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Magnetic molecularly imprinted polymers (MMIPs) have fused molecular imprinting technology with magnetic separation technology, emerging as an innovative material capable of recognizing specific molecules and efficiently separating target substances. Their application to the extraction and purification of mycotoxins has great potential, due to the toxicity and economic impact of these contaminants. In this work, MMIP has been proposed as a sample treatment for the determination of main four aflatoxins (B 1 , B 2 , G 1 and G 2 ) in pig feed. The MMIP was formed through the integration of magnetic material (Fe 3 O 4 ) with commercial molecularly imprinted polymers, avoiding the synthesis step and, therefore, simplifying the process. The analyses were carried out by high-performance liquid chromatography with fluorescence detection and the method was validated and limits of quantification (LOQs) between 0.09 and 0.47 ng/g were obtained, below the allowed or recommended levels by the European Union. Repeatability and intermediate precision showed relative standard deviations lower than 10% in all cases and trueness ranged from 92 to 111%. Finally, the proposed method was applied to 31 real pig feed samples, detecting aflatoxins with concentrations between 0.2 and 3.2 ng/g.

The co-occurrence of fungi and mycotoxins in various foods has been frequently reported in many countries, posing a serious threat to the health and safety of consumers. In this study, the mycobiota in five types of commercial bee pollen samples from China were first revealed by DNA metabarcoding. Meanwhile, the content of total aflatoxins in each sample was investigated by high-performance liquid chromatography with fluorescence detection. The results demonstrated that Cladosporium (0.16 %-89.29 %) was the most prevalent genus in bee pollen, followed by Metschnikowia (0-81.12 %), unclassified genus in the phylum Ascomycota (0-81.13 %), Kodamaea (0-73.57 %), and Penicillium (0-36.13 %). Meanwhile, none of the assayed aflatoxins were determined in the 18 batches of bee pollen samples. In addition, the fungal diversity, community composition, and trophic mode varied significantly among five groups. This study provides comprehensive information for better understanding the fungal communities and aflatoxin residues in bee pollen from different floral origins in China., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by the fungi of the Fusarium genera, and is a contaminant of cereals and plant products. ZEN and its metabolites are considered endocrine disruptors, and could have various toxic effects on animals and humans. In recent years, there has been a significant demographic increase in wild boar ( Sus scrofa ) in many mountainous and hilly areas of Italy, including the Campania region, mainly due to global climate change. The wild boar can be defined as a generalist and omnivorous species capable of varying its diet; therefore, it can play a role as an environmental bioindicator towards contaminants such as mycotoxins. This study was conducted to evaluate, for the first time, the concentrations of ZEN and its metabolites in the liver, kidney, and muscle of 82 wild boars shot in their habitat by hunters with hunting permits in different localities of Avellino province (Campania region, Southern Italy) from 2021 to 2022. The samples were collected and analyzed with an SPE clean-up and high-pressure liquid chromatography method with fluorescence detection. The results indicated that ZEN and α-Zearalenol were present in most of the samples, suggesting that a plan to monitor these mycoestrogens is essential to achieve the goals of "One Health".

Mycotoxins are bioaccumulative contaminants impacting animals and humans. The simultaneous detection of frequent active exposures and accumulated mycotoxin level (s) in exposed organisms would be the most ideal to enable appropriate actions. However, few methods are available for the purpose, and there is a demand for dedicated, sensitive, reliable, and practical assays. To demonstrate the issue, mice were exposed to a relevant agent Ochratoxin A (OTA), and accumulated OTA was measured by fine-tuned commercial assays. Quantitative high-performance liquid chromatography with fluorescence detection, enzyme-linked immunosorbent assay, and flow cytometry assays have been developed/modified using reagents available as commercial products when appropriate. Assays were performed on excised samples, and results were compared. Accumulated OTA could be detected and quantified; positive correlations (between applied doses of exposure and accumulated OTA levels and the results from assays) were found. Dedicated assays could be developed, which provided comparable results. The presence and accumulation of OTA following even a short exposure could be quantitatively detected. The assays performed similarly, but HPLC had the greatest sensitivity. Blood contained higher levels of OTA than liver and kidney. We demonstrate that specific but flexible and practical assays should be used for specific/local purposes, to measure the exposure itself and accumulation in blood or organs.

