recovery studies, and it was applied to survey commercial wines and musts for OTA contamination. Concerning LFIA, validation according to the Commission Regulation 519/2014 showed that samples exceeding 2 ng/mL were properly scored as uncompliant. More importantly, illegal samples provided a complete inhibition of the test signal, making this test an easy-to-use, rapid, and convenient screening method for in-house control of OTA in wineries.

Determination of Ochratoxin A and Ochratoxin B in Archived Tokaj Wines (Vintage 1959-2017) Using On-Line Solid Phase Extraction Coupled to Liquid Chromatography.

According to the EU legislation, ochratoxin A contamination is controlled in wines. Tokaj wine is a special type of sweet wine produced from botrytized grapes infected by “noble rot” Botrytis cinerea. Although a high contamination was reported in sweet wines and noble rot grapes could be susceptible to coinfection with other fungi, including ochratoxigenic species, no screening of Tokaj wines for mycotoxin contamination has been carried out so far. Therefore, we developed an analytical method for the determination of ochratoxin A (OTA) and ochratoxin B (OTB) involving online SPE coupled to HPLC-FD using column switching to achieve the fast and sensitive control of mycotoxin contamination. The method was validated with recoveries ranging from 91.6% to 99.1% with an RSD less than 2%. The limits of quantification were 0.1 and 0.2 µg L−1 for OTA and OTB, respectively. The total analysis time of the online SPE-HPLC-FD method was a mere 6 min. This high throughput enables routine analysis. Finally, we carried out an extensive investigation of the ochratoxin contamination in 59 Slovak Tokaj wines of 1959–2017 vintage. Only a few positives were detected. The OTA content in most of the checked wines did not exceed the EU maximum tolerable limit of 2 µg L−1, indicating a good quality of winegrowing and storing.