The contamination of food commodities with mycotoxins could be a serious health threat to humans and animals. Therefore, identification, quantification and reduction of mycotoxins in food commodities, particularly of aflatoxins (AFs) and ochratoxin A (OTA) in grain foods, is essentially required to guarantee safe food. This study determined the levels of AFs and OTA in 135 maize grains samples belonging to eight salient maize varieties cultivated in Pakistan, and evaluated the usefulness of radiations and adsorbents to reduce their levels. High performance liquid chromatography (HPLC)-based method was validated for the determination of AFs and OTA in maize grains. The results showed that 69 and 61% samples were positive for AFs and OTA, respectively and 54 and 22% of the respective samples had AFs and OTA above the permissible limits set by Pakistan Standards and Quality Control Authority. The concentration of AFs, AFB1and OTA in grains ranged from 14.5 to 92.4, 1.02 to 2.46 and 1.41 to 53.9 μg kg -1 , respectively. Among the varieties, Pearl had the highest level of total AFs and OTA, whereas YH-5427 had the highest AFB1 level. The lowest concentration of AFs and OTA was found in Malaka and 30Y87, respectively. The use of 15 kGy gamma irradiation for 24 h, sunlight-drying for 20 h and UV irradiation for 12 h almost completely degraded the mycotoxins. The microwave heating for 120 s resulted in 9-33% degradation of mycotoxins. Moreover, the treatment of grains' extract with activated charcoal (5% w/w) removed > 96% of total AFs and AFB1, and up to 43% of OTA. The use of bentonite at the same rate removed OTA, total AFs and AFB1 by 93, 73 and 92%, respectively. Thus, it is concluded that contamination of maize grains with mycotoxins was fairly high in the collected maize grain samples in Pakistan, and treatment with radiations and adsorbents can effectively reduce mycotoxins contamination level in maize grains., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Infestation of food crops by fungi has led to serious economic losses in the international market. There is a paucity of information on the multi-mycotoxin profile of cashew nuts from different geographical locations in Nigeria and South Africa. This work aims at comparing the fungal metabolite profile of cashew nut from two African countries using HPLC-FLD and LC-MS/MS. The differences in distribution of the fungal metabolites across the two countries were also examined. A total of 75 (39; Nigeria, 36; South Africa) roasted cashew nuts were purchased from 14 different locations in Nigeria and 10 major supermarkets in South Africa. The samples were subjected to HPLC analysis for aflatoxin and zearalenone and LC-MS/MS analysis for multiple metabolite profiling. Total aflatoxins (0.03-0.77 µg/kg and 0.01-0.28 µg/kg) and zearalenone (123.2 and 788.6 µg/kg) were quantified in nut samples from South Africa and Nigeria, respectively. In contrast, LC-MS/MS analysis showed that none of the major fungi metabolites except for one (citrinin) was present in detectable amount in Nigerian cashew nuts (31.3 μg/kg). Other microbial metabolites present in the nuts were; Aspergillus metabolites (< LOD to 398 × 10 4 μg/kg and < LOD to 1760 μg/kg); flavoglaucin (5.4-177.4 μg/kg and 45.5-18,368 μg/kg), 7-hydroxykaurenolide (5.5-45.8 μg/kg and 3.5-33.7 μg/kg), chloramphenicol (0.3-4.5 μg/kg) for South African and Nigerian cashew nuts, respectively. The incidence rate of the fusarium toxins beauvericin and bikaverin were higher in Nigerian cashew nut (100% and 17%) than the South African nuts (8% and 8%), 7-Hydroxykaurenolide and chloramphenicol were found in all the cashew nut samples. A total of 15 unspecific metabolites (17-100%) were recorded in the nuts from both countries. Information on the metabolite profile of cashew nuts will provide useful information for policy makers, border agencies and other health-related Institutes.

Background: Mycotoxins, which may contaminate peanut and peanut products, are responsible for many diseases to humans. Aflatoxin B1 (AFB1), aflatoxin G1 (AFG1), aflatoxin B2 (AFB2), aflatoxin G2 (AFG2), ochratoxin A (OTA) and zearalenone (ZEN) are considered the most relevant groups of mycotoxins found in food. This work aimed to develop a high-performance liquid chromatography method with a fluorescence detector (HPLC-FLD) combined with dispersive liquid-liquid microextraction (DLLME) method for the simultaneous determination of the six mycotoxins in peanuts. The six mycotoxins were simultaneously determined under their best wavelength by means of changing wavelength., Results: Under the optimum conditions, the linear ranges were 1-100 ng mL -1 for AFB1, AFG1 and OTA, 0.3-30 ng mL -1 for AFB2 and AFG2, 5-1000 ng mL -1 for ZEN, with the correlation coefficient (R 2 ) of 0.9969-0.9997. Limits of detection (LODs) were 0.10, 0.10, 0.30, 0.03, 0.03 and 1.0 µg kg -1 , respectively, and the mean recoveries were in the range of 83.1% to 99.3% with RSD < 10% (n = 6, independent analysis). Thirteen (46%) of these tested samples were contaminated with at least one mycotoxin., Conclusion: The proposed method was demonstrated to be simple, highly selective, accurate, reliable, and was successfully applied to simultaneously analyse the six mycotoxins in real peanut samples from China. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Twenty-four samples including 14 functional foods and 10 spices obtained from Chinese markets were examined for their mould profile. The mycotoxin contamination levels were also determined by an optimized HPLC-FLD method. 124 fungal isolates belonging to four different genera were recovered with Aspergillus and Penicillium as predominant fungi, with an incidence of 66.1% and 15.3%, respectively. In functional foods Aspergillus niger section (57.1%) was isolated more frequently, followed by Aspergillus flavi section (50.0%) and Aspergillus ochraceus section (21.4%), with the most contaminated samples being Coix seeds. Similar fungal presence and frequency were encountered in spice with A. niger section group (60.0%) and A. flavi section (40.0%) as main fungi. Cumin and Pricklyash peel samples showed the highest fungal contamination. Four functional foods and three spices were found to be positive at low levels for mycotoxins including aflatoxin B1 (up to 0.26μg/kg) and ochratoxin A (OTA) (5.0μg/kg). The more frequently detected mycotoxin was AFB1 (16.7%)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The natural co-occurrence of aflatoxins (AFs) and ochratoxin A (OTA) in liquorice roots obtained from different herbal outlets of Karachi, Pakistan, was evaluated. A total of 60 samples were obtained during 2019‒2021 and analysed using HPLC with fluorescence detection. The method was validated according to the European Union (EU) regulation. The incidence of AFs was 52 in all tested samples (87%), with a contamination range of 1.2‒6.4 µg kg−1 and an average of 2.1 ± 0.3 µg kg−1. In all samples, the AFs contamination was below the maximum limit (ML) of 10 µg kg−1 for total AFs as set by the EU. The OTA contamination in all samples ranged 1.5‒60.3 µg kg−1, with a mean of 18.9 ± 0.8 µg kg−1. In 36 samples (60%) the OTA contamination was below the ML of 20 µg kg−1 as set by the EU. These results led to the recommendation to test liquorice root on mycotoxins, as it comes to food quality standards. [ABSTRACT FROM AUTHOR]

Blatina is an autochthonous and most represented red wine variety in Herzegovina. Wine is the second source of daily mycotoxins intake for humans with domination of ochratoxin A (OTA) contamination of grapes. The aim of this paper was to determine occurrence of OTA in Blatina. Representative sampling was achieved by selecting twenty quality and premium wines from Mostar vineyard produced in 2018 and 2019. Quantification of OTA was performed by high performance liquid chromatography with a fluorescent detector (HPLCFLD). The research determined that the wines contained OTA. The values ranged from 0.205 μg/L to 0.933 μg/L in wines produced in 2018, and from 0.211 μg/L to 0.628 μg/L in wines produced in 2019. All samples had an OTA concentration of less than 1 μg/L. It can be concluded that the presence of ochratoxin A in Blatina wine is within acceptable limits of Regulation in the European Union (EU). The level of ochratoxin A in wine depends on the type of grape, wine region, agricultural practice, weather conditions and the process of wine production. In order to protect consumer health, it is necessary to monitor its values in wines on the market. [ABSTRACT FROM AUTHOR]

Nuts are an important food group that contributes to maintaining health; however, they can be a source of exposure to aflatoxins. This study was conducted from 2017 to 2021 to assess the incidence of aflatoxins in nuts and nut products imported to the UAE from 57 countries. Associations between container type and processing technique and aflatoxin levels were also analyzed. A total of 5401 samples of pistachios, peanuts, peanut butter, and mixed nuts were examined using HPLC-FLD analysis in conjunction with immunoaffinity cleanup. In nuts, non-conformity was detected in samples imported from 32 different countries. Mean aflatoxin values for the non-compliant samples ranged from 81.0 to 92.7 μg/kg in pistachios, peanuts, and mixed nuts. A significant difference (p

Milk and dairy products are the most important nutritional foods among all age groups. Aflatoxin M1 (AFM1) contaminates milk and makes its consumption potentially dangerous. Infants are mostly at risk because they are typically fed as many as six and more times per day, which is indeed a disquieting concern. This study aimed at evaluating AFM1 levels especially above international (European Food Safety Authority, EFSA) (0.05 µg/kg) and local (Ghana Standards Authority, GSA) (0.5 µg/kg) standards and cancer risks associated with the ingestion of raw cow milk (n = 120) sampled from Southern Ghana (Greater Accra, Volta, Western and Eastern Regions). AFM1 were measured with High-Performance Liquid Chromatography with a Fluorescence Detector (HPLC-FLD). Risk assessments were also conducted using models prescribed by the Joint FAO/WHO Expert Committee on Additives (JECFA). Out of the 120 samples analyzed for AFM1, 67 (55.8%) tested positive, 63 (52.5%) exceeded the limits of EFSA and were between the range 0.06 ± 0.001–3.52 ± 0.5 µg/kg whereas 50(41.7%) within the range of 0.50 ± 0.03–3.52.01 ± 0.5 µg/kg exceeded GSA limits. Risk assessments of AFM1 for infants, toddlers, children, adolescents, and adults ranged between 0.06 and 2.03 ng/kg bw/day, 197.04–6666.67, 0–0.0323 ng aflatoxins/kg bw/day and 1.94 × 10-3- 0.07 cases/100,000 person/yr respectively for Estimated Daily Intake (EDI), Margin of Exposure (MOE), Average Potency, and Cancer Risks. It was concluded that the consumption of raw milk posed adverse health effects on all age categories studied for the regions investigated. The use of raw cow milk may cause some problems and endanger the health of people of different age groups due to noncompliance with prescribed regulatory limits.

The current research was designed to examine the protective effect of probiotic-fortified ration against aflatoxin B1 (AFB1) toxicity and its residual level in broilers' edible tissues. Sensitive high-performance liquid chromatography coupled with a fluorescence detector (HPLC-FLD) was used for measuring the toxin. Ninety, one-day-old Cobb chicks were allocated into three equal groups (n=30) with three replicates per group. The first control group (G1) was fed a balanced basal diet only and the second group (G2) received AFB1 (2 mg/kg basal diet), while the third group (G3) received a combination of AFB1 (2 mg/kg basal diet) with Saccharomyces cerevisiae (SC; 1.5 g/Kg basal diet). Experimental birds were monitored for 6 weeks, their growth performance was then compared. AFB1 residue was assessed in the meat and liver sample. AFB1 resulted in a significant (P<0.05) reduction of growth performance parameters such as body weight and carcass yield in comparison to the control and SC supplemented groups. Moreover, AFB1 residue significantly (P<0.05) diminished in SC fortified group when compared with the AFB1 group. In conclusion, probiotics such as Saccharomyces cerevisiae could be considered as a potential feed additive and a growth promoter. Besides, its role in controlling AFB1 residue in the edible tissues of boiler chicken. [ABSTRACT FROM AUTHOR]

A different, reliable, and cost-effective strategy for the analysis of aflatoxins, ochratoxin A, and zearalenone in corn-based foods was proposed, including one multi-toxin immunoaffinity column (IAC) sample preparation and three different high-performance liquid chromatography fluorescence detection methods. The analytical procedures were tested and verified, keeping in mind their occurrence at trace levels in corn-based foods. With the validation of the proposed multi-toxin IAC methodology and comparison of the performance characteristics with methods using a single-toxin IAC, we confirmed the reliability of the multi-toxin IAC procedure versus the single-toxin IAC. The methods were validated by revealing satisfactory performance characteristics; for example, the obtained values of limit of detection were significantly lower than the maximum limits for all mycotoxins of concern. In addition, the recovery values were between 70.9 % and 106.1 % for all mycotoxins of interest, with precision values lower than 10.5 %.

A different, reliable, and cost-effective strategy for the analysis of aflatoxins, ochratoxin A, and zearalenone in corn-based foods was proposed, including one multi-toxin immunoaffinity column (IAC) sample preparation and three different high-performance liquid chromatography fluorescence detection methods. The analytical procedures were tested and verified, keeping in mind their occurrence at trace levels in corn-based foods. With the validation of the proposed multi-toxin IAC methodology and comparison of the performance characteristics with methods using a single-toxin IAC, we confirmed the reliability of the multi-toxin IAC procedure versus the single-toxin IAC. The methods were validated by revealing satisfactory performance characteristics; for example, the obtained values of limit of detection were significantly lower than the maximum limits for all mycotoxins of concern. In addition, the recovery values were between 70.9 % and 106.1 % for all mycotoxins of interest, with precision values lower than 10.5 %. The proposed methodology is an excellent candidate for future standardization due to its trustfulness and efficiency for mycotoxin analysis. [ABSTRACT FROM AUTHOR]

Mycotoxins are bioaccumulative contaminants impacting animals and humans. The simultaneous detection of frequent active exposures and accumulated mycotoxin level (s) in exposed organisms would be the most ideal to enable appropriate actions. However, few methods are available for the purpose, and there is a demand for dedicated, sensitive, reliable, and practical assays. To demonstrate the issue, mice were exposed to a relevant agent Ochratoxin A (OTA), and accumulated OTA was measured by fine-tuned commercial assays. Quantitative high-performance liquid chromatography with fluorescence detection, enzyme-linked immunosorbent assay, and flow cytometry assays have been developed/modified using reagents available as commercial products when appropriate. Assays were performed on excised samples, and results were compared. Accumulated OTA could be detected and quantified; positive correlations (between applied doses of exposure and accumulated OTA levels and the results from assays) were found. Dedicated assays could be developed, which provided comparable results. The presence and accumulation of OTA following even a short exposure could be quantitatively detected. The assays performed similarly, but HPLC had the greatest sensitivity. Blood contained higher levels of OTA than liver and kidney. We demonstrate that specific but flexible and practical assays should be used for specific/local purposes, to measure the exposure itself and accumulation in blood or organs.

Alternariol (AOH) and Alternariol monomethyl ether (AME) mycotoxins are found to be present naturally in various food commodities, such as barley, oats, pepper, rye, sorghum, sunflower seeds, tomatoes, and wheat. A few epidemiological studies have correlated the consumption of Alternaria‐contaminated cereal grains with higher occurrence of esophageal cancer in Chinese populations. In addition, several studies have reported the toxicological properties of Alternaria mycotoxins. However, surveillance data on AOH and AME occurrence are still limited. Therefore, the goal of this study was to determine the presence of AOH and AME in various commonly consumed, edible oils using HPLC‐FLD method. Thirty four percent of samples were found positive for AOH and 35% for AME. Moreover, AOH retained 80% stability, while AME retained 84% stability, after deep frying for 25 min, which is an important factor with respect to Indian cooking style. To the best of our knowledge, this is the first report on the presence of Alternaria mycotoxins in edible oils and their probable dietary intake in Indian population. This surveillance study may help in formulating guidelines for Alternaria mycotoxin levels in India, which are not yet implemented by Food Safety and Standards Authority of India. Practical Applications: At present, no safety guidelines exist for Alternaria mycotoxins in any part of the world. This study will help the regulatory bodies to set permissible levels of Alternaria mycotoxins to safeguard the health of consumers. This study shows that Alternaria mycotoxins are heat stable even after deep frying for 25 min. The data will also help to issue guidelines against exposure of these mycotoxins, keeping in the mind the heat stability factor. [ABSTRACT FROM AUTHOR]

Aflatoxin M1 (AFM1) is a well-known mycotoxin that can be found in the milk of animals that have ingested feed contaminated with aflatoxin B1 (AFB1). In Italy, the development of donkey farms is mainly due to growing request of donkey milk, which is considered an incomparable substitute for human mother's milk for its chemical composition and organoleptic characteristics. The aim of this study was to assess the occurrence of AFM1 in donkey milk produced in a farm in Northern Italy, also in view of the few data available about the presence of this mycotoxin in this type of milk. Therefore, 63 milk samples were collected and analyzed using a fast and sensitive HPLC and fluorescence detection (FLD) method previously optimized and validated. None of the milk samples collected were found to be contaminated at a level above the limit of quantification (LOQ) (0.0125 ng/mL), while only one sample showed traces of the mycotoxin at a concentration between the limit of detection (LOD) and LOQ (0.0044 ng/mL), well below the legal limit established for infant milk and follow-on milk (0.025 ng/mL). These results are in line with those of the few similar surveys carried out on donkey milk and seem to indicate a low risk of AFM1 contamination for this food. [ABSTRACT FROM AUTHOR]

This study presents the determination of aflatoxins B1, B2, G1, and G2 in rice, applying a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction followed by high-performance liquid chromatography-fluorescence detection (HPLC-FLD). The objective of the proposed method is a fast, inexpensive, and easy analysis of aflatoxins. The aflatoxins were separated on a C18 column by isocratic elution with water: methanol: acetonitrile (65 : 25 : 10, v/v/v), and detection was achieved at excitation/emission wavelengths of 360/450 nm, without derivatization. The calibration curves showed good linearity (R2 > 0.99), and the limits of detection and quantification were ≤ 6 and ≤ 8 µg·kg-1, respectively. Average intraday and interday recoveries were in the range 104 - 119 % and 104 - 113 % with RSD % ≤ 12 % for concentrations between 6 - 20 µg·kg-1. The proposed method was tested on rice samples sold on markets, of which none exhibited presence of aflatoxins.

Ochratoxin A (OTA) is a mycotoxin that may be present in various food and feed of plant and animal origin, including chicken meat. In Italy, backyard poultry farming is rather widespread. Animals are raised for meat, eggs and for ornamental purpose, and they are often fed with home-made diets not subject to official controls. The purpose of this study was to evaluate exposure of ornamental and backyard chickens to OTA using biliary ochratoxin A as a biomarker. Therefore, bile samples, in addition to kidney, liver and muscle, were collected from 102 chickens reared in 16 farms located in 6 Italian regions. High-performance liquid chromatography method and fluorimetric detection (HPLC-FLD) analysis were carried out firstly on bile from all animals, and OTA was detected in 12 chickens (concentration range 3.83-170.42 μg/L). Subsequently, the kidneys of these chickens were also analysed, and the mycotoxin was not detected. The analytical detection limits (LODs) of OTA in bile and kidney were 2.1 μg/L and 0.1 μg/kg, respectively. In conclusion, these animals were exposed to OTA but their meat can be considered safe, given that this mycotoxin, if present, concentrates highest in kidneys. Biliary ochratoxin A confirms its use as a valid biomarker to assess exposure of poultry to OTA. [ABSTRACT FROM AUTHOR]

This survey describes the occurrence and levels of AFs and OTA in chocolate products consumed in Turkey. A total of 130 samples, including bitter chocolate, milk chocolate and chocolate wafers were analysed for these mycotoxins by high-performance liquid chromatography coupled with fluorescence detection (HPLC-FLD). The values of recovery (81–92%) and precision (RSD < 9%) fulfilled the requirements of EC Regulation No. 401/2006. OTA was the most prevalent mycotoxin, with an incidence of 46.7% in bitter chocolate, 22.8% in milk chocolate and 17.4% in chocolate wafers, ranging from 0.18 to 0.75 μg kg−1. AFs were detected in 13.3% of bitter chocolate, in 19.6% of milk chocolate and in 8.7% of chocolate wafers, in concentrations ranging from 0.15 to 2.04 μg kg−1. [ABSTRACT FROM AUTHOR]

Aflatoxins (AFs) are type of mycotoxins produced by certain Aspergillus species. In the livers of cows, the aflatoxin В1 (AFB1) ingested with contaminated food is metabolized in carcinogenic aflatoxin М1 (AFM1) which is then excreted in the milk and poses a serious risk to the health of the consumers. In January of 2019, twenty five (25) samples of milk were collected from local farms in the surrounding area of Kumanovo, Republic of North Macedonia. The samples were analysed in the Centre for Public Health, Kumanovo. A method MKC EN ISO 14501:2007 for analysis of AFM1 by HPLC with fluorescence detection was applied. This method is selective, precise and applicative. The final analytical result has shown AFM1 below the LOD in any of the analyzed samples. Due to the well-known fact that AFM1 is toxic, i.e. carcinogenic and the fact that milk is widely used as staple food, continuous development as well as introduction of new, more sensitive methods for detection of AFM1 are required. [ABSTRACT FROM AUTHOR]

A total of 172 different salamis were purchased from farms and small salami factories located in four Italian regions (Piedmont, Veneto, Calabria, and Sicily) and analyzed for the presence of ochratoxin A (OTA). Analysis was performed by high-performance liquid chromatography coupled to a fluorimetric detector (HPLC-FLD). The detection limit (LOD) for the method used was 0.05 μg/kg, while the quantitation limit (LOQ) was 0.20 μg/kg; the average recovery rate was 89.1%. OTA was detected in 22 salamis, and 3 samples exceeded the Italian guidance value for OTA in pork meat (1 μg/kg). In particular, what emerges from this research is the high percentage of spicy salamis among positive samples (68.2%, 15 out of 22), although spicy salamis are only 27.3% of the total number of samples collected and analyzed. Red chili pepper contaminated by OTA could be responsible for the presence of the mycotoxin in these spicy salamis. It follow that, also the control of some ingredients used in the manufacture of these meat products, like spices, should not be neglected. [ABSTRACT FROM AUTHOR]

Locust is esteemed as a traditional Chinese medicine, as well as one of the most important nutritional foods especially in Asian countries. However, some toxic secondary metabolites such as mycotoxins are usually found in different parts of locust to affect its quality and safety. This study aimed to investigate the aflatoxins (AFs) contaminated parts by observing Aspergillus flavus, spores’ diameter, amount and distribution on head, tentacle, wing, belly and shank parts of the locust with scanning electron microscopy (SEM). Furthermore, to assess the residue levels of multi-mycotoxins in the locust, the high performance liquid chromatography with fluorescence detection (HPLC-FLD) was adopted. The technique was used to determine the contents of AFs, zearalenone (ZON) and a-zearalenol (a-ZOL) in locust and the positive samples were confirmed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The chromatographic conditions, MS/MS parameters and the method of sample extraction were carefully optimized. Results revealed that obvious differences of Aspergillus flavus strains and spores were found, while the spores’ diameter ranged from 3.0 to 13.0 mm in different contaminated parts of the locust samples. The HPLCFLD method for multi-mycotoxins analysis showed good selectivity, linearity, recovery and precision. Limits of quantification (LOQs) were lower than 27.6 mg/kg, while limits of detection (LODs) were in the range of 0.02-8.6 mg/kg. The accuracy of the developed method was validated regarding recoveries of 80.1-118.1% with relative standard deviation (RSD) ⩽ 11.4%. Finally, the developed multi-mycotoxin method was applied for screening of these mycotoxins in 11 commercial locust samples. Only AFB1 and AFB2 were found in six samples, and the contamination levels ranged from 0.12 to 4.4 mg/kg, which were lower than the maximum residue limit and can be used safely. This is the first report on the exploration of contamination parts and levels of multi-mycotoxins in medicinal and edible locust. The combined method of SEM and HPLC-FLD exhibited advantages of low cost, high sensitivity, rapid determination, convenience and especially intuitive judgment, which is proposed for contamination parts observation, for the large-scale quantification of multi-mycotoxins in other medicinal animal matrices. [ABSTRACT FROM AUTHOR]

Endometrial cancer is one of the most commonly diagnosed cancers in women. The search for factors that contribute to the development of cancer cells in reproductive organs should involve the detection of xenoestrogens, in particular zearalenone (ZEA) and its metabolites. Xenoestrogens are endocrine disruptors–ZEA and its metabolites are structurally similar to estrogens (macrocyclic lactone ring) and show high affinity for estrogen receptors. This study proposes a new method for the preparation of samples of human tissues with endometrial cancer by the use of the QuEChERS technique. Analytical parameters such as centrifugation temperature, extraction solvent, and adsorbents were modified to obtain satisfactory recovery for ZEA (R = 82.6%, RSD = 2.9%) and one of its metabolites, α-zearalenol (R = 50.1%, RSD = 3.2%). High-performance liquid chromatography (HPLC) with fluorescence detection and tandem mass spectrometry (LC-QTOF-MS) were used for the identification and quantitative determination of the analyzed compounds. The developed procedure was applied for analyses of human tissues with endometrial cancer. The presence of α-zearalenol was detected in 47 out of the 61 examined tissue samples.Methodology for isolation and identification of zearalenone and its major metabolites. [ABSTRACT FROM AUTHOR]

BACKGROUND Ochratoxin A (OTA) is a natural contaminant of food including tea with multiple toxic effects, which poses a threat to human health. In terms of lifestyle, the Turkish population is a frequent visitor of tearooms, and the traditional Turkish tea preparation is one of the most popular ways of preparing tea infusion. RESULTS The aim of this study was to investigate OTA transfer from raw black tea to the tea infusion prepared according to the Turkish tradition. A high-performance liquid chromatography method with a limit of quantification of 0.35 ng g−1 was used for OTA determination. The OTA amount in raw black teas from Turkey ranged from ≤0.35 ng g−1 up to 56.7 ng g−1. An homogenised sample of black tea naturally contaminated with 55.0 ng g−1 was used to prepare infusions. The OTA transfer from the black tea to the infusion was found to be 41.5% ± 7%. CONCLUSION These data are important for the realisation of a 'Total Diet study' (TDS). The TDS can be a complementary tool to estimate the population dietary exposure to OTA across the entire diet by analysing main foods prepared 'as consumed' (tea infusions) and not 'as purchased' (raw tea). © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Fumonisin B 1 was investigated in samples of rice intended for human consumption, including polished parboiled rice, whole grain rice and whole grain parboiled rice. Until the present, no studies on the occurrence of fumonisin B 1 have been performed on these types of rice that are commercially available in the south-eastern region of Brazil. A careful intralaboratory validation was carried out to demonstrate the fitness-of-purpose of the applied method for determining fumonisin B 1 in the three studied rice types. The performance criteria – selectivity, reliable limits of detection (50 μg kg −1 ) and quantification (100 μg kg −1 ), linearity (range 100–2500 μg kg −1 ), precision (RSD values ≤ 17.0%) and recovery (71.7–112.0 %) were evaluated, and the expanded measurement uncertainty was estimated by using the data obtained from precision and recovery experiments. Matrix-matched calibration standards were employed to quantify the mycotoxin levels in the rice samples, in which the residual normality, homoscedasticity and independence were confirmed. In addition, the measurement uncertainty values are consistent with the maximum acceptable uncertainty established by European Union regulation for analytical methods for controlling mycotoxins in foodstuffs. Among the thirty-one commercial samples of rice analysed in the present study, five samples presented detectable levels of the mycotoxin, and these levels ranged from 64.8 to 163.0 μg kg −1 . [ABSTRACT FROM AUTHOR]

A rapid and sensitive method was developed and validated for citrinin determination in red fermented rice products by liquid chromatography tandem mass spectrometry (LC-MS/MS) under the selected reaction monitoring mode. Sample preparation was especially focused, and the quantitative methods of LC-MS/MS and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) were compared. In red fermented rice samples, the limit of detection was 1.0 μg/kg for LC-MS/MS compared to 250 μg/kg for HPLC-FLD, the limit of quantification was 3.0 μg/kg for LC-MS/MS compared to 825 μg/kg for HPLC-FLD. High correlation coefficient was obtained ( R2 = 0.999) within the linear range (0.1 to 100 μg/L) in the MS method. The recoveries ranging from 80.9% to 106.5% were obtained in different spiking concentrations. The average intra- and inter-day accuracy ranged from 75.4% to 103.1%, and the intra- and inter-day precisions were from 3.3% to 7.9%. The developed method was applied to 12 commercial red fermented rice products, and citrinin was found in 10 samples ranging from 0.14 to 44.24 mg/kg. Compared to traditional qualitative and quantitative methods, the newly developed LC-MS/MS method for citrinin determination includes the merits of using a small amount of extraction solvent, simple preparation steps, and high sensitivity. [ABSTRACT FROM AUTHOR]

[Display omitted] • HPLC-FLD was adapted, optimized, and validated to quantify aflatoxins in merkén. • Interferents were adequately eliminated with Tween 20 (10%) • LLOQ (1 ng g−1) were lower than international regulations. • The validated method is reliable for routine aflatoxin analysis in merkén. • Concentration levels in the range of 1.00–176 ng g−1 were quantified in merkén. Mycotoxin contamination in agro-food systems has become a serious global concern over the last decade. Several methods have been developed in recent years to detect aflatoxins (AFs) in single spices, although none have been developed for mixed spices like merkén. The present study adapted, optimized, and validated an analytical method for the extraction, purification, and quantification of total aflatoxins (AFs; B1, B2, G1, and G2) in merkén by high performance liquid chromatography with fluorescence detection (HPLC-FLD) and post-column derivatization. The results provide satisfactory AFs separation in an elution time of 21 min with a 10-µL injection loop. The method was linear (r > 0.98) over the concentration ranges of 0.6–6.2 ng g−1 (B2 and G2) and 1.96–20 ng g−1 (B1 and G1). The lower limits of detection (LLOD) and lower limits of quantification (LLOQ) for B1, B2, G1, and G2 were 0.3 and 1.0 ng g−1, respectively. The recoveries of AFs in merkén ranged from 71% (G2) to 86% (B1) and were considered satisfactory for matrix fortifications of 0.99 (B1 and B2), 0.98 (G1), and 1.03 (G2), respectively. The intra-day precision ranged from 1.2 to 5.1% while the inter-day precision ranged from 3.7 to 14.9%. The uncertainty values calculated for the method at 1 ng g−1 (LLOQ) were 38, 25, 34, and 59% for B1, B2, G1, and G2, respectively. The method was successfully applied for AFs quantification in merkén (1.00–176 ng g−1). The calculated EDI values were lower than the limits proposed by the European Union (0.93–2.451 ng kg−1 bw day−1) and the United States (2.7 ng kg−1 bw day−1) and were not indicative of a health risk to consumers. This is the first study to describe the development and validation of a method for AFs quantification in merkén, which is important for future quality monitoring efforts of this mixed spice. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin that occurs in raw material and in several foods including meat and meat products. Cured ham may be contaminated by OTA directly forAspergillusandPenicilliumattack or indirectly as resulting from the carry-over of contaminated feeds to the edible tissues and organs of pigs. Due to OTA possible carcinogenic activity, the Italian Ministry of Health, took a precautionary measure, and a guideline value of 1 µg/kg of OTA in pork meat and derived products is in place. Analytical validated methods for the detection of OTA in meat products are required in order to comply with the guideline value and to respond to the official control requirements. Two validated methods are proposed, one based on high performance liquid chromatography fluorescence detection and one based on ultra performance liquid chromatography tandem mass spectrometry. The obtained data showed the strengths and also the limits of the two methods in terms of accuracy, limit of detection, and limit of quantification. The goal of this study was to identify the most suitable strategy for OTA determination in cured ham. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin produced by different Aspergillus and Penicillium species. Since its mechanism of action is not fully understood yet, it is important to gain further insight into different interactions of OTA at the molecular level. OTA is found worldwide in many foods and drinks. Moreover, it can also be detected in human and animal tissues and body fluids, as well. Therefore, the development of highly sensitive quantitative methods for the determination of OTA is of utmost importance. OTA most likely forms complexes with divalent cations, both in cells and body fluids. In the present study, the OTA-zinc interaction was investigated and compared to OTA-magnesium complex formation using fluorescence spectroscopy and molecular modeling. Our results show that zinc(II) ion forms a two-fold higher stable complex with OTA than magnesium(II) ion. In addition, based on the enhanced fluorescence emission of OTA in its magnesium-bound form, a novel RP-HPLC-fluorescence detector (FLD) method was also established. Our results highlight that the application of magnesium chloride in alkaline eluents results in an approximately two-fold increase in sensitivity using the HPLC-FLD technique. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a naturally occurring mycotoxin and widespread food contaminant which results in a probable human exposure. A total of 85 samples (50 dried grapes, 10 grape juices and 25pekmez(boiled and concentrated grape juices) were collected from different supermarkets and traditional bazaars in Istanbul during 2008–2009. An analytical method based on immuno-affinity column for clean-up and high-performance liquid chromatography equipped with fluorescence detection was used to determine the OTA. Contamination frequencies were 8%, 20% and 88% with mean concentrations of positive samples of 1.15, 1.40 and 2.04 µg/kg for dried grapes, grape juices andpekmezsamples, respectively. These levels were lower than the maximum levels as set by the European Commission (EC). However, 12 of 25pekmezsamples had higher levels than permitted by the European Union (EU) for safe consumption. [ABSTRACT FROM AUTHOR]

Abstract: Natural occurrence of citrinin in traditional Chinese food red yeast rice, medicinal plants and their related products has been investigated for the first time. Samples were extracted by methanol/water, cleaned-up with an immunoaffinity column (IAC) and quantified by HPLC–FLD. The mean recoveries, spiking with citrinin at levels ranging from 25 to 200μg/kg, were 73.4–92.5%, and the coefficients of variations (CVs) were 1.4–7.9%. The limit of detection (LOD) was 0.8μg/kg. Out of a total of 109 widely consumed samples analysed, citrinin was detected in 31 samples (28%) ranging from 16.6 to 5253μg/kg, all of them derived from 59 red yeast rice and related products. None of the remaining 50 medicinal plants samples was found to contain citrinin. The positive samples were further confirmed using LC–ESI–MS/MS. [Copyright &y& Elsevier]

Abstract: A new method for the determination and analysis of aflatoxins in noodles, based on the use of the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) procedure, enhanced with a high performance liquid chromatography–coupled to a fluorescence detector (HPLC–FLD) has been developed. The method excludes the use of dispersive solid-phase extraction (dSPE) extraction cleanup step to reduce time and cost of analysis. Separation of the four(4) aflatoxins with higher selectivity and sensitivity, and within reasonable limits of retention time, was performed by using an isocratic mobile phase of water/methanol/acetonitrile (65/25/10) at a column temperature of 30 °C. Optimization of sample preparation and analytical conditions gave recoveries in the range of 75%–107% at spike levels of 5.0–20.0 μg/kg, and relative standard deviations (RSDs) lower than 13% in all the cases. The repeatability and reproducibility results were in range of 2.0–12.3% and 3.4–16.5% respectively. Additionally, the limits of detection (LOD) (0.01–1.00 μg/kg) and the limits of quantification (LOQ) (0.05–1.80 μg/kg) of these aflatoxins are well within the legal limits set by the European Union (EU). [Copyright &y& Elsevier]

Abstract: A novel, cost-efficient method for the analytical extraction of the Fusarium mycotoxin zearalenone (ZON) from edible oils by dynamic covalent hydrazine chemistry (DCHC) was developed and validated for its application with high performance liquid chromatography-fluorescence detection (HPLC-FLD). ZON is extracted from the edible oil by hydrazone formation on a polymer resin functionalised with hydrazine groups and subsequently released by hydrolysis. Specifity and precision of this approach are superior to liquid partitioning or gel permeation chromatography (GPC). DCHC also extracts zearalanone (ZAN) but not α-/β-zearalenol or -zearalanol. The hydrodynamic properties of ZON, which were estimated using molecular simulation data, indicate that the compound is unaffected by nanofiltration through the resin pores and thus selectively extracted. The method''s levels of detection and quantification are 10 and 30μg/kg, using 0.2g of sample. Linearity is given in the range of 10–20,000μg/kg, the average recovery being 89%. Bias and relative standard deviations do not exceed 7%. In a sample survey of 44 commercial edible oils based on various agricultural commodities (maize, olives, nuts, seeds, etc.) ZON was detected in four maize oil samples, the average content in the positive samples being 99μg/kg. The HPLC-FLD results were confirmed by HPLC–tandem mass spectrometry and compared to those obtained by a liquid partitioning based sample preparation procedure. [Copyright &y& Elsevier]

The complete extraction of the analytes is the most important item during the analysis of matrix samples. Within the discussion, which of both extractants—aqueous methanol or aqueous acetonitrile—is the best for the extraction of the mycotoxins T-2 and HT-2, a recently published work pointed out, that the latter one resulted in incorrect high toxin concentrations. Due to water absorption of dry samples and salting out effects, a phase separation of the extractant occurs and the toxins accumulate in the organic layer. Based on these results, investigations were performed by comparing and evaluating the extraction efficiency of methanol/water and acetonitrile/water—the most frequently used extractants for T-2 and HT-2 toxin. Therefore, a reliable and routine capable method combining the extraction with acetonitrile/water and a cleanup with immunoaffinity columns was necessary. Due to the sensitivity of the antibodies against acetonitrile, the cleanup of extracts containing acetonitrile was critical. A modified workup based on the investigated antibodies’ tolerance of 5% of acetonitrile was established. The method was validated according to common guidelines (linearity, limit of detection/limit of quantitation, accuracy, method precision, recovery). Especially for more complex matrices a better extraction efficiency with acetonitrile/water was obtainable, simple or unprocessed matrices resulted in identical toxin concentrations independent from extraction solvent. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Immunoaffinity columns (IACs) are most popularly used for mycotoxin clean-up in complex matrices prior to chromatographic analysis. But, their high cost has limited their wide application and the regeneration of IACs for multiple instances of reuse is important. This study aimed to investigate the feasibility of regeneration and reuse of IACs for purification of ochratoxin A (OTA) in spiked raw malt and dried ginger samples followed by high performance liquid chromatography-fluorescence detection. After each use, the IACs were filled with phosphate buffer saline (PBS) as the preservation solution and stored at 8 °C overnight for regeneration and reuse until the recovery rate was <70%. The results showed that matrix type, preparation procedure, and pH value of sample extraction exhibited major effects on the reuse of IACs for OTA clean-up. While, after modifying the sample preparation procedure using water as the diluent and the solution at a pH of 7 to 8, the IACs could be used eight and three times for the spiked raw malt and dried ginger samples with OTA after regeneration. Regarding the traditional procedure recommended in Chinese Pharmacopoeia (2015 edition), the IACs could be used for three and two times for the spiked raw malt and dried ginger samples with OTA, respectively. Therefore, the corresponding experimental cost could be reduced to one-eighth and one-third of the original cost. This is the first study on the regeneration and reuse of IACs for OTA clean-up in complex Chinese herbal medicines, providing a green and economical tool for a large number of samples analysis with low cost. [ABSTRACT FROM AUTHOR]